A 6.1 kb 5' upstream region of the mouse tryptophan hydroxylase gene directs expression of E. coli lacZ to major serotonergic brain regions and pineal gland in transgenic mice.
Tryptophan hydroxylase (TPH) catalyzes the first step of serotonin biosynthesis in serotonergic neurons and neuroendocrine cells. Serotonin influences diverse vital physiological functions and is thought to play an important role in several human psychiatric disorders. To localize DNA element(s) important for serotonergic tissue-specific expression of TPH, 6.1 kb of the 5' flanking region of the mouse TPH gene was fused to the coding region of the E. coli lacZ gene, and expression of the resulting fusion gene was analyzed in transgenic mice. The 6.1 kb of 5' flanking sequence was able to direct the expression of a lacZ reporter gene to serotonergic tissues in six lines of transgenic mice. A high level of lacZ expression in transgenic mice carrying the fusion gene was detected in the pineal gland as well as a moderate level of lacZ expression in serotonergic brain regions such as the median and dorsal raphe nuclei, the nuclei raphe magnus and raphe pallidus. In contrast, a smaller 5' flanking sequence of 1.1 kb directed no detectable serotonergic tissue-specific lacZ expression in five lines of transgenic mice. These results presented in this paper suggest first that DNA elements critical to serotonergic tissue-specific expression reside between -6.1 kb and -1.1 kb of 5' flanking region of the mouse TPH gene, but second that this region confers a restricted tissue-specific expression.
Laboratory of Molecular Neurobiology, Cornell University Medical College, W.M. Burke Medical Research Institute, White Plains, NY 10605.
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D000818:Animals; D001483:Base Sequence; D001921:Brain / Q000378:metabolism*; D017931:DNA Primers; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics*; D005260:Female; D015971:Gene Expression Regulation, Enzymologic; D005798:Genes, Bacterial; D015698:Genomic Library; D007150:Immunohistochemistry; D008297:Male; D051379:Mice; D008822:Mice, Transgenic; D008969:Molecular Sequence Data; D009928:Organ Specificity; D010870:Pineal Gland / Q000378:metabolism*; D016133:Polymerase Chain Reaction; D011903:Raphe Nuclei / Q000378:metabolism; D012016:Reference Values; D012045:Regulatory Sequences, Nucleic Acid; D015183:Restriction Mapping; D012701:Serotonin / Q000378:metabolism*; D014365:Tryptophan Hydroxylase / Q000032:analysis / Q000096:biosynthesis / Q000235:genetics*; D001616:beta-Galactosidase / Q000032:analysis / Q000096:biosynthesis* / Q000235:genetics
D017931:DNA Primers; D012701:Serotonin; D014365:Tryptophan Hydroxylase; D001616:beta-Galactosidase