Albumin and collagen mRNA expression in normal and analbuminemic rodent liver: analysis by in situ hybridization using biotinylated probes.
We used in situ nucleic acid hybridization cytochemistry to examine cell types and subcellular sites expressing albumin (alb) or pro alpha 2 collagen (col) mRNA in livers from normal and analbuminemic rodents. Biotinylated cDNA or RNA probes were applied to aldehyde-fixed, non-frozen sections and the resulting DNA-RNA or RNA-RNA hybrids were subsequently visualized by enzymatic detection of either peroxidase or alkaline phosphatase conjugated to anti-biotin IgG or streptavidin. In normal rat liver, alb mRNA was expressed in all hepatocytes and was localized to discrete subcellular structures distributed as aggregates in the cytoplasm and in specific structures encircling the nucleus; these subcellular structures most likely represent the endoplasmic reticulum and nuclear envelope. In mouse liver, pro alpha 2 col mRNA was identified in a subpopulation of sinusoidal lining cells which have the morphological appearance of lipocytes. In liver from analbuminemic rats, a small number of hepatocytes, distributed throughout the hepatic lobule, expressed alb mRNA at high levels; the subcellular distribution of this alb mRNA was essentially identical to that observed in normal rat hepatocytes. Since non-radioactive in situ hybridization detected mRNA within the boundaries of individual cells and showed its precise subcellular location under conditions in which there was excellent preservation of tissue morphology, this procedure should be useful for a wide variety of histopathologic studies.
Saber
M A
MA
Department of Biochemistry, Bilharz Research Institute, Cairo, Egypt.
Novikoff
P M
PM
Shafritz
D A
DA
D000418:Albumins / Q000172:deficiency / Q000235:genetics*; D000818:Animals; D001710:Biotin; D002467:Cell Nucleus / Q000032:analysis; D003094:Collagen / Q000032:analysis / Q000235:genetics*; D004247:DNA / Q000032:analysis; D015342:DNA Probes / Q000032:analysis*; D004721:Endoplasmic Reticulum / Q000032:analysis; D007124:Immunoenzyme Techniques; D008099:Liver / Q000032:analysis / Q000378:metabolism*; D008297:Male; D051379:Mice; D009693:Nucleic Acid Hybridization; D015347:RNA Probes / Q000032:analysis*; D012333:RNA, Messenger / Q000032:analysis*; D051381:Rats; D011919:Rats, Inbred Strains; D012709:Serum Albumin / Q000172:deficiency*; D013347:Subcellular Fractions / Q000032:analysis; D014021:Tissue Preservation
D000418:Albumins; D015342:DNA Probes; D015347:RNA Probes; D012333:RNA, Messenger; D012709:Serum Albumin; D001710:Biotin; D003094:Collagen; D004247:DNA