Language：English   Publishing type：Research paper (scientific journal)  

Communication between osteoblasts and osteoclasts plays a key role in bone metabolism. We describe here an unexpected role for matrix vesicles (MVs), which bud from bone-forming osteoblasts and have a well-established role in initiation of bone mineralization, in osteoclastogenesis. We show that the MV cargo miR-125b accumulates in the bone matrix, with increased accumulation in transgenic (Tg) mice overexpressing miR-125b in osteoblasts. Bone formation and osteoblasts in Tg mice are normal, but the number of bone-resorbing osteoclasts is reduced, leading to higher trabecular bone mass. miR-125b in the bone matrix targets and degrades Prdm1, a transcriptional repressor of anti-osteoclastogenic factors, in osteoclast precursors. Overexpressing miR-125b in osteoblasts abrogates bone loss in different mouse models. Our results show that the MV cargo miR-125b is a regulatory element of osteoblast-osteoclast communication, and that bone matrix provides extracellular storage of miR-125b that is functionally active in bone resorption.

DOI： 10.1038/s42003-020-0754-2

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：MDPI  

Salinity critically limits rice metabolism, growth, and productivity worldwide. Improvement of the salt resistance of locally grown high-yielding cultivars is a slow process. The objective of this study was to develop a new salt-tolerant rice germplasm using speed-breeding. Here, we precisely introgressed the hst1 gene, transferring salinity tolerance from Kaijin into high-yielding Yukinko-mai (WT) rice through single nucleotide polymorphism (SNP) marker-assisted selection. Using a biotron speed-breeding technique, we developed a BC3F3 population, named YNU31-2-4, in six generations and 17 months. High-resolution genotyping by whole-genome sequencing revealed that the BC3F2 genome had 93.5% similarity to the WT and fixed only 2.7% of donor parent alleles. Functional annotation of BC3F2 variants along with field assessment data indicated that YNU31-2-4 plants carrying the hst1 gene had similar agronomic traits to the WT under normal growth condition. YNU31-2-4 seedlings subjected to salt stress (125 mM NaCl) had a significantly higher survival rate and increased shoot and root biomasses than the WT. At the tissue level, quantitative and electron probe microanalyzer studies indicated that YNU31-2-4 seedlings avoided Na+ accumulation in shoots under salt stress. The YNU31-2-4 plants showed an improved phenotype with significantly higher net CO2 assimilation and lower yield decline than WT under salt stress at the reproductive stage. YNU31-2-4 is a potential candidate for a new rice cultivar that is highly tolerant to salt stress at the seedling and reproductive stages, and which might maintain yields under a changing global climate.

DOI： 10.3390/ijms20102585

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：WILSON ORNITHOLOGICAL SOC  

The Japanese captive population of Crested Ibis (Nipponia nippon) has rapidly increased, and reintroduction programs were initiated on Sado Island in 2008. The Japanese captive population, however, was founded by only 5 individuals originating from the Chinese captive population. Because efforts were directed toward preserving genetic diversity in the captive population, it was appropriate to consider genetic diversity in the reintroduced population to evaluate its potential to survive in the wild, where environmental changes are more significant than in captivity. The purpose of this study was to determine the number of individuals to release in the future to preserve genetic diversity of the reintroduced population. We simulated the probability that the reintroduced population after 50 years would retain a rare allele existing in the captive population, based on demographic parameters obtained from either the Japanese captive or reintroduced populations, or Chinese wild populations. As a result, the 10-15 individuals currently released in the annual reintroduction would be the number needed to be released each year over 50 years to retain the rare allele with a frequency in the captive population of 0.05 with a >90% probability. Therefore, the current reintroduction program seems reasonable and should be continued.

DOI： 10.1676/1559-4491.130.4.874

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：ELSEVIER SCIENCE INC  

Management of dairy cow productivity requires monitoring of their nutritional status by visual observation. It has been suggested that changes in hair coat appearance are among the indicators of nutritional state in dairy cows. Temporal changes in the skin morphology in cows, however, have riot been reported. In this study, we examined the changes in the skin of dairy cows that occur during the peripartum period. Seven pluriparous cows were used. Skin samples were collected at 28 d before the due date and 28 d and 56 d after calving for morphological examination. Hair follicle width was 108.8 +/- 5.9 mu m (+/- SD) in the dry period, 95.5 +/- 5.5 at 28 d after calving, and 104.2 +/- 5.3 mu m at 56 d postpartum. The percentages of anagen hair follicles during these 3 periods were 41.4 +/- 3.4, 18.5 +/- 3.4, and 32.3 +/- 3.3%, respectively. The corresponding sebaceous gland sizes were 8,362.0 +/- 707.6, 7,800.0 +/- 831.4, and 9,186.8 +/- 962.6 mu m(2), respectively. Hair follicle width was positively correlated with percentage of anagen hair follicles. The thickness of epidermal and proliferation rate of epidermal cell were also correlated. However, the hair follicle width, sebaceous gland size arid cell proliferation rate, and thickness and proliferation rate of epidermal cells did not show any marked changes.

DOI： 10.3168/jds.2017-13831

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：NATURE PUBLISHING GROUP  

Medullary bone (MB), an estrogen-dependent reproductive tissue present in extant gravid birds, is texturally, histologically and compositionally distinct from other bone types. Phylogenetic proximity led to the proposal that MB would be present in non-avian dinosaurs, and recent studies have used microscopic, morphological, and regional homologies to identify this reproductive tissue in both theropod and ornithischian dinosaurs. Here, we capitalize on the unique chemical and histological fingerprint of MB in birds to characterize, at the molecular level, MB in the non-avian theropod Tyrannosaurus rex (MOR 1125), and show that the retention of original molecular components in fossils allows deeper physiological and evolutionary questions to be addressed.

DOI： 10.1038/srep23099

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：JAPAN POULTRY SCIENCE ASSOC  

The Japanese captive population of Japanese crested ibis (Nipponia nippon) was established using 5 founders derived from the Chinese captive population. Its size has increased rapidly, and the maintenance phase is about to start. Thus, this study was designed to perform genetic analyses in this population with pedigree information, considering the adoption of mean kinship strategy as the breeding strategy suited to the maintenance phase. Because the relationships among the 5 founders were unknown, different assumptions were set up ranging from 0 to 0.25 of kinship coefficients between the 5 founders. Assuming that the 5 founders were non-inbred in all the assumptions, the results showed that the gene diversity and the mean inbreeding coefficient would fluctuate largely from similar to 65% to similar to 82% and from similar to 0.07 to similar to 0.29, respectively. Moreover, the genetic importance of individuals based on mean kinship shifted largely. This study suggested that the Japanese captive population had low gene diversity and high mean inbreeding coefficient even under the assumption that the 5 founders were unrelated and non-inbred. In addition, the study also suggested that it became more effective to analyze the genetic status and to introduce mean kinship strategy into this population with more credible molecular evaluation of the relationships among founders.

DOI： 10.2141/jpsa.0150040

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：WILEY  

The calcium (Ca) demand on alligators in active reproduction is significant, yet the source of this additional Ca is unclear. Three possible sources for Ca mobilization are endolymphatic deposits, as in anurans and some lizards; short-lived skeletal medullary bone or analogous deposits, as in birds; or some other source such as the osteoderm layer of the integument or simply mobilization of structural bone. Here, we investigate possible extra-skeletal sources for labile Ca in the reproducing alligator, including endolymphatic Ca deposits, by analogy with anuran amphibian and some reptiles and integumentary osteodermal (scale) Ca deposits. We conducted X-ray image analyses of skulls for the presence of significant endolymphatic Ca deposits. We also examined dermal bone of scutes (osteoderm, scales) from the dorsal integument using both X-ray and histological analyses. Tissues from reproducing females containing mature but unovulated follicles were compared with those from specimens that had nested (laid eggs) or contained eggs within the oviduct at advanced stages of calcification. A small number of immature specimens and an adult male were also compared. No clear differences were observed in endolymphatic deposits between pre- and post-ovulatory specimens. Scute (osteoderm) X-ray density was significantly greater in females with ripe ovarian follicles compared with those that had recently laid (nested) or contained heavily calcified eggs within their oviducts. The latter groups also showed histological evidence of scute resorption compared with the former, suggesting that the scutes play a role in Ca storage during egglay.

DOI： 10.1111/jzo.12272

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：JAPAN SOC VET SCI  

The Spontaneously Diabetic Toni (SDT) fatty rat, a model for obese type 2 diabetes, shows bone quantitative abnormalities, namely low bone mineral density (BMD). The objective of this study was to evaluate bone morphological changes, in particular identifying the bone qualitative abnormalities, in the SDT fatty rat. Male SDT fatty rats showed increases in total trabecular area and trabecular number and decreases in trabecular thickness in cancellous bones of the proximal tibia, indicating trabecular miniaturization. The SDT fatty rat is useful for investigation of pathophysiological changes in bone quality in diabetic osteoporosis.

DOI： 10.1292/jvms.15-0154

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：DE GRUYTER OPEN LTD  

Marbling characterized by the amount and distribution of intramuscular fat (IMF) in longissimus muscle (LM) and measured as beef marbling score (BMS), is an economically important trait of beef cattle in Japan. The myosin binding protein C, slow type (MYBPC1) gene, involved in efficient energy metabolism and homeostasis during muscle contraction in slow skeletal muscle, has been previously shown to be expressed at different levels in the LM between high-marbled and low-marbled steer groups using differential-display PCR (ddPCR). In this study, we found that IMF area (%) in the sacrococcygeus muscle (SM) was positively correlated with BMS in the LM in Japanese Black steers (n=22, r=0.941, P<0.0001). This suggested that the IMF area (%) in the SM tends to equate marbling level in the LM. Furthermore, we showed that the MYBPC1 expression level in SM was significantly higher in the Japanese Black steers (n=5) with high BMS than in the Japanese Black steers (n=5) with low BMS (P<0.001). Moreover, correlation analyses showed that the expression level of the MYBPC1 gene was positively correlated with IMF area (%) (n=22, r=0.858, P<0.0001) and BMS (n=22, r=0.769, P<0.0001), indicating the association of MYBPC1 expression level with marbling trait. These results, together with the previous ddPCR result, suggested that high level of MYBPC1 expression may be associated with the development of marbling in Japanese Black beef cattle.

DOI： 10.1515/aoas-2015-0014

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：PUBLIC LIBRARY SCIENCE  

The major histocompatibility complex (MHC) is a highly polymorphic genomic region that plays a central role in the immune system. Despite its functional consistency, the genomic structure of the MHC differs substantially among organisms. In birds, the MHC-B structures of Galliformes, including chickens, have been well characterized, but information about other avian MHCs remains sparse. The Japanese Crested Ibis (Nipponia nippon, Pelecaniformes) is an internationally conserved, critically threatened species. The current Japanese population of N. nippon originates from only five founders; thus, understanding the genetic diversity among these founders is critical for effective population management. Because of its high polymorphism and importance for disease resistance and other functions, the MHC has been an important focus in the conservation of endangered species. Here, we report the structure and polymorphism of the Japanese Crested Ibis MHC class II region. Screening of genomic libraries allowed the construction of three contigs representing different haplotypes of MHC class II regions. Characterization of genomic clones revealed that the MHC class II genomic structure of N. nippon was largely different from that of chicken. A pair of MHC-IIA and -IIB genes was arranged head-to-head between the COL11A2 and BRD2 genes. Gene order in N. nippon was more similar to that in humans than to that in chicken. The three haplotypes contained one to three copies of MHC-IIA/IIB gene pairs. Genotyping of the MHC class II region detected only three haplotypes among the five founders, suggesting that the genetic diversity of the current Japanese Crested Ibis population is extremely low. The structure of the MHC class II region presented here provides valuable insight for future studies on the evolution of the avian MHC and for conservation of the Japanese Crested Ibis.

DOI： 10.1371/journal.pone.0108506

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：AMER ASSOC LABORATORY ANIMAL SCIENCE  

Many attempts have been made to culture germ cells in vitro by mimicking their development in vivo. The objective of this study was to establish an alternative method of xenotransplantation by developing a new approach for the rapid induction of spermatogenesis by using the chorioallantoic membrane of developing chicken embryos. Fertilized chicken eggs were incubated for 7 d, after which a small window was cut into the shell of the egg. We then transplanted testes from 7- to 8-d-old B6D2F1 mice onto the vessels of the chorioallantoic membrane and incubated them at 35.0 degrees C for 14 d or 37.5 degrees C for 12 d. After this in ovo CAM (iCAM) culture, the survival rates of the eggs and testes were assessed histologically and immunohistologically. The transplanted testes in the chicken embryos that survived were supported by the CAM, with an associated chronic vascularization response. The testes cultured at 35.0 degrees C had lower rates of generation and higher rates of death than did those cultured at 37.5 degrees C. Histologic examination of the testes cultured at 37.5 degrees C revealed the presence of spermatogonia and primary spermatocyte-like germ cells in the seminiferous tubules. The number of cells positive for synaptonemal complex protein 3 in the seminiferous tubules was significantly higher than that in the noniCAM-cultured testes from control mice. These results suggest that iCAM culturing of neonatal donor testis induces androcyte development. This method could be the foundation for a method that would enable in vitro spermatogenesis.

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：WILEY  

Japanese population of the Japanese crested ibis Nipponia nippon was founded by five individuals gifted from the People's Republic of China. In order to exactly evaluate genetic structure, we first performed development of novel genetic makers using 89 microsatellite primer pairs of related species for cross-amplification. Of these, only three primer pairs were useful for the genetic markers. Additionally, we sequenced allelic PCR products of these three markers together with 10 markers previously identified. Most markers showed typical microsatellite repeat units, but two markers were not simple microsatellites. Moreover, over half of the markers did not have the same repeat units as those of the original species. These results suggested that development of novel genetic markers in this population by cross-amplification is not efficient, partly because of low genetic diversity. Furthermore, the cluster analysis by STRUCTURE program using 17 markers showed that the five founders were divided into two clusters. However, the genetic relationships among the founders indicated by the clustering seemed to be questionable, because the analysis relied largely on a small number of triallelic markers, in spite of the addition of the three useful markers. Therefore, more efficient methods for identifying large numbers of single nucleotide polymorphisms are desirable.

DOI： 10.1111/asj.12155

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：Japanese Society of Animal Breeding and Genetics  

DOI： 10.5924/abgri.42.65

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Publishing type：Research paper (scientific journal)   Publisher：Japanese Society of Animal Breeding and Genetics  

DOI： 10.5924/abgri.42.3

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：PUBLIC LIBRARY SCIENCE  

The Japanese crested ibis is an internationally conserved, critically threatened bird. Captive-breeding programs have been established to conserve this species in Japan. Since the current Japanese population of crested ibis originates only from 5 founders donated by the Chinese government, understanding the genetic diversity between them is critical for an effective population management. To discover genome-wide single nucleotide polymorphisms (SNPs) and short tandem repeats (STRs) while obtaining genotype data of these polymorphic markers in each founder, reduced representation libraries were independently prepared from each of the founder genomes and sequenced on an Illumina HiSeq2000. This yielded 316 million 101-bp reads. Consensus sequences were created by clustering sequence reads, and then sequence reads from each founder were mapped to the consensus sequences, resulting in the detection of 52,512 putative SNPs and 162 putative STRs. The numbers of haplotypes and STR alleles and the investigation of genetic similarities suggested that the total genetic diversity between the founders was lower, although we could not identify a pair with closely related genome sequences. This study provided important insight into protocols for genetic management of the captive breeding population of Japanese crested ibis in Japan and towards the national project for reintroduction of captive-bred individuals into the wild. We proposed a simple, efficient, and cost-effective approach for simultaneous detection of genome-wide polymorphic markers and their genotypes for species currently lacking a reference genome sequence.

DOI： 10.1371/journal.pone.0072781

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：ZOOLOGICAL SOC JAPAN  

The Japanese crested ibis Nipponia nippon is a critically threatened bird. We assessed genetic diversity and structure in the Sado captive population of the Japanese crested ibis based on 24 and 50 microsatellite markers developed respectively for the same and related species. Of a total of 74 loci, 19 showed polymorphisms in the five founder birds of the population, and therefore were useful for the analysis of genetic diversity and structure. Genetic diversity measures, A, ne, He, Ho and PIC, obtained by genotyping of the 138 descendants were similar to those of other species with population bottlenecks, and thus considerably low. The low level of genetic diversity resulting from such bottlenecks was consistent with the results of lower genetic diversity measures for the Sado captive relative to the Chinese population that is the source population for the Sado group as determined using previously reported data and heterozygosity excess by Hardy-Weinberg equilibrium tests. Further, individual clustering based on the allele-sharing distance and Bayesian model-based clustering revealed that the founder genomes were equally at population in total, and with various admixture patterns at individual levels inherited by the descendants. The clustering results, together with the result of inheritance of all alleles of the microsatellites from the founders to descendants, suggest that planned mating in captive-breeding programs for the population has succeeded in maintaining genetic diversity and minimizing kinship. In addition, the Bayesian model-based clustering assumed two different components of genomes in the Sado captive Japanese crested ibis, supporting a considerably low level of genetic diversity.

DOI： 10.2108/zsj.30.432

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：WILEY  

The principal objective of this experiment was to evaluate the effect of 25-hydroxy-cholecalciferol (25-OH-D3) on the development of osteochondrosis in 6- to 110-kg castrated male pigs. The growth rate and serum calcium and inorganic phosphate levels neither increased nor decreased in response to supplementation of 25-OH-D3. However, supplemental 25-OH-D3 significantly increased serum levels of 25-OH-D3 and 1,25-hydroxy-cholecalciferol without any influence on bone mineral density. The 25-OH-D3-treated group had significant (P<0.05) reduced incidence of osteochondrotic lesions compared to the control group as evidenced by macroscopically examining the articular cartilage of the distal humerus (32.4% vs. 59.3%) and distal femur (47.1% vs. 87.5%). Likewise, supplemental 25-OH-D3 significantly reduced osteochondrotic lesions over the control when histologically examining humerus (20.6% vs. 43.8%) and femur (52.9% vs. 87.5%). The results of this experiment suggested that 25-OH-D3 supplementation in pig diets had a tendency to promote normal endochondral ossification, inhibit osteochondrosis progression and possibly regenerate destroyed cartilage tissue.

DOI： 10.1111/asj.12000

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Eggshell quality deterioration associated with heat stress is a well-known phenomenon. The involvement of the 28-kDa calcium-binding protein (calbindin, CaBP-D28k) localization in this failure is not clearly understood. To test a possible direct effect of ambient temperature on calbindin-D28k localization, 40 White Leghorn laying hens were housed in individual cages and exposed to high ambient temperature (30-33°C) and thermoneutral temperature (20-22°C) which served as a control. Eggshell quality characteristics and immunohistochemical localization of all intestinal segments and eggshell gland calbindin-D28k were performed under both environmental conditions. As expected, egg weight, eggshell thickness, eggshell percentage, and eggshell density were negatively affected by high ambient temperature (P ≤ 0.01). Immunohistochemistry showed that calbindin was localized in the intestinal enterocyte cytoplasm and glandular cell cytoplasm under thermoneutral conditions. However, the calbindin intensity was prominently decreased in ileum, cecum, colon, and eggshell gland under heat stress conditions. Therefore, it could be concluded that calbindin-D28k localization in intestinal segments and eggshell gland is negatively affected by high ambient temperature which might be related to the deterioration of eggshell quality characteristics under heat stress conditions.

DOI： 10.3382/ps.2011-01898

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：KOREAN SOC VETERINARY SCIENCE  

The temporal expression of estrogen receptor (ER)-alpha and ER-beta mRNA was examined in male Japanese quails. Femurs of quails receiving 17 beta-estradiol underwent RT-PCR and histochemical analysis 1 to 15 days after treatment. Untreated quails were used as controls (day 0). Between days 0 and 5, cells lining the bone endosteal surface differentiated into osteoblasts, which in turn formed medullary bone. Expression of ER-alpha was already observed on day 0 and increased slightly during bone formation whereas ER-beta was hardly detected throughout this process. After osteoclasts appeared on the medullary bone surface, this type of bone disappeared from the bone marrow cavity (days 7 similar to 15). ER-alpha expression simultaneously decreased slightly and ER-beta levels remained very low. These results suggest that estrogen activity mediated by ER-alpha not only affects medullary bone formation but also bone resorption.

DOI： 10.4142/jvs.2012.13.3.223

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：WILEY-BLACKWELL  

Medullary bone is formed reticularly in the bone marrow cavity of the long bones of female birds. Although this bone matrix contains fewer collagen fibers and more acid mucopolysaccharides than cortical bone, it is not clear that the expression pattern of osteoblast phenotypic genes during bone remodeling. Therefore, 17 beta-estradiol (E2)-treated male Japanese quails were used to examine the temporal expression patterns of osteoblast phenotypic genes, and to simultaneously confirm the morphological changes occurring in the bone marrow cavity during medullary bone formation and resorption. After E2 treatment, bone lining cells proliferated and developed into mature osteoblasts that had intense alkaline phosphatase (ALP) activity. These cells began to form medullary bone that contained acid mucopolysaccharides and tartrate-resistantacid phosphatase. Runt-related gene 2 (Runx2) mRNA was stably expressed throughout the process. The expression of both ALP and type I collagen mRNAs increased initially, and then rapidly decreased after day 7, while osteoclasts began to resorb medullary bone at day 5. The expression of bone matrix-related genes peaked at day 5, and suddenly decreased at day 7, except for osteopontin. Taken together with these results, the expression patterns of bone matrix-related genes during the later stages might be related to osteoclast activity. Additionally, the constant expression of Runx2 during bone formation and resorption suggested that osteoprogenitor cells always exist in the bone marrow cavity. Therefore, the expression patterns of these genes and the characteristics of bone matrix might extremely be related to the quick remodeling of medullary bone. J. Exp. Zool. (Mol. Dev. Evol.) 318B:344-352, 2012. (c) 2012 Wiley Periodicals, Inc.

DOI： 10.1002/jez.b.22451

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The Japanese crested ibis Nipponia nippon is a critically threatened bird. Accurate sexing is necessary to perform effective management of captive breeding toward a national project for a tentative release of the Japanese crested ibis on Sado Island. A PCR-based sexing method targeting a 0.6 kb EcoRI fragment (EE0.6) sequence on W chromosome with AWS03 and USP3 primers has been developed for the Japanese crested ibis. However, the primers were selected from the EE0.6 sequences from bird species other than the Japanese crested ibis. In this study, we determined the W- and Z-linked EE0.6 sequences in the Japanese crested ibis, and clarified Japanese crested ibis sequence mismatch in the binding sites of the primers. Further, we found no polymorphism in the primer binding sites among five founder birds for the Sado captive Japanese crested ibis population. These findings validated the PCR-based sexing method with the AWS03 and USP3 as accurate molecular sexing methods of captive Japanese crested ibis on the Sado Island. Additionally, we designed a primer set for a novel PCR-based sexing, based on the EE0.6 sequences obtained in this study. This novel sexing method may be useful for future ecological research following the release of Japanese crested ibis on Sado Island. This is the first report to show the EE0.6 sequences in Japanese crested ibis.

DOI： 10.1111/j.1740-0929.2011.00971.x

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：MEDWELL ONLINE  

Researchers have previously showed that the septin 7 (CDC10) gene involved in cellular proliferation possesses expression differences in musculus longissimus muscle between low-marbled Holstein and high-marbled Japanese Black steer groups. In the present study, researchers found that a marker (BM6437) close to the CDC10 was polymorphic between low-marbled Holstein and high-marbled Japanese Black steer groups and exhibited significantly different allelic distribution between Japanese Black sires with extremely high predicted breeding value for marbling and with extremely low one. Further, researchers detected Single Nucleotide Polymorphism (SNP) in the promoter region of the CDC10 gene between low-marbled Holstein and high-marbled Japanese Black steer groups. The allelic distribution of the SNP, referred to as g.63629097G>C in the CDC10 was indistinguishable between Japanese Black sires with extremely high predicted breeding value for marbling and with extremely low one. The findings suggest that an unidentified true causal mutation which is in linkage disequilibrium with the BM6437 marker but not the g.636290970>C SNP may be related to changes in CDC10 gene expression and/or marbling.

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Osteoporosis is a bone disease in which Bone Mineral Density (BMD) is reduced with a consequent increase in the risk of bone fractures. P-cryptoxanthin (beta-CRP) is present in large amounts in Satsuma mandarins and was recently reported to stimulate bone formation. In this study, researchers investigated the effects of beta-CRP in Satsuma Mandarin. Pulp (SW; 2 mg g(-1) beta-CRP) on bone metabolism in an Ovariectomized (OVX) rat model of osteoporosis. Female rats (12 weeks of age) were ovariectomized and orally administered vehicle, 0.03 g day(-1) SMP or 0.3 g day(-1) SMP for 5 weeks. After that serum concentrations of osteocalcin (an osteoblastic bone formation marker) tended to be higher in the SMT groups than in the OVX vehicle group while those of collagen type I degradation products an osteoclastic bone resorption marker) tended to be lower in the SMP groups. By bone histomorphometry, bone trabecular volume/tissue volume ratios and trabecular numbers were significantly higher in the SNIP groups than in the OVX vehicle group while trabecular separation and osteoclast number/bone surface ratios were significantly lower in the SNIP groups. By immunohistochemistry, percentage areas of osteocalcun immunoreactivity on trabecular surface were significantly greater in the SNIP groups than in the OVX vehicle group. Dual-energy X-ray absorptiometry analyses revealed that BMDs of the lumbar vertebrae and femora, tibiae tended to increase as the dose of SMP increased in the OVX rats. In conclusion, oral Styli) administration stimulated osteoblastic bone formation and inhibited osteoclastic bone resorption in OVX rats, thereby preventing the bone loss associated with osteoporosis.

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：MEDWELL ONLINE  

Researchers have previously showed that the mitofusin 2 (MFN2) gene involved in energy balance through mitochondrial fusion and expressed in slow-twitch oxidative fiber that is observed in high-marbled muscle, possesses expression differences in muscles in musculus longissimus muscle between low-marbled Holstein and high-marbled Japanese Black steer groups. In the present study, researchers found that a marker (IDVGA-49) close to the MFN2 was polymorphic between low-marbled Holstein and high-marbled Japanese Black steer groups and exhibited significantly different allelic distribution between Japanese Black sires with extremely high predicted breeding value for marbling and with extremely low one. Further, researchers detected Single Nucleotide Polymorphism (SNP) in the MFN2 gene between low-marbled Holstein and high-marbled Japanese Black steer groups. The allelic distributions of the 6 SNPs in the MFN2 were indistinguishable between Japanese Black sires with extremely high predicted breeding value for marbling and with extremely low one. The findings suggest that an unidentified true causal mutation which is in linkage disequilibrium with the IDVA-49 marker but not the 6 SNPs may be related to changes in MFN2 gene expression and or marbling. The IDVA-49 marker may be useful for effective marker-assisted selection to increase the levels of marbling.

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：MEDWELL ONLINE  

Researchers have previously showed that the triadin (TRDN) gene involved in muscle contraction possesses expression differences in musculus longissimus muscle between low-marbled Holstein and high-marbled Japanese Black steer groups. In the present study, researchers found that a marker (BMS817) close to the TRDN was polymorphic between low-marbled Holstein and high-marbled Japanese Black steer groups and exhibited significantly different allelic distribution between Japanese Black sires with extremely high predicted breeding value for marbling and with extremely low one. Further, researchers detected Single Nucleotide Polymorphism (SNP) in the TRDN gene between low-marbled Holstein and high-marbled Japanese Black steer groups. The allelic distribution of the SNP referred to as g.3834941C>T in the TRDN was indistinguishable between Japanese Black sires with extremely high predicted breeding value for marble and with extremely low one. The findings suggest that an unidentified true causal mutation which is in linkage disequilibrium with the BMS17 marker but not the g.3834941C>T SNP may be related to changes in TRDN gene expression and/or marbling. The BMS817 marker may be useful for effective marker-assisted selection to increase the levels of marbling.

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：AMER CHEMICAL SOC  

The carotenoid beta-cryptoxanthin (beta-CRX) is abundant in Satsuma mandarins (Citrus unshiu Marc). Several studies have shown a relationship between Satsuma mandarin consumption and a low risk of several diseases, for example, diabetes, gout, and hypertension, suggesting beta-CRX involvement in disease prevention. We investigated the effect of beta-CRX on mildly obese males. beta-CRX administration reduced visceral adipose tissue, body weight, and abdominal circumference. However, the detailed mechanism by which beta-CRX mediates these changes remains unknown. To identify this mechanism, we used an obese model mouse (TSOD). Oral beta-CRX administration repressed body weight, abdominal adipose tissue weight, and serum lipid concentrations in TSOD; these results are identical to previous human trial results. beta-CRX administration significantly repressed adipocyte hypertrophy. Gene expression analysis strongly indicated that beta-CRX can alter cytokine secretion and cell proliferation. These results suggest that beta-CRX derived from Satsuma mandarins can help prevent obesity by repressing hypertrophy of abdominal adipocytes.

DOI： 10.1021/jf202821u

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：WILEY-BLACKWELL  

The Japanese Crested Ibis Nipponia nippon is a critically threatened bird. The post-hatch eggs of the current captive population of this species on Sado Island have been stored at room temperature for the long-term. In this study, we investigated the suitability of the vascularized chorioallantois membrane from the eggs as a non-invasive DNA source. Using microsatellite loci developed for the Japanese Crested Ibis, we performed three experiments for comparison of genotypes obtained among DNA. First, DNA from five different sites of the identical membrane showed the same genotypes at either of two loci examined. Second, DNA from the membrane of each full-sibling birds and blood of their parents showed the genotypes that were consistent with Mendelian parent-offspring relationships at any of eight loci examined. Third, DNA from the membrane and blood of the same bird showed the matched genotypes at any of eight loci examined. These results indicate that the vascularized chorioallantois membrane from post-hatch eggs stored at room temperature for the long-term can be used as a reliable DNA source of offspring that had hatched from the egg. This study will promote a molecular genetics study on genetic diversity of the current captive Japanese Crested Ibis population on Sado Island.

DOI： 10.1111/j.1740-0929.2011.00902.x

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Crossbred (Japanese Black male and Holstein female) steers were used to investigate the effects of feeding a large amount of high-protem milk replacer on growth performance, morphological change and myogenic regulation in skeletal muscle. Group HP (n = 7) was fed high-protein milk replacer (Crude Protein (CP): 29%, maximum intake 2.0 kg day-1) and group C (n = 14) was fed a standard milk replacer (CP: 26%, 0.5 kg day-1) from 56-147 days of age. Changes in Body Weight (BW), body frame measurements, plasma Insulin-like Growth Factor-1 (IGF-1) and plasma Alkaline Phosphatase (ALP) were investigated. M. Longissimus Thoracis (LT) samples were obtained by biopsy to investigate the myofiber type composition, diameter and the mRNA expression of Myogenic Regulatory related genes by quantitative real-time RT-PCR. A large amount of high protein milk replacer (Intensified Nursing) in group HP improved their synthesis of IGF-1 in the liver and accelerated skeletal development by maintaining high levels of plasma IGF-1 and ALP concentration, which was highly controlled by CP intake from diet. The myofiber type composition and size in the LT was not influenced by nutritional condition and aging during the nursing period. Intensified Nursing extends myofiber length rather than increasing the thickness of the myofiber, according to the increase in the size of the body frame during this phase. The expression of MyoD and IGF-1 receptors in the LT decreased at the early stage of the nursing period without the influence of nutritional condition. This suggested the change would control the level of differentiation from the myoblast to the myocyte. © Medwell Journals, 2010.

DOI： 10.3923/javaa.2010.1037.1047

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Publishing type：Research paper (scientific journal)   Publisher：The Japanese Society of Swine Science  

DOI： 10.5938/youton.47.136

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：ELSEVIER GMBH, URBAN & FISCHER VERLAG  

Medullary bone is a unique tissue in female birds and forms in the cavity of tong bones. This bone displays rapid remodeling in response to circulating estrogen levels, suggesting that the osteoblasts in this bone are highly sensitive to estrogen. The present study examined expression of two estrogen receptor (ER) mRNAs in osteogenic cells of medullary bone of white Leghorn hens in vitro. At day 3, isolated cells from the hen medullary bone expressed alkaline phosphatase activity. Using immunocytochemistry, ER protein was demonstrated in the nuclei of these cells. RT-PCR analysis revealed that ER-alpha mRNA was constantly expressed from day 3 to day 15 of culture, white ER-beta mRNA was not detected throughout the culture period. These results indicate that estrogen may act via ER-alpha, but not ER-beta, on osteogenic cells of the avian medullary bone. (C) 2008 Elsevier GmbH. All rights reserved.

DOI： 10.1016/j.acthis.2008.06.003

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We observed angiogenesis and endochondral ossification of the femora of chick embryos in vitro, on chorioallantoic membrane (CAM), and in vivo to clarify the cellular processes of avian endochondral ossification. We found that the inside of the in vitro femora was still filled with chondrocytes, despite being cultured for 10 days, while calcification of the diaphysis was not observed. As well, only the cartilage tissue of the epiphysis was enlarged. By contrast, blood vessels invaded into the diaphysis and the marrow cavity was formed in CAM-cultured femora. Cartilage canals extended from the marrow cavity and reached to the resting chondrocyte zone, with normal endochondral ossification occurring as in vivo. This study demonstrates that endochondral ossification occurs in femora in CAM culture similar to that in in vivo femora, but not in in vitro femora. The ossification is dependent on vascular invasion into the embryonic femora. In conclusion, for the endochondral ossification of long bones it is essential to supply the embryo with blood vessels. CAM culture system was found to be a superior endochondral ossification model of the embryonic femur. Copy right © 2008, Japan Poultry Science Association.

DOI： 10.2141/jpsa.45.51

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1. The aim of the present study was to investigate expression and localisation of a 28-kDa calcium-binding protein (CaBP-D28k) related to active calcium (Ca) absorption, in the entire intestine of egg-laying hens. 2. Western blotting analysis showed that the entire intestine expressed CaBP-D28k to the following degree: duodenum > jejunum > caecum > ileum > colon. Immunohistochemistry showed strong CaBP-D28k localisation in enterocytes along the villus tip-crypt axis in the duodenum and in villus tips in the caecum and colon. The jejunum and ileum had moderate localisation with respect to the number of immunoreactive cells and staining intensity. 3. These results suggest that laying hens actively absorb Ca in both the large and small intestines.

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DOI： 10.2141/jpsa.44.426

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：TAYLOR & FRANCIS LTD  

Changes in the localization of calpain in conditioned and pressurized bovine skeletal muscles were investigated by immunogold electron-microscopy. In the muscle immediately after thawing (control), the relative distributions of colloidal particles statistically calculated by counting the colloidal particles were about 65% and 35% in the I-band/Z-disk and A-band regions, respectively. In the muscle conditioned for 7 days, distribution of colloidal particles was more than two times greater in both the I-band/Z-disk and A-band regions than in the control muscle. Almost no change in either the absolute concentration or relative distribution of the colloidal particles was detectable during further storage. In the muscle exposed to a pressure of 100 MPa or, 200 MPa, slightly more immunogold was detected in the sarcomere than in that of the control muscle. Increasing pressure up to 300 MPa enabled high-density particles to be seen throughout the sarcomere. Conversely, few particles were detected in the sarcomere of the muscle exposed to 400 MPa. These electron-microscopic observations were confirmed from the statistical analysis as with the conditioned muscles. It was clear from the results obtained that the pressure-induced changes in calpain localization were much;more pronounced than those from conditioning.

DOI： 10.1271/bbb.70.1249

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The aim of this study was to observe the expression and localization of estrogen receptor (ER) α and ER β mRNA in the medullary bone of laying hens. First, medullary bone, liver, kidney, and shell gland of the oviduct tissues were dissected from laying hens. Then, the total cellular RNA was isolated from each tissue specimen, and the ER α and ER β mRNA expression was observed using semiquantitative RT-PCR. Second, the localization of ER α mRNA in the medullary bone was detected with in situ hybridization using digoxigenin-11-UTP-labeled cRNA probes. As a result, the expression of ER α mRNA was higher than that of ER β mRNA in the medullary bone, liver, and shell gland of the oviduct from laying hens. In the kidney, ER α mRNA expression was lower than that of ER β mRNA. The expression pattern of ER α and ER β mRNA of the medullary bone was similar to that of the shell gland of the oviduct. Moreover, ER α mRNA was intensively expressed in osteoblasts on the medullary bone surface and bone marrow stromal cells but was not expressed in osteoclasts. These results suggest that in medullary bone, estrogen action may be regulated not by ER β but by ER α. © 2006 Japanese Society of Animal Science.

DOI： 10.1111/j.1740-0929.2006.00341.x

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In this study, cells isolated from hen medullary bone were cultured to examine their matrix formation. Furthermore, we compared medullary bone cells with rat bone marrow cells regarding the temporal changes in osteoblast developmental markers. Medullary bone cells were positive for alkaline phosphatase (ALP) activity and formed bone nodules, apparent with Alcian blue and von Kossa staining. The intensity of these stains became stronger with the maturation of those bone nodules. In this developmental process, the expression patterns of osteoblast phenotypes of medullary bone cells differed from those of rat bone marrow cells. ALP mRNA was expressed at the maximum level in the proliferation stage and gradually decreased in medullary bone cells, but that expression showed the opposite pattern in rat bone marrow cells. Medullary bone cells strongly expressed two non-collagenous protein mRNAs from the early stages, but the expression of these mRNAs in rat bone marrow cells increased only in the later stages. These results suggest that the features of medullary bone osteoblasts differ from those of mammalian osteoblasts and are reflected in the characteristics of medullary bone in vivo. © 2005 Elsevier Inc. All rights reserved.

DOI： 10.1016/j.cbpb.2005.09.006

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In order to examine the effects of the calcium regulating hormones on the differentiation of hen medullary bone osteoclasts, isolated medullary bone marrow cells were cultured for 8 days in media including parathyroid hormone (PTH), 1,25-dihydroxyvitamin D3 [l,25(OH)2D3] or 17β-estradiol (E2), respectively. After 8 days of culture, tartrate-resistant acid phosphatase (TRAP) activity, a marker for osteoclasts, was enzymehistochemically detected on the cultured marrow cells and these cells were also stained with Mayer's hematoxylin. Thereafter, TRAP-positive multinuclear cells were counted as osteoclast-like cells, and the effects of the calcium regulating hormones on the osteoclast differentiation from medullary bone marrow cells were indicated as the percentage (%) of the controls when vehicle was added in place of the hormones. As a result, treatment with PTH or l,25(OH)2D3 significantly stimulated the differentiation of osteoclast-like cells, up to 218.3% or 159.2%, respectively. On the other hand, E2 treatment resulted in a moderate, but not significant, suppression of differentiation (85.8% of controls). These results indicate that PTH and l,25(OH)2D3 stimulate medullary bone osteoclast differentiation, but E2 is likely inhibitory. © 2004, Japan Poultry Science Association. All rights reserved.

DOI： 10.2141/jpsa.41.307

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DOI： 10.2141/jpsa.39.256

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：ELSEVIER SCI LTD  

Changes in the localization of cathepsin D in postmortem and pressurized rabbit muscles were investigated by immunoelectron microscopy. The anti-cathepsin D monoclonal antibody strongly labeled a large vesicle in a subsarcolemmal part of the cell, which strongly suggests that this is the primary lysosome. The liberation of the cathepsin D entrapped in the lysosomes in the muscle prepared immediately after death proceeded with the progress of the conditioning. The release of almost all cathepsin D from the lysosomes and its absorption on the myofibrils were observed in the muscle conditioned for 14 days. The accumulation of lysosomes having various volumes and shapes accompanied with the disruption of myofibrillar structure was also observed. The liberation of cathepsin D from the lysosomes can be attributed to the modification of membranes permeability of the lysosomes during conditioning. When the muscle was pressurized at 100 MPa, the modification of the round shape of the lysosome was observed. With the increase of the pressure applied to the muscle, the release of cathepsin D from the lysosome due to the disruption of membrane was accelerated, and absorption of the released cathepsin D on the myofibrils was observed. From the results obtained, it was clear that the changes in the localization of cathepsin D accompanied with the modification of lysosomes induced by the brief exposure to high pressure were drastic in comparison with that in the muscle during conditioning. (C) 2002 Elsevier Science Ltd. All rights reserved.

DOI： 10.1016/S0309-1740(01)00214-5

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DOI： 10.1046/j.1344-3941.2002.00071.x

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：ASIAN-AUSTRALASIAN ASSOC ANIMAL PRODUCTION SOCIETIES  

Avian calcium metabolism in an egg-laying period is extraordinary when compared with all other classes of vertebrates. The hard eggshell consists of calcium carbonate (CaCo3). About 60-75% of the calcium in the eggshell is derived from dietary sources and the remaining 25-40% from skeletal stores, called "medullary bone". Medullary bone is specifically developed in marrow cavities of long bones and plays an important role as a calcium reservoir for eggshell formation. An onset of medullary bone formation coincides with initiation of ovarian follicle maturation and the secretion of gonadal hormones, estrogen and androgen. Therefore their formation requires the combined influence of both estrogen and androgen. However, not only gonadal hormones but 1,25-dihydroxy vitamin D-3 is necessary for the formation of fully mineralized medullary bone. There are osteoblasts, bone-forming cells, and osteoclasts, bone-resorbing cells, on the hen medullary bone surface. In the domestic hen, medullary bone formation and resorption alternately occur during the 24-hour egg-laying cycle. These cyclic changes in medullary bone metabolism depend on the plasma levels of endogenous hormones fluctuating during the egg-laying cycle. Namely, when an egg is in the infundibulum, magnum or isthmus of the oviduct, osteoblasts actively form medullary bone with the induction of estrogen secreted by matured follicles. On the other hand, when an egg enters into the shell gland and begins to be calcified, the mobilization of calcium for eggshell formation causes a decrease of plasma ionized calcium concentrations. Consequently, to compensate the plasma calcium concentrations to be normal, parathyroid hormone is secreted from parathyroid glands to accelerate the osteoclastic bone resorption. With the end of eggshell formation, calcitonin is secreted from ultimobranchial glands thus inhibiting the osteoclastic bone resorption. In recent years, a number of manmade chemicals have shown to be able to mimic estrogen and androgen. It has been hypothesized that alterations in the normal pattern of endocrine systems, seen in some populations of wildlife, are linked with exposure to these chemicals. Interestingly, the medullary bone formation is induced by both estrogen and androgen, and the receptors of endocrine disruptors such as estrogen, androgen and aryl hydrocarbon (dioxin) receptors are highly expressed in the medullary bone. Therefore, it is sure that medullary bone would be a useful model to elucidate the effects of endocrine disruptors on wildlife and humans.

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：ELSEVIER SCI LTD  

Changes in the connectin/titin localization in post-mortem and pressurized chicken muscles were investigated by immunoelectron microscopy. The anti-connectin monoclonal antibody, ID11, strongly labeled the sides of thick filaments near the H-zone and weakly labeled the sides of Z-line in the sarcomere prepared immediately after death. With the development of the muscle contraction, the shortening of the sarcomere and the dispersion of the connectin epitope near the H-zone were observed. With the gradual increase of the sarcomere length during further storage, the apparent increase of the width of the epitope in the A-band region stained by the antibody was observed, but the distance from the epitope to M-line remained almost the same length. In the case of high pressure treatment, significant changes in the labeling pattern of the antibody were observed with the increase of the pressure applied. The increase of the distance from the epitope to M-line and dispersion of the epitope were observed in the fiber pressurized at 100 MPa. These phenomena were accelerated with the increase of the pressure applied. The discontinuous dense materials labeled by the antibody at the thick filament near the H-zone were observed in the fiber bundles pressurized at 200 MPa or more. This is probably due to the accumulation of connectin molecule from ordinary location in the sarcomere, because of the pressure-induced destruction of the thick and connectin filaments, In the fiber bundles pressurized at 300 MPa, a significant increase in the distance from the epitope to M-line accompanied with the increase of the sarcomere length was observed. From the results obtained, it was clear that the changes in the location of the connectin epitope induced by the brief exposure to high pressure were drastic in comparison with that in the sarcomere during post-mortem storage. (C) 2001 Elsevier Science Ltd. All rights reserved.

DOI： 10.1016/S0309-1740(01)00070-5

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Closed population of 6 generations Duroc pigs was selected for higher productivity and better leg soundness. The leg joint lesions of 174 slaughtered pigs at approximately 90kg live weight were scored from 0 (no lesion) to 4 (very severe lesion) in optical, radiographical and histological standpoints throughout the six generations. Moreover, one thousand and three hundred eighty three pigs were scored from 1 (very good) to 5 (very weak) for leg weakness. Significant differences in the leg weakness score and joint lesion scores were observed among selected generations. Significant differences were observed between gilt and boar/barrow group in 9 traits. The heritabilities of joint lesion scores were low through intermediate. The heritabilities of histological joint scores were also low through intermediate. The phenotypic and genetic correlations between the scores of front (humerus and ulna) and rear legs (femur) were low. The genetic correlations between optical, radiographical and histological scores of the same joint were generally high. The phenotypic correlations between the leg weakness score and joint lesion scores were positive although statistically not significant from zero. The genetic correlations between the leg weakness score and joint lesion scores were generally intermediately or highly positive. The result suggested that selection based on joint lesion scores could lead to a better leg joint condition optically and histologically. It also suggested that selection based on leg weakness score could lead to the reduction of leg joint lesion according to the correlated response.

DOI： 10.2508/chikusan.71.353

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：POULTRY SCIENCE ASSOC INC  

We present evidence that the polar, matrix-forming osteoblasts are connected laterally to form an impervious layer of cells. Next, the possible mechanisms by which calcium ions are translocated across the layer of cells into sites of mineralization are analyzed. Finally, mechanisms of attachment of bone-resorbing osteoclasts are considered. Osteoclasts adhere to matrix, in part, though an arginine-glycine-aspartic acid (RGD)-dependent mechanism. Adherence is under control of parathyroid hormone and 17 beta-estradiol.

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Language：English   Publisher：Japan Poultry Science Association  

The characteristics of the bone matrices formed by osteogenic cells isolated from both chick embryonic calvaria and hen medullary bones were examined. Isolated calvarial cells had developed organelles in the cytoplasm compared to isolated medullary bone cells. The cells isolated from both types of bones exhibited alkaline phosphatase (ALP) activity. Isolated calvarial cells formed a matrix that contained small amounts of acid mucopolysaccharide and densely-distributed thick collagen fibers. On the other hand, isolated medullary bone cells formed a matrix that contained a large amount of acid mucopolysaccharide and sparsely-distributed thin collagen fibers. Both matrices displayed calcium deposits. These results suggest that cells isolated from embryonic chick calvaria and hen medullary bones have the capacity for osteogenesis, forming cortical bone-like matrix and medullary bone-like matrix, respectively.

DOI： 10.2141/jpsa.35.228

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Language：English   Publisher：JAPANESE SOCIETY OF VETERINARY SCIENCE  

In the present study, osteoclasts were isolated from hen medullary bones at the formative and resorptive phases. The cells were cultured on glass culture dishes and bone slices. After culturing, the adhesion activity of the isolated osteoclasts with the substrates was estimated with a light microscope, and the surfaces of the bone slices were observed with a scanning electron microscope. The results showed that the adhesion activity of tartrate-resistant acid phosphatase (TRAP)-positive osteoclasts is higher at the bone resorptive phase than at the bone formative phase, and this tendency in isolated osteoclasts was observed more frequently on the bone slices than on the glass culture dishes. Furthermore, scanning electron microscopy showed that the isolated osteoclasts in the bone resorptive phase adhered to the bone surface with developed-cytoplasmic projections and formed broad pits where collagen fibrils were exposed. On the other hand, isolated osteoclasts in the bone formative phase adhered to the bone slice with board-shaped cytoplasmic projections and did not form any pits. These results suggest that isolated osteoclasts in the bone resorptive phase have a high level of adhesion activity and actively resorb the bone, whereas isolated osteoclasts in the bone formative phase have a low level of adhesion activity and cease bone resorption. The procedure reported here is useful for studying the bone-resorptive mechanism of authentic osteoclasts.

DOI： 10.1292/jvms.60.573

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The localizations of calcium-adenosine triphosphatase (Ca²⁺ -ATPase) and carbonic anhydrase II (CAII) in the shell gland of the hen oviduct were examined by immunohistochemical technique.<br>Immunoreactions showing Ca²⁺ -ATPase localization were observed strongly in the luminal surface of the tubular gland cells and in the excretory duct of the tubular gland, but not in the epithelial cells. On the other hand, Immunoreactions showing CAII localization were observed strongly in the cytoplasm of epithelial cells and moderately in the cytoplasm of tubular gland cells. In the shell gland, these immunoreactions showing Ca²⁺ -ATPase and CAII localization were not different at between 3 hours and 12 hours after oviposition.<br>These results suggest that Ca²⁺ is secreted by active transport of Ca²⁺ -ATPase into the shell gland lumen from the tubular gland and HCO₃^- is produced and secreted in the surface epithelium rather than in the tubular gland.

DOI： 10.2141/jpsa.33.371

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Other Link： http://agriknowledge.affrc.go.jp/RN/2010542236

Language：English   Publisher：Japanese Society of Animal Science  

Medullary bones dissected from hen femurs at 3 hours (when osteoclasts cease bone resorption) and 12 hours (when osteoclasts actively resorb bone) after oviposition were cultured in BGJb media containing synthetic human parathyroid hormone (PTH) and synthetic eel calcitonin (CT), respectively, and the effects of PTH and. CT on the distributional patterns of actin filaments in the medullary bone osteoclasts were observed. Three hours after oviposition, the osteoclasts displayed strong fluorescence of actin filaments in the form of amorphous bands in the apical region of the cytoplasm, corresponding to clear zones. There was no change in the distributional patterns of actin filaments in osteoclasts after culturing without PTH for 24 hours. However, after culturing with PTH for 24 hours, central parts of the actin bands in the osteoclasts became obscure, and the osteoclasts displayed actin filaments as linear forms perpendicular to the bone surface. On the other hand, 12 hours after oviposition, actin filaments appeared as lines perpen-dicular to the bone surface in the apical region of the cytoplasm, where ruffled borders had developed. There was no change in the distributional patterns of actin filaments in osteoclasts after culturing without CT for 24 hours. However, after culturing with PTH for 24 hours, the linear actin filaments disappeared and the osteoclasts displayed amorphous bands of actin filaments in the apical region of the cytoplasm. These results suggested that PTH and CT would regulate osteoclastic bone resorption by modifying the organization of actin filaments in osteoclasts.

DOI： 10.2508/chikusan.67.526

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：ELSEVIER SCIENCE INC  

Distributions of actin filaments in osteoclasts on chicken medullary bone were examined during the egg-laying cycle. When an egg was in the magnum or isthmus of the oviduct at 6 h after oviposition, the osteoclasts lacked ruffled borders and attached to medullary bone surface via clear zones. Actin filaments were localized as amorphous bands in the bone site of the cytoplasm, corresponding to clear zones in the osteoclasts. When an egg was in the shell gland of the oviduct at 15 h after oviposition, the osteoclasts had long and slender, well-developed ruffled borders. Actin filaments were localized as lines perpendicular to the bone surface at the bone site of the cytoplasm. In medullary bone osteoclasts, actin filaments changed during the egg-laying cycle. The relationship between the actin filaments and the ultrastructure of the osteoclasts are discussed in this report.

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DOI： 10.2141/jpsa.31.392

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：CARFAX PUBL CO  

1. The ultrastructure of osteodasts on hen medullary bone during the egg-laying cycle was observed by electron microscopy.
2. At 0 to 6 h after oviposition, osteoclasts lacking ruffled borders were attached to the bone via the clear zone and appeared to have ceased bone resorption. Small vacuoles were scattered throughout this cytoplasm.
3. At 9 to 21 h after oviposition, most of the osteoclasts had ruffled borders and appeared to be resorbing bone. The ruffled borders at 15 h were well developed, whereas at 9, 18 and 21 h they were poorly developed or showed similarities in structure to the clear zone. The small vacuoles were concentrated under the ruffled borders at 9 h and then decreased.
4. These results demonstrate cyclic changes in osteoclasts during the egg-laying cycle, indicate that ruffled borders form at the beginning of bone resorption and suggest that they are derived from the clear zone and the small vacuoles. The results also indicate that the ruffled borders fuse into the clear zone and disappear at the completion of bone resorption.

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DOI： 10.2141/jpsa.30.16

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：Japanese Society of Animal Science  

Ultrastructural features of osteoclasts and osteoblasts on the medullary bone surface were observed in the cultured bones of laying hens via an electron microscope. The medullary bones were excised from the femur during the formative phase when the eggs were in the magnum of the oviduct and during the resorptive phase when the eggs were in the shell gland of the oviduct. During the formative phase, osteoclasts had abundant cytoplasm but no ruffled border and osteoblasts showed well developed cytoplasm containing a large amount of rough-surfaced endoplasmic reticulum within 48 hours of culturing. These characteristics of osteoclasts and osteoblasts did not differ from those before culturing. During the resorptive phase, on the other hand, osteoclasts had a ruffled border of finger-like cell processes within 72 hours and osteoblasts had poorly developed cytoplasm within 24 hours. These features of osteoclasts and osteoblasts did not differ from those before culturing. These results indicate that the function of osteoclasts and osteoblasts on the medullary bone surface is maintained for a short period of culturing.

DOI： 10.2508/chikusan.63.468

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