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M Ekhlasi-Hundrieser

Publications and source records attributed to M Ekhlasi-Hundrieser.

7 recordsLinked to original sources
PubMed2006

Structural, biochemical and functional aspects of sperm-oocyte interactions in pigs.

Polyspermic fertilization is still a major issue in porcine IVF systems. New information is available to characterize the zona pellucida (ZP) at different developmental stages by scanning electron microscopy (SEM) and by confocal microscopy to show the distribution of ZP glycoproteins. SEM images indicated no differences between in vivo and in vitro matured oocytes; however a change in the surface structure between immature and matured oocytes, as well as between mature oocytes and preimplantation embryos was obvious. In addition, spermatozoa were more tightly fixed in the ZP of in vivo produced compared to the ZP of in vitro produced embryos. The ZP undergoes biochemical changes during maturation prior to fertilization. The acidity of the ZP increases during maturation as indicated by a shift of 1.3 pl units for ZPB/ZPC and 0.8 pl units for ZPA in 2D gel electrophoresis, which is based on increasing sulfation of the oligosaccharides during maturation. Mass spectrometry in combination with in-gel deglycosylation allowed the mapping of new glycosylation sites. Functionality of the ZP also depends on its maturation status. Induction of the acrosome reaction was delayed when capacitated spermatozoa were exposed to immature oocytes.

Animals↗
PubMed2004

Role of volume-stimulated osmolyte and anion channels in volume regulation by mammalian sperm.

The ability to maintain cellular volume is an important general physiological function. Swelling induced by hypotonic stress results in the opening of channels, through which ions exit with accompanying water loss (regulatory volume decrease, RVD). RVD has been shown to occur in mammalian sperm, primarily through the opening of quinine-sensitive potassium channels. However, as yet, direct evidence for the participation of anion channels in sperm RVD has been lacking. The chloride channel type ClC-3 is believed to be involved in RVD in other cell types. Using electronic cell sizing for cell volume measurement, the following results were obtained. (i) The anion channel blockers 5-nitro-2-(3-phenylpropylamino)-benzoic acid (NPPB), tamoxifen and 4,4'-diisothiocyanostilbene-2,2'-disulphonic acid (DIDS) increased hypotonic swelling in concentration-dependent fashion, whereas verapamil (P-glycoprotein inhibitor) had little effect. The most potent, NPPB and DIDS, blocked RVD without affecting cell membrane integrity at effective concentrations. (ii) When gramicidin was included to dissipate Na+/K+ gradients, major secondary swelling was observed under hypotonic conditions. This secondary swelling could be reduced by NPPB, and suppressed completely by replacing chloride in the medium with sulphate, an ion which does not pass through chloride channels. It was deduced that the initial hypotonic swelling activated an anion channel through which chloride ions could then enter freely down a concentration gradient, owing to the lack of a counter-gradient of potassium. (iii) Taurine, an osmolyte often involved in RVD, does not appear to play a role in sperm RVD because lengthy preincubation with taurine did not alter sperm RVD response. Our observations provide direct evidence that a chloride channel (possibly ClC-3) is involved in the process of volume regulation in mammalian sperm.

4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid↗
PubMed2003

Fate of lactadherin P47 during post-testicular maturation and capacitation of boar spermatozoa.

Polyclonal avian antibody was used partially to characterize the pig sperm lactadherin P47. P47 is a mosaic protein, composed of two epidermal growth factor (EGF)-like domains and two C1/C2 domains. P47 is homologous to the bovine mammary gland protein MGP 53/57 and mouse milk fat globule protein. Expression of P47 along the male genital tract and its localization on spermatozoa during post-testicular maturation and capacitation were studied. P47 was detected in the testis and in all parts of the epididymis by immunohistochemistry and by western blots of tissue extracts. By indirect immunocytochemistry, P47 was localized at the apical ridge of the sperm head in testicular, epididymal and ejaculated spermatozoa. The fluorescence intensity progressed during sperm transit from caput to cauda epididymis, probably caused by the ongoing expression and subsequent accumulation of P47 on the sperm surface. During the time course of capacitation, P47 appears to be unmasked by the release of coating proteins and appears to migrate from the apical ridge onto the entire acrosomal region, showing an intensive fluorescence pattern after 3 h capacitation in vitro. The kinetics of signal changes during in vitro capacitation were different in epididymal and ejaculated spermatozoa, indicating accelerated capacitational plasma membrane destabilization in epididymal spermatozoa.

Animals↗
PubMed2001

Sperm adhesion molecules: structure and function.

Fertilisation is a unique event in which the morphologically disparate gametes recognise, bind and fuse with each other. This event follows a highly regulated schedule of biochemical interactions, in which molecules are involved that mediate cell adhesion, signal transduction and the initiation of metabolic pathways. A plethora of molecules has been found on the male gamete and with regard to the different protein structures it is almost impossible to overlook the structures involved. Even more, carbohydrate structures cause an additional diversity with regard to the generation of surface structures. In this communication we try to elucidate the structures of proteins that have been known so far. We have focussed on spermadhesins, the zonadhesin, proacrosin and the PH-20 antigen. The variety of structures and also the common features among them as well as the presence of redundant systems are attributable to the evolutionary force of intraspecific sperm competition. This evolutionary force is assumed to be also responsible for the species selectivity observed in these adhesion molecules, which explains the preferential binding of gametes in a homologous system.

Animals↗
PubMed2000

Oocyte-sperm interactions.

The penetration of the zona pellucida is a crucial step during fertilization. Spermatozoa that are unable to recognize and bind to the zona pellucida glycoproteins or respond to the zona pellucida by undergoing the acrosome reaction fail to fertilize the egg. In most mammalian species, after entering the fallopian tube sperm are stored in the isthmic part of the oviduct under conditions that maintain sperm viability and synchronize both sperm transport and the process of acquisition of fertilizing ability, called capacitation. Only capacitated sperm are enabled to recognize the oocyte and respond to the oocyte signals in an appropriate manner. Close to time of ovulation sperm are released from the oviductal epithelium and swim to site of fertilization. The oviduct and the oocyte itself appear to coordinate sperm function and gamete interaction. The gamete recognition and the next levels of interaction are probably granted by the carbohydrate-protein interactions. Upon binding the signal cascade leading to acrosomal exocytosis is activated, eventually initiated by aggregation of zona pellucida receptor molecules. These signal transducing mechanisms are primed during the capacitation process. Tyrosine phosphorylation, tightly connected to the cholesterol efflux from the plasma membrane, and hyperpolarization seem be involved in this priming by activation of Ca(2+) pathways. Further preparational steps of the acrosome reaction may be mediated by osmosensitive signal transducing mechanisms. The current perspective focuses on the molecules involved in the complex hierarchy of sperm-egg interactions and regulative events priming sperm cell during capacitation for the acrosome reaction.

Acrosome↗
PubMedDate not supplied

Spermadhesins: a new protein family. Facts, hypotheses and perspectives.

Spermadhesins are a novel family of secretory proteins expressed in the male genital tract of pig, horse and bull. They are major products of the seminal plasma and have been found to be peripherally associated to the sperm surface. The structure and function of spermadhesins have been thoroughly investigated in the pig, which exhibits the greatest diversity of members: AWN, AQN-1, AQN-2, PSP-I and PSP-II and its glycosylated isoforms. They are multifunctional proteins showing a range of ligand-binding abilities, e.g. carbohydrates, sulfated glycosaminoglycans, phospholipids and protease inhibitors, suggesting that they may be involved in different steps of fertilization. Isolated porcine spermadhesins bind the zona pellucida glycoproteins in a cation-dependent manner with a Kd in a low micromolar range, and AWN, AQN-1 and AQN-3 display similar binding affinity for glycoproteins containing Gal beta(1-3)-GalNAc and Gal beta(1-4)-GlcNAc sequences in O-linked and N-linked oligosaccharides, respectively. During sperm passage through the epididymis AQN-3 and AWN have been shown to bind tightly to the sperm surface by interaction with the phospholipids of the membrane bilayer. At ejaculation the spermadhesins form a protective coat around the sensitive acrosomal region of the sperm head, thus possibly preventing premature acrosome reaction. During in vitro capacitation most of these aggregated sperm adhesins are lost, with the exception of phospholipid-bound spermadhesins. AWN and AQN-3 may now serve as a primary receptor for the oocyte zona pellucida, thus contributing to initial binding and recognition between sperm and egg. The amino acid sequence of spermadhesins does not show any discernible similarity with known carbohydrate recognition domains (CRD). However, they belong to the superfamily of proteins with a CUB domain with a predicted all-beta structure. The crystal structure of the heterodimeric complex of the spermadhesins PSP-I/PSP-II has been solved, showing that the overall structure of both spermadhesins consists of a beta-sandwich with five (parallel and antiparallel) beta-strands. It is the first three-dimensional structure of a zona pellucida-binding protein and reveals the architecture of the CUB domain. The spermadhesins represent a novel class of lectins that may be involved in sequential steps of fertilization, at least in the pig.

Animals↗