Molecular Cloning of Genes Related to Apoptosis in Spermatogenic Cells of Mouse.

An apoptotic model of spermatogenic cells of mouse has been established by the method of operative cryptorchidism. By using light microscope and electronmicroscope, as well as TUNEL-stain (terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling method), it had been confirmed that operative cryptorchidism could successfully induce apoptosis of spermatogenic cells of mouse.On the basis of this research, 24 ESTs (expressed sequence tags) of the genes related to apoptosis in spermatogenic cells have been isolated and cloned by the method of suppression subtractive hybridization(SSH). After being confirmed with PCR and digestion by restriction enzyme PstI, they were sequenced and searched in GenBank for homology analysis. All of those were novel ESTs and were banked into GenBank with accession numbers. With the probes derived from three arbitrary cDNA sequences, Northern hybridization confirmed that there was obvious differential expression between cryptorchid testis and contralateral testis. These results suggest that experimental cryptorchidism is a suitable model for studying gene expression in the process of hypospermatogenesis and spermatogenic cell apoptosis, and ESTs isolated from this work may be involved in this apoptosis.