Gene expression pattern of spermatogenic cell‐specific type 1 hexokinase splicing variants in mouse spermatogenic cells microdissected by laser caputure microdissection (LCM)

Abstract

Hexokinase is the first enzyme in glycolysis. We have cloned cDNAs representing spermatogenic cell specific type 1 hexokinase that has three splicing variants; Hk1_v1, Hk1_v2 and Hk1_v3. They share a spermatogenic cell specific region in N-terminus. In addition to the glycolysis, they seem to have an important role in spermatogenesis. However, the information on their function is limited. In the present study, we examined their gene expression patterns in spermatogenesis, respectively. Samples used were testes from adult C57BL/6 mice. Using LCM, the spermatogonia, pachytene spermatocyte, early spermatid, and late spermatid were microdissected. The purification of the spermatogonia and spermatocyte was confirmed using c-kit and hsp70-2, respectively, by RT-PCR analysis. The expression levels of Hk1_v1, Hk1_v2 and Hk1_v3genes in these cells were investigated by real-time RT-PCR analysis. Real-time RT-PCR analysis revealed that the expression levels of Hk1_v1, Hk1_v2 and Hk1_v3 had a tendency to be higher in early and late spermatids than in pachytene spermatocyte. Moreover, different expression patterns of three splicing variants of Hk1 genes were also shown in spermatids. These results suggest that their respective functions are not the same and might play cooperatively an important role in spermatogenesis.