The Effects of Glycogen Synthase Kinase-3beta in Serotonin Neurons
Figure 1
Characterization of snGSK3β-KO mice.
(A) Tail DNA genotyping of GSK3β-flox and Cre for selecting snGSK3β-KO and littermate WT mice. Lane 1, GSK3β-flox homozygote and ePet1-Cre (+) = GSK3β knockout (KO); Lane 2, GSK3β-flox homozygote and ePet1-Cre (-) = GSK3β wildtype (WT); Lane 3, GSK3β-flox heterozygote and ePet1-Cre (-); Lane 4, GSK3β-flox heterozygote and ePet1-Cre (+). Mice with genotypes in Lane 1 and Lane 2 were used for experiments. (B) Immunostain of TpH2-positive cells (red), GSK3β-containing cells (green), and GSK3β/TpH2 co-stained cells in the midbrain (4x magnification) and the dorsal raphe (DRN) (63x magnification), and immunostain of GSK3β-containing cells (green) in the cerebral cortex (4x magnification). (C) quantification of immunostained TpH2-positive cells (upper panel) and GSK3β/TpH2 co-stained cells (lower panel) in the ventral raphe (VRN), dorsal raphe (DRN), and dorsal lateral raphe (DLRN) areas. Upper panel: TpH2-positive neurons were counted to represent the number of serotonin neurons in each area. No statistically significant difference (p>0.05, n = 4–5 per genotype) in Student's t-test when each area of the two genotypes was compared. Lower panel: GSK3β in GSK3β/TpH2 co-immunostained cells were calculated as percent of total TpH2-positive cells in each area. *p<0.05, n = 4–5 per genotype, in Student's t-test when each area of the two genotypes was compared. (D) Phenotypic comparisons of body weight, body temperature, defecation, and Rotarod movement between WT and snGSK3β-KO mice. Mean ± SEM; Mixed model analysis showed no significant difference between the two genotypes in body weight or body temperature (p>0.05, n = 26–30 per genotype); Student's t-test showed no significant difference between the two genotypes in defecation (p>0.05, n = 7 per genotype) and Rotarod movement (p>0.05, n = 5 per genotype).