Intracellular free calcium oscillations in normal and cleavage-blocked embryos and artificially activated eggs of Xenopus laevis.
We have measured levels of intracellular free calcium ([Ca2+]i) in albino Xenopus laevis embryos using recombinant aequorin and a photon-counting system. We observed sinusoidal oscillations in [Ca2+]i that had the same frequency as cleavage, with cleavage occurring when [Ca2+]i was lowest. An increase in calcium was seen to precede first cleavage. The cyclic changes in calcium were superimposed on a secondary pattern that increased, peaked between third and fifth cleavages and then slowly declined to a level similar to that measured before first cleavage. The amplitude of the oscillations was small during the first few cleavages but became larger with each cycle, with the largest oscillations occurring when the secondary pattern peaked (between third and fifth cleavage). As the secondary pattern declined, the amplitude of the oscillations also became smaller. The oscillations are due to release of calcium from intracellular stores, since the signal was the same in calcium-free solution as in normal medium. When cleavage was blocked with the microtubule-disrupting drugs colchicine or nocodazole, the [Ca2+]i oscillations persisted. Calcium oscillations of a similar magnitude and frequency were also present in artificially activated eggs. The secondary pattern was different in cleavage-blocked embryos and artificially activated eggs, the baseline increasing until about the third cycle and then remaining elevated for the rest of the recording (> 8 hours). By fixing embryos at various points in the calcium cycle, we determined that mitosis began shortly after calcium levels reached their peak and was complete before the calcium level dropped to its lowest point.(ABSTRACT TRUNCATED AT 250 WORDS)
Keating
T J
TJ
Department of Biological Sciences, Purdue University, West Lafayette, IN 47907-1392.
Cork
R J
RJ
Robinson
K R
KR
D000331:Aequorin / Q000235:genetics / Q000378:metabolism; D000417:Albinism; D000818:Animals; D002118:Calcium / Q000378:metabolism*; D002453:Cell Cycle / Q000502:physiology*; D004625:Embryo, Nonmammalian / Q000502:physiology; D005306:Fertilization; D008163:Luminescent Measurements; D008839:Microelectrodes; D010063:Ovum / Q000502:physiology*; D010507:Periodicity; D017785:Photons; D011994:Recombinant Proteins / Q000378:metabolism; D014982:Xenopus laevis / Q000196:embryology
D011994:Recombinant Proteins; D000331:Aequorin; D002118:Calcium