Characterizing the relationship between gonadotropin releasing hormone (GnRH), kisspeptin, and RFamide related peptide 3 (RFRP-3) neurons in the equine hypothalamus across the estrous cycle and in the anovulatory seasons.

To understand better the role that kisspeptin plays in regulating seasonal and estrous cycle changes in the mare, this study investigated the number, location and interactions between GnRH, kisspeptin and RFRP-3 neurons in the equine hypothalamus. Hypothalami were collected from mares during the non-breeding season, vernal transition and various stages of the breeding season. Fluorescent immunohistochemistry was used to label the neuropeptides of interest. GnRH cells were observed primarily in the arcuate nucleus (ARC), while very few labeled cells were identified in the pre-optic area (POA). Kisspeptin cells were identified primarily in the ARC, with a small number of cells observed dorsal to the ARC, surrounding the third ventricle (3V). The mean number of kisspeptin cells varied between animals and typically showed no pattern associated with season or stage of estrous cycle, but a seasonal difference was identified in the ARC population. Small numbers of RFRP-3 cells were observed in the ARC, ventromedial hypothalamus (VMH) and dorsomedial hypothalamus (DMH). The mean number of RFRP-3 cells appeared higher in pre-ovulatory animals compared to all other stages. The percentage of GnRH cell bodies with kisspeptin appositions did not change with season or stage of estrous cycle. The percentage of kisspeptin cells receiving inputs from RFRP-3 fibers did not vary with season or stage of estrous cycle. These interactions suggest the possibility of the presence of an ultra-short loop feedback system between these three peptides. The changes in RFRP-3 neurons suggest the possibility of a role in the regulation of reproduction in the horse, but it is unlikely to be as a gonadotropin inhibitory factor.

Insulin-like growth factor levels during pregnancy in the cow are affected by protein supplementation in the maternal diet.

To determine if dietary protein supplementation in early pregnancy alters total circulating insulin-like growth factor (IGF) levels, genetically similar heifers were fed diets containing different levels of protein in the first and second trimesters of gestation. The groups were: low/low (L/L), fed a diet containing 7% crude protein (CP) per kg/DM (low protein) in the first and second trimesters; high/high (H/H), fed a diet containing 14% CP per kg/DM (high protein) in the first and second trimesters; low/high (L/H), fed low protein in the first trimester and high in the second trimester and vice versa for the high/low (H/L) group. At day 62 of gestation, there was a significant difference (P<0.01) in IGF I concentrations between the high and low protein groups (149 versus 119 ng/ml, S.E. 5.9). There was a strong effect (P<0.001) of protein levels in the second trimester on IGF I levels on days 119, 153, and 183 of gestation but not at day 257. Mean IGF I levels for high and low nutrition in the second trimester were 157 and 97 (S.E. 6.6) for days 119, 191, and 88 (S.E. 12.6) for days 153 and 160, and 67 (S.E. 7.7) for day 183. At day 257, there was a significant interaction (P<0.01) between treatments with the means being 98(ab), 110(b), 116(b) and 79(a gamma) (means followed by a letter in common do not differ significantly, P<0.05) (S.E. 7.5) for H/H, H/L, L/H, and L/L, respectively. There was a significant (P<0.05) effect of protein supplementation in the first trimester on calf IGF I levels at birth with means being 42 and 25 (S.E. 5.2) for high and low protein supplementation, respectively. There was a significant (P<0.01) effect of protein supplementation in second trimester upon IGF II levels and a significant (P<0.05) negative correlation between calf birth weight and IGF II levels.