External Mg2+ triggers oscillations and a subsequent sustained level of intracellular free Ca2+, correlated with changes in membrane conductance in the oocyte of the prawn Palaemon serratus.
We have provided evidence that external Mg2+ induces correlated changes in [Ca2+]i and membrane current or potential in prawn oocytes without any requirement of fertilization, using the fluorescent Ca2+ indicator Ca green dextran and voltage and current clamp methods. Replacement of Mg2+-free ASW with standard (40 mM Mg2+)ASW triggered a diphasic [Ca2+]i response consisting of an oscillation period followed by a second state of sustained [Ca2+]i level devoid of oscillation, lasting about 70 min and up to 3 hr, respectively. In contrast, oocytes maintained for a long time in Mg2+-free ASW showed no changes in [Ca2+]i which remained at a basal concentration of about 0.2 microM. The simultaneous records of [Ca2+]i and membrane or current changes showed that the oscillation period started with a first [Ca2+]i peak (about 1.7 microM) and was correlated with an initial transient membrane hyperpolarization or a transient initial outward current peak. The first [Ca2+]i peak was followed by a slow decrease in [Ca2+]i followed by a series of [Ca2+]i transients, concurrent with a slow depolarization of the membrane or a related inwardly directed current. The oscillation period ended with an oscillatory plateau of [Ca2+]i (about 0.6 microM) lasting 49.6 +/- 4.5 min, the onset of which was concomitant with a final membrane hyperpolarization or a related final outward current. A continuous contact between external Mg2+ and the oocyte membrane was necessary to maintain the program of [Ca2+]i and membrane conductance changes. The sources of Ca2+ mobilization are of internal origin for both the first peak and the subsequent series of [Ca2+]i oscillations, and are mainly of external origin for the oscillation plateau and for the second state of sustained [Ca2+]i level. The first and second step of the cortical reaction occurred during the oscillatory plateau and the second state of sustained [Ca2+]i level, respectively.
Goudeau
M
M
Observatoire Océanographique et de Biologie marine de Roscoff, UPR CNRS, Roscoff, 9042, France.
Goudeau
H
H
D000818:Animals; D002118:Calcium / Q000378:metabolism*; D002462:Cell Membrane / Q000187:drug effects / Q000502:physiology*; D005453:Fluorescence; D066298:In Vitro Techniques; D007473:Ion Channels / Q000378:metabolism; D008274:Magnesium / Q000494:pharmacology / Q000502:physiology*; D008564:Membrane Potentials / Q000187:drug effects / Q000502:physiology*; D009865:Oocytes / Q000187:drug effects / Q000502:physiology*; D033541:Palaemonidae / Q000378:metabolism*; D018408:Patch-Clamp Techniques / Q000379:methods; D012623:Seawater / Q000737:chemistry; D015398:Signal Transduction / Q000187:drug effects / Q000502:physiology
D007473:Ion Channels; D008274:Magnesium; D002118:Calcium