Detection of full-length and truncated neurokinin-1 receptor mRNA expression in human brain regions.
We have applied a newly developed SYBR green-based real-time RT-PCR assay for quantification of full-length and truncated neurokinin-1 receptor (NK1R) mRNA expression in nine regions of human brain tissues obtained from 23 subjects who died with no evidence of neurological or neurodegenerative disease. The following brain regions were examined: cingulate cortex, cerebellum, nucleus accumbens, caudate nucleus, putamen, pons, hippocampus, locus coeruleus, and basal ganglia. The SYBR green-based real-time PCR was more sensitive than TaqMan probe-based real-time PCR in amplifying both full-length and truncated NK1R mRNA. The real-time RT-PCR assay had excellent specificity and sensitivity, with a dynamic range of detection between 100 and 1,000,000 copies of the NK1R cDNA per reaction. The truncated NK1R mRNA levels were more abundant than those of the full-length NK1R in most of the regions examined and there was no significant difference in the truncated NK1R mRNA levels among the nine regions studied. There was, however, a significant difference in the expression of full-length NK1R mRNA levels among the nine regions (P=0.0024), and the putamen region expressed the highest full-length NK1R mRNA. Further studies are needed in order to examine the differences between full-length and truncated NK1R in signal transduction and functional consequences in order to delineate the significance of the co-presence of the two forms of NK1R in the human brain.
Lai
Jian-Ping
JP
Division of Allergy and Immunology, Joseph Stokes Jr. Research Institute at the Children's Hospital of Philadelphia, Department of Pediatrics, University of Pennsylvania Medical School, Philadelphia, PA 19104, USA.
Cnaan
Avital
A
Zhao
Huaqing
H
Douglas
Steven D
SD
D000328:Adult; D000368:Aged; D052160:Benzothiazoles; D001921:Brain / Q000378:metabolism* / Q000473:pathology; D001927:Brain Diseases / Q000473:pathology; D003959:Diamines; D005260:Female; D015870:Gene Expression / Q000502:physiology*; D006801:Humans; D008297:Male; D008875:Middle Aged; D009154:Mutation; D009930:Organic Chemicals; D016133:Polymerase Chain Reaction; D011804:Quinolines; D012333:RNA, Messenger / Q000378:metabolism*; D018040:Receptors, Neurokinin-1 / Q000235:genetics* / Q000378:metabolism
D052160:Benzothiazoles; D003959:Diamines; D009930:Organic Chemicals; D011804:Quinolines; D012333:RNA, Messenger; D018040:Receptors, Neurokinin-1; C098022:SYBR Green I