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Journal of Neuroscience, Vol 12, 4460-4467, Copyright © 1992 by Society for Neuroscience
5' flanking sequences of the rat tyrosine hydroxylase gene target accurate tissue-specific, developmental, and transsynaptic expression in transgenic mice
SA Banerjee, P Hoppe, M Brilliant and DM Chikaraishi
Department of Microbiology and Molecular Biology, Tufts University School of Medicine, Boston, Massachusetts 02111.
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.
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