TY - JOUR
T1 - Sex Differences in the Role of CNIH3 on Spatial Memory and Synaptic Plasticity
AU - Frye, Hannah E.
AU - Izumi, Yukitoshi
AU - Harris, Alexis N.
AU - Williams, Sidney B.
AU - Trousdale, Christopher R.
AU - Sun, Min Yu
AU - Sauerbeck, Andrew D.
AU - Kummer, Terrance T.
AU - Mennerick, Steven
AU - Zorumski, Charles F.
AU - Nelson, Elliot C.
AU - Dougherty, Joseph D.
AU - Morón, Jose A.
N1 - Funding Information:
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).
Funding Information:
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). Experiments utilizing the Zeiss LSM 880 Confocal with Airyscan and Zeiss AxioScan Z1 microscopes and image analyses utilizing Bitplane Imaris 3D visualization software were performed through the use of the Washington University Center for Cellular Imaging supported by Washington University School of Medicine, the Children's Discovery Institute of Washington University and St. Louis Children's Hospital (CDI-CORE-2015-505 and CDI-CORE-2019-813), and the Foundation for Barnes-Jewish Hospital (3770 and 4642). Viral vector packaging was performed by the Hope Center Viral Core at Washington University in St. Louis. We thank Susan Maloney, Ph.D. Rebecca Ouwenga, Ph.D. Carla Yuede, Ph.D. Nicolas Massaly, Ph.D. Grace Yuan, Krishna Vaidyanathan, Tania Lintz, Tamara Markovic, Ph.D. Justine Mae, Azra Zec, and Justin Meyer for technical assistance. A previous version of this article was published as a preprint on bioRxiv: https://www.biorxiv.org/content/10.1101/724104v2. CFZ serves on the Scientific Advisory Board and owns stock in Sage Therapeutics. All other authors report no biomedical financial interests or potential conflicts of interest.
Funding Information:
Experiments utilizing the Zeiss LSM 880 Confocal with Airyscan and Zeiss AxioScan Z1 microscopes and image analyses utilizing Bitplane Imaris 3D visualization software were performed through the use of the Washington University Center for Cellular Imaging supported by Washington University School of Medicine, the Children’s Discovery Institute of Washington University and St. Louis Children’s Hospital (CDI-CORE-2015-505 and CDI-CORE-2019-813), and the Foundation for Barnes-Jewish Hospital (3770 and 4642). Viral vector packaging was performed by the Hope Center Viral Core at Washington University in St. Louis.
Publisher Copyright:
© 2021 Society of Biological Psychiatry
PY - 2021/12/1
Y1 - 2021/12/1
N2 - 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.
AB - 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.
KW - AMPA receptors
KW - Hippocampus
KW - Memory
KW - Sex differences
KW - Super-resolution microscopy
KW - Synaptic plasticity
UR - http://www.scopus.com/inward/record.url?scp=85115194472&partnerID=8YFLogxK
U2 - 10.1016/j.biopsych.2021.07.014
DO - 10.1016/j.biopsych.2021.07.014
M3 - Article
C2 - 34548146
AN - SCOPUS:85115194472
SN - 0006-3223
VL - 90
SP - 766
EP - 780
JO - Biological Psychiatry
JF - Biological Psychiatry
IS - 11
ER -