The Journal of Cell Biology, Vol 95, 1-7, Copyright © 1982 by The Rockefeller University Press
Messenger RNA in the cytoskeletal framework: analysis by in situ hybridization
WR Jeffery
The possibility of an association of mRNA with the cytoskeletal framework
(CF) of ascidian (Styela plicata) follicle cells was examined in this
study. The approach was to extract the follicle cells with Triton X-100 and
determine whether mRNA persisted in the insoluble residue by two methods,
in situ hybridization with poly(U) and actin DNA probes and the
incorporation of radioactive isotopes into RNA. Triton X-100 extraction of
follicle cells yielded a filamentous CF containing approximately 70% of the
total poly (A) but only 9% of the total lipid, 23% of the total protein,
and 28% of the total RNA. In situ hybridization with a poly (U) probe
indicated that approximately 70% of the poly (A) was associated with the
CF. In situ hybridization with a cloned actin DNA probe indicated that
approximately 60% of the actin mRNA was associated with the CF.
Autoradiography of detergent- extracted follicle cells, which had been
labeled with [3H]uridine or [3H]adenosine, indicated that greater than 90%
of the newly synthesized poly (A)+RNA was preserved in the CF. Thus more
newly synthesized mRNA than steady-state mRNA may be present in the Triton
X-100 insoluble fraction. It is concluded that a significant proportion of
the mRNA complement of ascidian follicle cells is associated with the CF.
CiteULike 
Complore 
Connotea 
Del.icio.us 
Digg 
Reddit 
Technorati What's this?
