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During reproductive senescence in females, the function of GnRH neurons becomes compromised, and this may play a role in the transition from normal estrous cycles to acyclicity. One hypothalamic component of this dysregulation is an alteration in the stimulatory effects of glutamate, acting via N-methyl-D-aspartate receptors (NMDARs), on GnRH release. The present study examined whether GnRH neurons express the subunits necessary to make functional NMDARs, and how subunit expression may change during aging in association with compromised reproductive physiology. Colocalization of the three NMDAR subunits that are most abundant in the hypothalamus (NR1, NR2A, or NR2B) with GnRH perikarya was determined in female rats at different stages of the reproductive life cycle: young (3-4 months) rats with regular estrous cycles, middle-aged (8-10 months) rats with regular estrous cycles, middle-aged rats with irregular estrous cycles, and middle-aged acyclic rats in persistent estrus. The number, percent, and localization of GnRH perikarya expressing NR1, NR2A, or NR2B were mapped and quantified by double label immunofluorescence microscopy. Overall, each of the NMDAR subunits was present in a majority of GnRH neurons. There were no age- or reproductive status-related changes in coexpression of NR1 or NR2A subunits in GnRH neurons. However, coexpression of the NR2B subunit, which affects several functional channel characteristics, was significantly lower in young compared with middle-aged rats, irrespective of reproductive status. This may result in an age-related increase in the ratio of the NR2B to the NR1 and NR2A subunits on GnRH neurons. These data indicate that the majority of GnRH neurons express the proteins needed to receive direct NMDAR-mediated glutamatergic input, and that a change in the stoichiometry of the NMDAR pentamer occurs during aging that precedes, and may have consequences for, altered neuroendocrine function.

N-Methyl-d-Aspartate Receptor Subunit Expression in GnRH Neurons Changes during Reproductive Senescence in the Female Rat