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N-methyl-N-nitrosourea-induced neuronal cell death in a large animal model of retinal degeneration in vitro

Taylor, Linnéa LU ; Arnér, Karin LU and Ghosh, Fredrik LU (2016) In Experimental Eye Research 148. p.55-64
Abstract

N-methyl-N-nitrosourea (MNU) has been reported to induce photoreceptor-specific degeneration with minimal inner retinal impact in small animals in vivo. Pending its use within a retinal transplantation paradigm, we here explore the effects of MNU on outer and inner retinal neurons and glia in an in vitro large animal model of retinal degeneration. The previously described degenerative culture explant model of adult porcine retina was used and compared with explants receiving 10 or 100 μg/ml MNU (MNU10 and MNU100) supplementation. All explants were kept for 5 days in vitro, and examined for morphology as well as for glial and neuronal immunohistochemical markers. Rhodopsin-labeled photoreceptors were present in all explants. The number... (More)

N-methyl-N-nitrosourea (MNU) has been reported to induce photoreceptor-specific degeneration with minimal inner retinal impact in small animals in vivo. Pending its use within a retinal transplantation paradigm, we here explore the effects of MNU on outer and inner retinal neurons and glia in an in vitro large animal model of retinal degeneration. The previously described degenerative culture explant model of adult porcine retina was used and compared with explants receiving 10 or 100 μg/ml MNU (MNU10 and MNU100) supplementation. All explants were kept for 5 days in vitro, and examined for morphology as well as for glial and neuronal immunohistochemical markers. Rhodopsin-labeled photoreceptors were present in all explants. The number of cone photoreceptors (transducin), rod bipolar cells (PKC) and horizontal cells (calbindin) was significantly lower in MNU treated explants (p <0.001). Gliosis was attenuated in MNU10 treated explants, with expression of vimentin, glial fibrillary protein (GFAP), glutamine synthetase (GS), and bFGF comparable to in vivo controls. In corresponding MNU100 counterparts, the expression of Müller cell proteins was almost extinguished. We here show that MNU causes degeneration of outer and inner retinal neurons and glia in the adult porcine retina in vitro. MNU10 explants display attenuation of gliosis, despite decreased neuronal survival compared with untreated controls. Our results have impact on the use of MNU as a large animal photoreceptor degeneration model, on tissue engineering related to retinal transplantation, and on our understanding of gliosis related neuronal degenerative cell death.

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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
Gliosis, N-methyl-N-nitrosourea, Neuronal degeneration, Porcine retina, Retinal explant culture, Retinal transplantation
in
Experimental Eye Research
volume
148
pages
10 pages
publisher
Elsevier
external identifiers
  • scopus:84973129455
  • wos:000379374900008
ISSN
0014-4835
DOI
10.1016/j.exer.2016.05.023
language
English
LU publication?
yes
id
ce1abd41-b559-433b-9f48-2e1223086dc6
date added to LUP
2016-07-04 10:47:26
date last changed
2017-03-05 04:35:07
@article{ce1abd41-b559-433b-9f48-2e1223086dc6,
  abstract     = {<p>N-methyl-N-nitrosourea (MNU) has been reported to induce photoreceptor-specific degeneration with minimal inner retinal impact in small animals in vivo. Pending its use within a retinal transplantation paradigm, we here explore the effects of MNU on outer and inner retinal neurons and glia in an in vitro large animal model of retinal degeneration. The previously described degenerative culture explant model of adult porcine retina was used and compared with explants receiving 10 or 100 μg/ml MNU (MNU10 and MNU100) supplementation. All explants were kept for 5 days in vitro, and examined for morphology as well as for glial and neuronal immunohistochemical markers. Rhodopsin-labeled photoreceptors were present in all explants. The number of cone photoreceptors (transducin), rod bipolar cells (PKC) and horizontal cells (calbindin) was significantly lower in MNU treated explants (p &lt;0.001). Gliosis was attenuated in MNU10 treated explants, with expression of vimentin, glial fibrillary protein (GFAP), glutamine synthetase (GS), and bFGF comparable to in vivo controls. In corresponding MNU100 counterparts, the expression of Müller cell proteins was almost extinguished. We here show that MNU causes degeneration of outer and inner retinal neurons and glia in the adult porcine retina in vitro. MNU10 explants display attenuation of gliosis, despite decreased neuronal survival compared with untreated controls. Our results have impact on the use of MNU as a large animal photoreceptor degeneration model, on tissue engineering related to retinal transplantation, and on our understanding of gliosis related neuronal degenerative cell death.</p>},
  author       = {Taylor, Linnéa and Arnér, Karin and Ghosh, Fredrik},
  issn         = {0014-4835},
  keyword      = {Gliosis,N-methyl-N-nitrosourea,Neuronal degeneration,Porcine retina,Retinal explant culture,Retinal transplantation},
  language     = {eng},
  month        = {07},
  pages        = {55--64},
  publisher    = {Elsevier},
  series       = {Experimental Eye Research},
  title        = {N-methyl-N-nitrosourea-induced neuronal cell death in a large animal model of retinal degeneration in vitro},
  url          = {http://dx.doi.org/10.1016/j.exer.2016.05.023},
  volume       = {148},
  year         = {2016},
}