5' flanking sequences of the rat tyrosine hydroxylase gene target accurate tissue-specific, developmental, and transsynaptic expression in transgenic mice

Abstract

Transgenic mice were generated in which sequences that flank the rat tyrosine hydroxylase (TH) gene were linked to the bacterial chloramphenicol acetyl transferase (CAT) gene. Mice bearing 4.8 kilobases (kb) of 5' flanking DNA exhibited correct tissue-specific expression in the CNS and periphery. Expression was more robust in the CNS than in the periphery, although CAT activity was clearly detected in sympathetic ganglia (superior cervical ganglia) and the adrenal, the two peripheral tissues that contain TH-positive cells. Within the brain, CAT expression was seen in all the expected areas containing TH-positive cell bodies, with little or no expression in other regions. In the olfactory bulb, which contains the majority of the CNS TH cells, developmental expression of CAT was quantifiable and was found to parallel the postnatal rise in endogenous TH, with both TH and CAT reaching adult levels by postnatal day 21. Since TH activity in the olfactory bulb requires afferent input, the dependence of CAT activity on transsynaptic input was also assayed in transgenic mice. Like the endogenous TH activity, CAT levels were also reduced by deafferentation, in parallel with loss in endogenous dopamine levels. While previous experiments demonstrated that shorter 5' flanking regions (2.5 kb and 3.5 kb of 5' upstream sequences of the human and mouse TH gene, respectively) failed to direct accurate tissue-specific expression, our data demonstrate that 4.8 kb of 5' flanking sequence of the rat TH gene contains sufficient regulatory information to mediate appropriate tissue-specific expression in all CNS and PNS tissues, as well as to mediate developmental and transsynaptic expression in the olfactory bulb.

Transgenic zebrafish that express tyrosine hydroxylase promoter in inner retinal cells

“…The same TH promoter was characterized previously in other vertebrate species. In mice, this promoter is expressed in a tissue-specific manner (Banerjee et al, 1992(Banerjee et al, , 1994. In transgenic mice, however, this promoter drove gene expressions in both type 1 (colocalization with TH antibodies) and type 2 (no colocalization with TH antibodies) catecholaminergic cells (Gustincich et al, 1997;Zhang et al, 2004).…”

“…A 4.5-kb rat TH promoter (Banerjee et al, 1992(Banerjee et al, , 1994 was cloned into the pEGFP-1 vector (Clonetech). Plasmid DNA was prepared by using the Qiagen plasmid kit.…”

Orphan nuclear receptor Nurr1 directly transactivates the promoter activity of the tyrosine hydroxylase gene in a cell‐specific manner

“…reporter gene in the catecholaminergic neurons (Banerjee et al 1992;Min et al 1994), we employed various reporter constructs containing 9.0 kb or shorter upstream sequences of the TH gene. As shown in Fig.…”

“…Transcriptional regulation of the TH gene has been extensively investigated in numerous laboratories. In vivo transgenic mice experiments have been performed to address whether different promoter regions of the human, rat, and mouse TH gene can confer cell type-specific gene expression (Gandelman et al 1990;Kaneda et al 1991;Banerjee et al 1992;Min et al 1994;Wong et al 1995;Morgan et al 1996). Despite some variations between such studies, in general they established that 5¢ upstream sequences (> 5 kb) of the TH gene are able to drive reporter gene expression in most catecholaminergic (CA) neurons and endocrine cells, with some ectopic expressions.…”

Visualization of β-Galactosidase by Enzyme and Immunohistochemistry in the Olfactory Bulb of Transgenic Mice Carrying the LacZ Transgene

“…Some of the transgenic mouse lines express ␤ -galactosidase in the OB (Banerjee et al 1992;Smeyne et al 1992;Morita et al 1993;Arai et al 1994;Hoyle et al 1994;Masters et al 1994;Ponte et al 1994;RinconLimas et al 1994;Vanselow et al 1994;Erickson et al 1995;Morgan et al 1996), but in none of these cases was the intracellular distribution of ␤ -galactosidase determined.…”