The activating effects of bicarbonate on sperm motility and respiration at ejaculation.
ABSTRACT Mature porcine sperm preserved in the cauda epididymis are quiescent. At ejaculation, they are mixed with the seminal vesicle fluid containing HCO3- and are rapidly activated. The role of HCO3- on the sperm activation process at ejaculation was studied in vitro. HCO3- quickly increased the motility, respiration rate and cAMP content of the porcine epididymal sperm. The extent of activation was proportional to the pCO2 in the medium. The activating effect of HCO3- on the motility was observed even in the absence of fructose as well as in the presence of KCN. 8-Bromoadenosine 3',5'-cyclic monophosphate and theophylline showed similar activating effects to that of HCO3-. However, HCO3(-)-free seminal plasma, Ca2+, amino acids, intermediates of the Krebs cycle, substrates of respiration and increases in the intracellular pH, extracellular pH or ionic strength of the medium had no effect. Fructose sustained the active state of the sperm and gradually increased both the motility and respiration rate when the dose of HCO3- was low. The anion channel blocker enhanced the activating effect of HCO3-. These results suggest that, upon ejaculation, HCO3- is a unique activator in vivo which makes the quiescent sperm motile via the HCO3(-)-adenylate cyclase-cAMP system, to which an endogenous HCO3- derived from metabolic CO2 may be related.
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ABSTRACT: The solute carrier 26 (SLC26) family emerges as a distinct class of anion transporters with its members SLC26A3 (Slc26a3) and SLC26A6 (Slc26a6) reported to be electrogenic Cl(-) /HCO3 (-) exchangers. While it is known that uterine fluid has high HCO3 (-) content and that HCO3 (-) is essential for sperm capacitation, the molecular mechanisms underlying the transport of HCO3 (-) across uterine epithelial cells and sperm have not been fully investigated. The present review re-examines the results from early reports studying anion transport, finding clues for the involvement of Cl(-) /HCO3 (-) anion exchanges in electrogenic HCO3 (-) transport across endometrial epithelium. We also summarize recent work on Slc26a3 and Slc26a6 in uterine epithelial cells and sperm, revealing their functional role in working closely with the cystic fibrosis transmembrane conductance regulator (CFTR) for HCO3 (-) transport in these cells. The possible involvement of these anion exchangers in other HCO3 (-) dependent reproductive processes and their implications for infertility are also discussed.Cell Biology International 01/2014; 38(1). DOI:10.1002/cbin.10183 · 1.64 Impact Factor
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ABSTRACT: Numerous reports have appeared on the occurrence of undefined protein factors in male reproductive fluids that promote motility of mature sperm and initiate forward motility in the immature (immotile) caput-epididymal sperm. This study reports for the first time purification to apparent homogeneity of a motility initiating protein (MIP) from epididymal plasma and its characterization using the caprine sperm model. It is a 125 kDa (approximately) dimeric protein made up of two subunits: 70 and 54 kDa. MIP is an acidic protein with an isoelectric point of 4.75. The motility protein at 30 microg/ml (240 nM) level showed nearly maximal motility-promoting activity. MIP is heat stable and it is maximally active at pH 8. It is a glycoprotein that binds with high affinity to concanavalin A and it contains mannose, galactose, and N-acetyl glucosamine approximately in the ratios of 6:1:6. It is sensitive to the actions of alpha-mannosidase and beta-N-acetylglucoseaminidase thereby demonstrating that the sugar side chain of the glycoprotein is essential for its biological activity. Epididymal plasma is its richest source. It is also capable of enhancing forward motility of mature cauda-sperm. Its antibody markedly inhibits sperm motility. MIP antibody is highly immunospecific and it recognizes both the subunits. MIP causes significant increase of the intrasperm level of cyclic AMP. MIP: the physiological motility-activating protein has potential for use as a contraceptive vaccine and for solving some of the problems of human infertility and animal breeding.Journal of Cellular Physiology 01/2010; 222(1):254-63. DOI:10.1002/jcp.21947 · 3.87 Impact Factor
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ABSTRACT: The HCO3(-) anion activates sperm motility, an important early step in capacitation, by increasing flagellar beat frequency through a pathway that requires the atypical adenylyl cyclase SACY and the sperm-specific C alpha2 catalytic subunit of PKA. Here we show that the accelerating action of HCO3(-) also requires the continued presence of external Ca2+ (EC50 approximately 0.5 mM), and find that Ca2+ can be replaced by Sr2+ but not by Mn2+. Ca2+ is required for HCO3(-) to elevate cAMP, but not for cAMP-AM to increase beat frequency, indicating that external Ca2+ acts before rather than after stimulation of SACY by HCO3(-). With external Ca2+ present, HCO3(-) does not alter cytosolic or near-membrane [Ca2+]. Removal of external Ca2+ initiates a slow decline in intracellular [Ca2+] and rapid block of the HCO3(-)-evoked acceleration that is not relieved upon increasing internal [Ca2+] by rapid photolysis of caged Ca2+. We also find that the rapid (t(1/2) approximately 10 s) accelerating action of HCO3(-) is slowed more than three-fold by the carbonic anhydrase inhibitor acetazolamide. It is unaltered by the broad spectrum anion transport inhibitor SITS, and is not accompanied by detectable changes in intracellular pH. We propose that external Ca2+ binds an unidentified extracellular protein that is required for HCO3(-) to engage cAMP-mediated activation of motility.Developmental Biology 01/2008; 312(1):183-92. DOI:10.1016/j.ydbio.2007.09.017 · 3.64 Impact Factor