Journal: Frontiers in Endocrinology
Article Title: CLK2 in GABAergic neurons is critical in regulating energy balance and anxiety-like behavior in a gender-specific fashion
doi: 10.3389/fendo.2023.1172835
Figure Lengend Snippet: Deletion of CLK2 in GABAergic neurons leads to increased adiposity in female mice. (A) PCR analysis of mice tails genomic DNA. Clk2: the 238 bp and 324 bp bands correspond to the wild type (WT) or the floxed (loxP) alleles, respectively. Vgat: the 323 bp and ~200 bp bands correspond to the WT or the Vgat-Cre (Vgat Cre ) alleles, respectively. (B, C) Body weight (g) throughout the study (females (B) n = 16 for Vgat-Cre; Clk2 loxP/loxP , n = 12 for Clk2 loxP/loxP ; males (C) n = 6 for Vgat-Cre; Clk2 loxP/loxP , n = 15 for Clk2 loxP/loxP ); (D, F) Total fat mass (g) at DEXA analysis day at a mean age of 23 weeks (females (D) : n = 9 for Vgat-Cre; Clk2 loxP/loxP , n = 8 for Clk2 loxP/loxP ; males (F) n = 6 for Vgat-Cre; Clk2 loxP/loxP , n = 9 for Clk2 loxP/loxP ); (E, G) Fat weight composition (g/100 g of BW), represented by mesenteric (MAT), gonadal/epidydimal (GAT/EAT) and brown (BAT) adipose tissue at 25-28 weeks of age (females (E) n = 14 for Vgat-Cre; Clk2 loxP/loxP , n = 12 for Clk2 loxP/loxP ; males (G) n = 5-6 for Vgat-Cre; Clk2 loxP/loxP , n = 16 for Clk2 loxP/loxP ); (H, J) Average daily food intake (average 5-days; g/day) at 16-18 weeks of age in both genders (females (H) n = 9 for Vgat-Cre; Clk2 loxP/loxP , n = 6 for Clk2 loxP/loxP ; males (J) n = 6 for Vgat-Cre; Clk2 loxP/loxP , n = 9 for Clk2 loxP/loxP ); (I, K) mRNA levels of neuropeptides ( Agrp – Agouti-related peptide , Crh – a corticotropin-releasing hormone , Npy – neuropeptide Y, Pomc – proopiomelanocortin) determined by RT-PCR in hypothalamus of mice at 26-28 weeks of age (females (I) : n = 5 for Vgat-Cre; Clk2 loxP/loxP , n = 5 for Clk2 loxP/loxP ; males (K) n = 5 for Vgat-Cre; Clk2 loxP/loxP , n = 5 for Clk2 loxP/loxP ). Two-way ANOVA analyzed data from (B) and (C) with the Bonferroni post-test. Two-tailed unpaired Student’s t-test analyzed data from (D–K) . All bars and errors represent mean ± SEM. *P < 0.05 versus Clk2 loxP/loxP .
Article Snippet: Mice were anesthetized with a mixture of ketamine (100 mg/kg of BW) and xylazine (10 mg/kg of BW), and the body composition (whole-body fat and lean masses) was assessed using Dual-energy X-ray Absorptiometry (DEXA) system (Discovery Wi 83901 QDR Series; Hologic Apex Software v13.3:5, Hologic Inc) at 22-25 weeks.
Techniques: Reverse Transcription Polymerase Chain Reaction, Two Tailed Test
