The isoform type of gamma subunits of GABA(A) receptor is a molecular determinant of its pharmacological characteristics. At present, the existence of GABA(A) receptor in mammalian sperm is still a controversy. By using degenerate primers designed according to highly conserved region in all three gamma (gamma1, gamma2 and gamma3) subunits cloned in rat brain, we performed reverse transcription polymerase chain reaction (RT-PCR) to examine the expression pattern of gamma subunits of GABA(A) receptor in rat testis. Only one 370 bp fragment was obtained from RT-PCR in rat testis and sequencing results showed that it represented gamma1 subunit, but not gamma2 or gamma3 subunit. Using the cloned fragment as probe, a 3.8 kb transcript which in size as same as gamma1 subunit in rat brain was detected in rat testis mRNA by performing Northern blot assay. Furthermore, results of in situ hybridization assay confirmed that gamma1 subunit was expressed in round spermatids and spermatozoa, maybe also in secondary spermatocyte. These evidences proved that gamma1 subunit of GABA(A) receptor is exclusively expressed in rat testis and this feature may be the structural basis of the specific function of GABA(A) receptors in sperm acrosome reaction.