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AAV vector-mediated secretion of chondroitinase provides a sensitive tracer for axonal arborisations

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AAV vector-mediated secretion of chondroitinase provides a sensitive tracer for axonal arborisations. / Alves, Joao Nuno; Muir, Elizabeth M; Andrews, Melissa Renee; Ward, Anneliese; Michelmore, Nicholas; Dasgupta, Debayan; Verhaagen, Joost; Moloney, Elizabeth B; Keynes, Roger J; Fawcett, James W; Rogers, John H.

In: Journal of Neuroscience Methods, Vol. 227, 30.04.2014, p. 107-120.

Research output: Contribution to journalArticle

Harvard

Alves, JN, Muir, EM, Andrews, MR, Ward, A, Michelmore, N, Dasgupta, D, Verhaagen, J, Moloney, EB, Keynes, RJ, Fawcett, JW & Rogers, JH 2014, 'AAV vector-mediated secretion of chondroitinase provides a sensitive tracer for axonal arborisations', Journal of Neuroscience Methods, vol. 227, pp. 107-120. https://doi.org/10.1016/j.jneumeth.2014.02.010

APA

Alves, J. N., Muir, E. M., Andrews, M. R., Ward, A., Michelmore, N., Dasgupta, D., ... Rogers, J. H. (2014). AAV vector-mediated secretion of chondroitinase provides a sensitive tracer for axonal arborisations. Journal of Neuroscience Methods, 227, 107-120. https://doi.org/10.1016/j.jneumeth.2014.02.010

Vancouver

Alves JN, Muir EM, Andrews MR, Ward A, Michelmore N, Dasgupta D et al. AAV vector-mediated secretion of chondroitinase provides a sensitive tracer for axonal arborisations. Journal of Neuroscience Methods. 2014 Apr 30;227:107-120. https://doi.org/10.1016/j.jneumeth.2014.02.010

Author

Alves, Joao Nuno ; Muir, Elizabeth M ; Andrews, Melissa Renee ; Ward, Anneliese ; Michelmore, Nicholas ; Dasgupta, Debayan ; Verhaagen, Joost ; Moloney, Elizabeth B ; Keynes, Roger J ; Fawcett, James W ; Rogers, John H. / AAV vector-mediated secretion of chondroitinase provides a sensitive tracer for axonal arborisations. In: Journal of Neuroscience Methods. 2014 ; Vol. 227. pp. 107-120.

Bibtex - Download

@article{dcf01635fd7f4bf7bd18b00af56cbc9f,
title = "AAV vector-mediated secretion of chondroitinase provides a sensitive tracer for axonal arborisations",
abstract = "As part of a project to express chondroitinase ABC (ChABC) in neurons of the central nervous system, we have inserted a modified ChABC gene into an adeno-associated viral (AAV) vector and injected it into the vibrissal motor cortex in adult rats to determine the extent and distribution of expression of the enzyme. A similar vector for expression of green fluorescent protein (GFP) was injected into the same location. For each vector, two versions with minor differences were used, giving similar results. After 4 weeks, the brains were stained to show GFP and products of chondroitinase digestion. Chondroitinase was widely expressed, and the AAV-ChABC and AAV-GFP vectors gave similar expression patterns in many respects, consistent with the known projections from the directly transduced neurons in vibrissal motor cortex and adjacent cingulate cortex. In addition, diffusion of vector to deeper neuronal populations led to labelling of remote projection fields which was much more extensive with AAV-ChABC than with AAV-GFP. The most notable of these populations are inferred to be neurons of cortical layer 6, projecting widely in the thalamus, and neurons of the anterior pole of the hippocampus, projecting through most of the hippocampus. We conclude that, whereas GFP does not label the thinnest axonal branches of some neuronal types, chondroitinase is efficiently secreted from these arborisations and enables their extent to be sensitively visualised. After 12 weeks, chondroitinase expression was undiminished.",
keywords = "AAV , Chondroitinase, Vibrissal motor cortex, Whisker movements, Pathway tracing, Green fluorescent protein, Corticospinal tract, Thalamus",
author = "Alves, {Joao Nuno} and Muir, {Elizabeth M} and Andrews, {Melissa Renee} and Anneliese Ward and Nicholas Michelmore and Debayan Dasgupta and Joost Verhaagen and Moloney, {Elizabeth B} and Keynes, {Roger J} and Fawcett, {James W} and Rogers, {John H}",
year = "2014",
month = "4",
day = "30",
doi = "10.1016/j.jneumeth.2014.02.010",
language = "English",
volume = "227",
pages = "107--120",
journal = "Journal of Neuroscience Methods",
issn = "0165-0270",
publisher = "Elsevier",

}

RIS (suitable for import to EndNote) - Download

TY - JOUR

T1 - AAV vector-mediated secretion of chondroitinase provides a sensitive tracer for axonal arborisations

AU - Alves, Joao Nuno

AU - Muir, Elizabeth M

AU - Andrews, Melissa Renee

AU - Ward, Anneliese

AU - Michelmore, Nicholas

AU - Dasgupta, Debayan

AU - Verhaagen, Joost

AU - Moloney, Elizabeth B

AU - Keynes, Roger J

AU - Fawcett, James W

AU - Rogers, John H

PY - 2014/4/30

Y1 - 2014/4/30

N2 - As part of a project to express chondroitinase ABC (ChABC) in neurons of the central nervous system, we have inserted a modified ChABC gene into an adeno-associated viral (AAV) vector and injected it into the vibrissal motor cortex in adult rats to determine the extent and distribution of expression of the enzyme. A similar vector for expression of green fluorescent protein (GFP) was injected into the same location. For each vector, two versions with minor differences were used, giving similar results. After 4 weeks, the brains were stained to show GFP and products of chondroitinase digestion. Chondroitinase was widely expressed, and the AAV-ChABC and AAV-GFP vectors gave similar expression patterns in many respects, consistent with the known projections from the directly transduced neurons in vibrissal motor cortex and adjacent cingulate cortex. In addition, diffusion of vector to deeper neuronal populations led to labelling of remote projection fields which was much more extensive with AAV-ChABC than with AAV-GFP. The most notable of these populations are inferred to be neurons of cortical layer 6, projecting widely in the thalamus, and neurons of the anterior pole of the hippocampus, projecting through most of the hippocampus. We conclude that, whereas GFP does not label the thinnest axonal branches of some neuronal types, chondroitinase is efficiently secreted from these arborisations and enables their extent to be sensitively visualised. After 12 weeks, chondroitinase expression was undiminished.

AB - As part of a project to express chondroitinase ABC (ChABC) in neurons of the central nervous system, we have inserted a modified ChABC gene into an adeno-associated viral (AAV) vector and injected it into the vibrissal motor cortex in adult rats to determine the extent and distribution of expression of the enzyme. A similar vector for expression of green fluorescent protein (GFP) was injected into the same location. For each vector, two versions with minor differences were used, giving similar results. After 4 weeks, the brains were stained to show GFP and products of chondroitinase digestion. Chondroitinase was widely expressed, and the AAV-ChABC and AAV-GFP vectors gave similar expression patterns in many respects, consistent with the known projections from the directly transduced neurons in vibrissal motor cortex and adjacent cingulate cortex. In addition, diffusion of vector to deeper neuronal populations led to labelling of remote projection fields which was much more extensive with AAV-ChABC than with AAV-GFP. The most notable of these populations are inferred to be neurons of cortical layer 6, projecting widely in the thalamus, and neurons of the anterior pole of the hippocampus, projecting through most of the hippocampus. We conclude that, whereas GFP does not label the thinnest axonal branches of some neuronal types, chondroitinase is efficiently secreted from these arborisations and enables their extent to be sensitively visualised. After 12 weeks, chondroitinase expression was undiminished.

KW - AAV

KW - Chondroitinase

KW - Vibrissal motor cortex

KW - Whisker movements

KW - Pathway tracing

KW - Green fluorescent protein

KW - Corticospinal tract

KW - Thalamus

U2 - 10.1016/j.jneumeth.2014.02.010

DO - 10.1016/j.jneumeth.2014.02.010

M3 - Article

VL - 227

SP - 107

EP - 120

JO - Journal of Neuroscience Methods

JF - Journal of Neuroscience Methods

SN - 0165-0270

ER -

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