RESEARCH GRANTS

National Institutes of Health Academic Research Enhancement Award (AREA Grant, 1999-2002).

Title: "Galanin Regulation of Pituitary Reproductive Hormones"

Amount of award: $74,222.

Summary of proposed research activities:

A substantial body of evidence now exists which implicates a role for the hypothalamic neuropeptide galanin in the regulation of pituitary reproductive hormone secretion. Galanin neurons regulate pituitary luteinizing hormone (LH) secretion at two distinct levels: 1). at the level of the hypothalamus, where galanin indirectly regulates LH by stimulating the production of the hypothalamic releasing factor gonadotropin-releasing hormone (GnRH), and 2). at the level of the pituitary gland, where galanin directly stimulates the secretion of LH from gonadotropes and enhances GnRH-stimulated LH secretion. While the hypothalamic effects of galanin on GnRH secretion are relatively well-characterized, little is known about galanin regulation of LH secretion directly at the level of the pituitary gland. Experiments outlined in this proposal were designed to characterize the pituitary effects of galanin on reproductive hormone secretion in both males and females, using the rat as an animal model. Specific aim #1 of this proposal is to determine whether gonadal steroids alter gonadotrope sensitivity to galanin stimulation. This hypothesis will be tested in a series of in vivo infusion experiments examining LH secretory responses to galanin in 1) intact, castrated, and castrated + testosterone-replaced male rats; and 2) female rats at different timepoints during the four day estrous cycle. Studies in females will specifically address whether gonadotrope sensitivity to galanin stimulation is upregulated within the context of the proestrous (i.e. preovulatory) endocrine milieu. Experiments of Specific aim #2 are designed to characterize the galanin receptor subtype that is expressed in gonadotropes in the anterior pituitary gland. Double-label in situ hybridization experiments will be performed to determine if galanin type 2 receptor (GalR2) messenger RNA (mRNA) expression is present in anterior pituitary cells that respond to GnRH, and if so, to determine the relative levels of GalR2 mRNA expression in gonadotropes vs. other anterior pituitary cell types that respond to galanin. Specific Aim #3 of this proposal is to investigate the effects of galanin on reproductive hormone gene expression in the anterior pituitary gland. An in vitro perifusion system will be used to determine if galanin, in addition to stimulating the secretion of LH, also has an effect on the mRNA expression of the LHb subunit. Furthermore, the effects of galanin on GnRH receptor mRNA expression will be assessed. Results from these studies will provide important information regarding the role of galanin in regulating LH synthesis and secretion from pituitary gonadotropes, events that are critically important for maintaining reproductive competence in both males and females. Additionally, participation in the execution of experiments outlined in this proposal will provide a unique opportunity for undergraduate students to learn fundamental techniques in reproductive physiology and molecular biology, experiences that will serve them well as they pursue their graduate studies.

 

National Science Foundation Course, Curriculum and Laboratory Improvement Grant (CCLI Grant; 1999-2001).

Title of Grant : "Improvement of Molecular Biology Components of the Human Biology Curriculum: Incorporation of a Research-Based Learning Approach".

Amount of award: $44,701 from NSF, with a match of $44,701 from UW-Green Bay.

Summary of proposed research activities:

The Biology and Human Biology Programs at the University of Wisconsin (UW)- Green Bay wish to incorporate modern methods of molecular and cell biology into the undergraduate curriculum. Specifically, funds were obtained to purchase equipment that will allow students to perform in situ analyses of protein and/or gene expression. The Developmental Biology Laboratory Course will utilize the equipment to conduct laboratory exercises that utilize a research project approach in order to assess whether this learning approach enhances student scientific reasoning and analytical skills. Developmental Biology students will design experiments to analyze the tissue-specific expression of a gene during embryogenesis, perform labeling experiments for visualization of this expression in embryo sections generated with the requested cryostats, record and analyze their data using the requested image analysis station, and disseminate their scientific data at a student poster session at the end of the semester and possibly at a UW - System Student Research Conference at the end of the academic year. Cell Biology students will perform a similar laboratory project involving enzyme cytochemistry. Finally, other courses (Molecular Biology, Immunology, Microscopic Anatomy and Molecular Genetics) as well as a students conducting independent research projects will utilize the requested equipment for performing related laboratory projects or learning activities during the course of the academic year. The implementation of these techniques into the Biology and Human Biology curricula will provide students with sophisticated training in modern methods of molecular / cell biology, including tissue cryosectioning, immunocytochemistry, enzyme cytochemistry and in situ hybridization for the analysis of gene expression. Furthermore, while conducting these methods within the context of a research project and not a "canned" laboratory exercise, it is anticipated that student scientific reasoning / analytical skills will be sharpened.