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Identification, Cloning, and Initial Characterization of a Novel Mouse Testicular Germ Cell-Specific Antigen¹

^a Department of Immunology & Parasitology, Yamagata University School of Medicine, Yamagata 990-9585, Japan ^b Section of Bioprocess Engineering, Faculty of Agriculture, Yamagata University, Tsuruoka 997-8555, Japan ^c Departments of Anatomy and ^d Obstetrics & Gynecology, Jichi Medical School, Tochigi 329-0498, Japan ^e Department of Obstetrics & Gynecology, Center for Reproductive Biology Research, Vanderbilt University School of Medicine, Nashville, Tennessee 37232-2633 ^f Department of Cell Biology, Institute for Virus Research, Kyoto University, Kyoto 606-8507, Japan

ABSTRACT

A monoclonal antibody, designated TES101, was raised by immunizing BALB/c mice with an allogenic mouse testicular homogenate followed by immunohistochemical selection as the initial screening method. By searching the expressed sequence tag (EST) database with the N-terminal amino acid sequence of TES101 reactive protein, we found that the predicted amino acid sequence encoded by a mouse testicular EST clone matched the TES101 protein sequence. Sequence analysis of the clone revealed no homologous molecule in the DNA/protein database. Based on data obtained from N-terminal amino acid analysis of the TES101 protein, the derived amino acid sequence contained a signal peptide region of 25 amino acids and a mature protein region of 225 amino acids, which translated into a protein with a molecular weight of 24 093. Northern blot analysis showed that mRNA of the TES101 protein was found in testis but not in any other mouse tissues examined. Western blot analysis revealed that TES101 reacted with a 38-kDa band on SDS-PAGE under nonreducing conditions, and this reactivity was abrogated under reducing conditions. Immunoelectron microscopic studies demonstrated that the molecule was predominantly located on the plasma membrane of spermatocytes and spermatids but not in Sertoli cells or interstitial cells, including Leydig cells. Thus, the TES101 protein is a novel molecule present primarily on the surface of developing male germ cells. TES101 protein may play a role in the processes underlying male germ cell formation.

FOOTNOTES

First decision: 19 June 2000.

¹ This work was supported in part by Grant-in-Aid for General Scientific Research 11671593 and by project grants from the Center for Molecular Medicine of Jichi Medical School and the Ministry of Education, Science, Sports and Culture, Japan.

² Correspondence: Yoshihiko Araki, Department of Obstetrics & Gynecology, Center for Reproductive Biology Research, Vanderbilt University School of Medicine, 1161 21st Avenue South, Room U3305, Medical Center North, Nashville, TN 37232-2633. FAX: 615 343 7797; yoshihiko.araki{at}mcmail.vanderbilt.edu

³ The authors contributed equally to this work.

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