The past two years of NIH funding have allowed us to explore the effects of ethanol (ETOH) both in vivo and in vitro on hypothalamic luteinizing hormone-releasing hormone (LHRH) release, and pituitary luteinizing hormone (LH) secretion (both basal and stimulated), microheterogeneity and bioactivity. The current proposal is an indepth extension of these studies. The effect of in vivo ETOH feeding on the synthesis of LHRH will be studied by measuring prepro-LHRH mRNA levels. In addition to synthesis, the post- translational processing of prepro-LHRH and LHRH. Next, the bioactivity of LHRH will be investigated with the use of pituitary cells from normal, non-ethanol exposed animals in culture studying LH release after the addition of LHRH obtained from the hypothalami of the ethanol-fed and control animals. The degradation of LHRH in both the hypothalamus and pituitary from ETOH-exposed and control animals will also be explored. While the above described studies investigate in vivo ethanol effects, the secretion of LHRH in vitro will be examined both basally and when stimulated by a variety of physiologically-relevant secretagogues including dopamine, norepinephrine, naloxone, prostaglandin E2, and potassium in the presence of different doses of ETOH. In addition to the hypothalamic studies, the effect of ETOH feeding on the pituitary will be further characterized. LHRH receptor number and affinity on pituitary membranes obtained from ETOH and control animals will be determined. Next, the effect of ETOH on the second messenger calcium system in pituitary cells will be examined in an attempt to quantitate changes in intracellular free calcium levels. In addition, the impact of ETOH on calmodulin translocation in the gonadotrope will be explored. Lastly, LH synthesis in the in vivo ETOH-fed animal will be examined by measuring LH mRNA. We believe that the above described studies will physiologically dissect the exact impact of ETOH on the LHRH-LH reproductive unit in an organized and concise manner. The proposed project will take approximately for years to complete.

Agency
National Institute of Health (NIH)
Institute
National Institute on Alcohol Abuse and Alcoholism (NIAAA)
Type
Research Project (R01)
Project #
2R01AA006755-04A1
Application #
3110087
Study Section
Alcohol Biomedical Research Review Committee (ALCB)
Project Start
1989-04-01
Project End
1992-03-31
Budget Start
1989-04-01
Budget End
1990-03-31
Support Year
4
Fiscal Year
1989
Total Cost
Indirect Cost
Name
Edward Hines Jr VA Hospital
Department
Type
DUNS #
City
Hines
State
IL
Country
United States
Zip Code
60141
Uddin, S; Wilson, T; Emanuele, M A et al. (1996) Ethanol-induced alterations in the posttranslational processing, but not secretion of luteinizing hormone-releasing hormone in vitro. Alcohol Clin Exp Res 20:556-60
Uddin, S; Emanuele, M A; Emanuele, N V et al. (1994) The effect of in vitro ethanol exposure on luteinizing hormone and follicle stimulating hormone mRNA levels, content, and secretion. Endocr Res 20:201-17
Halloran, M M; Emanuele, M A; Draski, L et al. (1993) Failure of ethanol to induce changes in gonadotropin gene expression in selectively bred ethanol-sensitive rats. Endocr Res 19:317-29
Kelley, M R; Jurgens, J K; Tentler, J et al. (1993) Coupled reverse transcription-polymerase chain reaction (RT-PCR) technique is comparative, quantitative, and rapid: uses in alcohol research involving low abundance mRNA species such as hypothalamic LHRH and GRF. Alcohol 10:185-9
Wilson 3rd, D M; Emanuele, N V; Jurgens, J K et al. (1992) Prolactin message in brain and pituitary of adult male rats is identical: PCR cloning and sequencing of hypothalamic prolactin cDNA from intact and hypophysectomized adult male rats. Endocrinology 131:2488-90
Emanuele, M A; Tentler, J J; Kirsteins, L et al. (1992) The effect of ""binge"" ethanol exposure on growth hormone and prolactin gene expression and secretion. Endocrinology 131:2077-82
Emanuele, M A; Tentler, J J; Halloran, M M et al. (1992) The effect of acute in vivo ethanol exposure on follicle stimulating hormone transcription and translation. Alcohol Clin Exp Res 16:776-80
Emanuele, M A; Tentler, J; Emanuele, N V et al. (1991) In vivo effects of acute EtOH on rat alpha and beta luteinizing hormone gene expression. Alcohol 8:345-8
Azad, N; Agrawal, L; Emanuele, M A et al. (1991) Neuroimmunoendocrinology. Am J Reprod Immunol 26:160-72
Emanuele, M A; Tentler, J; Reda, D et al. (1990) The effect of in vitro ethanol exposure on LHRH release from perifused rat hypothalami. Endocr Res 16:313-21

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