Neuronal COUP-TFII Regulation Figure 1. In vivo COUP-TFII expression in the hypothalamus during the fasting refeeding cycle. C57BL/6J mice are either fasted for 24 h (%) or fasted 24 h and refed on a regular diet (&). (A) RT-qPCR analysis of POMC (upper left panel), AgRP mRNA levels (upper right panel), NPY mRNA (lower left panel), Neuro D mRNA levels (lower right panel) (B) RT-qPCR analysis of COUP-TFII mRNA levels. Data are expressed as mean 6 SEM. (n = 5 mice/group) *P#0.05; **P#0.01 when compared to fasted conditions. (C) Autoradiogram of a western blot analysis of COUP-TFII protein expression in the hypothalamus. Upper panel: COUP-TFII protein, lower panel: b actin protein. doi:10.1371/journal.pone.0013464.g001 Recently a transcriptome analysis of the VMH has allowed the identification of some specific markers of the different cell subpopulations [12]. Among them, COUP-TFII had one of the highest levels of expression [12]. In order to delineate if COUP- TFII was more specifically expressed in neurons or astrocytes, we undertook a co-localization study with an astrocyte specific marker, the Glial Fibrilary Acidic Protein (GFAP). No overlapping expression with COUP-TFII protein was observed, suggesting that COUP-TFII was not expressed in astrocytes (data not shown). Among the identified VMH markers, the nuclear receptor steroidogenic factor 1 (SF1) is expressed in a large subpopulation of neurons involved in the central control of glucose homeostasis [21,22]. We addressed the possible overlap between COUP-TFII and SF1 expression within the VMH with a double immuno- PLoS ONE | www.plosone.org 3 October 2010 | Volume 5 | Issue 10 | e13464