Synchronized synthesis and intracellular transport of serum albumin and apolipoprotein B in cultured rat hepatocytes as studied by double immunofluorescence.
Synthesis and intracellular transport of two secretory proteins, serum albumin (SA) and apolipoprotein B (apo B) have been synchronized in primary cultures of normal rat hepatocytes to make possible immunocytochemical study of the transport pathway. Under appropriate conditions of cycloheximide treatment, synthesis of new protein was inhibited and, by double immunofluorescent labeling, the cells were found to be largely depleted of the SA and apo B previously synthesized. Re-initiation of protein synthesis led to sequential appearance of SA and apo B, first in the endoplasmic reticulum, then in the Golgi complex, and finally at the cell surface. These results indicate that it should be feasible to use this cell system for high-resolution investigation of the sequence of structures involved in intracellular transport of SA and apo B by corresponding immunolabeling experiments as observed by electron microscopy.
D000818:Animals; D001055:Apolipoproteins B / Q000096:biosynthesis* / Q000378:metabolism; D001692:Biological Transport / Q000187:drug effects; D001826:Body Fluids / Q000378:metabolism*; D002478:Cells, Cultured; D003513:Cycloheximide / Q000494:pharmacology; D005455:Fluorescent Antibody Technique; D006056:Golgi Apparatus / Q000378:metabolism; D007424:Intracellular Fluid / Q000187:drug effects / Q000378:metabolism*; D008099:Liver / Q000166:cytology / Q000378:metabolism*; D051381:Rats; D012709:Serum Albumin / Q000096:biosynthesis* / Q000378:metabolism; D013997:Time Factors
D001055:Apolipoproteins B; D012709:Serum Albumin; D003513:Cycloheximide