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. 2021 Nov 23;12:749628. doi: 10.3389/fphar.2021.749628

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Copyright © 2021 Pilarzyk, Farmer, Porcher and Kelly.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Social isolation selectively decreases PDE11A4 protein expression in ventral hippocampal membrane fractions of adult mice. Western blots were used to measure PDE protein expression in fractionated samples from hippocampi of adult group-housed mice (GH) and adult mice that were single-housed for 1 h (SH), 1 day (SD), 1 week (SW) or 1 month (SM). Samples from the hippocampi of a Pde11a wildtype (WT) and knockout (KO) mouse are included as a positive and negative control, respectively, for the PDE11A4 antibody. Cerebellum is run as a negative control on PDE2A blots (not shown—see Supplementary Data 1) and striatum is run as a positive control on PDE10A blots (not shown—see Supplementary Data 1). (A) Both acute and chronic isolation decreases PDE11A4 (∼97kD) protein expression in the VHIPP membrane fraction (n = 9–10 samples/time point/sex; effect of group: F (4,65) = 5.95, p < 0.001; Post hoc: each group vs GH, P = 0.007 to p < 0.001), (B) but not the VHIPP soluble fraction (effect of group: F (4,66) = 0.991, P = 0.42) (C) nor the VHIPP nuclear fraction (effect of group: F (4,66) = 1.50, P = 0.21). Relative to group housing, social isolation does not significantly change PDE11A4 expression in the dorsal hippocampal (DHIPP) membrane fraction. A 2-way RM ANOVA for all groups fails equal variance; however, separate analyses for GH vs SH, SD, and SM as well as GH vs SW each pass normality and equal variance (GH vs SH, SD, and SM effect of group: F (3,51) = 4.76, FDR-P = 0.011 with Post hoc for each group vs GH, P = 0.057-0.868; GH vs SW effect of group: F (1,34) = 0.36, FDR-P = 0.552), soluble fraction (effect of group: F (4,64) = 1.74, P = 0.152), or nuclear fraction (effect of group: F (4,69) = 2.85, P = 0.03 with Post hoc for each group vs GH, P = 0.057-0.866). (G) To determine whether the isolation-induced decreases in VHIPP membrane PDE11A4 are specific to this PDE family, protein expression of both PDE2A (∼98 kDA) and PDE10A (∼80 kDa) were measured in VHIPP membrane samples. Neither (H) PDE2A (effect of group: F (4,65) = 1.64, P = 0.18) nor (I) PDE10A (effect of group: F (4,67) = 1.54, P = 0.20) differed in expression among the groups. Post hoc *vs GH, P = 0.007 to <0.001. Data are plotted as means ± SEMs. Brightness and contrast of blot images adjusted for graphical clarity.