No abstract

A highly sensitive blot-assay was developed for glycosaminoglycans (GAGs) and proteoglycans (PGs) utilizing a precipitation reaction by a cationic dye Cuprolinic Blue. The precipitates were deposited into 1-2 mm2 spots on nitrocellulose membrane by using a 96-well filtration apparatus. The dried sheet was digitized by a flat bed scanner and the intensity of the dots was quantitated by an image analysis software. The working range for chondroitin sulfate was 10-300 ng. The response of various GAGs differed according to the number of anionic groups, both sulphate and carboxyl groups being able to bind the dye. The sensitivity of the assay was decreased by high concentrations of GuC, CsC and protein, but not by nonionic detergents, common buffers and 8 M urea. Contact exposure to autoradiography film enabled quantitation of 25-250 DPM, and 1-10 DPM, of 35SO4-radioactivity in precipitated PGs after overnight and 14 days' exposures, respectively.

The bovine seminal plasma is formed mainly by secretions of epididymis and the glandular epithelia in ampulla, seminal vesicles, prostate and Cowper's glands. The contribution of each organ to the hydrolytic enzyme activities (glycosidases, exopeptidases, phospholipases) of the bull seminal plasma has been analyzed and is reviewed in this paper with special emphasis on the role of the accessory glands. Seminal vesicles seem to have a major role in the secretion of seminal. plasma acid a-glucosidase, acid a-mannosidase and P-N-acetylhexosaminidase, aminopeptidase A, dipeptidyl peptidase I1 and IV and y-glutamyl transpeptidase as well as Ca2+-dependent and Ca2+-independent phospholipases A, with distinct substrate specificities, a choline-specific phospholipase C and a Co2+ (Mn2+)-activated sphingomyelinase. The enzyme pattern in the ampulla closely resembled that of the seminal vesicles and obviously contributes to the seminal plasma level of these hydrolases. The bull prostate and Cowper's glands contained a strong CaZ+dependent phospholipase A, activity. However, these glands may not contribute to the seminal plasma PLA, activity. At ejaculation the epididymal spermatozoa are exposed to these enzymes. They may have a specific affinity to sugar, peptide or phospholipid residues at distinct sites of the sperm surface. These enzymes may also participate in the digestion of various other semen components to create a suitable milieu for the emitted spermatozoa.Zusammmenfassung. Das Seminalplasma des Bullen wird hauptsachlich von den Sekreten des Nebenhodens und der Driisenepithelien der Ampulle, Blaschendruse, Prostata und Cowperschen Druse gebildet. Der Beitrag eines jeden Organs zu den hydrolytischen Enzymaktivitaten (Glycosidasen, Exopeptidasen, Phospholipasen) des Bullen-Seminalplasmas wurde analysiert und in diesem Ubersichtsartikel besonders im Hinblick auf die Rolle der akzessorischen Geschlechtsdriisen dargestellt. Die Blaschendrusen haben offensichtlich eine groRere Bedeutung fur das Seminalplasma bei der Sekretion der a-Glucosidase, sauren a-Mannosidase und P-N-Acetylhexosaminidase, Aminopeptidase A, Dipeptidylpeptidase I1 und IV und der y-Glutamyltranspeptidase sosie CaZ+-abhangigen und Ca2+-unabhangigen Phospholipasen A, mit verschiedenen Substratspezifizitaten einer Cholin-spezifischen Phospholipase C und einer Co2+ (Mn2+)-aktivierten Sphingomyelinase. Das Enzymmuster der Ampulle ahnelt dem der Blaschendriise sehr stark und tragt Hydrolases in bull accessory glands 11 offensichtlich zum Seminalplasmaspiegel dieser Hydrolasen bei. Die Bullenprostata und Cowpersche Druse enthalten eine starke Ca2+-abhangige Phospholipase A,-Aktivitat. Diese Drusen scheinen jedoch nichts zur PLA,-Aktivitat des Seminalplasmas beizutragen. Bei der Ejakulation werden die Nebenhodenspermien diesen Enzymen ausgesetzt. Sie durften eine spezifische Affinitat fur Zucker, Peptide oder Phospholipidreste an bestimmten Domanen der Spermienoberflache besitzen. AuSerdem konnten diese Enzyme bei der Zerlegung verschiedener anderer Kompone...