Single cell RT-PCR analysis of tyrosine kinase receptor expression in adult rat retinal ganglion cells isolated by retinal sandwiching
(2002) In Brain Research Protocols 10(2). p.75-83- Abstract
We describe a protocol for analysis of gene expression in single, acutely dissociated adult rat retinal ganglion cells using RT-PCR. Retrograde tracing of retinal ganglion cells from the superior colliculi was conducted using Fluorogold. Retinas were dissected and ganglion cells isolated using retinal layer separation (sandwiching). Single, fluorescently labelled retinal ganglion cells were aspirated using a micropipette and used for PCR. Two PCR protocols are described where single cell cDNA was analysed for TrkB and GAPDH or TrkB, TrkC, Ret, Met, ErbB2 and Beta-actin by multiplex-PCR. All five tyrosine kinase receptors were amplified from single retinal ganglion cells. The method will prove useful for the molecular characterization of... (More)
We describe a protocol for analysis of gene expression in single, acutely dissociated adult rat retinal ganglion cells using RT-PCR. Retrograde tracing of retinal ganglion cells from the superior colliculi was conducted using Fluorogold. Retinas were dissected and ganglion cells isolated using retinal layer separation (sandwiching). Single, fluorescently labelled retinal ganglion cells were aspirated using a micropipette and used for PCR. Two PCR protocols are described where single cell cDNA was analysed for TrkB and GAPDH or TrkB, TrkC, Ret, Met, ErbB2 and Beta-actin by multiplex-PCR. All five tyrosine kinase receptors were amplified from single retinal ganglion cells. The method will prove useful for the molecular characterization of adult retinal ganglion cells.
(Less)
- author
- Lindqvist, Niclas LU ; Vidal-Sanz, Manuel and Hallböök, Finn
- publishing date
- 2002-10
- type
- Contribution to journal
- publication status
- published
- keywords
- Fluorogold (FG), Polymerase chain reaction (PCR), Retinal ganglion cell (rgc), Reverse transcription (RT)
- in
- Brain Research Protocols
- volume
- 10
- issue
- 2
- pages
- 9 pages
- publisher
- Elsevier
- external identifiers
-
- pmid:12431706
- scopus:0036808209
- ISSN
- 1385-299X
- DOI
- 10.1016/S1385-299X(02)00184-8
- language
- English
- LU publication?
- no
- id
- 9fae4c88-f5fd-4714-b60e-01822ac6c734
- date added to LUP
- 2017-06-02 15:18:55
- date last changed
- 2024-01-13 22:14:13
@article{9fae4c88-f5fd-4714-b60e-01822ac6c734,
abstract = {{<p>We describe a protocol for analysis of gene expression in single, acutely dissociated adult rat retinal ganglion cells using RT-PCR. Retrograde tracing of retinal ganglion cells from the superior colliculi was conducted using Fluorogold. Retinas were dissected and ganglion cells isolated using retinal layer separation (sandwiching). Single, fluorescently labelled retinal ganglion cells were aspirated using a micropipette and used for PCR. Two PCR protocols are described where single cell cDNA was analysed for TrkB and GAPDH or TrkB, TrkC, Ret, Met, ErbB2 and Beta-actin by multiplex-PCR. All five tyrosine kinase receptors were amplified from single retinal ganglion cells. The method will prove useful for the molecular characterization of adult retinal ganglion cells.</p>}},
author = {{Lindqvist, Niclas and Vidal-Sanz, Manuel and Hallböök, Finn}},
issn = {{1385-299X}},
keywords = {{Fluorogold (FG); Polymerase chain reaction (PCR); Retinal ganglion cell (rgc); Reverse transcription (RT)}},
language = {{eng}},
number = {{2}},
pages = {{75--83}},
publisher = {{Elsevier}},
series = {{Brain Research Protocols}},
title = {{Single cell RT-PCR analysis of tyrosine kinase receptor expression in adult rat retinal ganglion cells isolated by retinal sandwiching}},
url = {{http://dx.doi.org/10.1016/S1385-299X(02)00184-8}},
doi = {{10.1016/S1385-299X(02)00184-8}},
volume = {{10}},
year = {{2002}},
}