Figure 3

Dopamine-related biochemical markers indicate puberty as the starting point of the appearance of COMT-by-sex developmental dichotomy. Densitometric analysis of tyrosine hydroxylase (TH) in the PFC of COMT +/+, +/− and −/− male and female mice at three different stage of development, (a) pre-puberty: P15, (e) puberty: P35 and (i) post-puberty: 5–6 months old. Actin was used as a loading control and did not differ between groups in each developmental stage. Expression levels were normalized to the male wild-type group. Pre-pubertal COMT+/+ males N=6, females, N=6; COMT+/− males N=8, females N=7; COMT−/− males N=3, females N=3. Pubertal COMT+/+ males N=8, females, N=8; COMT+/− males N=13, females N=12; COMT−/− males N=5, females N=9. Post-pubertal COMT+/+ males N=5, females, N=6; COMT+/− males N=5, females N=9; COMT−/− males N=6, females N=5. *P<0.05 versus COMT +/+ within the same sex, ^#P<0.05 versus male within the same genotype. Densitometric analysis of total Akt levels (b, f and j), phosphoAkt (pAkt, THr308) (c, g and k), and pAkt (Ser473) (d, h and l) in PFC of COMT +/+, +/− and −/− male and female mice at three different stage of development, (b–d) pre-puberty: P15, (f–h) puberty: P35, and (j–l) post-puberty: 5–6 months old. For the measurement of total protein levels, actin was used as loading controls; whereas, for the measurement of phospho-protein levels, total protein levels were used as loading controls. Expression levels were normalized to the male wild-type group. Pre-pubertal COMT+/+ males N=6, females, N=6; COMT+/− males N=6, females N=6; COMT−/− males N=4, females N=3. Pubertal COMT+/+ males N=7, females, N=5; COMT+/− males N=5, females N=6; COMT−/− males N=6, females N=7. Post-pubertal COMT+/+ males N=4, females, N=5; COMT+/− males N=3, females N=3; COMT−/− males N=4, females N=6. *P<0.05 versus COMT +/+ within the same sex, ^#P<0.05 versus male within the same genotype, ^$P<0.05 versus males. PFC, prefrontal cortex.