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. Author manuscript; available in PMC: 2021 Feb 28.
Published in final edited form as: Nat Neurosci. 2020 Aug 31;23(10):1240–1252. doi: 10.1038/s41593-020-0695-6

Figure 1. Juvenile social isolation impairs activation of mPFC->pPVT projection neurons upon social exposure in adulthood.

Figure 1.

(A) (Left) Timeline showing weaning at p21, and subsequent 2 weeks of juvenile social isolation (jSI), followed by re-housing or continued control group housing (GH), and subsequent In vivo fiber photometry calcium imaging of GCaMP6f-expressing mPFC->pPVT neurons in behaving adult mice. (right) Selective viral expression of GCaMP6f in mPFC->pPVT projection neurons were achieved by injecting AAV1-DIO-GCaMP6f in mPFC and retrograde rAAV2-Cre in pPVT. (B) Representative localization of optic fiber and GCaMP6f expression in mPFC. Scale; 500μm. Experimental images were obtained from 42 mice, three images per mouse, with similar results obtained. (C) During fiber photometry imaging, mice were exposed to a novel mouse or novel object (order of object and social exploration was counterbalanced). (D) Optic fiber placement and proximity to injection sites were confirmed from all mice (GH: open circles, jSI: red circles). (E-F) Heat maps of GCaMP6f signals (red-blue: high-low) for each trial f trials (one trial per mouse) from all mice (upper panel) and averaged traces of GCaMP6f signals from mPFC->pPVT neurons (lower panel) of (E) GH mice (n=23 biologically independent mice) and (F) jSI mice (n=19 biologically independent mice). (right) Quantification of social- or object-evoked normalized GCaMP6f signals (mean Z score of 30 sec post stimulus introduction subtracted with mean Z score of 30sec baseline) from mPFC->pPVT neurons in adult GH or jSI mice. Social exposure evoked higher responses than object exposure in GH mice (two tailed paired t-test, t22=4.759, ****P=0.946×10−4) but not in jSI mice (two tailed paired t-test, t18=0.757, P=0.459). (G) jSI mice showed less difference in mPFC->pPVT neuron activity between social and object exposure (S-O) than that of GH mice (two tailed t-test, t40=2.913, **P=0.006, n=23 (GH), 19 (jSI) biologically independent mice). Data in E-G are presented as mean +/− s.e.m.