Outer dense fibers are specific subcellular components of mammal sperm flagella, apart from the axoneme and its associated proteins. As one of the outer dense fibers components, ODF3 is essential for the formation of ...Outer dense fibers are specific subcellular components of mammal sperm flagella, apart from the axoneme and its associated proteins. As one of the outer dense fibers components, ODF3 is essential for the formation of mammal sperm flagella. In the present study, we isolated the Odf3 gene, 1033 bp in length, from Zhikong scallop Chlamys farreri, which was named as Cf-Odf3. It was predicted that Cf-Odf3 encodes a protein of 240 amino acids which contains 5 discontinuous Pro-Gly-Pro repeats. The Cf-Odf3 transcripts were detected in both testis and ovary of C. farreri at various stages through q RT-PCR. The Cf-ODF3 protein synthesized by prokaryotes was purified using Ni^(2+)-NTA affinity chromatography and used to produce its polyclonal antibody. The antibody product was effective and specific as was examined by Western blotting. The Cf-Odf3 transcript and Cf-ODF3 protein were located in intragonadal somatic cells(ISCs), spermatogonia and spermatocytes of C. farreri testes, while no transcription of Cf-Odf3 and synthesis of Cf-ODF3 were detected in spermatids and spermatozoa. The location of Odf3 expression is different from that of vertebrates where ODF3 is synthesized specifically in spermatids and sperm flagella. Moreover, the Cf-Odf3 expression was also detected in ISCs, oogonia and oocytes of the scallop ovaries. Our finding revealed a different characteristic of Odf3 expression between scallop and vertebrates, which implied that Cf-Odf3 played a role in the gametogenesis of C. farreri.展开更多
基金supported by the National High Technology Research and Development Program of China(863 Program)(No.2012AA10A402)
文摘Outer dense fibers are specific subcellular components of mammal sperm flagella, apart from the axoneme and its associated proteins. As one of the outer dense fibers components, ODF3 is essential for the formation of mammal sperm flagella. In the present study, we isolated the Odf3 gene, 1033 bp in length, from Zhikong scallop Chlamys farreri, which was named as Cf-Odf3. It was predicted that Cf-Odf3 encodes a protein of 240 amino acids which contains 5 discontinuous Pro-Gly-Pro repeats. The Cf-Odf3 transcripts were detected in both testis and ovary of C. farreri at various stages through q RT-PCR. The Cf-ODF3 protein synthesized by prokaryotes was purified using Ni^(2+)-NTA affinity chromatography and used to produce its polyclonal antibody. The antibody product was effective and specific as was examined by Western blotting. The Cf-Odf3 transcript and Cf-ODF3 protein were located in intragonadal somatic cells(ISCs), spermatogonia and spermatocytes of C. farreri testes, while no transcription of Cf-Odf3 and synthesis of Cf-ODF3 were detected in spermatids and spermatozoa. The location of Odf3 expression is different from that of vertebrates where ODF3 is synthesized specifically in spermatids and sperm flagella. Moreover, the Cf-Odf3 expression was also detected in ISCs, oogonia and oocytes of the scallop ovaries. Our finding revealed a different characteristic of Odf3 expression between scallop and vertebrates, which implied that Cf-Odf3 played a role in the gametogenesis of C. farreri.
