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Mass Isolation of Muscle Lineage Blastomeres from Ascidian Embryos
Author(s) -
MITAMIYAZAWA IZUMI,
SATOH NORIYUKI
Publication year - 1986
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.1986.00483.x
Subject(s) - aphidicolin , embryo , cytochalasin b , percoll , acetylcholinesterase , blastomere , biology , microbiology and biotechnology , aché , dna replication , dna , centrifugation , cell , enzyme , biochemistry , embryogenesis
The aim of this investigation was to establish an experimental system for studying the causal relationship between DNA replication and tissue‐specific enzyme development in ascidian embryos. Blastomeres were dissociated from 44∽64‐ceIl Halocynthia roretzi embryos and fractionated by centrifugation through a discontinuous Percoll density gradient. When cells harvested from the fraction at the bottom of the tube were division‐arrested with cytochalasin B (an inhibitor of cytokinesis) soon after their isolation, more than 70% of them developed histochemically‐detectable muscle‐specific acetylcholinesterase (AChE) activity, suggesting that they were almost all blastomeres of muscle lineage. When these cells were arrested with aphidicolin (an inhibitor of DNA replication) and cytochalasin B immediately after their isolation, however, none of them showed AChE activity. When they were allowed to divide once and then arrested with the inhibitors, nearly 40 % of them developed AChE activity, and when they were allowed to divide twice before arrest, about 70% of them showed AChE activity.