Mapping GABA and Glutamate inputs to gonadotropin‐releasing hormone (GnRH) neurones in male and female mice

Abstract

Gonadotropin releasing hormone (GnRH) neurone function is dependent upon gonadal steroid hormone feedback, which is communicated in large part through an afferent neuronal network. The classical neurotransmitters gamma‐aminobutyric acid (GABA) and glutamate are important regulators of GnRH neurone activity and are implicated in mediating feedback signals. We aimed here to determine whether GABAergic or glutamatergic input to GnRH neurones differs between males and females and/or exhibits morphological plasticity in response to steroid hormone feedback in females. Tissue collected from GnRH‐green fluorescent protein (GFP) male and female mice in diestrus underwent immunofluorescent labelling of GFP and either the vesicular GABA transporter (VGAT) or the vesicular glutamate transporter 2 (VGLUT2). No differences in the densities or absolute numbers of VGAT‐immunoreactive (‐ir) or VGLUT2‐ir puncta apposed to GnRH neurones were identified between males and females. The most significant input from either neurotransmitter was to the proximal dendritic region, and 80% of VGAT‐ir puncta apposed to GnRH neurones co‐localised with synaptophysin. Putative inputs were also assessed in ovariectomised (OVX) female mice treated with negative (OVX+E) or positive (OVX+E+E) feedback levels of oestrogen, and OVX+E+E mice were killed during the expected GnRH/luteinising hormone surge. No differences in VGLUT2‐ir contacts to GnRH neurones were identified between animals under the negative feedback influence of oestrogen (OVX+E) or the positive influence of oestrogen (OVX+E+E), regardless of cFos activation status. In contrast, a significant elevation in putative GABAergic inputs to GnRH neurones at the time of the preovulatory surge was found in the cFos‐negative subset of GnRH neurones, both at the level of the soma and at the proximal dendrite. Together, these data suggest that although GABAergic and glutamatergic innervation of GnRH neurones is not sexually differentiated, cyclic fluctuations in steroid hormone feedback over the female estrous cycle results in plastic changes in GABAergic inputs to a subpopulation of GnRH neurones.

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