Keiko O. Yanagisawa

Effects of the Brachyury (T) mutation on morphogenetic movement in the mouse embryo

TT embryos have been first distinguishable at 8 days post coitum by their gross morphological abnormalities. By quantitative morphometry of histological sections, anomalies in the homozygotes were expressed numerically. At 8 days p.c., morphologically identifiable T-homozygotes had an increased number of ectodermal and a reduced number of mesodermal cells compared to the wild type. At 7 days p.c., embryos with a low mesoderm/ectoderm ratio were found only in litters of T+ × T+ matings at the expected frequency. At 6 days p.c., one-fourth of the embryos in T+ × T+ litters showed a delay in the transition from cuboidal to squamous endoderm. No such embryos were found in the +/+ × +/+ matings. In 6-, 7-, and 8-day mutant embryos, cells proliferated at statistically normal rates. Therefore, it may be said that advanced morphological irregularities of 8-day homozygotes cannot be accounted for by anomalies in cell proliferation. When the total cell number was 5 × 104/embryo (8 days), a sudden change was observed in the regional distribution of mesodermal and ectodermal cells along the anteroposterior axis of TT embryos. Since no regional difference in the cell cycle time was observed, these abnormalities may best be explained by anomalies in cell migration. These results strongly suggest abnormal morphology of TT mutants resulting from defects in morphogenetic movement.

Identification of sperm antigenic determinants with phylogenetically diverse and limited distribution using monoclonal antibodies

Mouse monoclonal antibodies capable of recognizing mouse sperm antigens were produced and the inter-species cross-reactivity of the antigenic determinants was examined. Of the seven clones independently established, it was found by indirect immunofluorescence staining that one was bound to the tail, one could recognize the head and five other antibodies reacted with the crescent-shaped anterior acrosome. Ultrastructural studies showed that the anti-head antibody recognized the sperm surface, and the anti-tail antibody reacted with the fibrous sheath. Five acrosomal antigenic determinants were found in the acrosome. These antibodies reacted with sperm from the male reproductive tract (testis, cauda epididymis, vas deferens), but not with somatic tissues. The anti-tail antibody reacted with sperm from four other mammalian species (musk shrew, rat, boar and human). The anti-head antibody and one of the anti-acrosome antibodies were bound only to mouse sperm. The other four acrosomal antigenic determinants showed various degrees of inter-species cross-reactivity. None of these antibodies reacted with chicken sperm.