Insulin increases the Na(+)-K(+)-ATPase alpha 2-subunit in the surface of rat skeletal muscle: morphological evidence
| Autor: | C. Ackerley, L. Lavoie, J. Krischer, J. L. Carpentier, Amira Klip, André Marette |
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| Rok vydání: | 1993 |
| Předmět: |
Macromolecular Substances
Physiology medicine.medical_treatment Protein subunit Immunoelectron microscopy Alpha (ethology) Diaphragm pump Biology Rats Sprague-Dawley Immunoglobulin Fab Fragments Sarcolemma medicine Animals Insulin Na+/K+-ATPase Microscopy Immunoelectron Cellular localization Muscles Cell Membrane Antibodies Monoclonal Skeletal muscle Cell Biology Molecular biology Recombinant Proteins Rats medicine.anatomical_structure Biochemistry Enzyme Induction Immunoglobulin G Sodium-Potassium-Exchanging ATPase |
| Zdroj: | American Journal of Physiology-Cell Physiology. 265:C1716-C1722 |
| ISSN: | 1522-1563 0363-6143 |
| Popis: | The cellular localization of the alpha 2-subunit of the Na(+)-K(+)-ATPase was defined by immunoelectron microscopy, and the effect of insulin on the amount of alpha 2-immunoreactive subunits on the cell surface was quantitated. Two protocols were used for tissue fixation and immunolocalization. Protocol 1 was characterized by fixation with 2% paraformaldehyde, use of a monoclonal antibody, and detection with 3-nm-diameter gold-labeled Fab fragments or 10-nm gold-labeled immunoglobulin G. Protocol 2 was characterized by fixation with 4% paraformaldehyde plus 0.1% glutaraldehyde, use of a polyclonal antibody, and detection with 10-nm gold-labeled protein A. In control muscle, the alpha 2-subunit of the Na(+)-K(+)-ATPase was present at the plasma membrane and in intracellular tubular and vesicular structures located in subsarcolemmal and triadic regions. Acute insulin stimulation increased the number of immunolabeled alpha 2-subunits in the plasma membrane after both fixation protocols. The gain in the plasma membrane ranged from 1.5- to 3.7-fold and was significant at the level of P < 0.005. These results provide morphological quantitative evidence that the alpha 2-subunit of the Na(+)-K(+)-ATPase is present both at the plasma membrane and intracellularly in mammalian skeletal muscle and that insulin acutely increases its abundance in the muscle surface. |
| Databáze: | OpenAIRE |
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