Effects of Selective Deletion of Tyrosine Hydroxylase from Kisspeptin Cells on Puberty and Reproduction in Male and Female Mice

Abstract

The neuropeptide kisspeptin, encoded by Kiss1, regulates reproduction by stimulating GnRH secretion. Kiss1-syntheizing neurons reside primarily in the hypothalamic anteroventral periventricular (AVPV/PeN) and arcuate (ARC) nuclei. AVPV/PeN Kiss1 neurons are sexually dimorphic, with females expressing more Kiss1 than males, and participate in estradiol (E2)-induced positive feedback control of GnRH secretion. In mice, most AVPV/PeN Kiss1 cells coexpress tyrosine hydroxylase (TH), the rate-limiting enzyme in catecholamine synthesis (in this case, dopamine). Dopamine treatment can inhibit GnRH neurons, but the function of dopamine signaling arising specifically from AVPV/PeN Kiss1 cells is unknown. We generated a novel TH flox mouse and used Cre-Lox technology to selectively ablate TH specifically from Kiss1 cells. We then examined the effects of selective TH knock-out on puberty and reproduction in both sexes. In control mice, 90% of AVPV/PeN Kiss1 neurons coexpressed TH, whereas in mice lacking TH exclusively in Kiss1 cells (termed Kiss THKOs), TH was successfully absent from virtually all Kiss1 cells. Despite this absence of TH, both female and male Kiss THKOs displayed normal body weights, puberty onset, and basal gonadotropin levels in adulthood, although testosterone (T) was significantly elevated in adult male Kiss THKOs. The E2-induced LH surge was unaffected in Kiss THKO females, and neuronal activation status of kisspeptin and GnRH cells was also normal. Supporting this, fertility and fecundity were normal in Kiss THKOs of both sexes. Thus, despite high colocalization of TH and Kiss1 in the AVPV/PeN, dopamine produced in these cells is not required for puberty or reproduction, and its function remains unknown.

Stress rapidly suppresses in vivo LH pulses and increases activation of RFRP-3 neurons in male mice

“…Double-label ISH for cfos+Kiss1 and cfos+Rfrp were performed on an additional set of brain sections in order to assess neuronal activation. Both single- and double-label ISH assays were performed using well-published protocols and validated P33 or DIG-labeled riboprobes, as previously reported (Poling, et al 2017; Semaan and Kauffman 2015; Stephens, et al 2016; Stephens, et al 2017). After ISH slides were developed and coverslipped, a custom automated image processing and quantification software program (Dr. Don Clifton, University of Washington) was used to analyze ISH slides under computer-assisted microscopy, by an investigator blinded to treatment groups.…”

GABAergic input through GABAB receptors is necessary during a perinatal window to shape gene expression of factors critical to reproduction such as Kiss1

“…Puberty onset was determined in females by the age of vaginal opening (VO) (89) and males by the age of preputial separation (PS) (53). Females were examined daily starting on PND21 and males starting on PND24.…”

“…In the AVPV/PeN, the only gene that showed a dramatic effect on its expression due to the GABA B R antagonist was tyrosine hydroxylase (Th), the limiting synthesizing enzyme for dopamine (19,89). CGP induced a sharp decrease in Th expression in females, thus promoting loss of the typical sexual dimorphism (female Ͼ male).…”

GLUT1-mediated glycolysis supports GnRH-induced secretion of luteinizing hormone from female gonadotropes

“…To induce the LH surge 43 , 67 , 68 , female C57BL/6 mice underwent bilateral ovariectomy. LH surge was induced through one of two paradigms.…”