We used preparative freeze-fracture to crush glutaraldehyde fixed tissues after impregnation in cryoprotective solution. The fragments were thawed and labeled with wheat germ agglutinin permeated with concanavalin A. They were also fracture- permeated with native ferritin and with cationized ferritin. In autonomic ganglia, primary sensory ganglia and peripheral nerves, fracture-label provides access to all plasma and intracellular membranes as well as extracellular matrices. Con A, WGA and cationized ferritin intensely labels the basal membrane. Furthermore, con A and WGA appears to be localized to synaptic vesicles and the synaptic complex. We worked also with rat glomeruli where freeze-fracture was used as a means to get generalized access to the cell surfaces. This new approach by-passes successive permeability barriers (endothelial cells, basement membranes) that had to be passed in previous cytochemical studies of the glomerulus. In human lymphocytes we used monoclonal anti-T3 and anti-T4 antibodies to study the partition of the transmembrane proteins which contain these antigens. During freeze-fracture, the T4 antigen partitions completely with the protoplasmic half of the membrane, gave an indication of its association with components of the membrane skeleton or of the cytoskeleton.

Agency
National Institute of Health (NIH)
Institute
Division of Cancer Biology And Diagnosis (NCI)
Type
Intramural Research (Z01)
Project #
1Z01CB008376-04
Application #
3939299
Study Section
Project Start
Project End
Budget Start
Budget End
Support Year
4
Fiscal Year
1987
Total Cost
Indirect Cost
Name
Cancer Biology and Diagnosis
Department
Type
DUNS #
City
State
Country
United States
Zip Code