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H J Glander

Publications and source records attributed to H J Glander.

101 records · Page 6Linked to original sources
PubMedDate not supplied

Influence of cryopreservation on the microelectrophoretic motility (EPM) of human spermatozoa.

The net surface charge of spermatozoa represents a characteristic of the plasmamembrane. It can be evaluated indirectly, by the measurement of the sperm electrophoretic motility (EPM). The EPM of washed human spermatozoa was determined in the microelectrophoresis to investigate three problems: (a) intra- and interindividual variation coefficients of EPM of cryopreserved spermatozoa, (b) effects of cryopreservation medium, temperature shock and cryoprotection on the EPM, (c) relationship between the EPM and the cryotolerance of the spermatozoa. The inter- and intrain-dividual variation coefficients of the sperm EPM of 24 cryopreserved semen samples of 6 fertile semen donors amounted to 7% and to 6%, respectively. The cryopreservation did not cause a significant change of sperm EPM in contrast to the temperature shock in liquid nitrogen. Temporary incubation of spermatozoa with egg yolk decreased their EPM. A significant correlation between EPM and the cryotolerance ascertained by the alteration of sperm motile efficiency could be found. After contact with egg yolk spermatozoa achieved a behaviour that corresponded to a better cryotolerance.

Cell Membrane↗
PubMedDate not supplied

Lack of HLA-molecules on human spermatozoa and in seminal plasma.

The expression of human leukocyte antigens (HLA) on ejaculated spermatozoa and on lymphocytes was compared by flow cytometry using monoclonal antibodies towards HLA class I (pan-HLA-A, -B, -C) and class II (DR) antigens. Soluble antigens of HLA class I (s HLA-A, -B, -C) in seminal plasma and in blood plasma were monitored with an ELISA technique. Lymphocytes showed specific fluorescence after incubation with the antibodies against HLA class I and class II (DR), whereas, on spermatozoa no positive immunofluorescence could be detected. No antibodies were bound to any significant extent either after modifications of sperm preparation (density gradient centrifugation, swim up-technique, addition of azide, foetal calf serum or benzamidine chloride) or after treatment of spermatozoa with detergents. Furthermore, different concentrations of soluble HLA-A, -B, -C in seminal plasma and in blood plasma were detected. The latter one showed soluble HLA about four-fold more concentrated than the seminal plasma (means +/- SD: 262.5 +/- 144.4 nmol l-1 vs. 62.5 +/- 27.1 nmol l-1). These results suggest, that the HLA-expression differs between human spermatozoa and somatic cells.

Flow Cytometry↗
PubMedDate not supplied

Disintegration of human spermatozoal membranes in seminal plasma decreases the binding capacity of integrins.

The in vitro binding capacity of spermatozoal integrins to matrix components after disintegration of sperm membranes was evaluated. The percentage of spermatozoa with functionally-relevant integrins was determined before and after devitalization of spermatozoa, which were resuspended in seminal plasma or in culture medium. The devitalization was performed by cryoshock or by incubation of spermatozoa with triton X-100 in a concentration ranging from 0.01 to 1.0%. The spermatozoal integrins were detected by the binding of anti-integrin antibodies and flow cytometry and the functional activity was monitored by the binding of the spermatozoa to the matrix components in a cell attachment assay. The seminal plasma decreased the binding of anti-integrin antibodies to the spermatozoal surface and the binding of spermatozoa to ligands and matrix components. respectively. In contrast, the expression of fibronectin and laminin on spermatozoa was increased. Not all spermatozoa, which expressed integrins on their surface bound to the ligands in the cell attachment assay. These results suggest that the detectable integrins only partially exert functional relevance. It can be concluded that the spermatozoa with fragile plasma membranes are more prone to functional inactivation of their integrins by the seminal plasma.

Cell Adhesion↗
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Localization of adhesion molecules on human spermatozoa by fluorescence microscopy.

The expression of adhesion molecules on human spermatozoa of healthy probands was analysed. The localization patterns of adhesion molecules (AM) on the spermatozoal surface were documented by fluorescence microscopy. Spermatozoa were incubated with antibodies against alpha 1 (CD49a), alpha 2 (CD49b), alpha 3 (CD49c), alpha 4 (CD49d), alpha 5 (CD49e), alpha 6 (CD49f) chains of beta 1 integrins, beta 1 (CD29), beta 2 (CD18), alpha V (CD51), beta 3 (CD61) and beta 4 integrin chains, the LFA-3 (Lymphocyte function antigen, CD58) from the immunoglobulin superfamily and the extra-cellular matrix proteins laminin, fibronectin and collagen IV. For collagen IV, alpha 1 and alpha 2 chains no expression could be noticed. Laminin was detected at the acrosomal membrane, fibronectin and beta 4 chain mainly at the equatorial membrane. The fibronectin receptors alpha 3, alpha 4 and alpha 5 chains of the beta 1 integrins were mainly located on acrosomal and equatorial membrane areas. Laminin receptor alpha 6 chain was located postacrosomal and less frequently acrosomal. beta 2 chain and vitronectin receptors alpha V and beta 3 chains had a mainly postacrosomal localization pattern. LFA-3 was found constantly on postacrosomal membrane areas. Double staining technique was used to prove the simultaneous occurrence of fibronectin and its integrin receptors alpha 3, alpha 4 and alpha 5 chains and of alpha V and beta 2 chains on spermatozoa. The localization patterns of integrins on double stained spermatozoa were similar to the patterns described for single stained spermatozoa. The localization of fibronectin appeared to be influenced by the presence of integrins: the typical equatorial fibronectin band disappeared in case of an equatorial localization of integrins.

Antigens, CD↗
PubMedDate not supplied

Effects of pure human follicle-stimulating hormone (pFSH) on sperm quality correlate with the hypophyseal response to gonadotrophin-releasing hormone (GnRH)

In 41 men with idiopathic infertility, the effect of pure follicle-stimulating hormone (pFSH) therapy on semen parameters was evaluated in relation to the results of the gonadotrophin-releasing hormone (GnRH) stimulation test in an open study. The patients showed a mean (+/-standard error) sperm concentration of 15.84 (+/-2.87) Mill. ml-1, 41.02 (+/-4.19)% of the spermatozoa were motile and 46.16 (+/-2.36)% normomorph before treatment. All patients were administered 150 IU pFSH subcutaneously three times weekly for 10 weeks. After therapy with pFSH, no significant differences compared with the pre-treatment sperm characteristics were observed (P > 0.05) except for the velocity average path, VAP (P = 0.029). However, the stimulation factors of FSH in the GnRH test showed a significantly negative correlation with the pFSH mediated improvement of sperm concentration (r = -0.662, P = 0.000002), of percentage of normomorph spermatozoa (r = -0.480, P = 0.0015) and total count of motile spermatozoa per ejaculate (r = -0.567, P = 0.00014). In consequence, the patients were divided into two groups depending on the stimulation factor of hypophyseal FSH secretion, FSH-SF. The cut-off point of the FSH-SF was set at 1.7 because the correlation analyses detected a 1.5-fold improvement of sperm parameters at this hypophyseal response on average. Pure FSH treatment increased the concentration of spermatozoa (P = 0.0007), the total count of motile spermatozoa in the ejaculate (P = 0.015) as well as the computer-aided sperm motion parameters VAP (P = 0.029) and velocity curve linear, VCL (P = 0.049) in patients with FSH-SF < 1.7, whereas in the patient group with FSH-SF > 1.7 no improvement of semen parameters was found. Insufficient stimulation of hypophyseal FSH secretion may be a prerequisite but not a guarantee for responsiveness to pFSH treatment. The results of the present investigation suggest that FSH stimulation in the GnRH test should be taken into account in idiopathic infertile men before pFSH therapy.

Adult↗
PubMedDate not supplied

Adhesion molecules and matrix proteins on human spermatozoa.

Ejaculated spermatozoa and spermatogenic cells express alpha- and beta-chains of beta 1, 3 and 4 integrins as well as their ligands fibronectin and laminin in an extended intra- and interindividual variation and in different patterns of location. The mRNA transcripts of these molecules were detectable by nested polymerase chain reaction in the spermatozoa. The conclusion of a functional competence of these adhesion molecules (AM) was supported by their relation to the results of the zona-free hamster oocyte penetration (HOP) test, the in vitro fertilization of human oocytes and cell attachment assays. AM labelling was influenced by the disintegration of the sperm plasma membrane, especially in seminal plasma, by progesterone, human follicular fluid and microorganisms, but was barely modified by sperm cryopreservation. Despite substantial advances in the knowledge about sperm adhesion molecules, many questions remain to be answered.

Animals↗
PubMedDate not supplied

Selected sperm differ from nonselected cells in their electrophoretic mobility (EPM) after demembranation.

Human semen samples were first treated by swim-up migration or by Percoll density gradient centrifugation for selection of spermatozoa. The net surface charges of spermatozoa in the resulting compartments were indirectly evaluated by the electrophoretic mobility (EPM) during spermatozoal microelectrophoresis. Significantly different electrophoretic mobilities were measured between the compartments after demembranation with 0.1% (v/v) Triton X-100. However, without intermediate treatment by the detergent no significant differences were detected between the spermatozoal EPM. In addition, spermatozoa were divided into migrated and nonmigrated spermatozoal populations by the swim-up technique, and then treated with urea to remove spermatozoal coats. No significant differences in EPM were detected between these two groups. This investigation demonstrates that selected spermatozoa from semen exhibit an additional dimension of superiority, in EPM, to nonselected spermatozoa, which may better equip them for the process of fertilization. Measurements of EPM, therefore, may provide yet another, perhaps valid, means of developing a clinical tool for evaluating the potential fertilizing capacity of a semen sample in infertility studies.

Cell Membrane↗
PubMedDate not supplied

Acrosome reaction changes the pattern of cyclic nucleotide phosphodiesterases in human sperm.

Three categories of cyclic nucleotide phosphodiesterases (cNPDE) are distinguished at present: type 1 with high affinity to cyclic GMP; type 2 with low affinity to cyclic AMP and to cyclic GMP; and type 3 with high affinity to cyclic AMP. For the evaluation of normal values in human spermatozoa 50 semen samples with normal classical semen parameters were investigated. The activities (means +/- SD) of the cNPDE types (10(-11) mol/10 min x 10(8) spermatozoa) of washed human spermatozoa amounted to 47 +/- 22 (type 1), 3350 +/- 1537 (type 2), and 70 +/- 38 (type 3). A significant inhibition of type 3 by cyclic GMP could not be detected. One milligram protein of the spermatozoa hydrolyzed about 20-fold the amount of cyclic nucleotides compared with 1 mg protein of the seminal plasma. Furthermore, the cNPDE of the spermatozoa and of the seminal plasma differed in the influence of type 3 by cyclic GMP and in the pattern of activities. The acrosome reaction (AR) induced by the cold shock method led to an activation of type 2 and 3 unlike the initiation of the AR by the digitonin method. The latter did not cause significant differences of the cNPDE activities before and after the AR.

3',5'-Cyclic-AMP Phosphodiesterases↗
PubMedDate not supplied

In vitro fertilization: increased VLA (very late antigen) integrins and fibronectin after acrosome reaction.

The pattern of detectable adhesion molecules (AM) on human ejaculated spermatozoa used for in vitro fertilization (IVF) of human oocytes was evaluated. The percentage of spermatozoa with the alpha chains 3, 4, 5, and 6 of the integrins (also called very late antigens, VLA alpha 3, 4, 5, 6), the alpha V and the beta 3 chains, as well as the matrix proteins laminin and fibronectin, were determined by flow cytometry using monoclonal antibodies against integrins and matrix proteins before and after the acrosome reaction. This reaction was induced by the low-temperature method according to P. Sanchez, E. Töpfer-Petersen, R. J. Aitken, and W. B. Schill (1991, Andrologia 23:197-203) and monitored by the binding of FITC-conjugated Pisum sativum agglutinin. Twenty-three of 34 investigated semen samples fertilized at least one oocyte (fertile group) and 11 samples did not induce a pronucleus formation or cleavage of an oocyte (infertile group). Significant differences in the classical semen parameters between both semen sample groups could be shown only for the percentage of progressively motile spermatozoa (p = .046, U test). The spermatozoa of the fertile group showed a significantly higher expression of VLA alpha 3 (p = .045) and fibronectin (p = .048). Additionally, after the loss of the acrosome a significantly higher expression of alpha 4, alpha 5, and alpha 6 chains of integrins (p < .05) was detected in the fertile semen group. In contrast, the inducibility of the acrosome reaction differed only to a nonsignificant extent (p = .094). These data suggest that spermatozoa showing a higher expression of the alpha chains of the integrins after the acrosome reaction have a better fertilization rate in vitro.

Acrosome↗
PubMedDate not supplied

Monosaccharide transport across membranes of human spermatozoa. II. Basic properties of D-fructose and D-glucose uptake.

D-glucose uptake by human spermatozoa shows like D-fructose uptake a hyperbolic kinetic. The affinities to the transport system are similar, but the maximal capacity of D-glucose transport is greater than the D-fructose transport. Both transport processes are inhibited by monoiodoacetate. Therefore these transports can be regarded as carrier-mediated active transports. The D-glucose transport moreover is inhibitable by ouabain and is influenced by extracellular Na+. These differences could be the expression that for both monosaccharides different transport systems exist. But the dependence of D-glucose and D-fructose uptake on the extracellular D-fructose-D-glucose-relation permits the conclusion that both systems interact with another.

Biological Transport, Active↗