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Endocrinology Vol. 140, No. 9 4070-4080
Copyright © 1999 by The Endocrine Society

ARTICLES

Discovery and Characterization of Endometrial Epithelial Messenger Ribonucleic Acids Using the Ovine Uterine Gland Knockout Model1

Thomas E. Spencer, Allison G. Stagg, Margaret M. Joyce, Guido Jenster, Christopher G. Wood, Fuller W. Bazer, Anne A. Wiley and Frank F. Bartol

Center for Animal Biotechnology and Genomics, Albert B. Alkek Institute of Biosciences and Technology, Texas A&M University System Health Science Center, and Department of Animal Science, Texas A&M University (T.E.S., A.G.S., M.M.J., F.W.B.), College Station, Texas 77843-2471; the Department of Urology, M.D. Anderson Cancer Center, University of Texas (C.A.W., G.J.), Houston, Texas 77030-4095; and the Department of Animal and Dairy Science, Auburn University (A.A.W., F.F.B.), Auburn, Alabama 36849-5415

Address all correspondence and requests for reprints to: Dr. Thomas E. Spencer, Center for Animal Biotechnology and Genomics, 444 Kleberg Center, Texas A&M University, College Station, Texas 77843-2471. E-mail: tspencer{at}ansc.tamu.edu

Prolonged exposure of the developing neonatal ovine uterus to a progestin from birth prevents uterine gland development and creates an adult endometrial phenotype characterized by the absence of glandular epithelium, the uterine gland knockout (UGKO) phenotype. This study used endometrium from normal and UGKO sheep to identify messenger RNAs (mRNAs) expressed differentially in the endometrial epithelium using the molecular techniques of mRNA differential display PCR (DD-PCR) and suppression subtractive complementary DNA (cDNA) hybridization (SSH). Sequence analyses of DD- and SSH-identified and cloned cDNAs indicated similarity of some to known mRNAs, including ß-lactoglobulin, alkaline phosphatase, type B and D endogenous sheep retroviruses, gp330/megalin, matrix Gla protein, and others. Other cDNAs were not similar to any known sequences and are considered novel, although some of these match human expressed sequence tags. In situ hybridization analyses of uteri from cyclic and pregnant ewes indicated that all DD-PCR- and SSH-identified mRNAs were expressed in either the endometrial lumenal and/or glandular epithelium, although some were also expressed in other uterine cell types. Northern and in situ hybridization analyses revealed that patterns of mRNA expression for most clones were affected by the day of the estrous cycle and pregnancy in a manner consistent with regulation by progesterone. Studies demonstrate the utility of the ovine UGKO model as a tool with which to identify known and novel uterine epithelial-specific genes. Cloned cDNAs identified here are expressed sequence tags useful for comparative and physical genetic mapping and may be used to reveal new factors and pathways regulating endometrial function.




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