Characterization of the relaxin family peptide receptor 3 system in the mouse bed nucleus of the stria terminalis

Apr 5, 2019The Journal of comparative neurology

Description of the relaxin receptor system in the mouse stress and anxiety control area

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Abstract

Approximately 48% of BNST RXFP3+ neurons are GABAergic.

BNST RXFP3+ cells are located in the dorsal BNST and include both inhibitory and excitatory neurons.
About a quarter of the GABAergic BNST RXFP3+ neurons also express the calcium-binding protein calbindin.
Approximately 41% of BNST RXFP3+ neurons co-express CaMKIIα, indicating they may be excitatory.
Around 35% of BNST RXFP3+ cells show expression of vVGluT2 mRNA, suggesting a glutamatergic subpopulation.
RXFP3+ neurons demonstrate direct hyperpolarization in response to a selective RXFP3 agonist, RXFP3-A2.
Chronic swim stress leads to a decrease in the proportion of RXFP3+ neurons expressing Type II calcium channels.

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The bed nucleus of the stria terminalis (BNST) is a critical node involved in stress and reward-related behaviors. Relaxin family peptide receptor 3 (RXFP3) signaling in the BNST has been implicated in stress-induced alcohol seeking behavior. However, the neurochemical phenotype and connectivity of BNST RXFP3-expressing (RXFP3+) cells have yet to be elucidated. We interrogated the molecular signature and electrophysiological properties of BNST RXFP3+ neurons using a RXFP3-Cre reporter mouse line. BNST RXFP3+ cells are circumscribed to the dorsal BNST (dBNST) and are neurochemically heterogeneous, comprising a mix of inhibitory and excitatory neurons. Immunohistochemistry revealed that ~48% of BNST RXFP3+ neurons are GABAergic, and a quarter of these co-express the calcium-binding protein, calbindin. A subset of BNST RXFP3+ cells (~41%) co-express CaMKIIα, suggesting this subpopulation of BNST RXFP3+ neurons are excitatory. Corroborating this, RNAscope® revealed that ~35% of BNST RXFP3+ cells express vVGluT2 mRNA, indicating a subpopulation of RXFP3+ neurons are glutamatergic. RXFP3+ neurons show direct hyperpolarization to bath application of a selective RXFP3 agonist, RXFP3-A2, while around 50% of cells were depolarised by exogenous corticotrophin releasing factor. In behaviorally naive mice the majority of RXFP3+ neurons were Type II cells exhibiting Iand T type calcium mediated currents. However, chronic swim stress caused persistent plasticity, decreasing the proportion of neurons that express these channels. These studies are the first to characterize the BNST RXFP3 system in mouse and lay the foundation for future functional studies appraising the role of the murine BNST RXFP3 system in more complex behaviors. h

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Characterization of the relaxin family peptide receptor 3 system in the mouse bed nucleus of the stria terminalis

Abstract

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