An efficient intrathecal delivery of small interfering RNA to the spinal cord and peripheral neurons

Persistent Link:
http://hdl.handle.net/10150/610210
Title:
An efficient intrathecal delivery of small interfering RNA to the spinal cord and peripheral neurons
Author:
Luo, Miaw-Chyi; Zhang, Dong-Qin; Ma, Shou-Wu; Huang, Yuan-Yuan; Shuster, Sam; Porreca, Frank; Lai, Josephine
Affiliation:
Department of Pharmacology, The University of Arizona Health Sciences Center, Tucson, AZ 85724, USA; Neuromics, Minneapolis, MN 55438, USA
Issue Date:
2005
Publisher:
BioMed Central
Citation:
Molecular Pain 2005, 1:29 doi:10.1186/1744-8069-1-29
Journal:
Molecular Pain
Rights:
© 2005 Luo et al; licensee BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0)
Collection Information:
This item is part of the UA Faculty Publications collection. For more information this item or other items in the UA Campus Repository, contact the University of Arizona Libraries at repository@u.library.arizona.edu.
Abstract:
We have developed a highly effective method for in vivo gene silencing in the spinal cord and dorsal root ganglia (DRG) by a cationic lipid facilitated delivery of synthetic, small interfering RNA (siRNA). A siRNA to the delta opioid receptor (DOR), or a mismatch RNA, was mixed with the transfection reagent, i-FectTM (vehicle), and delivered as repeated daily bolus doses (0.5 mug to 4 mug) via implanted intrathecal catheter to the lumbar spinal cord of rats. Twenty-four hours after the last injection, rats were tested for antinociception by the DOR selective agonist, D-Ala2, Glu4]deltorphin II (DELT), or the mu opioid receptor (MOR) selective agonist, D-Ala2, N-Me-Phe4, Gly-ol5]enkephalin (DAMGO). Pretreatment with the siRNA, but not the mismatch RNA or vehicle alone, blocked DELT antinociception dose-dependently. The latter was concomitant with a reduction in the spinal immunoreactivity and receptor density of DOR, and in DOR transcripts in the lumbar DRG and spinal dorsal horn. Neither siRNA nor mismatch RNA pretreatment altered spinal immunoreactivity of MOR or antinociception by spinal DAMGO, and had no effect on the baseline thermal nociceptive threshold. The inhibition of function and expression of DOR by siRNA was reversed by 72 hr after the last RNA injection. The uptake of fluorescence-tagged siRNA was detected in both DRG and spinal cord. The low effective dose of siRNA/i-FectTM complex reflects an efficient delivery of the siRNA to peripheral and spinal neurons, produced no behavioral signs of toxicity. This delivery method may be optimized for other gene targets.
EISSN:
1744-8069
DOI:
10.1186/1744-8069-1-29
Keywords:
rat; antinociception; sensory neurons; spinal cord; knockdown; RNA interference
Version:
Final published version
Additional Links:
http://mpx.sagepub.com/content/1/1744-8069-1-29.full

Full metadata record

DC FieldValue Language
dc.contributor.authorLuo, Miaw-Chyien
dc.contributor.authorZhang, Dong-Qinen
dc.contributor.authorMa, Shou-Wuen
dc.contributor.authorHuang, Yuan-Yuanen
dc.contributor.authorShuster, Samen
dc.contributor.authorPorreca, Franken
dc.contributor.authorLai, Josephineen
dc.date.accessioned2016-05-20T09:01:10Z-
dc.date.available2016-05-20T09:01:10Z-
dc.date.issued2005en
dc.identifier.citationMolecular Pain 2005, 1:29 doi:10.1186/1744-8069-1-29en
dc.identifier.doi10.1186/1744-8069-1-29en
dc.identifier.urihttp://hdl.handle.net/10150/610210-
dc.description.abstractWe have developed a highly effective method for in vivo gene silencing in the spinal cord and dorsal root ganglia (DRG) by a cationic lipid facilitated delivery of synthetic, small interfering RNA (siRNA). A siRNA to the delta opioid receptor (DOR), or a mismatch RNA, was mixed with the transfection reagent, i-FectTM (vehicle), and delivered as repeated daily bolus doses (0.5 mug to 4 mug) via implanted intrathecal catheter to the lumbar spinal cord of rats. Twenty-four hours after the last injection, rats were tested for antinociception by the DOR selective agonist, D-Ala2, Glu4]deltorphin II (DELT), or the mu opioid receptor (MOR) selective agonist, D-Ala2, N-Me-Phe4, Gly-ol5]enkephalin (DAMGO). Pretreatment with the siRNA, but not the mismatch RNA or vehicle alone, blocked DELT antinociception dose-dependently. The latter was concomitant with a reduction in the spinal immunoreactivity and receptor density of DOR, and in DOR transcripts in the lumbar DRG and spinal dorsal horn. Neither siRNA nor mismatch RNA pretreatment altered spinal immunoreactivity of MOR or antinociception by spinal DAMGO, and had no effect on the baseline thermal nociceptive threshold. The inhibition of function and expression of DOR by siRNA was reversed by 72 hr after the last RNA injection. The uptake of fluorescence-tagged siRNA was detected in both DRG and spinal cord. The low effective dose of siRNA/i-FectTM complex reflects an efficient delivery of the siRNA to peripheral and spinal neurons, produced no behavioral signs of toxicity. This delivery method may be optimized for other gene targets.en
dc.language.isoenen
dc.publisherBioMed Centralen
dc.relation.urlhttp://mpx.sagepub.com/content/1/1744-8069-1-29.fullen
dc.rights© 2005 Luo et al; licensee BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0)en
dc.subjectraten
dc.subjectantinociceptionen
dc.subjectsensory neuronsen
dc.subjectspinal corden
dc.subjectknockdownen
dc.subjectRNA interferenceen
dc.titleAn efficient intrathecal delivery of small interfering RNA to the spinal cord and peripheral neuronsen
dc.typeArticleen
dc.identifier.eissn1744-8069en
dc.contributor.departmentDepartment of Pharmacology, The University of Arizona Health Sciences Center, Tucson, AZ 85724, USAen
dc.contributor.departmentNeuromics, Minneapolis, MN 55438, USAen
dc.identifier.journalMolecular Painen
dc.description.collectioninformationThis item is part of the UA Faculty Publications collection. For more information this item or other items in the UA Campus Repository, contact the University of Arizona Libraries at repository@u.library.arizona.edu.en
dc.eprint.versionFinal published versionen
All Items in UA Campus Repository are protected by copyright, with all rights reserved, unless otherwise indicated.