Abstract:Objective: To explore the regulatory effect of selfrenewal and differentiation of germline stem cells for CG5033 gene in the Drosophila testis. Methods: This study uses the UAS/Gal4 system to drive the expression of UASCG5033 RNAi in germline stem cells (nosGal4), differentiated spermatogonial cells (bamGal4) and cyst cells (tjGal4). Single adult F1 male fly with genotype of nos>CG5033 RNAi, bam>CG5033 RNAi or tj>CG5033 RNAi was hybridized with three wildtype (WT) virgin flies to observe whether they could obtain offspring flies. To understand the function of CG5033 gene in the stem cell niche, the expression patterns of Vasa, eyes absent(Eya), DEcad, Zn finger homeodomain 1(Zfh1) and 1B1 were observed by light microscopy and immunofluorescence staining in WT and CG5033 RNAi testes. Results: Nos>CG5033 RNAi and tj>CG5033 RNAi flies were male infertile when compared with WT flies, while bam>CG5033 RNAi flies do not affect male fertility. NosGal4 driven UASCG5033 RNAi testes were significantly smaller than that of the normal group. Immunofluorescence staining showed that nosGal4 driven UASCG5033 RNAi testes led to germ cell lost and compensatory increase of cyst cells when compared with the normal group. However, tjGal4 driven UASCG5033 RNAi testes resulted in the formation of testicular germ cell tumor; there was no observable abnormality in bam>CG5033 RNAi testes. Conclusion: CG5033 encoded protein can influence the selfrenewal ability of germline stem cells in the Drosophila testis, and CG5033 can affect the differentiation process of germline stem cells by somatic stem cells, eventually leading to the formation of germ cell tumor.
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