CNTD1 showed expression in oocytes, and Cntd1 knockout mice displayed prematurely depleted follicular pools (IMAGE)
Caption
(A) The quantitative PCR in human germinal vesicle (GV) oocytes and human granulosa cells (hGCs) obtained from patients who underwent IVF treatment showed that CNTD1 was highly expressed in oocytes. (B) Immunofluorescence analysis showed that the CNTD1 was expressed in follicles. Immunofluorescence staining for CNTD1 was shown in green, and the cell nuclei were counterstained with DAPI and shown in blue from the mouse ovary sections. (C) Strategy for the generation of Cntd1 knockout mice; it caused the deletion of Cntd1 exon 2 and a frameshift mutation by CRISPR/Cas9 technology. Wild-type Cntd1 mice were denoted as Cntd1+/+. Heterozygous Cntd1 knockout mice were denoted as Cntd1+/−. Homozygous Cntd1 knockout mice were denoted as Cntd1−/−. Sanger sequencing confirmed the creation of the deletion from the mouse genomic DNA. The primers used for genotyping the wild-type and Cntd1 knockout mice are denoted. (D) Agarose gel electrophoresis of the PCR products obtained from genomic DNA of wild-type and Cntd1 knockout mice. (E) The size of ovaries from Cntd1−/− females was dramatically reduced compared with the control ovaries at postnatal day 60 (PD60). (F) The follicles at different developmental stages were observed in Cntd1+/+ ovaries. Cntd1−/− mouse ovaries showed that there were normal follicles at different developmental stages during PD1 to PD30, but no developing follicle was observed at PD60 when compared with the ovaries from Cntd1+/+ and Cntd1+/− mice, which indicated prematurely depleted follicular pools in Cntd1−/− mice.
Credit
Liwei Sun, Yi Chen, Keya Tong, Weiwei Liu, Bei Liu, Yifan Wang, Guoning Huang, Jingyu Li
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