Kinetochore detection in micronuclei: an alternative method for measuring chromosome loss.
Whole chromosomes within micronuclei (MN) in cytokinesis-blocked human lymphocytes were detected by using anti-kinetochore antibodies obtained from the serum of scleroderma patients. The primary antibody was localized using a peroxidase-labelled second antibody followed by a nickel chloride modification of the diamino-benzidine reaction to give a permanent slide preparation. Between 82 and 92% of colchicine-induced MN were shown to be kinetochore-positive. Results for spontaneously occurring micronuclei in young (20-35 years) and elderly (greater than 65 years) subjects indicated that 42 (+/- 6) and 50 (+/- 6)%, respectively, contained kinetochores. Perhaps the more novel observation was that approximately 12% of X-ray-induced micronuclei were kinetochore-positive and thus could have been the result of whole chromosome loss events. Kinetochore-detection in micronuclei provides a new approach to measure chromosome loss and therefore may be important in identifying aneuploidy-inducing agents.
Fenech
M
M
MRC Cell Mutation Unit, University of Sussex, Falmer, Brighton, UK.
Morley
A A
AA
D000328:Adult; D000368:Aged; D000906:Antibodies; D002455:Cell Division / Q000187:drug effects / Q000528:radiation effects; D002872:Chromosome Deletion; D002875:Chromosomes / Q000187:drug effects / Q000276:immunology / Q000528:radiation effects / Q000648:ultrastructure; D003078:Colchicine; D005821:Genetic Techniques; D006801:Humans; D007124:Immunoenzyme Techniques; D008214:Lymphocytes / Q000187:drug effects / Q000528:radiation effects / Q000648:ultrastructure; D015162:Micronucleus Tests; D014965:X-Rays
D000906:Antibodies; D003078:Colchicine