THE ZYMOGRAM TECHNIQUE AS A TOOL FOR STUDY OF GENOTYPIC DIFFERENCES

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Abstract
Combination of zone electro-phoresis in agar and histochemical techniques reveals genotypic enzymic differences between homologous organs of different inbred strains of mice and mutants versus normals. In choosing neuro-muscular mutants mostly patterns of hydrolytic enzyme activities from brain and brain regions have been obtained. "All or none "enzymic differences have occurred in five mutants compared with normals and using a variety of substrates for esterases. In one, "reeler," with 100 to 200% increase in (eserine sensitive) cholinesterase activity of the cerebellum, the enzyme has been shown to be present in a cryptic form; it is either tightly bound to a particulate fraction that becomes unmasked, is dissociated from an inactive complex by repeated freeze-thawing, or is blocked by an inhibitor that is differentially absorbed by Whatman filter paper No 4. Minute genotypic enzyme differences are detectable more easily in brains than livers because of lesser amounts of hydrolytic enzyme activity; also to avoid specific organ and tissue contaminations by enzymes in the blood mice have been heparinized and perfused prior to extraction.

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