INTRODUCTION In rodent, molecular markers of spermatogonia, spermatocyte, spermatid and sperm have been identified. It has been reported that DBA, PGP 9.5 and NanpG can be the markers for spermatogonia in pig. For further understanding the spermatogenesis of the pig on morphological and molecular level, we report identification of testicular cells in neonatal and pubertal pig testis, and a putative marker for spermatogonia and spermatid in pig testis. MATERIALS AND METHODS Neonatal (3 day) and pubertal testis (150 day) was cut and fixed in Bouin’s solution for immunohistochemistry (IHC). Gonocytes were isolated from neonatal testis for immunocytochemistry (ICC). Based on references (Phillips et al., 2010), thirteen antibodies (VASA, Oct4, NanoG, PGP 9.5, DAZL, SCF, GFR-alpha 1, PLZF, c-kit, integrin-beta1, Thy1, Sohlh1 and CD9) were used for IHC and ICC. Paraffin section was performed for IHC. Gonocytes were attached to the APS-coated slides for ICC. HRP-conjugated and florescent-labeled secondary antibody was used for IHC and ICC, respectively. RESULTS In histological analysis, spermatogonia and Sertoli cells, which are enclosed by seminiferous tubule and connective tissue, were observed in neonatal testis. However, complete spermiogenesis, including spermatocyte, spermatid and spermatozoa, was not observed in neonatal testis. Spermatocyte, spermatid and elongated spermatid were observed in pubertal testis but matured spermatozoa were not observed. As a result, two antibodies (PGP 9.5 and GFR-alpha1) of thirteen antibodies were available for IHC and ICC. As reported in other studies, PGP 9.5 protein was detected in spermatogonia of ne-onatal in IHC. In addition, it was observed in spermatogonia of pubertal testis. GFR- alpha1 protein was detected in spermatogonia of neonatal testis and spermatid of pubertal testis. In ICC, PGP 9.5 protein was detected in gonocytes as reported in other studies. GFR-alpha1 was also observed in gonocyte isolated from pig testis. In this study, we found that 1) only spermatogonia exist in neonatal pig testis (3 day), 2) GFR-alpha1 is a new marker for spermatogenesis in pig testis.

• Kyung Hoon Lee(Faculty of Natural Science, Division of Food Bioscience, Konkuk University, Chungju, Chungcheongbukdo, South Korea, 380-701)
• Hee Chan Kim(Faculty of Natural Science, Division of Food Bioscience, Konkuk University, Chungju, Chungcheongbukdo, South Korea, 380-701)
• Won Young Lee(Faculty of Natural Science, Division of Food Bioscience, Konkuk University, Chungju, Chungcheongbukdo, South Korea, 380-701)
• Woo Tae Ha(Faculty of Natural Science, Division of Food Bioscience, Konkuk University, Chungju, Chungcheongbukdo, South Korea, 380-701)
• Ran Lee(Faculty of Natural Science, Division of Food Bioscience, Konkuk University, Chungju, Chungcheongbukdo, South Korea, 380-701)
• Hyuk Song(Faculty of Natural Science, Division of Food Bioscience, Konkuk University, Chungju, Chungcheongbukdo, South Korea, 380-701)