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A selective method for transfection of retinal ganglion cells by retrograde transfer of antisense oligonucleotides against kynurenine aminotransferase II

Standard

A selective method for transfection of retinal ganglion cells by retrograde transfer of antisense oligonucleotides against kynurenine aminotransferase II. / Thaler, Sebastian; Rejdak, Robert; Dietrich, Karen; Ladewig, Thomas; Okuno, Etsuo; Kocki, Tomasz; Turski, Waldemar Andrzej; Junemann, Anselm; Zrenner, Eberhart; Schuettauf, Frank.

In: MOL VIS, Vol. 12, 22.02.2006, p. 100-7.

Research output: SCORING: Contribution to journal › SCORING: Journal article › Research › peer-review

Harvard

Thaler, S, Rejdak, R, Dietrich, K, Ladewig, T, Okuno, E, Kocki, T, Turski, WA, Junemann, A, Zrenner, E & Schuettauf, F 2006, 'A selective method for transfection of retinal ganglion cells by retrograde transfer of antisense oligonucleotides against kynurenine aminotransferase II', MOL VIS, vol. 12, pp. 100-7.

APA

Thaler, S., Rejdak, R., Dietrich, K., Ladewig, T., Okuno, E., Kocki, T., Turski, W. A., Junemann, A., Zrenner, E., & Schuettauf, F. (2006). A selective method for transfection of retinal ganglion cells by retrograde transfer of antisense oligonucleotides against kynurenine aminotransferase II. MOL VIS, 12, 100-7.

Vancouver

Thaler S, Rejdak R, Dietrich K, Ladewig T, Okuno E, Kocki T et al. A selective method for transfection of retinal ganglion cells by retrograde transfer of antisense oligonucleotides against kynurenine aminotransferase II. MOL VIS. 2006 Feb 22;12:100-7.

Bibtex

@article{594e38c34a2f4466a3280ad4a1273b72,
title = "A selective method for transfection of retinal ganglion cells by retrograde transfer of antisense oligonucleotides against kynurenine aminotransferase II",
abstract = "PURPOSE: Intravitreal administration of specific antisense oligonucleotides (ODNs) effectively downregulates gene expression in the retina but does not modulate it exclusively in retinal ganglion cells (RGCs). Expression of kynurenine aminotransferase II (KAT II) in RGCs has been well described in the literature. We describe a new method for downregulating cellular KAT II expression via transfection of RGC by retrograde transfer of ODN.METHODS: Fluorescently labeled, specific ODNs against KAT II were injected into rats either intravitreally or into the superior colliculi. Fluorescence microscopy of retinal flat-mounts and radial sections was used to compare the location, duration, and degree of transfection for both methods of delivery. The effects of both methods on KAT II expression in RGCs were studied immunohistochemically with unlabeled ODN. Retinal kynurenic acid (KYNA) contents were measured using high pressure liquid chromatography (HPLC).RESULTS: After intravitreal injection, fluorescently labeled ODN reached all retinal layers, whereas injections into the superior colliculus resulted in transfection of the RGC layer alone. Immunohistochemistry showed that both methods of ODN application had a similar effect on downregulation of KAT II expression in RGC. Retinal KYNA content decreased significantly 4 days after both types of ODN administration.CONCLUSIONS: This study demonstrated that retrograde transfer of specific ODN into RGC is feasible and induces downregulation of KAT II cellular expression. This may become a useful tool for modulating gene expression in the retinal ganglion cell layer in vivo without direct transfer of ODN to other retinal cell layers.",
keywords = "Animals, Biological Transport, Active, Carbocyanines, Chromatography, High Pressure Liquid, Down-Regulation, Fluorescent Dyes, Immunohistochemistry, Injections, Microscopy, Confocal, Microscopy, Fluorescence, Oligonucleotides, Antisense/administration & dosage, Rats, Rats, Inbred BN, Retinal Ganglion Cells/metabolism, Staining and Labeling, Stilbamidines, Superior Colliculi, Time Factors, Transaminases/genetics, Transfection/methods, Vitreous Body",
author = "Sebastian Thaler and Robert Rejdak and Karen Dietrich and Thomas Ladewig and Etsuo Okuno and Tomasz Kocki and Turski, {Waldemar Andrzej} and Anselm Junemann and Eberhart Zrenner and Frank Schuettauf",
year = "2006",
month = feb,
day = "22",
language = "English",
volume = "12",
pages = "100--7",
journal = "MOL VIS",
issn = "1090-0535",
publisher = "Molecular Vision",

}

RIS

TY - JOUR

T1 - A selective method for transfection of retinal ganglion cells by retrograde transfer of antisense oligonucleotides against kynurenine aminotransferase II

AU - Thaler, Sebastian

AU - Rejdak, Robert

AU - Dietrich, Karen

AU - Ladewig, Thomas

AU - Okuno, Etsuo

AU - Kocki, Tomasz

AU - Turski, Waldemar Andrzej

AU - Junemann, Anselm

AU - Zrenner, Eberhart

AU - Schuettauf, Frank

PY - 2006/2/22

Y1 - 2006/2/22

N2 - PURPOSE: Intravitreal administration of specific antisense oligonucleotides (ODNs) effectively downregulates gene expression in the retina but does not modulate it exclusively in retinal ganglion cells (RGCs). Expression of kynurenine aminotransferase II (KAT II) in RGCs has been well described in the literature. We describe a new method for downregulating cellular KAT II expression via transfection of RGC by retrograde transfer of ODN.METHODS: Fluorescently labeled, specific ODNs against KAT II were injected into rats either intravitreally or into the superior colliculi. Fluorescence microscopy of retinal flat-mounts and radial sections was used to compare the location, duration, and degree of transfection for both methods of delivery. The effects of both methods on KAT II expression in RGCs were studied immunohistochemically with unlabeled ODN. Retinal kynurenic acid (KYNA) contents were measured using high pressure liquid chromatography (HPLC).RESULTS: After intravitreal injection, fluorescently labeled ODN reached all retinal layers, whereas injections into the superior colliculus resulted in transfection of the RGC layer alone. Immunohistochemistry showed that both methods of ODN application had a similar effect on downregulation of KAT II expression in RGC. Retinal KYNA content decreased significantly 4 days after both types of ODN administration.CONCLUSIONS: This study demonstrated that retrograde transfer of specific ODN into RGC is feasible and induces downregulation of KAT II cellular expression. This may become a useful tool for modulating gene expression in the retinal ganglion cell layer in vivo without direct transfer of ODN to other retinal cell layers.

AB - PURPOSE: Intravitreal administration of specific antisense oligonucleotides (ODNs) effectively downregulates gene expression in the retina but does not modulate it exclusively in retinal ganglion cells (RGCs). Expression of kynurenine aminotransferase II (KAT II) in RGCs has been well described in the literature. We describe a new method for downregulating cellular KAT II expression via transfection of RGC by retrograde transfer of ODN.METHODS: Fluorescently labeled, specific ODNs against KAT II were injected into rats either intravitreally or into the superior colliculi. Fluorescence microscopy of retinal flat-mounts and radial sections was used to compare the location, duration, and degree of transfection for both methods of delivery. The effects of both methods on KAT II expression in RGCs were studied immunohistochemically with unlabeled ODN. Retinal kynurenic acid (KYNA) contents were measured using high pressure liquid chromatography (HPLC).RESULTS: After intravitreal injection, fluorescently labeled ODN reached all retinal layers, whereas injections into the superior colliculus resulted in transfection of the RGC layer alone. Immunohistochemistry showed that both methods of ODN application had a similar effect on downregulation of KAT II expression in RGC. Retinal KYNA content decreased significantly 4 days after both types of ODN administration.CONCLUSIONS: This study demonstrated that retrograde transfer of specific ODN into RGC is feasible and induces downregulation of KAT II cellular expression. This may become a useful tool for modulating gene expression in the retinal ganglion cell layer in vivo without direct transfer of ODN to other retinal cell layers.

KW - Animals

KW - Biological Transport, Active

KW - Carbocyanines

KW - Chromatography, High Pressure Liquid

KW - Down-Regulation

KW - Fluorescent Dyes

KW - Immunohistochemistry

KW - Injections

KW - Microscopy, Confocal

KW - Microscopy, Fluorescence

KW - Oligonucleotides, Antisense/administration & dosage

KW - Rats

KW - Rats, Inbred BN

KW - Retinal Ganglion Cells/metabolism

KW - Staining and Labeling

KW - Stilbamidines

KW - Superior Colliculi

KW - Time Factors

KW - Transaminases/genetics

KW - Transfection/methods

KW - Vitreous Body

M3 - SCORING: Journal article

C2 - 16518307

VL - 12

SP - 100

EP - 107

JO - MOL VIS

JF - MOL VIS

SN - 1090-0535

ER -