M. Carmen Vílchez, Marina
Morini, David S. Peñaranda, Víctor Gallego, Juan F. Asturiano, Luz Pérez
Abstract
The role of potassium from the seminal plasma and/or
the activation media was examined by selectively removing K+ from
this media, and by testing the use of K+ channel inhibitors and a
K-ionophore. Sperm motility was measured using a CASA system, intracellular K+
and pH were measured by flow cytometry, and sperm head area was measured by ASMA: Automated Sperm Morphometry Analyses. Sperm motility was notably
inhibited by the removal of K+ from the seminal plasma and by
treatment with the K+ ionophore valinomycin. This therefore indicates
that a reduction of K+ levels in the quiescent
stage inhibits further motility. The normal decrease in sperm head area induced
by seawater activation was altered by the removal of K+ from the seminal
plasma, and an increase in the pHi in the quiescent stage was also
induced. Intracellular pH (pHi) was quantitatively measured for
the first time in European eel spermatozoa, being 7.2 in the quiescent stage
and 7.1 post-activation. Intracellular and external pH levels influenced sperm
motility both in the quiescent stage and at activation. The alkalinization of
the pHi (by NH4Cl) inhibited sperm motility activation,
while acidification (by Na-acetate) did not have any effect. Our results
indicate that a pH gradient between the sperm cell and the seminal plasma is
necessary for sperm motility activation. The presence of the ion K+
in the seminal plasma (or in the extender medium) is necessary in order to
maintain sperm volume, intracellular pH and sperm motility.
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