Abstract
BackgroundIt has been proven that c-kit is crucial for proliferation, migration, survival and maturation of spermatogenic cells. A periodic expression of c-kit is observed from primordial germ cells (PGCs) to spermatogenetic stem cells (SSCs), However, the expression profile of c-kit during the entire spermatogenesis process is still unclear. This study aims to reveal and compare c-kit expression profiles in the SSCs before and after the anticipated differentiation, as well as to examine its relationship with retinoic acid (RA) stimulation.ResultsWe have found that there are more than 4 transcripts of c-kit expressed in the cell lines and in the testes. The transcripts can be divided into short and long categories. The long transcripts include the full-length canonical c-kit transcript and the 3′ end short transcript. Short transcripts include the 3.4 kb short transcript and several truncated transcripts (1.9-3.2 kb). In addition, the 3.4 kb transcript (starting from intron 9 and covering exons 10 ~ 21) is discovered to be specifically expressed in the spermatogonia. The extracellular domain of Kit is obtained in the spermatogonia stage, but the intracellular domain (50 kDa) is constantly expressed in both SSCs and spermatogonia. The c-kit expression profiles in the testis and the spermatogonial stem cell lines vary after RA stimulation. The wave-like changes of the quantitative expression pattern of c-kit (increase initially and decrease afterwards) during the induction process are similar to that of the in vivo male germ cell development process.ConclusionsThere are dynamic transcription and translation changes of c-kit before and after SSCs’ anticipated differentiation and most importantly, RA is a significant upstream regulatory factor for c-kit expression.
Highlights
It has been proven that c-kit is crucial for proliferation, migration, survival and maturation of spermatogenic cells
Transcription of c-kit in cell lines and testes Northern blots revealed at least 4 transcripts in the cells and testes (Figure 1B and C)
Even more c-kit transcripts expressed in c18-4, CRL-2053 and 5 dpp, 10 dpp and 60 dpp testes were assayed by Rapid amplification of cDNA ends (RACE) (Figure 2A, B, C, D)
Summary
It has been proven that c-kit is crucial for proliferation, migration, survival and maturation of spermatogenic cells. A periodic expression of c-kit is observed from primordial germ cells (PGCs) to spermatogenetic stem cells (SSCs), the expression profile of c-kit during the entire spermatogenesis process is still unclear. This study aims to reveal and compare c-kit expression profiles in the SSCs before and after the anticipated differentiation, as well as to examine its relationship with retinoic acid (RA) stimulation. Spermatogenesis starts from diploid spermatogonial stem cells (SSCs). The SSCs, known as type A single (As) Spg, are located on the basement membrane of the seminiferous tubules. The As Spg can self-renew or produce the type A paired (Ap) Spg. After successive divisions, Ap Spg differentiates, forms chains of 4, 8 or 16 aligned Spg (Aal) and migrates along the basement membrane.
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