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Masatomi HOSOI Shoko HOSOI-TANABE Hideki SAWADA Masahiro UENO Haruhiko TOYOHARA Isao HAYASHI 《Fisheries Science》2004,70(4):629-637
ABSTRACT: One of the biggest and long-standing difficulties in investigation of larval ecology in the field is species-level identification. In the present study, we developed polymerase chain reaction–restriction fragment length polymorphism (PCR-RFLP) analysis based on the large subunit (LSU) rRNA gene (rDNA) D1/D2/D3 region for identification of multiple species of bivalve larvae using 14 species of bivalve collected from Maizuru Bay. The LSU rDNA D1/D2/D3 region of all analyzed species could be amplified by PCR using bvD1f/bvD3r primers, and RFLP analysis by Hae III digestion on the PCR products showed species-specific fragment patterns. Furthermore, this analysis applied to single bivalve larvae in Maizuru Bay revealed efficient amplification of the target region and the species-specific pattern from 80% of the larvae, 75% of which showed a pattern that matched a certain pattern of the adult bivalves. In addition, the analysis of inter- and intraspecies variation of the LSU rDNA D1/D2/D3 region using the sequence data of the genus Crassostrea from the DDBJ database showed high applicability of this RFLP analysis on closely related species. Because of the wide applicability and technical simplicity, this method can become the standard for the identification of bivalve larvae species once the sequence data of the LSU rDNA D1/D2/D3 region of many bivalve species have been accumulated. 相似文献
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Shu HASHIMOTO Nao SUZUKI Ami AMO Takayuki YAMOCHI Yoshihiko HOSOI Yoshiharu MORIMOTO 《The Journal of reproduction and development》2013,59(5):496-499
Effects of supporting materials during vitrification procedure on the morphologies
of preantral follicles of pig ovaries were assessed. Ovarian cortical sections of
prepubertal pigs were randomly allocated to 5 groups. The sections were vitrified
ultrarapidly with 5 different vitrification devices. The sections were put on 4 fine
needles (Cryosupport), on a thin copper plate, or on a carbon graphite sheet or were
sandwiched between copper plates or between carbon graphite sheets before cooling.
The cooling and warming rates with the graphite sheets were significantly higher than
those with the copper plates (P<0.05). A total of 3,064 follicles were analyzed
following HE staining after vitrification with 5 different devices. The morphologies
follicles vitrified on the Cryosupport or on the graphite sheet were well preserved
compared with those vitrified on the copper plate or between copper plates
(P<0.01). The morphologies of follicles vitrified between copper plates were
mostly damaged (P<0.05). Taken together, good thermally conducting material
supports follicle morphologies of ovaries cryopreserved with ultrarapid
vitrification. 相似文献
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MASATOMI HOSOI SATOSHI KUBOTA MASAKO TOYOHARA HARUHIKO TOYOHARA ISAO HAYASHI 《Fisheries Science》2003,69(2):395-400
ABSTRACT: In order to identify free amino acids (FAA) that are importantas intracellular osmolytes in Crassostrea gigas , we investigatedthe change in FAA content in the mantle exposed to an abrupt decreaseor increase in salinity. In hypo-osmotic adaptation, most FAA showedremarkable and synchronous decreases from 2 to 8 h, suggestingthat the non-selective efflux of FAA was mainly responsible forthe decrease in FAA. Taurine that accounted for approximately 80% oftotal FAA content contributed most significantly to the hypo-osmoticadaptation. In hyper-osmotic adaptation, significant increases inglycine, alanine, β-alanine, proline, arginine and taurinewere observed. Of these, alanine showed an immediate increase thatis important to short-term adaptation to hyper-osmolality, whiletaurine showed a slower and substantial increase that contributesto a long-term adaptation to hyper-osmolality. 相似文献
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Natsumi SHIMIZU Kimihiro UENO Ena KURITA Seung-Wook SHIN Takuji NISHIHARA Tomoko AMANO Masayuki ANZAI Satoshi KISHIGAMI Hiromi KATO Tasuku MITANI Yoshihiko HOSOI Kazuya MATSUMOTO 《The Journal of reproduction and development》2014,60(3):179-186
In the mammalian testis, the ubiquitin-proteasome system plays important roles in the process that promotes the formation of
mature sperm. We recently identified zygote-specific proteasome
assembly phaperone (ZPAC), which is specifically expressed in the mouse
gonads and zygote. ZPAC mediates a unique proteasome assembly pathway in the zygote, but the expression profile and function
of ZPAC in the testis is not fully understood. In this study, we investigated the possible role of ZPAC during mouse
spermatogenesis. First, we analyzed the expression of ZPAC and 20S proteasome subunit α4/PSMA7 in the adult mouse testis.
ZPAC and α4 were expressed in spermatogonia, spermatocytes, and round spermatids. In elongating spermatids, ZPAC was
expressed until step 10, whereas expression of α4 persisted until step 12. We then examined the expression profile of ZPAC
and α4 in a mouse model of experimental unilateral cryptorchidism. Consistent with appearance of morphologically impaired
germ cells following cryptorchidism, the ZPAC protein level was significantly decreased at 4 days post induction of
experimental cryptorchidism (D4) compared with the intact testis, although the amount of α4 protein persisted at least until
D10. Moreover, intense ZPAC staining was co-localized with staining of annexin V, an early indicator of apoptosis in
mammalian cells, in germ cells of cryptorchid testis, but ZPAC was also expressed in germ cells showing no detectable
expression of annexin V. These results suggest that ZPAC plays a role during spermatogenesis and raises the possibility that
20S proteasome mediated by ZPAC may be involved in the regulation of germ cell survival during spermatogenesis. 相似文献
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Masaya TAKAGI Takeshi KIKKO Masatomi HOSOI Isao HAYASHI Haruhiko TOYOHARA 《Fisheries Science》2004,70(4):682-687
ABSTRACT: We have cloned a cDNA encoding the matrix metalloproteinase (MMP) from oyster Crassostrea gigas . The clone contains a 1797 bp open reading frame encoding a protein of 599 amino acids. Oyster MMP (Cg-MMP) has a transmembrane domain at its C-terminal and a furin/prohormone convertase cleavage site at the end of a propeptide domain, which are commonly observed in membrane-type MMPs (MT-MMPs). This suggests that Cg-MMP is an MT-MMP. The deduced amino acid sequence of oyster MMP shares approximately 30% identity with human MT4-MMP and MMPs from fruit fly and hydra. Cg-MMP mRNA was detected in the gill, mantle and adductor muscle, and more intense signals in the northern blot analysis were recognized in the gill and adductor muscle. Similar tissue distribution was observed for tissue inhibitor of metalloproteinase (Cg-TIMP) in oyster. In response to hypoxic stress, the abundance of Cg-MMP mRNA was elevated in the gill, while that of Cg-TIMP mRNA remained almost constant. These findings suggest that promotion of collagen metabolism may be implicated in the hypoxic adaptation in oyster. 相似文献
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Mai INOUE Atsuhiko HASEGAWA Yuta HOSOI Katsuaki SUGIURA 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2016,78(2):347-350
The association between breed, gender and age and cardiovascular disorders in the insured dog population in
Japan was investigated, using multiple logistic regression analysis and data from 299,555 dogs insured between
April 2010 and March 2011. The overall annual prevalence of cardiovascular disorder diagnosis was 2.1%. Using
the Miniature Dachshund as the reference breed, Cavalier King Charles Spaniel had the highest odds of
cardiovascular disorder with a ratio of 16.2 (95% confidence interval: 14.4–18.2), followed by Maltese,
Pomeranian, Chihuahua and Shih Tzu. Male dogs had increased odds of 1.2 (1.1–1.3). The dogs had increased odds
of having cardiovascular disorder by 1.5 times as their age increased by one year. 相似文献
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