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Volume 17, Number 17, Issue of September 1, 1997 pp. 6554-6564
Copyright ©1997 Society for Neuroscience

Differential Expression of -Bungarotoxin-Sensitive Neuronal Nicotinic Receptors in Adrenergic Chromaffin Cells: A Role for Transcription Factor Egr-1

Received Feb. 18, 1997; revised June 6, 1997; accepted June 13, 1997.

Manuel Criado^{1, 4}, Eduardo Domínguez del Toro^{1, 4}, Carmen Carrasco-Serrano^{1, 4}, Frazer I. Smillie^{1, 4}, José M. Juíz^{2, 4}, Salvador Viniegra^{1, 4}, and Juan J. Ballesta^{3, 4}

Departments of ¹ Neurochemistry, ² Histology, and ³ Pharmacology, and ⁴ Instituto de Neurociencias, Universidad Miguel Hernandez, 03550 San Juan, Alicante, Spain

Adrenomedullary chromaffin cells express at least two subtypes of acetylcholine nicotinic receptors, which differ in their sensitivity to the snake toxin -bungarotoxin. One subtype is involved in the activation step of the catecholamine secretion process and is not blocked by the toxin. The other is -bungarotoxin-sensitive, and its functional role has not yet been defined. The 7 subunit is a component of this subtype. Autoradiography of bovine adrenal gland slices with -bungarotoxin indicates that these receptors are restricted to medullary areas adjacent to the adrenal cortex and colocalize with the enzyme phenylethanolamine N-methyl transferase (PNMT), which confers the adrenergic phenotype to chromaffin cells. Transcripts corresponding to the 7 subunit also are localized exclusively to adrenergic cells. To identify possible transcriptional regulatory elements of the 7 subunit gene involved in the restricted expression of nicotinic receptors, we isolated and characterized its 5 flanking region, revealing putative binding sites for the immediate early gene transcription factor Egr-1, which is known to activate PNMT expression. In reporter gene transfection experiments, Egr-1 increased 7 promoter activity by up to sevenfold. Activation was abolished when the most promoter-proximal of the Egr-1 sites was mutated, whereas modification of a close upstream site produced a partial decrease of the Egr-1 response. Because Egr-1 was found to be expressed exclusively in adrenergic cells, we suggest that this transcription factor may be part of a common mechanism involved in the induction of the adrenergic phenotype and the differential expression of -bungarotoxin-sensitive nicotinic receptors in the adrenal gland.

Key words: ACh receptors; -bungarotoxin; adrenergic; chromaffin cell; 7 subunit; Egr-1

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