1. Loss of SORCS1 and SORCS3 increases hypothalamic expression of Agrp and Klf4
Loss of SORCS1 and SORCS3 increases hypothalamic expression of Agrp and Klf4 Expression of hypothalamic neuropeptides and hormones as assessed by quantitative (q)RT–PCR in overnight fasted and ad libitum fed mice at 21 weeks of age. Log2‐fold change expression in S1/3 KO relative to the expression in WT is shown. Agrp transcript levels are increased in S1/3 KO under both fasted and fed conditions. n = 8–11 (fasted) or 4–5 (fed) mice/group. Agrp, agouti‐related peptide; Npy, neuropeptide Y; Pomc, proopiomelanocortin; Crh, corticotropin‐releasing hormone; Trh, thyrotropin‐releasing hormone.Expression of hypothalamic neuropeptides and hormones as assessed by quantitative (q) RT–PCR in overnight fasted mice at 8 or 35 weeks of age. Log2‐fold change expression in S1/3 KO relative to the expression in WT is shown. n = 4–6 (8‐week‐old) or 14–16 (35‐week‐old) mice/group.Detection of NPY/AgRP neurons by native fluorescence of GFP (green) in the arcuate nucleus of Npy‐hrGFP mice either wild type (Npy‐GFP/WT) or homozygous deficient for SORCS1/3 (Npy‐GFP/S1/3 KO) at 10 weeks of age. Mice were fed ad libitum. Scale bar: 100 μm; 3V, third ventricle.The total number of NPY/AgRP neurons, as identified by GFP fluorescence, is not affected by loss of SORCS1/3. Six histological sections across the arcuate nucleus per Npy‐GFP/WT or Npy‐GFP/S1/3 KO mouse (as exemplified in panel C) were scored (n = 3 mice/group).Immunodetection of KLF4 (red) in the arcuate nucleus of Npy‐GFP/WT and Npy‐GFP/S1/3 KO mice. NPY/AgRP neurons were identified by GFP fluorescence (green). Mice were fed ad libitum (n = 6 mice/group). Scale bar: 100 μm; 3V, third ventricle.Quantification of the KLF4‐positive area in NPY/AgRP neurons on histological sections as exemplified in (E). Two sections per mouse in the middle part of the arcuate nucleus (the region 1.82–1.94 mm caudal to bregma) were analyzed. n = 3 (fasted) or 6 (fed) mice/group.Data information: Data are shown as mean ± SEM and were analyzed using a two‐tailed unpaired t‐test (A, B, D) or two‐way ANOVA with Bonferroni post‐test (F). *P < 0.05, **P < 0.01.
Aygul Subkhangulova et al. EMBO Rep. 2018;embr
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