Identification of Spermatogenic Cells Expressing Protamine mRNA in Japanese Quail by RT-PCR

Abstract

This study was conducted by reverse transcription-polymerase chain reaction (RT-PCR) analysis to determine when the protamine mRNA is expressed in testicular germ cells during spermatogenesis in quail. Ejaculated sperm were obtained from the female cloacal gland after mating. Adult male quail were used for collection of the testicular spermatogenic cells. Twenty cells of ejaculated sperm and those of each mechanically dispersed testicular cells including pachytene spermatocytes, round spermatids and elongated spermatids, were picked up using micropipette under phase-contrast microscope. Total RNA was extracted, reverse-transcribed, and amplified by using either quail protamine or chicken S17 ribosomal protein specific primers. The PCR products were run on TAE gel electrophoresis. There was no protamine mRNA expression in spermatocytes, but the expression was detected in round spermatids and elongated spermatids, as well as in ejaculated sperm. These results suggest that protamine mRNA is expressed at haploid stage of quail spermatogenic cells.