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Presence of immunoreactive insulin in neurons cultured from fetal rat brain (1983)

Weyhenmeyer, James A. ; Fellows, Robert E.

Springer

Cellular and molecular neurobiology 3 (1983), S. 81-86

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ISSN: 1573-6830

Keywords: insulin ; immunocytochemistry ; fetal rat brain ; primary cell culture

Source: Springer Online Journal Archives 1860-2000

Topics: Biology

Notes: Summary 1. Immunocytochemical studies have been performed which establish the presence of immunoreactive (ir-) insulin in primary cultures from fetal rat brain. Approximately 1% of the identified neurons in culture contained insulin immunoreactivity. Typically, a diffuse staining pattern was observed in the perikaryia and fiber profiles of peroxidase-anti-peroxidase-positive neurons. Varicosity-like structures containing ir-insulin were found on the majority of positively stained fiber profiles. A small number of the neurons appeared to have cytoplasmic granules that stained densely for insulin. No significant staining was observed in the background monolayer of cells predominantly of glial origin. 2. The substitution of primary antiserum absorbed with excess insulin or preimmune serum eliminated staining in the neurons. The observation of insulin immunoreactivity in neurons suggests that insulin may be a neuromodulator in the central nervous system.

Type of Medium: Electronic Resource

URL: http://dx.doi.org/10.1007/BF00735000

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Alteration of insulin binding and cytoskeletal organization in cultured fibroblasts by tertiary amine local anesthetics (1979)

Raizada, Mohan K. ; Fellows, Robert E.

New York, N.Y. : Wiley-Blackwell

Journal of Supramolecular Structure 11 (1979), S. 547-561

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ISSN: 0091-7419

Keywords: insulin receptors ; 125I-insulin binding ; microtubules and microfilaments ; cultured fibroblasts ; local anesthetics ; Life Sciences ; Molecular Cell Biology

Source: Wiley InterScience Backfile Collection 1832-2000

Topics: Biology , Chemistry and Pharmacology , Medicine

Notes: Tertiary amine local anesthetics cause a time- and dose-dependent, reversible increase in insulin binding sites in cultured chick embryo fibroblasts. Incubation of fibroblasts with 0.2 mM dibucaine for 3 h at 37°C results in a twofold to threefold increase in insulin binding, with an increase in average number of binding sites (Ka = 3.0 × 107M-1) from 9 × 103 to 29 × 103 per cell. Trypsin or ethylenegly coltetraacetic acid (EGTA) alone increases insulin binding twofold to threefold, but fails to further increase 125I-insulin binding in cells pretreated with dibucaine. Transformation of chick embryo fibroblasts with Rous sarcoma virus causes a threefold to fivefold increase in insulin binding, which is not further increased by incubation with dibucaine. As demonstrated by transmission electron microscopy, dibucaine and trypsin also induce changes in the cytoskeleton of chick embryo fibroblasts, characterized by disorganization and disappearance of microfilament and microtubule bundles. These alterations are accompanied by gross morphologic changes, including rounding of cells and appearance of numerous ruffles and blebs on the cell surface. These observations are consistent with the hypothesis that expression of surface receptors in cultured chick embryo fibroblasts is related to the organization and disorganization of cytoskeletal structures.

Additional Material: 5 Ill.