McHardy Smith - Combination of immunocytochemistry and radioligand receptor assay to identify beta-adrenergic receptor subtypes on astroglia in vitro.

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      Trimmer, PA, Evans, T, Smith, MM, Harden, TK, McCarthy, KD   4 1598-606-

      Abstract: There is an increasing need to assess   the distribution of receptors for neuroactive substances on   specific neural cell types. This study describes the   establishment of methodology that combines the quantification of   beta-adrenergic receptor subtypes by radioligand binding assays   with immunocytochemical analysis of the contribution of astroglia   (identified by the presence of glial fibrillary acidic protein)   and fibroblasts (identified by the presence of fibronectin) to   cultures prepared from neonatal rat cerebral cortex. The effects   of subtle changes in culture methodology on the cellular   composition of cerebral cortical cultures and the distribution of   beta-adrenergic receptor subtypes were examined. The data   indicate that (1) a decrease in the density of the initial   plating suspension, (2) an increase in the age of the animals, or   (3) supplementation of the cortical cell suspension with   meningeal fibroblasts all result in an increase in fibronectin   staining and a decrease in glial fibrillary acidic protein   antibody staining. This change in the cellular composition of the   cortical cultures correlated with an increase in the number of   beta 2-adrenergic receptors and a corresponding decrease in the   number of beta 1-adrenergic receptors. These observations point   out the care which must be exercised when preparing primary   astroglial cultures of sufficient purity for large-scale   biochemical and pharmacological studies.

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