|Budget Amount *help
¥2,000,000 (Direct Cost : ¥2,000,000)
Fiscal Year 1995 : ¥1,000,000 (Direct Cost : ¥1,000,000)
Fiscal Year 1994 : ¥1,000,000 (Direct Cost : ¥1,000,000)
(1) Monoclonal antibodies were raised against primary spermatocytes. Using indirect immunofluorescence assay and immunoelectron microscopy, it has confirmed that three antibodies recognize surface antigens of the different stages of spermatocytes in the mouse testis. Antibody 10B3 recognizes spermatocytes during middle to late pachytene stages (VI-X) , antibody 10D6 stains spermatocyte during leptotene to early pachytene stages (I-IV) and antibody 12F7 labeled spermatocytes during late stages(VII-VIII).
(2) Antibodies 10D6 and 12F7 also recognized specific sites of the surface of the sperm head. To see the effects of the antibodies on the fertilization, the antibodies were added to the culture medium of the in vitro fertilization system, but obvious effects were not obtained.
(3) Antigens recognized by the three antibodies were characterized by Western blotting. Antibodies 10D6 and 12F7 failed to detect any spot, whereas antibody 10B3 labeled one spot (MW 50kD,pI5.5) by two-dimensional W
(4) We tried to culture spermatogenic cells including spermatocytes and gonocytes. However, significant results were not obtained.
(5) Spermatocytes move from the basal to adluminal compartment in the seminiferous tubule during maturation, and cytoskeletal proteins such as actin filaments might influence the movement of the spermatocytes. Thus we investigated the distribution of actin filaments and actin-binding proteins in the testis. As a result, the distribution of actin filaments changed during postnatal development. Furthermore, in the cryptorchid testis, in which spermatogenesis was disturbad, the arrangement of actin filaments was also disrupted. These results suggest the relation between the distribution of the actin filaments and spermatogenic activity.