Complementary DNA cloning and characterization of rat spergen-3,a spermatogenic cell-specific gene-3,expressed in mammalian spermatogenic cells
| Project/Area Number |
15570179
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Developmental biology
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| Research Institution | KYUSHU UNIVERSITY |
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Principal Investigator |
IIDA Hiroshi Kyushu University, Faculty of Agriculture, Associate Professor, 農学研究院, 助教授 (70150399)
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| Project Period (FY) |
2003 – 2004
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| Project Status |
Completed (Fiscal Year 2004)
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| Budget Amount *help |
¥3,600,000 (Direct Cost: ¥3,600,000)
Fiscal Year 2004: ¥1,200,000 (Direct Cost: ¥1,200,000)
Fiscal Year 2003: ¥2,400,000 (Direct Cost: ¥2,400,000)
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| Keywords | spermatogenesis / spermatid / spergen / nuclear protein / ankyrin repeat / DNA-binding protein / 細子細胞 / spergen / DNA結合蛋白質 |
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| Research Abstract |
By use of differential display in combination with cDNA cloning approach, we isolated a novel gene termed spergen-3,which had an open reading frame of 1,500-length nucleotides encoding a protein of 500 amino acids. Spergen-3 contains ankyrin repeat motifs and a putative nuclear localization signal. We found that the expression of spergen-3 was developmentally up-regulated and that it was exclusively expressed in testis. In situ hybridization revealed that spergen-3 mRNA was expressed in spermatocytes and round spermatids (step 1〜6). Immunohistochemical analysis with confocal laser-scanning microscopy demonstrated that spergen-3 protein was predominantly expressed in nuclei of late spermatocytes (stage IX〜XIV) and spermatids (step 1〜11), indicating the restricted expression of spergen-3 during spermatogenesis. In nucleoplasm of spermatogenic cell's nuclei, spergen-3 tended to localize in tide interchromosome space with relatively low density of DNA. Extraction experiments of spergen-3 from isolated nuclei of seminiferous tubules demonstrated that spergen-3 is released from nuclei by DNase I in the presence of NaCl but not by RNase. Affinity column of DNA could bind spergen-3 in the presence of NaCl and it was released from the column by decreasing the concentration of NaCl. These data suggested that spergen-3 might be associated with chromatin DNA in spermatogenic cells. We interpreted these data as a potential role of spergen-3 in spermatogenesis, especially in cell differentiation from late pachytene spermatocytes to spermatids or in early spermati differentiation.
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Report
(3 results)
Research Products
(13 results)
