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. Author manuscript; available in PMC: 2015 Mar 28.
Published in final edited form as: Neuroscience. 2014 Jan 23;263:216–230. doi: 10.1016/j.neuroscience.2014.01.022

Figure 6.

Figure 6

Neonatal maternal separation (NMS) and vaginal balloon distension (VBD) significantly disrupted protein expression within the colon. RT-PCR (A) and Western blot (B–C) was performed on colon from naïve and NMS21 mice that either did not (Naïve-ND and NMS21-ND) or did undergo VBD (Naïve-VBD and NMS21-VBD). A) NMS only had a moderate effect on mRNA expression in the colon, with a trend towards increased IL10 mRNA expression in NMS21 colon. Neither NMS nor VBD had a significant effect on IL6, TNFα, Artemin (Art), or nerve growth factor (NGF) mRNA expression in the colon. B) Representative Western blots are shown for CRF1, CRF2, and corresponding GAPDH protein expression with bands at 55, 47, and 35kD, respectively. NMS and VBD had a significant interaction effect on CRF2 protein expression in the colon with NMS21-ND expressing higher CRF2 protein than both naïve-ND and NMS21-VBD colon. CRF1 protein expression in the colon was not significantly affected by either NMS or VBD. C) Representative Western blots are shown for TRPV1, TRPA1, and corresponding GAPDH protein expression with bands at 85, 127, and 35kD, respectively. Similar to its effects on vagina and bladder, VBD significantly decreased TRPV1 protein expression in the colon with naïve-VBD expressing significantly less TRPV1 than naïve-ND colon. NMS and VBD both separately induced a significant increase in TRPA1 protein expression in the colon. NMS21-ND colon expressed significantly more TRPA1 protein than naïve-ND colon. Housekeeping gene: GAPDH (A). Brackets indicate significant impact of NMS (*p < 0.05), VBD (#p < 0.05) or an NMS/VBD interaction (++p < 0.01), two-way ANOVA; *p < 0.05 vs. naïve, # p < 0.05 vs. baseline, Bonferroni posttest.