Calcium uptake by bovine epididymal spermatozoa is regulated by the redox state of the mitochondrial pyridine nucleotides.
Immature caput epididymal sperm accumulate calcium from exogenous sources at a rate 2- to 4-fold greater than mature caudal sperm. Calcium accumulation by these cells, however, is maximal in the presence of lactate as external substrate. This stimulation of calcium uptake by optimum levels of lactate (0.8-1.0 mM) is about 5-fold in caput and 2-fold in caudal sperm compared to values observed with glucose as substrate. Calcium accumulation by intact sperm is almost entirely mitochondrial as evidenced by the inhibition of uptake by rotenone, antimycin, and ruthenium red. The differences in the ability of the various substrates in sustaining calcium uptake appeared to be related to their ability to generate NADH (nicotinamide adenine dinucleotide). Previous reports have documented that mitochondrial calcium accumulation in several somatic cells is regulated by the oxidation state of mitochondrial NADH. A similar situation obtains for bovine epididymal sperm since calcium uptake sustained by site III oxidation of ascorbate in the presence of tetramethyl phenylenediamine and rotenone was also stimulated by NADH-producing substrates, including lactate, and inhibited by substrates generating NAD+ (nicotinamide adenine dinucleotide, oxidized form). Further, calcium uptake by digitonin-permeabilized sperm in the presence of succinate was stimulated when NADH oxidation was inhibited by rotenone. The compounds alpha-keto butyric, valeric, and caproic acids, which generate NAD+, inhibited the maximal calcium uptake observed in the presence of succinate and rotenone, and the hydroxy acids lactate and beta-hydroxybutyrate reversed this inhibition. These results document the regulation of sperm calcium accumulation by the physiological substrate lactate, emphasize the importance of mitochondria in the accumulation of calcium by bovine epididymal sperm, and suggest that the mitochondrial location of the isozyme LDH-X in mammalian sperm may be involved in the regulation of calcium accumulation.
Vijayaraghavan
S
S
Department of Reproductive Biology and Behavior, Oregon Regional Primate Research Center, Beaverton 97006.
Bhattacharyya
A
A
Hoskins
D D
DD
D020155:3-Hydroxybutyric Acid; D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D001205:Ascorbic Acid / Q000494:pharmacology; D002087:Butyrates / Q000494:pharmacology; D020148:Butyric Acid; D002118:Calcium / Q000378:metabolism*; D002208:Caproates / Q000494:pharmacology; D002417:Cattle; D004072:Digitonin / Q000494:pharmacology; D004822:Epididymis / Q000166:cytology*; D006885:Hydroxybutyrates / Q000494:pharmacology; D007773:Lactates / Q000494:pharmacology; D019344:Lactic Acid; D008297:Male; D008928:Mitochondria / Q000378:metabolism*; D009243:NAD / Q000378:metabolism*; D010084:Oxidation-Reduction; D010421:Pentanoic Acids / Q000494:pharmacology; D011773:Pyruvates / Q000494:pharmacology; D019289:Pyruvic Acid; D012402:Rotenone / Q000494:pharmacology; D013094:Spermatozoa / Q000187:drug effects / Q000378:metabolism* / Q000648:ultrastructure; D013772:Tetramethylphenylenediamine / Q000494:pharmacology
D002087:Butyrates; D002208:Caproates; D006885:Hydroxybutyrates; D007773:Lactates; D010421:Pentanoic Acids; D011773:Pyruvates; D012402:Rotenone; D009243:NAD; D020148:Butyric Acid; C001916:alpha-ketocaproic acid; D019344:Lactic Acid; D019289:Pyruvic Acid; D000255:Adenosine Triphosphate; C038780:n-pentanoic acid; D004072:Digitonin; D013772:Tetramethylphenylenediamine; D001205:Ascorbic Acid; D002118:Calcium; D020155:3-Hydroxybutyric Acid