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Biomedical subjects

A E Friess

Publications and source records attributed to A E Friess.

At least 37 records · Page 2Linked to original sources
PubMed1988

The acrosome reaction in boar spermatozoa.

In order to gain more insight into the molecular alterations of the acrosome, boar spermatozoa were incubated in a calcium-containing medium in the presence of the ionophore A23187. The time-course of the acrosome reaction was assessed by phase-contrast microscopy. Different stages of the acrosome reaction were studied by immunoelectron microscopy using a specific antibody directed against the complete outer acrosomal membrane. The introduction of monoclonal antibodies generated by immunization of Balb/c mice with the isolated outer acrosomal membrane permitted a study of the topography of distinct membrane proteins during the acrosome reaction. The exposure of activation of a fucose-binding protein important for sperm--zona attachment was studied using neo-glycoproteins labelled with colloidal gold for transmission electron microscopy.

Acrosome↗
PubMed1987

Electron microscopic localization of a fucose-binding protein in acrosome reacted boar spermatozoa by the fucosyl-peroxidase-gold method.

In this study we have examined the behaviour and the localization of the fucose-binding protein (FBP) in boar spermatozoa during ionophore induced acrosome reaction (AR) by means of normal TEM and specimen preparation in toto. During early stages of AR the FBP is first localized at the border between equatorial segment and anterior acrosome. With the propagation of the AR the FBP is dramatically expressed and visible over the entire surface of the acrosome and equatorial segment. TEM pictures of this stages show that the FBP is associated with the OAM. At later stages of AR, when acrosomal ghost formation occur, the FBP is associated with the acrosomal ghost, and equatorial segment and to a very low degree also with the IAM. It is concluded from this data that the FBP is responsible for the specific binding of the ghost-sperm unit to the zona pellucida.

Acrosome↗
PubMed1987

Fracture labelling of boar spermatozoa for the fucose-binding-protein (FBP).

Labelling of fractured boar spermatozoa with the FUC-HRP gold method for a fucose-binding-protein (FBP) gave evidence the FBP is localized in the acrosomal matrix. All fracture faces through the acrosome from the rostral end towards the equatorial segment show similar labelling pattern. This labelling is completely blocked by preincubation of the fractured tissue with focoidan.

Acrosome↗
PubMed1986

[Enzyme histochemistry of the pig placenta. III. Histotopics of enzymes in the uterine epithelium].

Within the uterine glands, the following enzymes were demonstrated by histochemical methods after 30, 58, 80, 100, and 110 d of pregnancy, respectively: beta-N-acetyl-hexosaminidase, beta-galactosidase, beta-glucuronidase, alpha-mannosidase, acid phosphatase, alkaline phosphatase, esterases, cytochrome oxidase, 5-nucleotidase, leucine aminopeptidase, adenosine triphosphatase, diaphorases (NADH, NADPH), glucose-6-phosphate dehydrogenase, 6-phosphogluconate dehydrogenase, succinate dehydrogenase, isocitrate dehydrogenase (NAD, NADP), beta-hydroxybutyrate dehydrogenase, glycero-3-phosphate dehydrogenase, NAD-glycero-3-phosphate dehydrogenase, glutamate dehydrogenase (NAD, NADP), lactate dehydrogenase. The results show that the activities of G-6-PDH, 6-PGDH, and cytochrome oxidase increase within secreting cells during the 2nd half of pregnancy. The activities of the other enzymes remained almost unchanged during the period of investigation. The description of our results distinguishes between gland neck, middle, and distal part of the secretory unit, respectively. In general, the enzyme activities are similar within the middle and distal gland segments, but lower in the epithelia of the neck region. The activity of dehydrogenases was medium to intensive within the middle and distal gland segments, but only low to medium within the neck portion. Of the hydrolases, the acid phosphatase, ATPase, leucine aminopeptidase, and beta-galactosidase demonstrated an intensive activity within activity secreting cells. The enzyme activities of the gland epithelia are compared with these of the uterine surface epithelia and the histochemical results are discussed in context with their significance in histiotrophic nutrition.

Animals↗
PubMed1985

Evidence for a fucose-binding protein in boar spermatozoa.

A fucose binding protein was detected in boar spermatozoa by means of a specifically developed modified enzyme-linked-lectin-assay using glycosylated peroxidase derivatives. The distribution of the fucose binding protein was assessed by means of fluorescence microscopy with fluoresceinyl-glycosylated peroxidase. Fucose binding was particularly prominent at the apical region of the sperm head. In order to gain more insight into the precise localization of the carbohydrate binding protein electron microscopical studies were performed using fucosyl peroxidase coupled to colloidal gold. In ultrathin sections as well as in specimens prepared in toto for TEM an intensive binding of fucosylperoxidase-colloidal gold was predominantly found at the apical part of the acrosome appearing as a crescent-like area. In some cases this binding pattern was replaced by a triangle-like intensive labelling at the equatorial segment as revealed clearly by specimens prepared in toto. By SDS-PAGE of the SDS-extractable sperm-proteins, followed by transblotting to nitrocellulose and visualization with the fucosylperoxidase by enzymatic amplification with 4-chloro-1-naphthol mainly one protein with the reduced molecular weight of approximately 53 kdal and some small proteins with apparent molecular weights less than 20 kdal was found to be responsible for the fucose-binding ability of porcine spermatozoa.

Acrosome↗
PubMed1985

Immunocytological characterization of the outer acrosomal membrane (OAM) during acrosome reaction in boar.

In order to study the acrosome reaction in boar, spermatozoa were incubated in a calcium-containing medium in the presence of the calcium ionophore A23187. The time course of the acrosome reaction was assessed by phase-contrast microscopy and correlated with the movement characteristics of the spermatozoa determined by means of multiple-exposure photography (MEP). Different stages of the acrosome reaction could be observed by indirect immunofluorescence using an antibody fraction raised in rabbits against the isolated outer acrosomal membrane (OAM). At the start of the acrosome reaction, a bright fluorescence located exclusively at the acrosomal cap of the sperm head could be observed, whereas after 60-120 min, the fluorescence vanished, indicating the complete loss of the OAM. However, to gain more insight into the stages of the plasma membrane and OAM during the acrosome reaction, immunoelectron-microscopical studies were performed using anti-OAM antibodies detected by the protein-A gold method. Ultrathin sections and total preparations in combination with transmission electron microscopy (TEM) confirmed, that boar spermatozoa start their acrosome reaction by a vesiculation of the plasma membrane, thus exposing the heavily labelled OAM, which is then lost as sheets or large vesicles. The newly exposed inner acrosomal membrane did not show any labelling with gold, thereby indicating clear differences in the antigenicity of both acrosomal membranes.

Acrosome↗
PubMed1985

[Enzyme-histochemical studies of the pig placenta. II. Histotopics of enzymes in the areolar placenta epithelium].

In porcine areolar placental epithelia, the following enzymes were demonstrated by histochemical methods after 30, 58, 80, 100, and 110 d of pregnancy, respectively: beta-N-acetyl-hexosaminidase, beta-galactosidase, beta-glucuronidase, alpha-mannosidase, acid phosphatase, alkaline phosphatase, nonspecific esterases, cytochrome oxidase, 5-nucleotidase, leucine aminopeptidase, adenosine triphosphatase, diaphorases (NADH, NADPH), glucose-6-phosphate dehydrogenase, 6-phosphogluconate dehydrogenase, succinate dehydrogenase, isocitrate dehydrogenase (NAD, NADP), beta-hydroxybutyrate dehydrogenase, glycero-3-phosphate dehydrogenase, NAD-glycero-3-phosphate dehydrogenase, glutamate dehydrogenase (NAD, NADP), lactate dehydrogenase. The results show that the enzyme activities remained almost unchanged during the period of investigation. Of the dehydrogenases, the diaphorases as well as succinate and lactate dehydrogenase demonstrated generally an intensive activity within the epithelia. The activity of the other dehydrogenases was only low. The activity of unspecific esterase was very intensive within the uterine epithelia but remarkably low within chorionic epithelia. Contrarily, the reaction of adenosine triphosphatase was more intensive within chorionic than uterine epithelia. All investigated glucosidases reacted distinctly positive within chorionic epithelia, but only beta-N-acetyl-hexosaminidase and beta-galactosidase in uterine epithelia. The high activity of acid phosphatase, especially within the chorionic epithelium, seems to be connected with uteroferrin, an iron-binding protein. The histochemical results are discussed in context with the function of the areolae in histiotrophic nutrition and iron transport.

Animals↗
PubMed1984

Con A- and WGA-binding sites on bovine epididymal spermatozoa: TEM of specimens in toto.

Binding sites for Con A and WGA were detected on bovine spermatozoa during epididymal maturation. We used colloidal gold as an EM-marker. The spermatozoa were treated according to a two-step method for lectin and colloidal gold, then adsorbed to lysine-coated nickel grids and subsequently examined by TEM in toto. Using this method we rapidly got information about the topographic distribution of lectin-binding sites. Major differences exist for WGA between caput and cauda spermatozoa. Conceding that cell-thickness poses some limitation, we consider this method to be practical and especially useful in studies concerning topographic distribution of cell surface components in single cell systems.

Animals↗
PubMed1984

Glycoproteins of bovine epididymal spermatozoa--a cytochemical study.

Modifications in bull sperm plasmamembrane during epididymal passage were investigated by the use of four different lectins: Concanavalin A (Con A); Ricinus communis I (RCA1); Wheat germ agglutinin (WGA); Ulex europaeus agglutinin I (UEA1). During sperm passage from caput to cauda epididymidis agglutination by RCA1 and WGA distinctly increased. Similar but somewhat less pronounced difference in the agglutinability was found for Con A. No agglutination was observed with UEA1. Ultrastructural examination of Con A binding sites on sperm plasma membrane with a Con A-horseradish peroxidase-gold technique (Con A-HRP-G) revealed a significant increase in the number of gold granules on the sperm tails during the epididymal passage of spermatozoa. No change in WGA-binding sites was observed between caput and cauda spermatozoa using a WGA-peroxidase method.

Agglutination↗
PubMed1984

[Enzyme histochemical studies of the swine placenta. Histoptics of enzymes in interareolar placental epithelia].

In porcine interareolar placental epithelia, the following enzymes were demonstrated by histochemical methods after 30, 58, 80, 100, and 110 d of pregnancy, respectively: beta-N-acetylhexosaminidase, beta-galactosidase, beta-glucuronidase, alpha-mannosidase, acid phosphatase, alkaline phosphatase, nonspecific esterases, cytochrome oxidase, 5-nucleotidase, leucine aminopeptidase, adenosine triphosphatase, diaphorases (NADH, NADPH), glucose-6-phosphate dehydrogenase, 6-phosphogluconate dehydrogenase, succinate dehydrogenase, isocitrate dehydrogenase (NAD, NADP), beta-hydroxybutyrate dehydrogenase, glycero-3-phosphate dehydrogenase, NAD-glycero-3-phosphate dehydrogenase, glutamate dehydrogenase (NAD, NADP), lactate dehydrogenase. The results show that most of the enzyme activities remained almost unchanged during the period of investigation. Only G-6-PDH and 6-PGDH activities increased within the uterine epithelium and nonspecific esterase activity within uterine as well as chorionic epithelia during the 2nd half of pregnancy. Within chorionic and uterine epithelia, hydrolases but not dehydrogenases demonstrated a higher activity at the bases of chorionic villi as compared to the apices and flanks of the latter. The action and influence of the demonstrated enzymes on metabolism, energy transfer, secretory, and resorptive activities of chorionic and uterine epithelia are discussed.

Animals↗
PubMed1983

Uterine glands of the pig during pregnancy. An ultrastructural and cytochemical study.

The ultrastructure of the porcine uterine glands is described from material taken from 11 pregnant pigs at exactly known stages of gestation (day 30; 58; 80; 100; 110). Fixation was performed by perfusion via a branch of the uterine artery and the tissue was routinely processed for electron microscopy. Additionally, cytochemical studies (phosphotungstic acid reaction for glycoproteins, according to Rambourg 1967; acid phosphatase reaction; ultrastructural localization of cellular iron, according to Parmley et al. 1978) were performed. On day 30 of pregnancy the uterine glands are coiled, simple tubular glands with a narrow lumen. The epithelial lining is simple columnar and consists basically of two cell types, ciliated cells and secretory cells. The secretory activity of the glandular epithelium is low; only a few secretory granules are present in the supranuclear cytoplasm. At midpregnancy the ultrastructure of the glands has significantly changed and the cells now show all the characteristics of high secretory activity: numerous parallel cisternae of rough endoplasmic reticulum, an extensively developed Golgi apparatus and many secretory granules which give a positive reaction for acid phosphatase and glycoproteins. The lumina of the glands are significantly enlarged and filled with a great amount of a granular, acid phosphatase-positive material. In the last third of pregnancy, only minor changes in the ultrastructure of the uterine glands are observed. The secretory activity is still high. The amount of rough endoplasmic reticulum has further increased and parallel arrays of cisternae occupy a considerable part of the supranuclear cytoplasm. The importance of the uterine secretion for embryonic nutrition and development is only partly understood. One of the secreted glycoproteins, uteroferrin, is believed to play an important role in the iron transfer from mother to fetus. From midpregnancy onward, a special cell type, the "granule laden cell" is found scattered between normal secretory cells of the uterine glands. Contrary to the opinion of Perry and Cromby (1982), we could demonstrate that these cells frequently extend to the lumen of the gland; hence the term "basal cell" seems inappropriate for this cell type.

Acid Phosphatase↗
PubMed1983

Localization of lectin receptors on bovine epididymal spermatozoa using a colloidal gold technique.

Mammalian spermatozoa that leave the testis are neither motile nor fertile. Maturation of spermatozoa occurs during epididymal transport. Among the changes during epididymal passage alterations of the surface properties of spermatozoa appear especially interesting. In this report we describe ultrastructural localization of Con A and WGA-binding sites on bovine spermatozoa from caput and cauda epididymides using a indirect lectin-horse-radish-peroxidase gold technique. With Con A the plasma-membrane covering the sperm head was heavily labelled with gold granules in caput as well as in cauda epididymal spermatozoa. A different distribution was observed for WGA-binding sites. The acrosomal region of caput and cauda spermatozoa was heavily labelled with gold, both in caput and cauda spermatozoa. The postacrosomal region was only sparsely marked in caput sperm whereas in sperm cells originating from the cauda a calix like membrane area displayed intense labelling. Some differences in the number of binding sites were also seen on sperm tails: those of caput spermatozoa show generally more Con A binding sites that those from cauda epididymal spermatozoa. No changes in the number of WGA-binding sites on sperm tails was observed during epididymal passage. The technical aspects of the indirect lectin-horse-radish-peroxidase gold technique are briefly discussed.

Animals↗