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Induction of Fertilization Membrane Formation and Cyanide‐insensitive Respiration in Sea Urchin Eggs by the Treatment with Dimethylsulfoxide Followed by an Incubation in an Ice Bath (dimethylsulfoxide/sea urchin egg/fertilization membrane/respiration/verapamil)
Author(s) -
FUJIWARA AKIKO,
ASAMI KOUICHI,
YASUMASU IKUO
Publication year - 1987
Publication title -
development, growth and differentiation
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.864
H-Index - 66
eISSN - 1440-169X
pISSN - 0012-1592
DOI - 10.1111/j.1440-169x.1987.00013.x
Subject(s) - hemicentrotus , sea urchin , human fertilization , incubation , chemistry , cyanide , egta , 2,4 dinitrophenol , respiration , sperm , botany , biophysics , biology , zoology , biochemistry , anatomy , calcium , ecology , inorganic chemistry , organic chemistry
In unfertilized eggs of the sea urchin Hemicentrotus pulcherrimus , fertilization membrane formation was induced by an incubation with dimethylsulfoxide (DMSO) for several min at 20°c followed by another incubation in an ice bath. The number of eggs with fertilization membrane, thus obtained, increased in relation to the concentration of DMSO between 1 and 3% (v/v) and was higher than 75% at concentrations above 3%. Fertilization membrane formation by this treatment occurred in Ca 2+ free‐ or Ca 2+ , Mg 2+ free‐ artificial sea water containing EGTA (50 mM) and was inhibited by verapamil. In the presence of DMSO, the membrane formation was also induced by 2, 4‐dinitrophenol or cyanide in considerable number of eggs at 20°c. Eggs remained fertilizable, even when they were kept with DMSO for 1 hr at 20°c. DMSO slightly enhanced respiratory rate in unfertilized eggs and substantially reduced it in fertilized eggs. DMSO‐treated eggs exhibited cyanide‐insensitive respiratory burst following chilling in an ice bath or by adding DNP or cyanide, in a similar manner to the burst induced by sperm.