Influence of Nutrition during the Juvenile Period on Gene Expression Within the Hypothalamic Arcuate Nucleus and on Age at Puberty in Heifers
Allen, Carolyn C.
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Developmental changes within the hypothalamus are necessary for maturation of the reproductive neuroendocrine axis. Recent reports have implicated several neuronal networks in this process, but genes involved in their regulation have not been elucidated. Using a well-established model for nutritional induction of precocious puberty, objectives were to 1) use microarray technology to examine changes in gene expression within the arcuate nucleus (ARC) of the hypothalamus in pre-pubertal heifers fed high or low-concentrate diets, and 2) determine if high-concentrate diets are required for nutritional induction of precocious puberty. In Experiment 1, early-weaned, cross-bred heifers were fed either a high-forage/low-gain (HF/LG; 0.45 kg/d) or a highconcentrate/ high-gain (HC/HG; 0.91 kg/d) diet for 91 d. Analysis of microarray data indicated that 346 genes were differentially expressed (P < 0.05) between HC/HG and HF/LG heifers. Expression of three key metabolic genes [neuropeptide Y (NPY), agoutirelated protein (AGRP), and growth hormone receptor (GHR)] observed to be differentially expressed in the microarray analysis was investigated further by quantitative PCR. Real-time RT-PCR indicated that expression of NPY, AGRP and GHR was lower (P < 0.05) in HC/HG compared to HF/LG heifers. In contrast, concentrations of insulin (P < 0.05), IGF-1 (P < 0.002) and leptin (P = 0.1) were greater in HC/HG compared to HF/LG. For Experiment 2, 48 heifers were used in 2 replicates (24 heifers/replicate) in a 2 x 2 factorial design to examine the roles of diet type (HF vs HC) and rate of gain (LG, 0.45 kg/d vs HG, 0.91 kg/d) on age at puberty. Heifers were fed HC/HG, HC/LG, HF/HG or HF/LG (n = 12/group) for 14 wk, and then switched to a common growth diet (0.68 kg/d) until puberty. Heifers in both HG groups reached puberty at a younger age (54.5 ? 1.8 wk) than heifers in both LG groups (60.2 ? 1.9 wk; P < 0.04). A marked increase (P < 0.01) in serum concentrations of leptin occurred in HC/HG heifers between 24 and 30 wk of age. This increase in circulating leptin was not observed in other groups. Overall, results indicate that nutritional regulation of reproductive neuroendocrine development involves the control of NPY, AGRP and GHR expression. The abrupt increase noted for circulating leptin in heifers fed HC/HG diets, if timed and sustained appropriately, could represent an important temporal cue for activation of the neuroendocrine system and the onset of puberty.