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Multiple receptor tyrosine kinases are expressed in adult rat retinal ganglion cells as revealed by single-cell degenerate primer polymerase chain reaction

Lindqvist, Niclas LU ; Lönngren, Ulrika; Agudo, Marta; Näpänkangas, Ulla; Vidal-Sanz, Manuel and Hallböök, Finn (2010) In Uppsala Journal of Medical Sciences 115(1). p.65-80
Abstract

BACKGROUND: To achieve a better understanding of the repertoire of receptor tyrosine kinases (RTKs) in adult retinal ganglion cells (RGCs) we performed polymerase chain reaction (PCR), using degenerate primers directed towards conserved sequences in the tyrosine kinase domain, on cDNA from isolated single RGCs univocally identified by retrograde tracing from the superior colliculi.

RESULTS: All the PCR-amplified fragments of the expected sizes were sequenced, and 25% of them contained a tyrosine kinase domain. These were: Axl, Csf-1R, Eph A4, Pdgfrbeta, Ptk7, Ret, Ros, Sky, TrkB, TrkC, Vegfr-2, and Vegfr-3. Non-RTK sequences were Jak1 and 2. Retinal expression of Axl, Csf-1R, Pdgfrbeta, Ret, Sky, TrkB, TrkC, Vegfr-2, and Vegfr-3,... (More)

BACKGROUND: To achieve a better understanding of the repertoire of receptor tyrosine kinases (RTKs) in adult retinal ganglion cells (RGCs) we performed polymerase chain reaction (PCR), using degenerate primers directed towards conserved sequences in the tyrosine kinase domain, on cDNA from isolated single RGCs univocally identified by retrograde tracing from the superior colliculi.

RESULTS: All the PCR-amplified fragments of the expected sizes were sequenced, and 25% of them contained a tyrosine kinase domain. These were: Axl, Csf-1R, Eph A4, Pdgfrbeta, Ptk7, Ret, Ros, Sky, TrkB, TrkC, Vegfr-2, and Vegfr-3. Non-RTK sequences were Jak1 and 2. Retinal expression of Axl, Csf-1R, Pdgfrbeta, Ret, Sky, TrkB, TrkC, Vegfr-2, and Vegfr-3, as well as Jak1 and 2, was confirmed by PCR on total retina cDNA. Immunodetection of Csf-1R, Pdgfralpha/beta, Ret, Sky, TrkB, and Vegfr-2 on retrogradely traced retinas demonstrated that they were expressed by RGCs. Co-localization of Vegfr-2 and Csf-1R, of Vegfr-2 and TrkB, and of Csf-1R and Ret in retrogradely labelled RGCs was shown. The effect of optic nerve transection on the mRNA level of Pdgfrbeta, Csf-1R, Vegfr-2, Sky, and Axl, and of the Axl ligands Gas6 and ProteinS, was analysed. These analyses show transection-induced changes in Axl and ProteinS mRNA levels.

CONCLUSIONS: The repertoire of RTKs expressed by RGCs is more extensive than previously anticipated. Several of the receptors found in this study, including Pdgfrbeta, Csf-1R, Vegfr-2, Sky, and Axl, and their ligands, have not previously been primarily associated with retinal ganglion cells.

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publication status
published
keywords
Animals, DNA Primers, DNA, Complementary, Female, Gene Expression Regulation, Enzymologic, Ligands, Male, Polymerase Chain Reaction, Protein Structure, Tertiary, Rats, Rats, Sprague-Dawley, Receptor Protein-Tyrosine Kinases, Retina, Retinal Ganglion Cells, Sex Factors
in
Uppsala Journal of Medical Sciences
volume
115
issue
1
pages
16 pages
publisher
Taylor & Francis
external identifiers
  • scopus:77749327686
ISSN
0300-9734
DOI
10.3109/03009731003597119
language
English
LU publication?
no
id
1ca86070-50e6-49fa-909b-0a110c3f5510
date added to LUP
2016-06-02 10:22:17
date last changed
2017-01-01 08:27:30
@article{1ca86070-50e6-49fa-909b-0a110c3f5510,
  abstract     = {<p>BACKGROUND: To achieve a better understanding of the repertoire of receptor tyrosine kinases (RTKs) in adult retinal ganglion cells (RGCs) we performed polymerase chain reaction (PCR), using degenerate primers directed towards conserved sequences in the tyrosine kinase domain, on cDNA from isolated single RGCs univocally identified by retrograde tracing from the superior colliculi.</p><p>RESULTS: All the PCR-amplified fragments of the expected sizes were sequenced, and 25% of them contained a tyrosine kinase domain. These were: Axl, Csf-1R, Eph A4, Pdgfrbeta, Ptk7, Ret, Ros, Sky, TrkB, TrkC, Vegfr-2, and Vegfr-3. Non-RTK sequences were Jak1 and 2. Retinal expression of Axl, Csf-1R, Pdgfrbeta, Ret, Sky, TrkB, TrkC, Vegfr-2, and Vegfr-3, as well as Jak1 and 2, was confirmed by PCR on total retina cDNA. Immunodetection of Csf-1R, Pdgfralpha/beta, Ret, Sky, TrkB, and Vegfr-2 on retrogradely traced retinas demonstrated that they were expressed by RGCs. Co-localization of Vegfr-2 and Csf-1R, of Vegfr-2 and TrkB, and of Csf-1R and Ret in retrogradely labelled RGCs was shown. The effect of optic nerve transection on the mRNA level of Pdgfrbeta, Csf-1R, Vegfr-2, Sky, and Axl, and of the Axl ligands Gas6 and ProteinS, was analysed. These analyses show transection-induced changes in Axl and ProteinS mRNA levels.</p><p>CONCLUSIONS: The repertoire of RTKs expressed by RGCs is more extensive than previously anticipated. Several of the receptors found in this study, including Pdgfrbeta, Csf-1R, Vegfr-2, Sky, and Axl, and their ligands, have not previously been primarily associated with retinal ganglion cells.</p>},
  author       = {Lindqvist, Niclas and Lönngren, Ulrika and Agudo, Marta and Näpänkangas, Ulla and Vidal-Sanz, Manuel and Hallböök, Finn},
  issn         = {0300-9734},
  keyword      = {Animals,DNA Primers,DNA, Complementary,Female,Gene Expression Regulation, Enzymologic,Ligands,Male,Polymerase Chain Reaction,Protein Structure, Tertiary,Rats,Rats, Sprague-Dawley,Receptor Protein-Tyrosine Kinases,Retina,Retinal Ganglion Cells,Sex Factors},
  language     = {eng},
  number       = {1},
  pages        = {65--80},
  publisher    = {Taylor & Francis},
  series       = {Uppsala Journal of Medical Sciences},
  title        = {Multiple receptor tyrosine kinases are expressed in adult rat retinal ganglion cells as revealed by single-cell degenerate primer polymerase chain reaction},
  url          = {http://dx.doi.org/10.3109/03009731003597119},
  volume       = {115},
  year         = {2010},
}