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. 2020 Jun 12;57(8):3439–3457. doi: 10.1007/s12035-020-01938-x

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© The Author(s) 2020

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 5 — Impact of combined neonatal PCP and isolation rearing on hippocampal calbindin and 5-HT immunoreactivity in the dorsal hippocampus. Representative a, b calbindin immunoreactivity throughout a all subfields and b part of CA1 (indicated in a by a solid border), together with e higher magnification images of typical 5-HT immunoreactivity in CA1 strata oriens (s.o.) and radiatum (s.r.) locations (indicated in a by dashed borders). Red represents a, b calbindin or e 5-HT, and blue in a and the top portion of each image in e represents DAPI nuclear counterstain; the bottom portion of each image in e is a duplicate presented without the nuclear counterstain. Scale bars are equivalent to 100 μm a, b or 10 μm e. Mean ± SEM c number of calbindin-positive cells in s.o., s.r., and stratum lacunosum-moleculare (s.l-m) of the dorsal hippocampal subiculum/fasciola cinereum (sub/fc), CA1, and CA2/3, and f intensity of 5-HT immunofluorescence in s.o. and s.r. of dorsal hippocampal CA1 and CA3 and molecular (mol) and polymorphic (pol) layers of the dentate gyrus (DG), plus correlation analyses of d calbindin and g 5-HT immunofluorescence intensity in CA1 versus the NOD choice trial discrimination ratio (time exploring novel/total choice trial object exploration) following acute SB-399885. Male Lister hooded rats that received saline (1 ml/kg s.c.; Veh) or PCP (10 mg/kg) on PND 7, 9, and 11 were housed in groups (Gr; Veh only) or isolation (Iso; Veh and PCP) from weaning on PND 21, then underwent NOD on three occasions (PND 57–80) before tissue collection (PND 78–80), to receive vehicle (0.5% methylcellulose 1% Tween 80; 1 ml/kg i.p. 30 min before the familiarization trial), SB-399885 (10 mg/kg), or MMPIP (10 mg/kg) on separate test days in a pseudorandom order and serve as their own controls (n = 13–14 per neurodevelopmental condition). Calbindin immunoreactivity a was strongest in the dentate gyrus and stratum pyramidale, and remaining cells matched the distribution of GABA interneurons. The b intensity of calbindin immunoreactivity in CA1 and c number of calbindin-positive cells throughout dorsal hippocampal subfields and cell layers were both influenced by neurodevelopmental condition (P < 0.05 and P < 0.01, respectively), and although there were some reductions in single-hit Veh-Iso, these were much more extensive in PCP-Iso and correlated with d NOD performance following acute administration of SB-399885. Patterns of e 5-HT immunoreactivity were consistent with labeling of fine varicose axons (marked by arrowheads) but labeling intensity f was not influenced by neurodevelopmental condition and g did not correlate with NOD flowing SB-399885 administration. ^†P < 0.05 Veh-Iso and ^†P < 0.05; ^††P < 0.01; ^††††P < 0.0001 PCP-Iso versus Veh-Gr; ^#P < 0.05 PCP-Iso versus Veh-Iso (three-way repeated measures ANOVA with Tukey post hoc)