We used suction-pipette recording and fluo-4 fluorescence to study
light-induced Ca2+ release from the visible double cones of
zebrafish. In Ringer, light produces a slow decrease in fluorescence
which can be fitted by the sum of two decaying exponentials with time
constants of 0.5 and 3.8 s. In 0Ca2+–0Na+
solution, for which fluxes of Ca2+ across the outer segment
plasma membrane are greatly reduced, light produces a slow increase in
fluorescence. Both the decrease and increase are delayed after
incorporation of the Ca2+ chelator BAPTA, indicating that
both are produced by a change in Ca2+. If the
Ca2+ pool is first released by bright light in
0Ca2+–0Na+ solution and the cone returned
to Ringer, the time course of Ca2+ decline is much faster
than in Ringer without previous light exposure. This indicates that the
time constants of 0.5 and 3.8 s actually reflect a sum of
Na+/Ca2+-K+ exchange and
light-induced release of Ca2+. The Ca2+ released
by light appears to come from at least two sites, the first comprising
66% of the total pool and half-released by bleaching 4.8% of the
pigment. Release of the remaining Ca2+ from the second site
requires the bleaching of nearly all of the pigment. If, after release,
the cone is maintained in darkness, a substantial fraction of the
Ca2+ returns to the release pool even in the absence of
pigment regeneration. The light-induced release of Ca2+ can
produce a modulation of the dark current as large as 0.75 pA
independently of the normal transduction cascade, though the rise time
of the current is considerably slower than the normal light response.
These experiments show that Ca2+ can be released within the
cone outer segment by light intensities within the physiological range
of photopic vision. The role this Ca2+ release plays remains
unresolved.