Neurotrophin-induced preprotachykinin-A gene promoter modulation in organotypic rat spinal cord culture.
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 [Sar9,Met(O2)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 [Sar9, Met(O2)11]-substance P (10 microm, 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.
Hilton
Kathryn J
KJ
Institute of Membrane and Systems Biology, University of Leeds, Leeds, UK.
Bateson
Alan N
AN
King
Anne E
AE
D000818:Animals; D019470:Biolistics / Q000379:methods; D019208:Brain-Derived Neurotrophic Factor / Q000494:pharmacology / Q000502:physiology; D005260:Female; D005786:Gene Expression Regulation / Q000502:physiology; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins / Q000096:biosynthesis / Q000235:genetics; D020932:Nerve Growth Factor / Q000494:pharmacology / Q000502:physiology; D009414:Nerve Growth Factors / Q000502:physiology*; D009924:Organ Culture Techniques; D011401:Promoter Regions, Genetic / Q000502:physiology*; D011498:Protein Precursors / Q000096:biosynthesis* / Q000235:genetics*; D051381:Rats; D017208:Rats, Wistar; D013116:Spinal Cord / Q000378:metabolism* / Q000502:physiology*; D015320:Tachykinins / Q000096:biosynthesis* / Q000235:genetics*
D019208:Brain-Derived Neurotrophic Factor; D009414:Nerve Growth Factors; D011498:Protein Precursors; D015320:Tachykinins; C485184:enhanced green fluorescent protein; C044206:preprotachykinin; D049452:Green Fluorescent Proteins; D020932:Nerve Growth Factor