NMD suppressed GRK6 expression in ARC. (a) The colorectal distention (CRD) threshold of NMD rats was significantly lower than that of control rats (n = 9 for each group, ***p < 0.001, two-tailed two- samples t-test). (b) The AWR score of NMD rats was significantly higher than the control rats (n = 9 for each group, *p < 0.05, **p < 0.01, two-way ANOVA). (c) GRK6 expression was significantly decreased in ARC of NMD rats (n = 6 for CON and n = 5 for NMD, *p < 0.05 two sample t-test). (d) The mRNA level of GRK6 was not altered in ARC of NMD rats (n = 4 for each group, p > 0.05, two sample t-test). There was no significant alteration in the expression of GRK6 in BLA (e) and IC (f) brain regions of NMD rats (n = 4 for each group, p > 0.05, two sample t-test). Compared with the age-matched control group (n = 6), there were no significant alterations in the thermal pain threshold ((g), p > 0.05, two-tailed two-sample t-test) and mechanical pain threshold ((h), p > 0.05, Mann–Whitney test) of hind limbs in NMD rats (n = 5). CON: control; NMD: neonatal maternal deprivation; ARC: arcuate nucleus; mRNA: messenger RNA; BLA: basolateral amygdala; IC: insular cortex; GAPDH: glyceraldehyde-3-phosphate dehydrogenase; GRK6: G protein-coupled kinase 6.