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Neurotrophin‐induced preprotachykinin‐A gene promoter modulation in organotypic rat spinal cord culture
Author(s) -
Hilton Kathryn J.,
Bateson Alan N.,
King Anne E.
Publication year - 2006
Publication title -
journal of neurochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.75
H-Index - 229
eISSN - 1471-4159
pISSN - 0022-3042
DOI - 10.1111/j.1471-4159.2006.03910.x
Subject(s) - substance p , neurotrophic factors , nerve growth factor , microbiology and biotechnology , neurotrophin , transfection , reporter gene , biology , brain derived neurotrophic factor , receptor , neurotrophin 3 , green fluorescent protein , spinal cord , endocrinology , chemistry , medicine , neuropeptide , gene expression , gene , neuroscience , biochemistry
To study regulation of the preprotachykinin‐A gene promoter, we utilised a biolistic gene transfer protocol to deliver a DNA construct that incorporates a portion of the preprotachykinin‐A gene promoter and an enhanced green fluorescent protein reporter gene into neonatal rat spinal cord organotypic slices. The ability of the neurokinin‐1 receptor agonist [Sar 9 ,Met(O 2 ) 11 ]‐substance P, nerve growth factor and brain derived neurotrophic factor to modulate positively preprotachykinin‐A gene promoter construct activity, as indicated by de novo enhanced green fluorescent protein expression, was determined. Treatment of organotypic slices with [Sar 9 , Met(O 2 ) 11 ]‐substance P (10 μ m , P < 0.05), nerve growth factor (200 ng/mL, P < 0.001) or brain derived neurotrophic factor (200 ng/mL, P < 0.02) significantly increased the proportion of cytomegaloviral promoter‐DsRed transfected cells (used to visualise total transfected cells) that co‐expressed enhanced green fluorescent protein. The distribution of enhanced green fluorescent protein/DsRed‐positive neurones across spinal laminae was broadly in line with the known distribution of spinal Trk and neurokinin‐1 receptors. These data suggest a modulated activity of the preprotachykinin‐A gene promoter in spinal neurones in vitro by substance P and/or neurotrophins. The functional consequences of such transcriptional changes within central peptidergic circuitry and their relevance to chronic pain are considered.