Binding of insulin and insulin-like growth factor I in bovine chromaffin cells in primary culture |
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| Institution: | 1. Department of Physiology, University of Bergen, Bergen, Norway;2. Department of Pediatrics, University of Bergen, Bergen, Norway;1. Department of Neurology, The Institute of Guangzhou Respiratory Disease, State Key Laboratory of Respiratory Disease, The First Affiliated Hospital of Guangzhou Medical University, Guangzhou 510120, PR China;2. Department of Biochemistry and Molecular Medicine, School of Medicine, University of California, Davis, CA 95817, USA;3. Institute for Pediatric Regenerative Medicine, Shriners Hospitals for Children, Sacramento, CA 95817, USA;1. Department of Molecular Neurobiology, Max Planck Institute of Experimental Medicine, 37075 Göttingen, Germany;2. Science Products GmbH, 65719 Hofheim am Taunus, Germany;3. Research Group Cellular Basis of Neural Circuit Plasticity, Max Planck Florida Institute for Neuroscience, Jupiter, FL 33458, USA;4. Max Planck Institute of Neurobiology, 82152 Martinsried, Germany;3. From the School of Biomedical Sciences, Hong Kong, China;4. State Key Laboratory of Brain and Cognitive Sciences, University of Hong Kong, Hong Kong, China;1. Department of Neurobiology, The University of Chicago, Chicago, IL 60637, USA;2. Department of Physiology, Feinberg School of Medicine, Northwestern University, Chicago, IL 60611, USA;3. Department of Neurological sciences, Rush University, Chicago, IL 60612, USA;4. Integrated Light Microscopy Facility, The University of Chicago, Chicago, IL 60637, USA;5. Department of Neurobiology, Weinberg College of Arts and Sciences, Northwestern University, Chicago, IL, USA;6. Department of Neurology, The University of Chicago, Chicago, IL 60637, USA;7. Department of Pathology, The University of Chicago, Chicago, IL 60637, USA;8. Department of Molecular Medicine and Byrd Alzheimer’s Institute, University of South Florida, Tampa, FL 33613, USA;1. Emeritus Laboratory of Membrane Biophysics, Max Planck Institute for Biophysical Chemistry, 37077 Göttingen, Germany;2. Department of Molecular Neurobiology, Max Planck Institute of Experimental Medicine, 37075 Göttingen, Germany;3. Cluster of Excellence “Multiscale Bioimaging: from Molecular Machines to Networks of Excitable Cells” (MBExC), University of Göttingen, Germany |
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| Abstract: | - 1.1. Insulin and insulin-like growth factor I (IGF-I) receptors were studied in bovine chromaffin cells isolated from the medulla by collagenase digestion and kept in primary culture.
- 2.2. Specific 125I-labelled insulin binding increased with time in culture with no significant change in the dissociation constant, Kd~0.3nM. Insulin was nearly 100-fold more potent than IGF-I in displacing 125I-labelled insulin.
- 3.3. Affinity crosslinking and SDS gel electrophoresis revealed increased binding of 125I-labelled insulin and 125I-IGF-I with time in culture, the densities of the labelling indicating relatively a much higher expression of IGF-I than insulin receptors in the cells. The apparent molecular weight of both the hormone binding subunits were 135,000, suggesting that the insulin and IGF-I receptors in the adrenal medulla are of the peripheral types.
- 4.4. Both receptors thus appeared to be affected by the collagenase treatment but with a subsequent recovery when cells were kept in culture.
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