Quantitative proteomic analysis of heifer serum at the peri-implantation period using two complementary methods.

Context: Systemic events resulting from mother-embryo relation at the peri-implantation period may result in specific changes to the protein composition of serum and thus provide a source of biomarkers for early detection of pregnancy.

Aims: We set out to use two different quantitative proteomic approaches to test this hypothesis by comparing heifer serum at the peri-implantation period to that from cycling heifers.

Methods: Two-dimensional electrophoresis (2-DE) and isobaric tags were used for relative and absolute quantitation (iTRAQ) of proteins.

Histological changes and transglutaminase 2 expression in the oviduct of advanced pregnant cows.

The oviduct is a dynamic organ that has not been assigned specific functions during advanced pregnancy. However, since changes in the oviductal epithelium during the estrous cycle are attributed mainly to variations in estradiol (E2) levels, and E2 levels increase along pregnancy, we hypothesized that advanced pregnant cows should present changes in the oviductal epithelium. In advanced pregnant cows, the oviducts showed higher leaf-like folds and lower mucosa width and epithelium height than those of cycling animals.

Hormone-regulated PKA activity in porcine oviductal epithelial cells.

The oviduct is a dynamic organ that suffers changes during the oestrous cycle and modulates gamete and embryo physiology. We analyse the possible existence of Protein kinase A (PKA)-dependent hormone-regulated pathways in porcine ampulla and primary cell cultures by 2D-electrophoresis/Western blot using anti-phospho PKA substrate antibodies. Differential phosphorylation was observed for ten proteins that were identified by mass spectrometry.

Recent advances in bovine sperm cryopreservation techniques with a focus on sperm post-thaw quality optimization.

Sperm cryopreservation facilitates the storage and transport of germplasm for its use in artificial insemination (AI) and other advanced reproductive technologies. The cryopreservation process can damage sperm and compromise functionality. Several cryobiological studies have found that the physical and biological factors that affect sperm survival at low temperatures during the cryopreservation process often involve the integrity of sperm membrane.

Expression profile and distribution of Annexin A1, A2 and A5 in human semen.

Annexins (ANXAs) have been identified in different seminal components mainly by proteomic methods. The presence and distribution of Annexin A1, A2 and A5 (ANXA1, ANXA2, ANXA5) in human semen was analysed and the corresponding mRNAs studied in spermatozoa. All three ANXAs were present in prostasomes and spermatozoa, but only ANXA1 in prostasomes-free seminal plasma.

Sperm binding to porcine oviductal cells is mediated by SRCR domains contained in DMBT1.

The oviduct is an organ in which a subpopulation of sperm is stored in a reservoir, preserving its fertilizing potential. In porcine, two oviductal proteins have been identified in relation to sperm binding, Annexin A2 and Deleted in Malignant Brain Tumor 1 (DMBT1). DMBT1 is a multifunctional, multidomain glycoprotein, and the characteristics of all of its domains, as well as its carbohydrates, make them candidates for sperm binding.

Annexin A2 and S100A10 in the mammalian oviduct.

In many mammals, upon entry into the female reproductive tract, a subpopulation of sperm is stored in the oviduct forming a functional reservoir. In the oviducts of pig and cow, Annexin A2 (AnxA2) has been linked to the binding of sperm. This protein may exist as a monomer or bound to S100A10 and both forms are associated with different biological functions.

Heparin binding analysis of boar sperm and its relation with farrowing capacity.

New methods for the evaluation of semen quality according to in vivo reproductive data are useful tools for identifying boars of lower fertility among individuals with standard semen parameters. In this study, indirect fluorescence microscopy was used to evaluate the heparin binding site distribution upon capacitation of sperm from eight boars arbitrarily distributed into Groups I and II according to differential farrowing rates (Group I: ≥ 70%, Group II: < 70%). Additionally, the ability of sperm to bind to solubilised zona pellucida (ZP) was assayed in the presence or absence of heparin.

S100A7 in the Fallopian tube: a comparative study.

The oviduct is a dynamic organ in which final gamete maturation, fertilization and early embryo development take place. It is considered to be a sterile site; however the mechanism for sterility maintenance is still unknown. S100A7 is an anti-microbial peptide that has been reported in human reproductive tissues such as prostate, testicle, ovary, normal cervical epithelium and sperm.

The effect of oviductal deleted in malignant brain tumor 1 over porcine sperm is mediated by a signal transduction pathway that involves pro-AKAP4 phosphorylation.

The interaction between sperm and oviduct results in the selection of sperm with certain qualities. Porcine oviductal deleted in malignant brain tumor 1, DMBT1 (previously called sperm-binding glycoprotein, SBG), has been proposed to be implicated in sperm selection through acrosome alteration and suppression of motility of a subpopulation of sperm that have begun capacitation prematurely. It produces in vitro acrosome alteration and decrease of motility of boar sperm, concomitant with tyrosine phosphorylation of a 97 kDa sperm protein (p97).

Apical membranes prepared by peeling from whole porcine oviducts interact with homologous sperm.

In mammals, interaction between sperm and oviductal epithelial cells provides the formation of a sperm reservoir and sperm selection at the isthmus of the oviduct. Several in vitro methods are used to study this interaction. Apical plasma membranes (APM) have been prepared by peeling from culture and differentiated kidney cells.

Molecular identification of the sperm selection involved porcine sperm binding glycoprotein (SBG) as deleted in malignant brain tumors 1 (DMBT1).

Porcine sperm binding glycoprotein (SBG) is involved in sperm-oviduct interaction. Here we use mass spectrometry to identify SBG, finding peptides corresponding to deleted in malignant brain tumors 1 (DMBT1), at scavenger receptor cysteine-rich (SRCR) and CUB domains. RT-PCR allowed the cloning of unique sequences, belonging to porcine DMBT1.

Porcine oviduct sperm binding glycoprotein and its deleterious effect on sperm: a mechanism for negative selection of sperm?

In their journey through the oviduct some subpopulations of sperm are preserved in a reservoir, while others are negatively selected. Sperm binding glycoprotein (SBG) is a pig oviductal epithelial cell glycoprotein that produces, under capacitating conditions, acrosome alteration, p97 tyrosine-phosphorylation and reduction of the motility of sperm. In this paper, we show that SBG is accessible at the extracellular surface of the oviductal epithelial cells, supporting a sperm interaction biological role in situ.

Dynamics of heparin-binding proteins on boar sperm.

The presence, topology and dynamics of heparin-binding proteins (HBP) on boar sperm were evaluated. HBP distribution was analyzed by subcellular parting, using biotinylated heparin followed by colorimetric detection. HBP were detected as peripherical and integral periacrosomal membrane proteins.

Annexin A2 is involved in pig (Sus scrofa)sperm-oviduct interaction.

The oviduct is a dynamic organ which modulates gamete physiology. Sperm-oviduct interaction provides the formation of a sperm storage reservoir and allows the selection of sperm with certain qualities in eutherian mammals. In sows, the oviductal sperm binding glycoprotein (SBG) has been proposed to be involved in sperm selection.

Sperm binding glycoprotein (SBG) produces calcium and bicarbonate dependent alteration of acrosome morphology and protein tyrosine phosphorylation on boar sperm.

The oviduct is a dynamic organ which modulates gamete physiology. Two subpopulations of sperm have been described in the oviduct of sows, a majority with normal appearance in the deep furrows and a minority, centrally located, and showing damaged membranes. Sperm-oviduct interaction provides the formation of a sperm storage and allows the selection of sperm with certain qualities.

Effect of heparin on in vitro capacitation of boar sperm.

Chlortetracycline (CTC) fluorescent pattern, the ability to undergo acrosome reaction (AR) upon exposure to 10 microM calcium ionophore A23187 and vitality estimation were used to investigate the effect of the sulfated glycosaminoglycan heparin on the in vitro capacitation of porcine spermatozoa. Sperm incubation in capacitating medium (CM) supplemented with 10 mM heparin for up to 120 min, showed an increase in the number of capacitated sperm (B pattern) and acrosome reacted sperm (AR pattern), without affecting their viability. In this condition, spermatozoa were incubated in CM depleted of albumin, calcium, bicarbonate or combinations, in the presence of heparin.

Sperm binding glycoprotein is differentially present surrounding the lumen of isthmus and ampulla of the pig's oviduct.

In several mammals a sperm reservoir is formed at the isthmus of the Fallopian tube, providing viable, potentially fertile sperm for an extensive period. In pig (Sus scrofa) the spermadhesin AQN-1 seems to be involved in the establishment of the sperm reservoir. The pig oviductal protein, sperm binding glycoprotein (SBG), binds to sperm and exposes carbohydrate groups that can be recognized by AQN-1.

One step purification and biochemical characterization of a spermatozoa-binding protein from porcine oviductal epithelial cells.

In pigs the binding of sperm to oviductal epithelial cells to form a sperm reservoir involves carbohydrate interactions. In the present study, we purify a sperm binding glycoprotein (SBG) from cells from the isthmus of the oviduct using an affinity column. This protein conjugated with FITC is able to bind to the heads of pig sperm.