Cell proliferation is increased in the hypothalamus of AgRP-Tfam mutant mice. Cell proliferation is increased in the hypothalamus of AgRP-Tfam mutant mice. A–D, Immunohistochemical analyses for endogenous cell proliferation markers Ki-67 (A, B) and PCNA (C, D) were carried out in 2.5- to 4-month-old male control and AgRP-Tfam mutant mice. E, F, A separate cohort of control (E) and mutant (F) mice was infused intracerebroventricularly with BrdU (3.2 μg/μl) via osmotic minipumps (flow rate of 0.15 μl/h for 6 weeks; Alzet model 2006; Durect). Immunofluorescence analysis of BrdU in the hypothalamus of controls and mutants is shown. G, Proportional increase of Ki-67-, PCNA-, or BrdU-positive cells in the mutants was compared with that of the controls. Detailed quantification of positive cells is shown in Table 1. Error bars represent SEM; **p ≤ 0.01. 3V, Third ventricle. Andrew A. Pierce, and Allison W. Xu J. Neurosci. 2010;30:723-730 ©2010 by Society for Neuroscience