Carrier-mediated gamma-aminobutyric acid uptake in human spermatozoa indicating the presence of a high-affinity gamma-aminobutyric acid transport protein.
Human spermatozoa are capable of a carrier-mediated gamma-aminobutyric acid (GABA) uptake. The uptake is dependent on the concentration of Cl- and Na+ in the external medium, and the kinetics of the carrier resembles high-affinity GABA transport proteins. The time-dependent uptake of GABA displays large interindividual differences and is not correlated to motility parameters or morphology in the individual sample. Incubation of human spermatozoa with radiolabeled GABA was performed. Swim-up preparations of human spermatozoa were incubated with [3H]GABA, and subsequent GABA uptake was measured at various times by scintillation counting. GABA was accumulated intracellularly, and the uptake could be inhibited by preincubation of the samples in 200 microM nipecotic acid. Addition of aminooxyacetic acid in the medium did not alter the results, indicating that the internalized GABA remained unmetabolized intracellularly throughout the observation period. Kinetic analysis of GABA uptake was performed, and the Km for GABA transport was 14 microM. GABA uptake was reduced by equimolar substitution of NaCl in the capacitating medium by KCl, choline chloride, LiCl, N-methyl-D-glucamine (HCl) or D-glucuronic acid (sodium salt). Maximal reduction of [3H]GABA uptake was observed when the Na+ fraction of the medium was replaced with KCl. The results indicate the presence of a high-affinity GABA transport protein in the plasma membrane of human spermatozoa. GABA uptake was subsequently measured in 30 individual semen samples from men of barren couples. Large interindividual differences in GABA uptake was observed, but GABA uptake was not correlated to motility parameters or to morphology in the individual samples analyzed.
Aanesen
A
A
Division for Obstetrics & Gynaecology, Karolinska Hospital, Stockholm, Sweden.
Fried
G
G
Andersson
E
E
Gottlieb
C
C
D002352:Carrier Proteins / Q000378:metabolism*; D002712:Chlorides / Q000378:metabolism; D005260:Female; D050485:GABA Plasma Membrane Transport Proteins; D006801:Humans; D066298:In Vitro Techniques; D007700:Kinetics; D008297:Male; D008565:Membrane Proteins / Q000378:metabolism*; D026901:Membrane Transport Proteins; D027361:Organic Anion Transporters; D012964:Sodium / Q000378:metabolism; D013081:Sperm Motility / Q000502:physiology; D013094:Spermatozoa / Q000378:metabolism* / Q000648:ultrastructure; D005680:gamma-Aminobutyric Acid / Q000378:metabolism* / Q000502:physiology
D002352:Carrier Proteins; D002712:Chlorides; D050485:GABA Plasma Membrane Transport Proteins; D008565:Membrane Proteins; D026901:Membrane Transport Proteins; D027361:Organic Anion Transporters; D005680:gamma-Aminobutyric Acid; D012964:Sodium