Evidence for a GABAergic System in Rodent and Human Testis: Local GABA Production and GABA Receptors

Abstract

The major neurotransmitter of the central nervous system, gamma-aminobutyric acid (GABA), exerts its actions through GABA_A, GABA_B and GABA_C receptors. GABA and GABA receptors are, however, also present in several non-neural tissues, including the endocrine organs pituitary, pancreas and testis. In the case of the rat testis, GABA appears to be linked to the regulation of steroid synthesis by Leydig cells via GABA_A receptors, but neither testicular sources of GABA, nor the precise nature of testicular GABA receptors are fully known. We examined these points in rat, mouse, hamster and human testicular samples. RT-PCR followed by sequencing showed that the GABA-synthesizing enzymes glutamate decarboxylase (GAD) 65 and/or GAD67, as well as the vesicular GABA transporter vesicular inhibitory amino acid transporter (VIAAT/VGAT) are expressed. Testicular GAD in the rat was shown to be functionally active by using a GAD assay, and Western blot analysis confirmed the presence of GAD65 and GAD67. Interstitial cells, most of which are Leydig cells according to their location and morphological characteristics, showed positive immunoreaction for GAD and VIAAT/VGAT proteins. In addition, several GABA_A receptor subunits (α1–3, β1–3, γ1–3), as well as GABA_B receptor subunits R1 and R2, were detected by RT-PCR. Western blot analysis confirmed the results for GABA_A receptor subunits β2/3 in the rat, and immunohistochemistry identified interstitial Leydig cells to possess immunoreactive GABA_A receptor subunits β2/3 and α1. The presence of GABA_A receptor subunit α1 mRNA in interstitial cells of the rat testis was further shown after laser microdissection followed by RT-PCR analysis. In summary, these results describe molecular details of the components of an intratesticular GABAergic system expressed in the endocrine compartment of rodent and human testes. While the physiological significance of this peripheral neuroendocrine system conserved throughout species remains to be elucidated, its mere presence in humans suggests the possibility that clinically used drugs might be able to interfere with testicular function.

Expression of multiple glutamate transporter splice variants in the rodent testis

“…These roles include the production of gamma-aminobutyric acid and glutathione. Since gamma-aminobutyric acid appears to be synthesized in both sperm 57 and interstitial glia-like Leylig cells, 58 this would be compatible with the view that GLAST1a and GLAST1b might supply some glutamate for this synthetic process in Leydig cells, while multiple glutamate transporters might contribute to substrate provision in sperm. Similarly, high levels of glutathione and the synthetic machinery to produce it are present in the testis, 59 particularly in cells such as spermatogonia, spermatocytes and Sertoli cells.…”

Non-Neuronal Release of Gamma-Aminobutyric Acid by Embryonic Pluripotent Stem Cells

“…GAD has been characterized as an indicator of GABAergic cells, which are present in GABAergic neurons and other non-neuronal GABAergic cells [7,22,23]. Parallel expression of GAD and VIAAT further validates GABAergic identification [24,25]. Our demonstration of colocalization of GAD, VIAAT, and Oct4 in a single cell suggests that the GABA synthesis and package components are assembled together within PS cells.…”

GABA exists as a negative regulator of cell proliferation in spermaogonial stem cells

“…This also provided the evidence for the spatial regulation of SSCs by GABA signaling. The expression of the GABA A receptor subunits α1, α5, β1, β2, β3 and γ3 in SSCs is authentic because these subunits have also been detected in the testes in other studies [14,28]. The GABA A receptor is a heteropentameric chlorideselective channel assembled from various combinations of subunits in the brain [29].…”