Monoclonal antibodies used in immunocytochemical localization by electron microscopy of carbamoyl phosphate synthetase I in liver from rats fed high-protein diets.
Carbamoyl phosphate synthetase I (CPS-I) is the most abundant protein of rat liver mitochondria. Biochemical measurements in liver homogenates have shown that the liver from rats fed a high-protein diet contains more CPS-I per gram tissue protein than controls. However, there is no information on changes in the intact tissue at the cellular and mitochondrial level. Therefore, monoclonal antibodies to beef liver CPS-I were produced by the hybridoma technique. Four clones, C-241/1A, B, C, and D secreted immunogammaglobulin (IgG) IgG1. Using C-241/C, we measured by electron microscopy immunogold procedures the labeling of CPS-I in mitochondria from liver of rats fed high protein (casein, 50 and 80% of total food intake) diets. CPS-I (expressed as gold particles/micron2 of mitochondrial cross-sectional area) was greater than in mitochondria from control rats (20% casein diet), whether the rats were fed for 1, 6, or 14 months on the high-protein diets. The immunocytochemical measurements shown here demonstrate that the increase in the level of CPS-I in high-protein diets is a reflection of both the larger number of CPS-I molecules per mitochondrial area and the larger proportion of the total hepatocyte volume occupied by mitochondria. Similar measurements were carried out with glutamate dehydrogenase (GDH) using previously characterized monoclonal antibodies. No differences in GDH labeling were found with high-protein diets. Interestingly, when mitochondria from hepatocytes of rats fed a high-protein diet were divided into two subpopulations on the basis of mitochondrial cross-sectional size (i.e., greater or less than 0.7 micron2), the large mitochondria had 1.2 times more CPS-I and 0.8 times less GDH than the small mitochondria nearby.
D000824:Animal Nutritional Physiological Phenomena; D000818:Animals; D000911:Antibodies, Monoclonal; D002222:Carbamoyl-Phosphate Synthase (Ammonia) / Q000378:metabolism*; D004044:Dietary Proteins / Q000378:metabolism*; D005969:Glutamate Dehydrogenase / Q000378:metabolism; D006046:Gold; D008025:Ligases / Q000378:metabolism*; D008854:Microscopy, Electron; D008930:Mitochondria, Liver / Q000378:metabolism*; D051381:Rats
D000911:Antibodies, Monoclonal; D004044:Dietary Proteins; D006046:Gold; D005969:Glutamate Dehydrogenase; D008025:Ligases; D002222:Carbamoyl-Phosphate Synthase (Ammonia)