PACAP is essential for the adaptive thermogenic response of brown adipose tissue to cold exposure

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   Figure 1

   Resting metabolic rate (RMR) in conscious, unrestrained Pacap^+/+ (females, n=10 and males, n=8) and Pacap^−/− (females, n=8 and males, n=6) mice housed at 24 and 4 °C. Data are expressed as mean±s.e.m. *P<0.05 indicates a temperature effect (24 vs 4 °C).
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   Figure 2

   Basal metabolic rate (BMR) and norepinephrine-induced maximal metabolic rate (MMR) under anesthesia in Pacap^+/+ (females, n=8 and males, n=7) and Pacap^−/− (females, n=5 and males, n=3) mice housed at 24 and 4 °C. Data are expressed as mean±s.e.m. *P<0.05 and **P<0.01 indicate a temperature effect (24 vs 4 °C within the same genotype).
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   Figure 3

   Norepinephrine-induced thermogenesis in Pacap^+/+ (females, n=8 and males, n=4) and Pacap^−/− (females, n=5 and males, n=2) mice housed at 24 and 4 °C. Data are expressed as mean±s.e.m. *P<0.05 and **P<0.01 indicate a temperature effect (24 vs 4 °C within the same genotype).
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   Figure 4

   Relative exchange ratio (RER) at 24 °C and after 3.5 weeks of cold exposure (4 °C) in Pacap^+/+ (females, n=8 and males, n=4) and Pacap^−/− (females, n=5 and males, n=2) mice. Data are expressed as mean±s.e.m. *P<0.05 indicates a temperature effect (24 vs 4 °C within the same genotype).
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   Figure 5

   Morphometric analysis of iBAT from Pacap^+/+ (n=6) and Pacap^−/− (n=5) mice following 3.5 weeks of cold exposure. (A) Hematoxylin and eosin-stained iBAT sections visualized at 60× magnification, along with their areal lipid proportions. (B) Visualization of vasculature in iBAT by fluorescence microscopy, along with vasculature quantification for both genotypes. *P<0.05 indicates a genotype effect (Pacap^+/+ vs Pacap^−/−).
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   Figure 6

   Representative images of hematoxylin and eosin-stained (A) gonadal (gWAT), 20× magnification and (B) inguinal ingWAT, 60× magnification, from Pacap^+/+ (n=6) and Pacap^−/− (n=5) mice following 3.5 weeks of cold exposure.
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   Figure 7

   Gene expression in iBAT of male Pacap^+/+ (n=9) and Pacap^−/− (n=8) mice after 3.5-week cold challenge. Target gene expression was normalized to three reference genes (18S (Rn18s), β-actin (Actb), and Gapdh). Data are expressed as mean±s.e.m. *P<0.05 indicates a genotype effect (Pacap^+/+ vs Pacap^−/−).
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   Figure 8

   Gene expression in gonadal white adipose tissue (gWAT) and inguinal white adipose tissue (ingWAT) of male Pacap^+/+ (n=9) and Pacap^−/− (n=8) mice after 3.5-week cold challenge. Target gene expression was normalized to three reference genes (18S (Rn18s), β-actin (Actb), and Gapdh). Data are expressed as mean±s.e.m. *P<0.05 indicates a genotype effect (Pacap^+/+ vs Pacap^−/−).

• © 2014 Society for Endocrinology