Abstract
Chromaffin cells have been recognized for their ability to transform into sympathetic ganglion-like cells in response to nerve growth factor (NGF) or to stimulation of other neurotrophic factors. Transforming growth factor β (TGFβ) family members have been shown to potentiate the effect of different trophic factors. The aim of this study was to investigate if TGFβ may influence NGF-induced neuronal transformation and regulation of NGF, TGFβ1, and their receptors in the adult rat chromaffin tissue after grafting. Intraocular transplantation of adult chromaffin tissue was employed and grafts were treated with TGFβ1 and/or NGF. Graft survival time was 18 days after which the grafts were processed for TGFβ luciferase detection assay, NGF enzyme immunoassay, or in situ hybridization. In grafts stimulated with NGF, increased levels of TGFβ1 and TGFβ1 mRNA were detected. When grafts instead were treated with TGFβ1, enhanced levels of NGF protein were found. Furthermore, a positive mRNA signal corresponding to the transforming growth factor II receptor (TβRII) was found in the chromaffin cells of the normal adrenal medulla as well as after grafting. No increase of TβRII mRNA levels was detected after transplantation or after TGFβ1 treatment. Instead a reduction of TβRII mRNA expression was noted after NGF treatment. NGF stimulation of grafts increased the message for NGF receptors p75 and trkA in the chromaffin transplants. Grafts processed for evaluations of neurite outgrowth were allowed to survive for 28 days and were injected weekly with NGF and/or TGFβ1. NGF treatment resulted in a robust innervation of the host irides. TGFβ1 had no additive effect on nerve fiber formation when combined with NGF. Combined treatment of NGF and anti-TGFβ1 resulted in a significantly larger area of reinnervation. In conclusion, it was found that NGF and TGFβ1 may regulate the expression of each other's protein in adult chromaffin grafts. Furthermore, TβRII mRNA was present in the adult rat chromaffin cells and became downregulated as a result of NGF stimulation. Although no synergistic effects of TGFβ1 were found on NGF-induced neurite outgrowth, it was found that TGFβ1 and NGF signaling are closely linked in the chromaffin cells of the adrenal medulla.
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