An autoradiographic study of macromolecular syntheses in the epithelium of the ductus epididymidis in the mouse. II. Incorporation of L-fucose-1-3H.

Glycoprotein dynamism in the mouse epididymis was studied by means ofhistoautoradiography after injection of L-fucose-1-3H. The label was detected, at thirty minutes p.i., in the area occupied by Golgi apparatus in the epithelial cells. At 4 h p.i. the label was already present inthe lumen of ductus epididymidis. At this time interval, the luminal labelling was highest in the initial segment of the epididymis and decreased against the more distal segments considerably. At ten days p.i. very high labelling was detected in the luminal contents in the terminal segment of the ductus epididymidis and in ductus deferens, the labelling in the proximal segments of the epididymis being much lower. These observations suggested a wave of labelled glycoprotein in epididymal plasma passing through the epididymis after a fucose pulse. Higher labelling was detected in so-called "clrial was seen in epididymal and uterine spermatozoa, mostly in sperm tail region.

Studies on acrosome labelling of mammalian spermatozoa by radioactive sugars.

The localization of glycoprotein synthesis and storage was studied during acrosome formation in guinea-pig using fine-structure autoradiography after (3H)-fucose incorporation. Three days after (3H)-fucose injection, labelling in spermatids was concentrated in the matrix of developing acrosomes, and it was evident that the fucosylation of acrosomal glycoproteins largely overshadowed the fucosylation of other spermatid glycoproteins. Acrosin labelling and its quantitative relation to labelling of other glycoproteins was examined in mature rabbit spermatozoa after incorporation of (14C)-fucose or (14C)-glucosamine during spermatogenesis. Cauda epididymis spermatozoa recovered 21 days after intratesticular application of (14C)-fucose or (14C)-glucosamine were analysed for acrosin specific labelling after acid extraction and gel filtration. In all the material examined, radioactivity was detected in the proacrosine fractions; radioactivity in purified proacrosin amounted to at least 2% of the total radioactivity in the epididymal sperm population. In addition to the peak with radioactive proacrosin, another radioactive peak in (14C)-glucosamine-labelled material was attributed to a glycoprotein intraacrosomal inhibitor of acrosin. It is concluded that (pro)acrosin (acrosin-inhibitor) complexes seem to contribute significantly to acrosomal glycoprotein labelling by radioactive sugars and that the distribution of these complexes may at least correspond to their cytochemically detectable component, acrosin. The superposition of the distribution of acrosin and of other acrosomal glycoproteins during acrosome reaction can be explained by the fact that the dispersal of most of the acrosomal content is linked to proacrosin activation.

The presence of blood group and lymphocyte antigens on porcine granulocytes.

Suspensions of highly viable (less than 95%) granulocytes minimally contaminated by other cell types were isolated from the peripheral blood of pigs by a single centrifugation with low molecular weight dextran and after preferential lysis of erythrocytes by hypotonic shock. A complement-dependent cytotoxic test showed the presence of antigens of the SLA major histocompatibility complex, the SLB leucocyte system and the A and E blood group systems on the granulocytes. Some SLA typing reagents against class I (SD) antigens did not react with granulocytes, however, or yielded dubious reactions. The findings showed that the reactivity of SLA sera resembles the reactivity fo human HLA sera. The results also show that compatibility in the SLA, SLB, A and E systems will have to be taken into account when preparing alloimmune sera for the determination of granulocyte-specific antigens of pigs.