Cleavage of Binder of Sperm Protein 3 (BSP3), Which Binds Bull Sperm to Oviductal Epithelium, by Sperm Surface Serine Protease.

2012 ◽  
Vol 87 (Suppl_1) ◽  
pp. 437-437
Author(s):  
Pei-hsuan Hung ◽  
Susan S. Suarez
Keyword(s):  
Serine Protease ◽  
Sperm Protein ◽  
Sperm Surface ◽  
Oviductal Epithelium ◽  
Bull Sperm

180. CHARACTERIZATION AND FUNCTION OF THE BULL SPERM PROTEIN Spam1: TWO DISTINCT ISOFORMS WITH DISTINCT ROLES

2009 ◽  
Vol 21 (9) ◽  
pp. 98
Author(s):  
G. Morin ◽  
R. Sullivan ◽  
I. Laflamme ◽  
C. Robert ◽  
P. Leclerc
Keyword(s):  
Transmembrane Domain ◽  
Inhibitory Effect ◽  
Anterior Region ◽  
Protein Orientation ◽  
Sperm Protein ◽  
Sperm Binding ◽  
Glycosylation Sites ◽  
Sperm Membrane ◽  
Bull Sperm ◽  
And Function

We identified an 80 kDa bull sperm protein (p80) that possesses homology with the Sperm adhesion molecule 1 (Spam1), a GPI-anchored glycoprotein conserved amongst mammals that is required for fertilization. Since bovine Spam1 had not been identified, the aim of this project was to determine if p80 is the bovine Spam1 and to test the hypothesis that it plays a role in gamete interaction during bovine fertilization. Amino acid sequence deduced from 3`/5`Race confirmed that homology between p80 and Spam1 in various species ranged from 47 to 61%.It also revealed specific differences including the absence of a GPI-anchor, the presence of a transmembrane domain, and N- and O- glycosylation sites. By generating and characterising antibodies against p80 N- and C-terminal domains, the protein orientation in the sperm membrane was evaluated. We identified two populations of p80 on the sperm head: one internalised in the anterior region and the second localised on the post-acrosomal region with its hyaluronidase domain exposed to the extracellular environment. Proteomic and immunologic analyses revealed that the p80 post-acrosomal population is a shorter isoform originating from the epididymis while the full length p80 located on the anterior region originates from the testis. Finally, the potential function of p80 during the sperm/zp interaction was evaluated by sperm/zona pellucida (zp) binding assay. The C-terminal extremity of p80 was implicated in sperm binding to the zp by antibody and native protein competition. Furthermore, glycosylation was not required during this interaction since deglycosylated p80 in the incubation medium had the same inhibitory effect on zona binding as the native p80. Collectively, the results demonstrated that p80 is the bovine Spam1, and that two isoforms are present on bull sperm. The hyaluronidase activity of the post-acrosomal isoform is required for cumulus penetration, while the other one participates in sperm/zp binding during fertilization.

Comprehensive mapping of the bull sperm surface proteome

PROTEOMICS ◽  
2012 ◽  
Vol 12 (23-24) ◽  
pp. 3559-3579 ◽  
Author(s):  
Keren Byrne ◽  
Tamara Leahy ◽  
Russell McCulloch ◽  
Michelle L. Colgrave ◽  
Michael K. Holland
Keyword(s):  
Sperm Surface ◽  
Bull Sperm ◽  
Surface Proteome ◽  
Comprehensive Mapping

scholarly journals Redox control of surface protein sulphhydryls in bovine spermatozoa reversibly modulates sperm adhesion to the oviductal epithelium and capacitation

Reproduction ◽  
2009 ◽  
Vol 138 (1) ◽  
pp. 33-43 ◽  
Author(s):  
Roberto Gualtieri ◽  
Valentina Mollo ◽  
Gennaro Duma ◽  
Riccardo Talevi
Keyword(s):  
Surface Protein ◽  
Surface Proteins ◽  
Sperm Surface ◽  
Sperm Suspension ◽  
Oviductal Epithelium ◽  
Bovine Spermatozoa ◽  
Sulphated Glycosaminoglycans ◽  
Oviductal Fluid ◽  
Adhesion Assays

Oviductal fluid molecules, such as sulphated glycosaminoglycans and disulphide-reductants, may represent periovulatory signals for the release of spermatozoa from the oviductal reservoir in the bovine species. Disulphide-reductants release spermatozoa through the reduction of sperm-surface disulphides to sulphhydryls (SH). Herein, we studied sperm-surface protein SH through labelling with maleimidylpropionyl biocytin in the initial sperm suspension, in the subpopulations able and unable to adhere to the in vitro cultured oviductal epithelium, and in spermatozoa released either through the disulphide-reductant penicillamine (PEN) or the sulphated glycosaminoglycan heparin (HEP). Adhesion assays were performed to study the ability of released spermatozoa to readhere to the oviductal epithelium. Results showed that the level of SH in sperm-surface proteins was: 1) low in adhering spermatozoa; 2) high in spermatozoa unable to adhere; and 3) markedly increased in released spermatozoa. Adhesion assays showed that: 1) PEN-released spermatozoa promptly recovered adhesion after removal of the disulphide-reductant and could be released again in response to PEN; 2) conversely, a limited number of HEP-released spermatozoa was able to readhere to the oviductal epithelium and this ability was not affected by HEP removal. Recovery of adhesion was associated to reoxidation of sperm-surface protein SH and to the reversal of capacitation. In conclusion, redox modulation of sperm-surface protein SH is involved in the release of spermatozoa adhering to the oviduct in vitro; the reversible action of disulphide-reductants might be responsible for intermittent phases of adhesions and releases; and the irreversible action of HEP indicates that it may represent a terminal releasing signal.

The endocannabinoid system in bull sperm and bovine oviductal epithelium: role of anandamide in sperm–oviduct interaction

Reproduction ◽  
2009 ◽  
Vol 137 (3) ◽  
pp. 403-414 ◽  
Author(s):  
María Gracia Gervasi ◽  
Maximiliano Rapanelli ◽  
María Laura Ribeiro ◽  
Mariana Farina ◽  
Silvia Billi ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Cannabinoid Receptors ◽  
Fatty Acid Amide Hydrolase ◽  
Acid Amide ◽  
Endocannabinoid System ◽  
Sperm Binding ◽  
Bovine Sperm ◽  
Sperm Release ◽  
Oviductal Epithelium ◽  
Bull Sperm

Anandamide binds to cannabinoid receptors and plays several central and peripheral functions. The aim of this work was to study the possible role for this endocannabinoid in controlling sperm–oviduct interaction in mammals. We observed that bull sperm and bovine oviductal epithelial cells express cannabinoid receptors, CB1 and CB2, and fatty acid amide hydrolase, the enzyme that controls intracellular anandamide levels. A quantitative assay to determine whether anandamide was involved in bovine sperm–oviduct interaction was developed. R(+)-methanandamide, a non-hydrolysable anandamide analog, inhibited sperm binding to and induced sperm release from oviductal epithelia. Selective CB1 antagonists (SR141716A or AM251) completely blocked R(+)-methanandamide effects. However, SR144528, a selective CB2 antagonist, did not exert any effect, indicating that only CB1 was involved in R(+)-methanandamide effect. This effect was not caused by inhibition of the sperm progressive motility or by induction of the acrosome reaction. Overall, our findings indicate for the first time that the endocannabinoid system is present in bovine sperm and oviductal epithelium and that anandamide modulates the sperm–oviduct interaction, by inhibition of sperm binding and induction of sperm release from oviductal epithelial cells, probably by activating CB1 receptors.

Sign in / Sign up

Export Citation Format

Share Document