Click
here to close Hello! We notice that
you are using Internet Explorer, which is not supported by Echinobase
and may cause the site to display incorrectly. We suggest using a
current version of Chrome,
FireFox,
or Safari.
Bio Protoc
2020 Oct 05;1019:e3778. doi: 10.21769/BioProtoc.3778.
Show Gene links
Show Anatomy links
Fluorescence Measurement and Calibration of Intracellular pH in Starfish Oocytes.
Hosoda E
,
Chiba K
.
???displayArticle.abstract???
Oocyte maturation is a process wherein an oocyte arrested at prophase of meiosis I resumes meiosis to become a fertilizable egg. In starfish ovaries, a hormone released from follicle cells activates the oocytes, resulting in an increase in their intracellular pH (pHi), which is required for spindle assembly. Herein, we describe a protocol for pHi measurement in living oocytes microinjected with the pH-sensitive dye BCECF. For in vivo BCECF calibration, we treated oocytes with artificial seawater containing CH3COONH4 to clamp pHi, injected pH-standard solutions, and converted the BCECF fluorescence intensity ratios to pHi values. Of note, if the actual pHi is higher or lower than the known pH of injected standard solutions, the BCECF fluorescence intensity ratio will decrease or increase, respectively. On the other hand, the pH of the injected solution displaying no change in fluorescence intensity should be considered the actual pHi. These methods for pHi calibration and clamping are simple and reproducible.
Anand,
Rise in intracellular pH is concurrent with 'start' progression of Saccharomyces cerevisiae.
1989, Pubmed
Anand,
Rise in intracellular pH is concurrent with 'start' progression of Saccharomyces cerevisiae.
1989,
Pubmed
Boyarsky,
Superiority of in vitro over in vivo calibrations of BCECF in vascular smooth muscle cells.
1996,
Pubmed
Chiba,
The primary structure of the alpha subunit of a starfish guanosine-nucleotide-binding regulatory protein involved in 1-methyladenine-induced oocyte maturation.
1992,
Pubmed
,
Echinobase
Chiba,
Induction of starfish oocyte maturation by the beta gamma subunit of starfish G protein and possible existence of the subsequent effector in cytoplasm.
1993,
Pubmed
,
Echinobase
Grillo-Hill,
Increased H⁺ efflux is sufficient to induce dysplasia and necessary for viability with oncogene expression.
2015,
Pubmed
Hamaguchi,
Regulation of intracellular pH in sea urchin eggs by medium containing both weak acid and base.
1997,
Pubmed
,
Echinobase
Harada,
Metaphase I arrest of starfish oocytes induced via the MAP kinase pathway is released by an increase of intracellular pH.
2003,
Pubmed
,
Echinobase
Harada,
Regulation of intracellular pH by p90Rsk-dependent activation of an Na(+)/H(+) exchanger in starfish oocytes.
2010,
Pubmed
,
Echinobase
Hiramoto,
A method of microinjection.
1974,
Pubmed
Hiraoka,
SGK phosphorylates Cdc25 and Myt1 to trigger cyclin B-Cdk1 activation at the meiotic G2/M transition.
2019,
Pubmed
,
Echinobase
Hosoda,
SGK regulates pH increase and cyclin B-Cdk1 activation to resume meiosis in starfish ovarian oocytes.
2019,
Pubmed
,
Echinobase
Johnson,
Intracellular pH and activation of sea urchin eggs after fertilisation.
1976,
Pubmed
,
Echinobase
Karagiannis,
Intracellular pH homeostasis during cell-cycle progression and growth state transition in Schizosaccharomyces pombe.
2001,
Pubmed
Moolenaar,
Na+/H+ exchange and cytoplasmic pH in the action of growth factors in human fibroblasts.
NULL,
Pubmed
Moriwaki,
Arrest at metaphase of meiosis I in starfish oocytes in the ovary is maintained by high CO2 and low O2 concentrations in extracellular fluid.
2013,
Pubmed
,
Echinobase
Murabe,
The Starfish Asterina pectinifera: Collection and Maintenance of Adults and Rearing and Metamorphosis of Larvae.
2021,
Pubmed
,
Echinobase
Rink,
Cytoplasmic pH and free Mg2+ in lymphocytes.
1982,
Pubmed
Roos,
Intracellular pH.
1981,
Pubmed
Sadler,
Components of the signaling pathway linking the 1-methyladenine receptor to MPF activation and maturation in starfish oocytes.
1998,
Pubmed
,
Echinobase
Schuldiner,
Na+/H+ antiport in Swiss 3T3 cells: mitogenic stimulation leads to cytoplasmic alkalinization.
1982,
Pubmed
Shen,
Direct measurement of intracellular pH during metabolic derepression of the sea urchin egg.
1978,
Pubmed
,
Echinobase
Tadenuma,
Properties of 1-methyladenine receptors in starfish oocyte membranes: involvement of pertussis toxin-sensitive GTP-binding protein in the receptor-mediated signal transduction.
1992,
Pubmed
,
Echinobase
Thomas,
Intracellular pH measurements in Ehrlich ascites tumor cells utilizing spectroscopic probes generated in situ.
1979,
Pubmed
Tilney,
Polymerization of actin. IV. Role of Ca++ and H+ in the assembly of actin and in membrane fusion in the acrosomal reaction of echinoderm sperm.
1978,
Pubmed
,
Echinobase