Patricia D. Finn

The Stimulatory Effect of Leptin on the Neuroendocrine Reproductive Axis of the Monkey

Leptin acts as a metabolic activator of the neuroendocrine reproductive axis in several rodent species, but whether leptin plays a similar role in primates is unknown. To explore this question, we examined the effects of leptin on gonadotropin and testosterone secretion in male rhesus monkeys that were fasted for 2 days. Mean plasma levels of LH and FSH, LH pulse frequency, and LH pulse amplitude were significantly higher in leptin-treated animals compared with saline-treated controls during the second day of the fast. To identify targets for leptin’s action, we used in situ hybridization and computerized imaging to map leptin receptor (Ob-R) messenger RNA (mRNA) distribution. Ob-R mRNA was observed in the anterior pituitary and several areas of the brain, including the arcuate and ventromedial nuclei of the hypothalamus. Ob-R mRNA was coexpressed in both POMC and neuropeptide Y neurons in the arcuate nucleus, whereas little or no coexpression of Ob-R mRNA was evident in GnRH neurons. These results sugges...

Galanin’s Functional Significance in the Regulation of the Neuroendocrine Reproductive Axis of the Monkey

Galanin stimulates the neuroendocrine reproductive axis in the rat, but whether galanin acts similarly in primate species is unknown. To test the hypothesis that galanin acts within the hypothalamo-hypophyseal axis to stimulate luteinizing hormone (LH) and gonadotropin-releasing hormone (GnRH) secretion in the primate, galanin was administered either systemically or directly into the arcuate nucleus-median eminence of ovariectomized macaques (pigtailed or rhesus, respectively) that were maintained on estradiol. The mean plasma levels of LH were significantly elevated in pigtailed macaques after peripheral injection of galanin (2 mg) as compared with vehicle treatment. In rhesus monkeys, galanin (80 µM) administered by push-pull perfusion into the arcuate nucleus-median eminence did not significantly alter either GnRH or LH release. To determine whether in the monkey, as in the rat, subpopulations of medial forebrain GnRH neurons coexpress galanin mRNA, we used single- and double-label in situ hybridization and computerized imaging techniques. GnRH mRNA-containing cells were identified in both the medial and lateral forebrain of the female pigtailed macaque. No galanin mRNA expression was detectable in GnRH neurons located in either the medial preoptic area or mediobasal hypothalamus; however, within the substantia innominata a subset of GnRH mRNA-expressing neurons did coexpress galanin mRNA. Taken together, these results suggest that galanin induces LH release in primates, but galanin may not act directly on hypothalamic GnRH neurons. Presently, we have confirmed in another primate species the existence of GnRH gene expression in the lateral forebrain and discovered that a small subset of these neurons coexpress galanin. These particular cells may have a unique and as of yet undefined physiological function that is distinct from those GnRH neurons serving a hypophysiotropic function.

The regulation of galanin gene expression in gonadotropin-releasing hormone neurons.

In rats, galanin modulates luteinizing hormone (LH) secretion, and gonadotropin-releasing hormone (GnRH) neurons provide a possible source of this galanin. To understand galanins physiological role in GnRH neurons, we used double-label in situ hybridization and computerized image analysis to examine the regulation of galanin message in GnRH neurons. We found that galanin gene expression in GnRH neurons is regulated by sex steroids, induced coincident with the LH surge, and persists well after the completion of the LH surge, and that the induction of galanin message in GnRH neurons coincident with the LH surge is sexually differentiated neonatally. We postulate that the rise in galanin gene expression in GnRH neurons at the time of the LH surge serves to replenish galanin released with GnRH that is needed for the production of the LH surge, or that galanin is involved in physiological events that occur subsequent to the LH surge.

Discovery and characterization of an inhibitor of glucosylceramide synthase.

Targeting glycosphingolipid synthesis has emerged as a novel approach for treating metabolic diseases. 32 (EXEL-0346) represents a new class of glucosylceramide synthase (GCS) inhibitors. This report details the elaboration of hit 8 with the goal of achieving and maintaining maximum GCS inhibition in vivo. 32 inhibited GCS with an IC(50) of 2 nM and achieved maximum hepatic GCS inhibition after four or five daily doses in rodents. Robust improvements in glucose tolerance in DIO mice and ZDF rats were observed after 2 weeks of q.d. dosing. Four weeks of dosing resulted in decreased plasma triglycerides and reduced hepatic fat deposition. Thus, 32 provides insight into the amount of metabolic regulation that can be restored following achievement of maximal target knockdown.

CHAPTER 3 – Regulation of the Hypothalamic–Pituitary–Gonadal Axis: Role of Gonadal Steroids and Implications for the Menopause

Menopause is characterized by a marked decrease in estrogen production from the ovaries. The estrogen decrease that occurs not only results in a loss of reproductive function, but is also associated with a number of conditions that reduce the quality of life of this population of women. It has become clear over the past 10-15 years that the ubiquitous presence of the estrogen receptor (ER) in body tissues and organ systems provides the substrate for estrogenic regulation or modulation of multiple body functions. That knowledge has run in parallel with ever increasing evidence of the beneficial effects of estrogen replacement therapy after the menopause. What started originally as an approach to treat vasomotor symptoms and vaginal dryness has evolved into a comprehensive therapeutic approach that can span several decades after the menopause and is intended to prevent and/ or treat osteoporosis, cardiovascular disease, cognitive dysfunction, and possibly other neurodegenerative disorders such as Alzheimer disease. Central to this expansive approach to hormone replacement therapy is not only a more refined understanding of the mechanisms that lead to estrogens beneficial effects on many tissues and functions, but also to an understanding of the undesired side effects that result from estrogenic activity at sites that are not the intended therapeutic targets (i.e., uterus and breast). The concept of tissue selectivity in estrogen action has emerged in recent years and provides the cellular and molecular basis to understand the diversity of estrogen activity and the distinct mode of action of full versus partial estrogen agonists.

Design of Gut-Restricted Thiazolidine Agonists of G Protein-Coupled Bile Acid Receptor 1 (GPBAR1, TGR5)

Bile acid signaling and metabolism in the gastrointestinal tract have wide-ranging influences on systemic disease. G protein-coupled bile acid receptor 1 (GPBAR1, TGR5) is one of the major effectors in bile acid sensing, with demonstrated influence on metabolic, inflammatory, and proliferative processes. The pharmacologic utility of TGR5 agonists has been limited by systemic target-related effects such as excessive gallbladder filling and blockade of gallbladder emptying. Gut-restricted TGR5 agonists, however, have the potential to avoid these side effects and consequently be developed into drugs with acceptable safety profiles. We describe the discovery and optimization of a series of gut-restricted TGR5 agonists that elicit a potent response in mice, with minimal gallbladder-related effects. The series includes 12 (TGR5 EC50: human, 143 nM; mouse, 1.2 nM), a compound with minimal systemic availability that may have therapeutic value to patients with type 2 diabetes mellitus, nonalcoholic steatohepatitis, or inflammatory bowel disease.