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178 117 P Separatration of early stage acrosome reacted sperm and analyses of the proteins Kenji Yamatoya, Chizuru Ito, Cheng Chen, Mamiko Maekawa, Yoshiro Toyama, Kiyotaka Toshimori Dept. Anatomy and Developmental Biology, Grad. Sch. Med., Univ. Chiba During the acrosome reaction, the contents of sperm acrosome are released from the original location to the final destination and working place. The molecular distribution and nature are posttranslationally modified. We reported that flow cytometric analyses using antiacrosomal membrane protein antibodies anti IZUMO1 and antiSPACA1 can discriminate the early stage of acrosome reaction, classifying into 7 populations. In this study, we analyzed the 7 populations and found that the populations number 4 and 5 contain acrosomeswelling sperm. Acrosomeswelling occurs at a very early stage of acrosome reaction which starts before the exposure of inner acrosomal membrane. One of the acrosomal membrane proteins, SPACA1, was cleaved during this swelling stage. Western blot showed that SPACA1 was 35 kDa before acrosome reaction and became 21 kDa in acrosomeswelling sperm. Thus, this flow cytometric method is useful to analyze the detailed acrosome reaction process and the molecular changes during the acrosome reaction which are the last preparatory process for the spermegg interaction. P Cell adhesion molecule Nectin Kannika Adthapanyawanich Cell adhesion molecule1 Cadm1 Nectin3 Cadm1 Poliovirus receptor Nectin3 Nectin2 Cadm1 Nectin3 Cadm1 Nectin3 Cadm1//Nectin3/ Cadm1/ Nectin3/ Cadm1 Nectin3 P SF SF1 2 anti Müllerian hormone type 2 receptor Cre CreloxP SF1 0 21 SF1 7 14 21 14 21 SF1 AMH p27 WT1SOX9GATA4androgen receptor SF1 SF1 P Autoimmune responses induced by immunization with xenogenic testicular germ cells alone Experimental autoimmune orchitis EAO is one of the models of immunological male infertility. Classically, the immunization of mice with a testicular homogenate emulsified in complete Freund's adjuvant followed by intravenous injections of Bordetella pertussis is necessary for the induction of murine EAO. Later, we previously established a mouse EAO model that can be induced by two subcutaneous injections of viable syngeneic testicular germ cells TGC alone, and the autoantigens involved in the mouse EAO have been analyzed using two dimensional gel systems and western blotting. In the present study, to examine whether the xenogenic TGC can induce a mouse EAO, we immunized mice with viable TGC taken from the rat or the guinea pig. The results showed that mouse EAO was also inducible by the rat TGC but not the guinea pig TGC. Therefore, the results suggest that the autoantigens that induce the mouse EAO are present in not only the mouse TGC but also xenogenic TGC = the rat TGC. Analyses of the common autoantigens responsible for EAO induction are now in progress. P Diethylhexyl phthalate Di2ethylhexyl phthalate=DEHP DEHP A/J 8 0%0.01%0.1%DEHP 8 0.01%DEHP 0.1%DEHP 0.01% 0.1%DEHP DEHP MHC Class RTPCR DEHP IL10IFNγ DEHP P The effect of cadmium on immunoenvironment in the testis Yuki Ogawa 1,2 , Masahiro Itoh 1 , Shuichi Hirai 1 , Munekazu Naito 1 , Ning Qu 1 , Hayato Terayama 1 , Hidenobu Miyaso 2 , Yoshiharu Matsuno 2 , Masatoshi Komiyama 2 , Chisato Mori 2 1 Dept. Anat., Tokyo Med. Univ., 2 Dept. Bioenv. Med., Grad. Sch. Med., Chiba Univ. Cadmium, one of various environmental toxicants, is known to suppress systemic immunity and to injure the testicular capillary endothelia with resultant necrosis of testicular tissues in mice and rats treated with high doses. Recently, it also became evident that cadmium can affect the integrity of the bloodtestis barrier BTB, the endocrine function of Leydig cells, apoptosis of germ cells and systemic immunity, even on treatment with a low dose that does not induce spermatogenic disturbance. Experimental autoimmune orchitis EAO, i.e., an organ specific autoimmunity of the testis, can be induced by repeated immunization with testicular antigens, and its pathology is characterized by lymphocytic inflammation and spermatogenic disturbance. In the present study, we investigated the morphological and functional changes of testes in mice treated with a low dose of cadmium chloride CdCl 2 and also examined its toxicity as to susceptibility to EAO.
117 179 P TIRF ECM ECM 1 I Col1FN2 VI Col6fib3 IV Col4TIRF ECM ECM ECM Col1FN fib Col4 Col1FN Col6 fib Col4 Col6 Col6 P PACAP TAC 1 1 2 3 4 1 2 3 4 2 PACAP PACAP PAC1R PACAP PACAP PACAP 2 PACAP PACAP PACAP DNA 3 40 P A TAC1 6 P in vitro 1 2 2 3 3 2 2 3 1 1 2 3 10 3 3 in vitro 1 mm endometrial glands in vitro P Localization of fatty acid binding protein in mouse placenta and its possible role of fatty acid transport through trophoblasts Ariful Islam, Nobuko Tokuda, Yasuhiro Adachi, Tomoo Sawada, Yuji Owada Department of Organ Anatomy, Yamaguchi University Graduate School of Medicine Background: Malnutrition of mother during pregnancy is now thought to contribute to the etiology of various metabolic and/or neural disorders that manifest throughout life. Placenta is the key organ for regulating the transfer of nutrients including fatty acids from mother to embryo. In this study, we investigated the functional role of fatty acid binding protein 3 FABP3 after determining the localization of FABPs, cellular fatty acid chaperons, in the mouse placenta. Results: In RTPCR, gene expression of FABP3, 4 & 5 was detected. The protein expression of FABP 3, 4 & 5 was also confirmed by western blot analysis. In immunohistochemistry, FABP3 was highly expressed in the labyrinthine transport zone of mouse placenta; FABP4 was highly expressed in deciduas basalis zone; FABP5 was weakly and widely distributed in the labyrinthine, decidua and spongiotrophoblast zone. Discussion: FABPs were expressed in the mouse placenta with spatial differences. FABP3 and 5 were suggested to be involved in regulation of cellular transfer of fatty acids in trophoblasts of placenta. The functional significance of FABP expression in trophoblasts is now under investigation. P Kardasewitsch Verocay 1,2 2,3 2,3 4 4 1 1 2 3 4 Kardasewitsch Verocay Kardasewitsch Verocay 2 HE Verocay Kardasewitsch P MAP 1 2 1 2 MAP MAP in situ MAP 13 17 ERK1/2 JNK1/2 ERK1/2 p38 ERK5 MAP ERK1/2 JNK1/2
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