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4 and 7.0; Bitplane), Adenylyl cyclase using the function spot to count nuclei and surface to measure the area or the volume of the different objects. The number of positive neurons was presented as means?��?SEM. Statistical analysis Data sets with more than two groups were analyzed using one-way analysis of variance (ANOVA) followed by Tukey's post hoc test. For statistical analyses of experiments involving insulin injections and inflammatory response, we performed two-way ANOVA followed by Tukey's post hoc test. Two-tailed Student's t-tests were used for comparisons involving only two groups. All data were presented as mean?��?SEM. P?click here trigger persistent changes in the function of hypothalamic neurocircuits that regulate energy and glucose homeostasis (Bouret 2009). We therefore estimated mRNA expression of hypothalamic neuropeptides critically involved in energy and glucose balance. We found no significant differences between CL8 control and CL12 mice in the expression of mRNA for ARC neuropeptide orexigenic and anorexigenic genes Agrp, NPY, and Pomc at 4 or 24?weeks of age, in response to fasting. We did note, however, slight but significant reduction in Agrp expression in CL 15, as compared to control mice, tested at 24?weeks of age (Fig.?(Fig.11A). Figure 1 Effects of crowded litter (CL) on hypothalamic neurocircuits. (A) Quantitative real-time PCR analysis of hypothalamic agouti-related peptide (Agrp), neuropeptide Y (NPY), and proopiomelanocortin (POMC) mRNA expression upon 18?h fasting at 4 and ... Hypothalamic axonal projections change buy GS-7340 rapidly in the first 3?weeks of postnatal development in mice (Bouret 2009). We paid particular attention to the development of projections from the ARC to the paraventricular nucleus of the hypothalamus (PVH), because of its well-established importance in the neural control of energy balance (Elmquist et?al. 2005). We analyzed the immunoreactivity of AgRP and ��-MSH containing fibers in PVH (Fig.?(Fig.1B).1B). Both parvo- and magnocellular regions of the PVH contained a high density of AgRP and ��-MSH fibers.