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Regular Article |
a Departments of Biomedical Sciences and
b Clinical Sciences, Tuskegee University, Tuskegee, Alabama 36088
c Department of Animal and Dairy Sciences, Program in Cell and Molecular Biosciences, Auburn University, Auburn, Alabama 36849
ABSTRACT
The fact that male estrogen receptor alpha (ER
) knockout mice are infertile indicates a role for this receptor in male reproduction. Here, objectives were to evaluate ER expression in male goat reproductive tissues at the transcriptional level using RNase protection assay (RPA) and in situ hybridization (ISH), and to clone a partial cDNA for caprine ER using reverse transcription-polymerase chain reaction (RT-PCR). For RPA and ISH procedures, a radiolabeled antisense cRNA probe, generated in vitro from the ovine oER8 cDNA template, was employed. Evaluations were made on individual samples obtained from adult goats. Labeled cRNA sense probe was used as a negative control in ISH. A 530-base pair amplicon was generated by RT-PCR from efferent ductules (EDs), epididymis (EP), and testis, cloned from the ED and EP, and sequenced. The caprine ER (cER) cDNA displayed 81%–96% sequence identity with that of other species. A signal indicative of ER mRNA was identified by both RT-PCR and RPA in all tissues, but was strongest in the ED. Compared with ED, ER signal was sixfold lower in the EP, and 66-fold lower in the testis. Similarly, strong ER expression was observed in ED epithelium, whereas little or no signal was detected in EP or testis by ISH. Thus, among different segments of the male reproductive tract and testis, the highest level of ER mRNA expression was found in epithelium of the ED.
First decision: 30 November 2000.
1 This work was supported by National Institutes of Health grants MBRS-5-S06-GM-08091 (to H.O.G.) and RCMI-5-G12RR03059-08; and by U.S. Department of Agriculture grant CSR-EES-ALX-TU-CTIF.
2 Correspondence: H.O. Goyal, Department of Biomedical Sciences, School of Veterinary Medicine, Tuskegee University, Tuskegee, AL 36088. FAX: 334 727 8177; goyalho{at}acd.tusk.edu
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