Immunoelectron microscopy of the outer membrane of rat hepatocyte nuclear envelopes in relation to the rough endoplasmic reticulum

Abstract

Prefixed nuclei were isolated from rat liver after perfusion with dilute glutaraldehyde. These nuclei sometimes were associated with the rough endoplasmic reticulum (ER), and occasionally continuity of the outer nuclear membrane with rough ER membrane was found. When these prefixed nuclei were incubated with ferritin antibody conjugates against cytochrome P-450, the cytoplasmic face of the outer nuclear membrane was labeled with ferritin similar to the labeling of the rough ER membrane. In the nuclear pore region, ferritin particles were present on the outer membrane up to the outer annuli of the pore complexes. When unfixed nuclei were incubated at 0-4 degrees C or at 20 degrees C, some outer nuclear membranes were detached from the nuclear surface. In this case the nuclear pore complexes remained associated with the inner nuclear membrane, and small inside-out vesicles were formed at these pore regions. We also found some rough vesicles heavily labeled with ferritin particles within the nuclear matrix. They probably were derived from the rough ER or from the outer nuclear envelope which was internalized artificially during incubation.