Kisspeptin, c-Fos and CRFR type 2 expression in the preoptic area and mediobasal hypothalamus during the follicular phase of intact ewes, and alteration after LPS

Increasing estradiol concentrations during the late follicular phase stimulate sexual behavior and the GnRH/LH surge, and it is known that kisspeptin signaling is essential for the latter. Administration of LPS can block these events, but the mechanism involved is unclear. We examined brain tissue from intact ewes to determine: i) which regions are activated with respect to sexual behavior, the LH surge and LPS administration, ii) the location and activation pattern of kisspeptin cells in control and LPS treated animals, and iii) whether CRFR type 2 is involved in such disruptive mechanisms. Follicular phases were synchronized with progesterone vaginal pessaries and control animals were killed at 0 h, 16 h, 31 h or 40 h (n = 4-6/group) after progesterone withdrawal (time zero). At 28 h, other animals received endotoxin (LPS; 100 ng/kg) and were subsequently killed at 31 h or 40 h (n = 5/group). LH surges only occurred in control ewes, during which there was a marked increase in c-Fos expression within the ventromedial nucleus (VMN), arcuate nucleus (ARC), and medial preoptic area (mPOA), as well as an increase in the percentage of kisspeptin cells co-expressing c-Fos in the ARC and mPOA compared to animals sacrificed at all other times. Expression of c-Fos also increased in the bed nucleus of the stria terminalis (BNST) in animals just before the expected onset of sexual behavior. However, LPS treatment increased c-Fos expression within the VMN, ARC, mPOA and diagonal band of broca (dBb), along with CRFR type 2 immunoreactivity in the lower part of the ARC and median eminence (ME), compared to controls. Furthermore, the percentage of kisspeptin cells co-expressing c-Fos was lower in the ARC and mPOA. Thus, we hypothesize that in intact ewes, the BNST is involved in the initiation of sexual behavior while the VMN, ARC, and mPOA as well as kisspeptin cells located in the latter two areas are involved in estradiol positive feedback only during the LH surge. By contrast, disruption of sexual behavior and the LH surge after LPS involves cells located in the VMN, ARC, mPOA and dBb, as well as cells containing CRFR type 2 in the lower part of the ARC and ME, and is accompanied by inhibition of kisspeptin cell activation in both the ARC and mPOA.

► We examined ewe brain tissue at various times in the follicular phase and after LPS. ► Kisspeptin cells are activated in the ARC and mPOA, during the LH surge. ► LPS was accompanied by failure of kisspeptin cells to become activated. ► LPS activated the VMN, ARC, mPOA and dBb. ► LPS increased CRFR type 2 immunoreactivity in the lower part of the ARC and ME.