Concentration and Distribution of Neuropeptide Y, Galanin, beta-endorphin, Vasoactive Intestinal Peptide and Gonadotrophin-Releasing Hormone in the Hypothalamus of Gilts during Oestrogen-Induced Surge Secretion of Luteinizing Hormone

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Contents The relationship of neuropeptide Y (NPY), galanin (GAL), β-endorphin (β-END) and vasoactive intestinal peptide (VIP) to GnRH neurons were determined during the estradiol-induced LH surge. In experiment 1, 16 ovariectomized (OVX) gilts received 15 μg estradiol benzoate (EB)/kg BW at 0800 h and were slaughtered at either 24 h (n = 5), 48 h (n = 6) or 72 h (n = 5) later and five were injected with corn oil vehicle (0 h controls). Concentrations of neuropeptides were determined in tissue extracts by RIA. In experiment 2, nine OVX gilts were injected with EB as in experiment 1 and killed at either 24, 48 or 72 h (n = 3) later and three were not injected with EB (0 h controls). Frozen sections were processed to localize neuropeptides. In experiment 1, all measured neuropeptides were highest in pituitary stalk median eminence (SME). The GnRH concentration was not different at any time point in medial basal hypothalamus (MBH), preoptic area (POA) or SME. The NPY content in MBH was lower at 24, 48 and 72 h after EB than at 0 h (p < 0.001), and lower in SME at 48 and 72 h than at 24 h (p < 0.05) and 0 h (p < 0.01), respectively. Concentration of GAL in SME was four times higher at 72 h than at 0, 24 or 48 h (p < 0.001). The VIP concentration increased in POA (p < 0.05) and MBH (p < 0.001) at 24 h and 72 h (p < 0.05). Concentration of VIP in SME was lower at 24 and 48 h than at 0 h (p < 0.05) and increased to more than twice (p < 0.05) by 72 h. Concentrations of β-END were not different at any time point in POA and MBH but the highest content of β-END in SME occurred at 24 h (p < 0.001). In experiment 2 a moderate number of GnRH-immunoreactive (IR) fibres were found in the periventricular area of the POA and in organum vasculosum of the laminae terminalis (OVLT). The GnRH-IR fibres formed networks in the external and internal layer of the median eminence (ME). At 24 h, GnRH-IR neurons and fibres in the POA and ME were more numerous and noticeable differences were found in the arcuate nucleus (ARC) and ventromedial nucleus (NVM). At 48 and 72 h, numbers of IR neurons and fibres were higher in the ARC and NVM, but no changes occurred in the POA and ME. The ARC contained a moderate number of NPY-IR fibres, but less numerous small cell bodies. Only a few NPY-IR perikarya and fibres were in the NVM and fibre density was similar at all times after EB injection. VIP-IR fibres were scarcely distributed mostly in the posterior POA and the internal layer of ME. The number of VIP-IR fibres was similar at all time points and regions. A moderate number of varicose β-END fibres supplied the POA, and they were especially dense near the OVLT, but the cell bodies were moderate in number and did not show strong immunoreactivity. In ME, ARC and NVM, the number of β-END immunoreactive structures was greater at 24 and 48 h than at 0 h. The number of β-END-IR nerve fibres in POA was higher at 72 h than at 0 h. Levels of all neuropeptides studied were similar in the POA and MBH and content of NPY, GAL and β-END was very high in the SME of the pig forebrain. The dynamic changes of NPY, GAL, VIP and β-END content in pig hypothalamus during the oestrogen-induced negative and positive feedback phases of LH secretion indicate their potential role in modulating GnRH release from the median eminence.

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