Transfer of spermatogenesis-related cDNAs into eel testis germ-somatic cell coculture pellets by electroporation: methods for analysis of gene function

Molecular Reproduction and Development
Chiemi I MiuraTakeshi Miura

Abstract

Genes encoding spermatogenesis-related substance (eSRSs) show unique expression patterns during spermatogenesis. To analyze their function, we developed a new assay system using gene transfer techniques combined with coculture of the eel germ-somatic cells. First, we investigated the efficacy of in vitro electroporation transfer of gene into germ-somatic cell pellets using green fluorescent protein (GFP) gene. Second, in order to define the function of the eSRSs, we electrophoretically transferred eel spermatogonial stem cell renewal factor (eSRS34) and eel spermatogenesis-preventing substance (eSRS21) genes into germ-somatic cell pellets. Presence of the transferred cDNA was examined by reverse transcription-polymerase chain reaction (RT-PCR). Furthermore, proliferating cells were detected histologically, after labeling with BrdU. Transfer of the eSRS34 gene induced spermatogonial stem cell renewal in the pellets. Moreover, 11-ketotestosterone (11-KT) treatment stimulated the proliferation of spermatogonia, which resulted in the appearance of late type B spermatogonia in the pellets. The proliferation of spermatogonia by 11-KT stimulation was suppressed by transfer of the eSRS21 gene. These results indicate that the transferre...Continue Reading

References

Dec 1, 1991·Molecular Reproduction and Development·C L BrazolotA M Verrinder Gibbins
Jul 1, 1991·Proceedings of the National Academy of Sciences of the United States of America·Takeshi MiuraY Nagahama
Feb 11, 1994·Science·M ChalfieD C Prasher
Aug 31, 1999·Development, Growth & Differentiation·Chiemi I MiuraKohei Yamauchi
Oct 21, 1999·Biochemical and Biophysical Research Communications·Takeshi MiuraK Yamauchi
Jan 5, 2000·The Journal of Experimental Zoology·Y YamazakiK Imoto
Feb 25, 2000·Molecular and Cellular Biology·Q LinA I Skoultchi
Oct 23, 2001·Genesis : the Journal of Genetics and Development·R Sussman
Jan 17, 2002·Journal of Immunological Methods·Zhi ChenRiitta Lahesmaa
Feb 21, 2002·Journal of Controlled Release : Official Journal of the Controlled Release Society·Nathalie DujardinVeronique Préat
Jan 28, 2004·Proceedings of the National Academy of Sciences of the United States of America·Kayoko KuritaNoriyoshi Sakai

Citations

Dec 20, 2015·ACS Applied Materials & Interfaces·Hongfei YuHaiyan Li
Oct 3, 2015·General and Comparative Endocrinology·Frank PfennigHerwig O Gutzeit
Aug 14, 2013·PloS One·Shin KobayashiFumitoshi Ishino
Mar 5, 2011·Annual Review of Analytical Chemistry·Adam Johan Bergren, Richard L McCreery
Dec 28, 2017·Human Reproduction Update·João Pedro Alves-Lopes, Jan-Bernd Stukenborg

Related Concepts

Freshwater eels
Spermiogenesis
Spermatogonia
Testis
Gene Transfer Techniques
cDNA Probes
Electroporation
Coculture Techniques
Green Fluorescent Proteins

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