Abstract
Krüppel-like factor 4 (KLF4) is a kind of zinc finger transcription factor, which is involved in terminal differentiation of epithelial cells and reprogramming of somatic cells to induced pluripotent stem (iPS) cells in mammals. In the present study, we identified a full-length cDNA of Klf4 in Zhikong scallop Chamys farreri (Cf-Klf4) and found that Cf-Klf4 presented a sexual dimorphic expression characteristic in C. farreri gonads. Cf-Klf4 expression was significantly higher in testes than in ovaries from growing stage to mature stage detected by quantitative real-time PCR, and was located in male gametes, except for spermatozoa during spermatogenesis through in situ hybridization and immunohistochemistry, while no positive signal was visible in female gametes during oogenesis. Furthermore, the knockdown of Cf-Klf4 in testes by means of in vivo RNA interference led to an obviously developmental retardance, lower gonadosomatic index, less male gametes and more apoptotic spermatocytes. Interestingly, we found that two out of eight scallops showed a hermaphroditic phenotype characteristic of male-to-female sex reversal when the Klf4 mRNA and protein levels were knocked down in males. These results verified that Klf4 plays an important role in testis functional maintenance and is necessary in spermatogenesis of C. farreri.
Highlights
Kruppel-like factor 4 (KLF4), a gut-enriched Kruppel-like factor (GKLF) or epithelial zinc finger (EZF), was firstly identified from the mouse serum-deprivation fibroblast cell cDNA library [1]
We identified a full-length cDNA of Klf4 in the Zhikong scallop C. farreri (Cf-Klf4), which is a dioecism with stable sex composition and important commercial shellfish in Northern China, and revealed that Cf-Klf4 was expressed in germ cells of C. farreri testes
We firstly reported the characteristics of Cf-Klf4 sexual dimorphic expression in C. farreri testes and ovaries, which is different from mammals
Summary
Kruppel-like factor 4 (KLF4), a gut-enriched Kruppel-like factor (GKLF) or epithelial zinc finger (EZF), was firstly identified from the mouse serum-deprivation fibroblast cell cDNA library [1]. It has the highly conversed domains of KLF family, the DNA-binding domains with three classical Cys2/His zinc fingers at the C-terminus binding GC-box or CACCCbox of the target DNA [2, 3], and two Kruppel-links, TGEKP(Y/F) X which connect zinc fingers [4]. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript
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