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A method for introducing non-silencing siRNA into the guinea pig cochlea in vivo.
J Neurosci Methods. 2008 Jan 30; 167(2):237-45.JN

Abstract

The aim of this preliminary study was to establish the methodology by which siRNA can be introduced into the adult guinea pig cochlea in vivo whilst preserving auditory function with a view to using targeted siRNAs to knockdown genes essential for auditory transduction. Initially a fluorescently tagged non-silencing siRNA complexed with a lipid-based transfection reagent was introduced into the perilymphatic compartment of the cochlea. Although auditory function was fully preserved, siRNA uptake was only observed in cells lining the perilymphatic space that are not critically involved in auditory transduction and therefore of little interest. Another approach was therefore adopted, in which siRNA was introduced directly into the scala media (endolymphatic compartment) of the apical (fourth) cochlear turn by slow pressure injection. During endolymphatic perfusion, the endocochlear potential (EP) and compound action potential (CAP) thresholds for basal turn frequencies from 6 to 20 kHz could be preserved, while CAP thresholds for 1-4 kHz were often elevated by 10-20 dB. CAP thresholds and EP were preserved 24 and 48 h after perfusion in some animals but reduced in others. siRNA uptake was observed predominantly in marginal and intermediate cells of the stria vascularis in all cochlear turns but not in cells of the organ of Corti.

Authors+Show Affiliations

The Auditory Laboratory, Discipline of Physiology, School of Biomedical, Biomolecular and Chemical Sciences, The University of Western Australia, Australia.No affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

17945347

Citation

Sellick, P, et al. "A Method for Introducing Non-silencing siRNA Into the Guinea Pig Cochlea in Vivo." Journal of Neuroscience Methods, vol. 167, no. 2, 2008, pp. 237-45.
Sellick P, Layton MG, Rodger J, et al. A method for introducing non-silencing siRNA into the guinea pig cochlea in vivo. J Neurosci Methods. 2008;167(2):237-45.
Sellick, P., Layton, M. G., Rodger, J., & Robertson, D. (2008). A method for introducing non-silencing siRNA into the guinea pig cochlea in vivo. Journal of Neuroscience Methods, 167(2), 237-45.
Sellick P, et al. A Method for Introducing Non-silencing siRNA Into the Guinea Pig Cochlea in Vivo. J Neurosci Methods. 2008 Jan 30;167(2):237-45. PubMed PMID: 17945347.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - A method for introducing non-silencing siRNA into the guinea pig cochlea in vivo. AU - Sellick,P, AU - Layton,M G, AU - Rodger,J, AU - Robertson,D, Y1 - 2007/09/06/ PY - 2007/06/27/received PY - 2007/08/21/revised PY - 2007/08/21/accepted PY - 2007/10/20/pubmed PY - 2008/4/12/medline PY - 2007/10/20/entrez SP - 237 EP - 45 JF - Journal of neuroscience methods JO - J Neurosci Methods VL - 167 IS - 2 N2 - The aim of this preliminary study was to establish the methodology by which siRNA can be introduced into the adult guinea pig cochlea in vivo whilst preserving auditory function with a view to using targeted siRNAs to knockdown genes essential for auditory transduction. Initially a fluorescently tagged non-silencing siRNA complexed with a lipid-based transfection reagent was introduced into the perilymphatic compartment of the cochlea. Although auditory function was fully preserved, siRNA uptake was only observed in cells lining the perilymphatic space that are not critically involved in auditory transduction and therefore of little interest. Another approach was therefore adopted, in which siRNA was introduced directly into the scala media (endolymphatic compartment) of the apical (fourth) cochlear turn by slow pressure injection. During endolymphatic perfusion, the endocochlear potential (EP) and compound action potential (CAP) thresholds for basal turn frequencies from 6 to 20 kHz could be preserved, while CAP thresholds for 1-4 kHz were often elevated by 10-20 dB. CAP thresholds and EP were preserved 24 and 48 h after perfusion in some animals but reduced in others. siRNA uptake was observed predominantly in marginal and intermediate cells of the stria vascularis in all cochlear turns but not in cells of the organ of Corti. SN - 0165-0270 UR - https://www.unboundmedicine.com/medline/citation/17945347/A_method_for_introducing_non_silencing_siRNA_into_the_guinea_pig_cochlea_in_vivo_ L2 - https://www.lens.org/lens/search/patent/list?q=citation_id:17945347 DB - PRIME DP - Unbound Medicine ER -