Microsurgical removal of centrosomes blocks cell reproduction and centriole generation in BSC-1 cells.
We have removed the centrosome from cultured BSC-1 cells by microsurgery, leaving enough cytoplasm with the nucleated cell fragment (karyoplast) to ensure survival and growth. In each experiment, we followed the fate of the karyoplast as well as the anucleate cell fragment (cytoplast) containing the original pair of centrioles. Experimental karyoplasts reestablish a juxtanuclear microtubule-organizing center, an astral array of microtubules, and a compact Golgi apparatus. They enter and presumably complete S phase, and they grow beyond the size of an average BSC-1 cell. However, they do not regenerate centrioles in time periods equivalent to more than 10 cell cycles and do not undergo cell division. Control-operated cells with centrosomes left in the karyoplast progress through the cell cycle, duplicate the centrosome, and form clonal cell colonies. We conclude that the removal of centrioles uncouples cell growth from cell reproduction and impedes centriole biogenesis and centrosome duplication.
Maniotis
A
A
Department of Molecular and Cell Biology, University of California, Berkeley 94720.
Schliwa
M
M
D000818:Animals; D001973:Bromodeoxyuridine / Q000378:metabolism; D002453:Cell Cycle; D002455:Cell Division; D002467:Cell Nucleus / Q000502:physiology; D002502:Centrioles / Q000502:physiology* / Q000648:ultrastructure; D002522:Chlorocebus aethiops; D003593:Cytoplasm / Q000502:physiology* / Q000648:ultrastructure; D005455:Fluorescent Antibody Technique; D006056:Golgi Apparatus / Q000648:ultrastructure; D066298:In Vitro Techniques; D008866:Microsurgery; D013997:Time Factors
D001973:Bromodeoxyuridine