NDLI: 5′-Flanking Sequences from the Rat Preprotachykinin Gene Direct High-Level Expression of a Reporter Gene in Adult Rat Sensory Neurons Transfected in Culture by Microinjection

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5′-Flanking Sequences from the Rat Preprotachykinin Gene Direct High-Level Expression of a Reporter Gene in Adult Rat Sensory Neurons Transfected in Culture by Microinjection

Content Provider	Semantic Scholar
Author	Mulderry, P. K. Chapman, Karen E. Lyons, Val Harmar, Anthony J.
Copyright Year	1993
Abstract	The rat preprotachykinin-A gene, which encodes substance P, is expressed in response to nerve growth factor in a subpopulation of dorsal root ganglion sensory neurons. To investigate mechanisms regulating preprotachykinin-A transcription, we transfected adult rat sensory neurons in culture by microinjection of plasmids containing genomic DNA sequences linked to a lacZ (beta-galactosidase) reporter gene. Expression of beta-galactosidase was seen in 10-15% of neurons receiving injections of prPPT-betaGAL1, which contained the preprotachykinin transcription start site and 3356 bp of 5'-flanking DNA. Deletion analysis showed that expression was directed by 865 bp lying immediately upstream of the transcription start site. Extension of the prPPT-betaGAL1 sequence to include the first intron of preprotachykinin increased beta-galactosidase two- to threefold. Functional promoter and enhancer sequences from the rat prolactin gene failed to direct expression in sensory neurons, indicating neuronal selectivity for preprotachykinin sequences. Expression of prPPT-betaGAL1, measured relative to a construct containing the Rous sarcoma virus promoter, was approximately fivefold higher in neurons than in nonneuronal cells. This suggests selectivity by preprotachykinin 5'-flanking sequences for neuronal expression. However, prPPT-betaGAL1 expression was not restricted to the neuronal subpopulation containing immunoreactive substance P nor was it dependent upon nerve growth factor. Therefore, it does not share all the characteristics of endogenous preprotachykinin expression implying the need for additional regulatory sequences or the involvement of post-transcriptional regulation. Our results show that transfection of differentiated neurons in culture by microinjection has considerable potential in studies of neuron-specific gene expression.
Starting Page	164
Ending Page	172
Page Count	9
File Format	PDF HTM / HTML
DOI	10.1006/mcne.1993.1020
PubMed reference number	19912919
Journal	Medline
Volume Number	4
Alternate Webpage(s)	https://api.elsevier.com/content/article/pii/S1044743183710201
Alternate Webpage(s)	https://www.sciencedirect.com/science/article/pii/S1044743183710201?dgcid=api_sd_search-api-endpoint
Alternate Webpage(s)	https://doi.org/10.1006/mcne.1993.1020
Journal	Molecular and Cellular Neuroscience
Language	English
Access Restriction	Open
Content Type	Text
Resource Type	Article

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5′-Flanking Sequences from the Rat Preprotachykinin Gene Direct High-Level Expression of a Reporter Gene in Adult Rat Sensory Neurons Transfected in Culture by Microinjection

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