Kisspeptin signaling is required for the Luteinizing hormone response in anestrous ewes following the introduction of males

Show simple item record De Bond, Julie-Ann P. Li, Qun Millar, Robert P. Clarke, Iain J. Smith, Jeremy T.
dc.contributor.editor Vaudry, Hubert 2013-04-16T11:13:34Z 2013-04-16T11:13:34Z 2013-02-28
dc.description Conceived and designed the experiments: JTS IJC. Performed the experiments: JPD QL JTS. Analyzed the data: JPD QL JTS. Contributed reagents/materials/analysis tools: RPM. Wrote the paper: JPD JTS IJC. en_US
dc.description Figure S1 Male exposure in anestrous ewes increased the number of kisspeptin cells in the rostral ARC. Sections representing the rostral, middle, and caudal regions of the ARC (as above) were chosen from each ewe and mounted on SuperFrost slides. Fluorescent immunocytochemistry was performed as previously described [20]. The primary kisspeptin antibody (AC566) was used at a concentration of 1:2000 and was visualized with a goat anti-rabbit secondary antibody (Alexa 448, 1:400; Molecular Probes Inc., Eugene, OR). Kisspeptin-ir cells were identified under fluorescent illumination, with a single observer counting the total number of cells. For each ewe, the number of kisspeptin-ir cells per section in each region was averaged to produce a mean (6SEM). A, Representative photomicrographs of the rostral ARC showing kisspeptin immunoreactive neurons (green). 3V, Third ventricle. Scale bar, 200 mm. B-C, The number of detectable kisspeptin neurons in the rostral ARC (B) was higher (P,0.05) in ewes exposed to males compared to control aCSF treated ewes. The number of kisspeptin neurons did not differ in the Mid (C), or Caudal (D) ARC. Data are the mean 6 SEM, n= 4 per group. en_US
dc.description.abstract The introduction of a novel male stimulates the hypothalamic-pituitary-gonadal axis of female sheep during seasonal anestrus, leading to the resumption of follicle maturation and ovulation. How this pheromone cue activates pulsatile secretion of gonadotropin releasing hormone (GnRH)/luteinizing hormone (LH) is unknown. We hypothesised that pheromones activate kisspeptin neurons, the product of which is critical for the stimulation of GnRH neurons and fertility. During the non-breeding season, female sheep were exposed to novel males and blood samples collected for analysis of plasma LH profiles. Females without exposure to males served as controls. In addition, one hour before male exposure, a kisspeptin antagonist (P-271) or vehicle was infused into the lateral ventricle and continued for the entire period of male exposure. Introduction of a male led to elevated mean LH levels, due to increased LH pulse amplitude and pulse frequency in females, when compared to females not exposed to a male. Infusion of P-271 abolished this effect of male exposure. Brains were collected after the male effect stimulus and we observed an increase in the percentage of kisspeptin neurons co-expressing Fos, by immunohistochemistry. In addition, the per-cell expression of Kiss1 mRNA was increased in the rostral and mid (but not the caudal) arcuate nucleus (ARC) after male exposure in both aCSF and P-271 treated ewes, but the percell content of neurokinin B mRNA was decreased. There was also a generalized increase in Fos positive cells in the rostral and mid ARC as well as the ventromedial hypothalamus of females exposed to males. We conclude that introduction of male sheep to seasonally anestrous female sheep activates kisspeptin neurons and other cells in the hypothalamus, leading to increased GnRH/LH secretion. en_US
dc.description.librarian am2013 en_US
dc.description.sponsorship Supported by the National Health and Medical Research Council of Australia (NHMRC) Project Grants 606538 and 1024346 and the Australian Research Council (ARC) Discovery Project DP120100521 and Future Fellowship FT0990986. en_US
dc.description.uri en_US
dc.identifier.citation De Bond J-AP, Li Q, Millar RP, Clarke IJ, Smith JT (2013) Kisspeptin Signaling Is Required for the Luteinizing Hormone Response in Anestrous Ewes following the Introduction of Males. PLoS ONE 8(2): e57972. DOI: 10.1371/journal.pone.0057972 en_US
dc.identifier.issn 1932-6203
dc.identifier.other 10.1371/journal.pone.0057972
dc.language.iso en en_US
dc.publisher Public Library of Science en_US
dc.rights © 2009 [Cameron et al]. This is an open-access article distributed under the terms of the Creative Commons Attribution License en_US
dc.subject Males en_US
dc.subject Kisspeptin signaling en_US
dc.subject Luteinizing Hormone Response en_US
dc.subject Anestrous Ewes en_US
dc.subject Female sheep en_US
dc.title Kisspeptin signaling is required for the Luteinizing hormone response in anestrous ewes following the introduction of males en_US
dc.type Article en_US

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