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Morphological assessment of GABA and glutamate inputs to GnRH neurons in intact female mice using expansion microscopy.

Published version
Peer-reviewed

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Abstract

The roles GABAergic and glutamatergic inputs in regulating the activity of the gonadotrophin-releasing hormone (GnRH) neurons at the time of the preovulatory surge remain unclear. We used expansion microscopy to compare the density of GABAergic and glutamatergic synapses on the GnRH neuron cell body and proximal dendrite in dioestrous and pro-oestrous female mice. An evaluation of all synapses immunoreactive for synaptophysin revealed that the highest density of inputs to rostral preoptic area GnRH neurons occurred within the first 45 µm of the primary dendrite (approximately 0.19 synapses µm-1 ) with relatively few synapses on the GnRH neuron soma or beyond 45 µm of the dendrite (0.05-0.08 synapses µm-1 ). Triple immunofluorescence labelling demonstrated a predominance of glutamatergic signalling with twice as many vesicular glutamate transporter 2 synapses detected compared to vesicular GABA transporter. Co-labelling with the GABAA receptor scaffold protein gephyrin and the glutamate receptor postsynaptic density marker Homer1 confirmed these observations, as well as the different spatial distribution of GABA and glutamate inputs along the dendrite. Quantitative assessments revealed no differences in synaptophysin, GABA or glutamate synapses at the proximal dendrite and soma of GnRH neurons between dioestrous and pro-oestrous mice. Taken together, these studies demonstrate that the GnRH neuron receives twice as many glutamatergic synapses compared to GABAergic synapses and that these inputs preferentially target the first 45 µm of the GnRH neuron proximal dendrite. These inputs appear to be structurally stable before the onset of pro-oestrous GnRH surge.

Description

Keywords

GnRH, VGAT, VGLUT2, expansion microscopy, gephyrin, homer, oestrous cycle, synaptophysin, Animals, Dendrites, Female, Glutamic Acid, Gonadotropin-Releasing Hormone, Mice, Mice, Inbred C57BL, Mice, Transgenic, Microscopy, Neurons, Synapses, Vesicular Glutamate Transport Protein 2, Vesicular Inhibitory Amino Acid Transport Proteins, gamma-Aminobutyric Acid

Journal Title

J Neuroendocrinol

Conference Name

Journal ISSN

0953-8194
1365-2826

Volume Title

33

Publisher

Wiley
Sponsorship
Wellcome Trust (212242/Z/18/Z)
Health Research Council of New Zealand (hrc_0318d926)
New Zealand Health Research Council