Elsevier

Biological Psychiatry

Volume 90, Issue 11, 1 December 2021, Pages 766-780
Biological Psychiatry

Archival Report
Sex Differences in the Role of CNIH3 on Spatial Memory and Synaptic Plasticity

https://doi.org/10.1016/j.biopsych.2021.07.014Get rights and content

Abstract

Background

CNIH3 is an AMPA receptor (AMPAR) auxiliary protein prominently expressed in the dorsal hippocampus (dHPC), a region that plays a critical role in spatial memory and synaptic plasticity. However, the effects of CNIH3 on AMPAR-dependent synaptic function and behavior have not been investigated.

Methods

We assessed a gain-of-function model of Cnih3 overexpression in the dHPC and generated and characterized a line of Cnih3−/− C57BL/6 mice. We assessed spatial memory through behavioral assays, protein levels of AMPAR subunits and synaptic proteins by immunoblotting, and long-term potentiation in electrophysiological recordings. We also utilized a super-resolution imaging workflow, SEQUIN (Synaptic Evaluation and Quantification by Imaging of Nanostructure), for analysis of nanoscale synaptic connectivity in the dHPC.

Results

Overexpression of Cnih3 in the dHPC improved short-term spatial memory in female mice but not in male mice. Cnih3−/− female mice exhibited weakened short-term spatial memory, reduced dHPC synapse density, enhanced expression of calcium-impermeable AMPAR (GluA2-containing) subunits in synaptosomes, and attenuated long-term potentiation maintenance compared with Cnih3+/+ control mice; Cnih3−/− males were unaffected. Further investigation revealed that deficiencies in spatial memory and changes in AMPAR composition and synaptic plasticity were most pronounced during the metestrus phase of the estrous cycle in female Cnih3−/− mice.

Conclusions

This study identified a novel effect of sex and estrous on CNIH3’s role in spatial memory and synaptic plasticity. Manipulation of CNIH3 unmasked sexually dimorphic effects on spatial memory, synaptic function, AMPAR composition, and hippocampal plasticity. These findings reinforce the importance of considering sex as a biological variable in studies of memory and hippocampal synaptic function.

Section snippets

Animals

Procedures were approved by the Institutional Animal Care and Use Committee at Washington University in St. Louis. Adult (8–16 weeks) male and female C57BL/6 wild-type (WT) (Cnih3+/+), Cnih3tm1a(KOMP)Wtsi, and Cnih3 knockout (KO) (Cnih3−/−) mice were kept in climate-controlled facilities with a 12-hour light/dark cycle and ad libitum access to food and water.

β-Galactosidase Staining

To visualize the anatomical expression of the Cnih3tm1a(KOMP)Wtsi gene in the brain, we stained the brains of homozygous mutant mice for

LacZ-Tagged Cnih3 Prominently Expressed in the Hippocampus

To identify anatomical expression of Cnih3 in the brain, we used a β-galactosidase staining assay to visualize the lacZ cassette contained within the Cnih3tm1a(KOMP)Wtsi gene of Cnih3tm1a(KOMP)Wtsi mice (51) (Figure 1A). No lacZ staining was observed in WT lacZ brains (Figure 1B). In lacZ+ brains, lacZ reporter expression was strongest in the prefrontal cortex, hypothalamus, cortical regions, amygdala, and hippocampus (Figure 1C–E, males and females) (19). Within the hippocampus, lacZ reporter

Discussion

Learning and memory play a pivotal role in animal behavior, and dysregulation of the memory mechanisms are found in a wide array of psychiatric disorders. In this study, overexpression of Cnih3 in the dHPC improved short-term spatial memory only in female mice, whereas short-term spatial memory was attenuated only in female Cnih3−/− mice. Impairments in spatial memory of female Cnih3−/− mice were particularly apparent during metestrus, a stage characterized by low circulating levels of

Acknowledgments and Disclosures

This work was funded by the National Institutes of Health (Grant Nos. DA041781, DA042581, DA042499, and DA045463 [to JAM]; Grant No. DA041883 [to JAM, ECN, and JDD]; Grant Nos. DA046436 and DA042620 [to ECN]; Grant No. I01BX005204 [to TTK]; and Grant No. GM008151 [to Washington University and HEF]), the BrightFocus Foundation (Grant No. A2017084S [to TTK]), the Brain Research Foundation (Grant No. BRFSG-2017-05 [to TTK]), and the McDonnell Center for Cellular and Molecular Neurobiology (to TTK).

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