Confocal laser scanning microscopy of calcium dynamics in living cells

Stricker, Stephen A.; Whitaker, Michael

doi:10.1002/(sici)1097-0029(19990915)46:6<356::aid-jemt4>3.0.co;2-6

Cited by 42 publications

(11 citation statements)

References 182 publications

(174 reference statements)

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“…The OGB-1 dextran is co-injected into the cell with Texas Red (TR) dextran, which is insensitive to Ca 2+ levels, and fluorescence of the cell in both channels is recorded, such that a ratio of OGB-1 fluorescence to TR fluorescence can be determined. These ratiometric readings allow the precise quantification of Ca 2+ -dependent fluorescence as they normalize for potential variation in injected dye quantities, or photobleaching during fluorescence recordings (Stricker and Whitaker, 1999). The recording parameters in these experiments required 10-second intervals between frames, precluding us from reliably detecting the Percentage of fertilized eggs forming normal FEs was determined for each treatment (A, Gαs-inhibiting reagents; B, Gαq-inhibiting reagents; C, Gαi-inhibiting reagents).…”

Section: + Release?supporting

confidence: 75%

βγ subunits of heterotrimeric G-proteins contribute to Ca2+ release at fertilization in the sea urchin

Voronina

Wessel

2004

Journal of Cell Science

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A cytoplasmic Ca2+ transient is required for egg activation at fertilization in all animals. The pathway leading to release of Ca2+ from the endoplasmic reticulum in echinoderms includes activation of a SRC homolog, followed by phospholipase Cγ activation, and formation of inositol trisphosphate. However, the upstream activators or modulators of this signaling pathway are not known. We recently identified four Gα subunits of heterotrimeric G-proteins present in the sea urchin egg, and here we find that activation of G-proteins of the Gαs and Gαq type, but not Gαi or Gα12 type, is required for normal Ca2+ dynamics at fertilization. The effects of these G-proteins are mediated by the Gβγ subunits, occur upstream of the cytoplasmic Ca2+ release, and influence both the amplitude of Ca2+ release and the duration of the lag phase. We propose integration of the G-protein input into the framework of signaling at sea urchin fertilization.

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Section: + Release?supporting

confidence: 75%

βγ subunits of heterotrimeric G-proteins contribute to Ca2+ release at fertilization in the sea urchin

Voronina

Wessel

2004

Journal of Cell Science

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“…Confocal imaging was also used to monitor fertilization-induced Ca 2+ signaling in specimens injected with Calcium-Green-and Rhodamine-B dextrans (Stricker and Whitaker, 1999;Stricker, 2000). Graphs were expressed as uncalibrated changes in fluorescence intensity relative to either the pre-stimulus CG/RB ratio (Ro) or the initial non-ratioed CG fluorescence (Fo).…”

Section: Morphology and Ca 2+ Imagingmentioning

confidence: 99%

Endoplasmic reticulum reorganizations and Ca2+ signaling in maturing and fertilized oocytes of marine protostome worms: the roles of MAPKs and MPF

Stricker

Smythe

2003

Development

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“…A variety of calcium signals are required for egg activation; these reflect both the plasticity of the signaling machinery as well as the distinct requirements for egg activation in different species. Species typically either display a single increase in calcium concentration (e.g., in sea urchins, starfish, frogs, and fish) or show calcium oscillations (e.g., in nemertian worms, ascidians, and mammals) (Stricker 1999;Stricker and Whitaker 1999;Miyazaki and Ito 2006). The mechanism(s) that mediate calcium influx remains poorly characterized in mammalian eggs.…”

Section: Fusion and Egg Activation Leading To Fertilizationmentioning

confidence: 99%

Vertebrate Reproduction

Kornbluth¹,

Fissore²

2015

Cold Spring Harb Perspect Biol

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Vertebrate reproduction requires a myriad of precisely orchestrated events-in particular, the maternal production of oocytes, the paternal production of sperm, successful fertilization, and initiation of early embryonic cell divisions. These processes are governed by a host of signaling pathways. Protein kinase and phosphatase signaling pathways involving Mos, CDK1, RSK, and PP2A regulate meiosis during maturation of the oocyte. Steroid signals-specifically testosterone-regulate spermatogenesis, as does signaling by G-protein-coupled hormone receptors. Finally, calcium signaling is essential for both sperm motility and fertilization. Altogether, this signaling symphony ensures the production of viable offspring, offering a chance of genetic immortality.

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Confocal laser scanning microscopy of calcium dynamics in living cells

Cited by 42 publications

References 182 publications

βγ subunits of heterotrimeric G-proteins contribute to Ca2+ release at fertilization in the sea urchin

βγ subunits of heterotrimeric G-proteins contribute to Ca2+ release at fertilization in the sea urchin

Endoplasmic reticulum reorganizations and Ca2+ signaling in maturing and fertilized oocytes of marine protostome worms: the roles of MAPKs and MPF

Vertebrate Reproduction

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