Molecular Cloning of
Genes Related to Apoptosis in Spermatogenic Cells of Mouse
JIANG Hong, LI Lu-Yun, LU Guang-Xiu*
( Human Reproductive Engineering Research Institute, Hunan Medical
University, Changsha 410078, China )
Abstract 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.
Key words cryptorchidism； spermatogenic cell； apoptosis； suppression subtractive
hybridization； differential expression
*Corresponding author： Tel, 86-731-4497661； Fax, 86-731-4497661