Pituitary and hypothalamic mRNA analysis. A, Male (left) and female (right) control and deletion Adipocyte-lep-null pituitary mRNA levels. For all mRNA results, levels were determined using qRT-PCR and Δ-Δ-cycle threshold analysis, with cyclophilin A as an internal control. Relative quantification values are expressed with ± SEM, with control mRNA levels set at 1. A Student's t test (two tailed) was used to determine differences within sexes, with significance set at P < .05. Target pituitary mRNAs included FSH, GH, ghrelin, GHRH, GHS-R, LH, POMC, prolactin, and TSH. At least five pituitaries were used per experimental group per sex. B, mRNA analysis for male (left) and female (right) control and Adipocyte-lep-null mutant hypothalami, and at least five whole hypothalamus samples were analyzed per group per sex. Target hypothalamus mRNAs were GHRH, ghrelin, leptin receptor (all isoforms), and somatostatin. C, mRNA analysis for male (left) and female (right) control and Global-lep-null hypothalami, and n is at least five per group per sex.