Advanced

DNA fragmentation characteristic of apoptosis and cell loss induced by kainic acid in rabbit retinas

Perez, M T LU ; Arnér, K LU and Håkansson, Anders P LU (1997) In Neurochemistry International 31(2). p.251-60
Abstract

We have examined whether in vivo exposure to the glutamate analogue, kainic acid, induces cell loss through apoptosis and/or through necrosis. The vulnerability of rabbit retinal cells was evaluated by routine histopathology. The DNA fragmentation was examined using an in situ method (TUNEL: TdT-mediated biotin-dUTP nick-end labelling) and agarose gel electrophoresis of extracted retinal DNA. Retinas were examined at 30 min, and 4, 16, 24 and 36 h, and 2-5 days following the intraocular administration of 140 nmol kainic acid. Although pyknotic cells could be seen already at 30 min post-injection, TUNEL-labelled nuclei were first observed 4 h after the injection. A relatively large number of pyknotic cells and of TUNEL-labelled nuclei... (More)

We have examined whether in vivo exposure to the glutamate analogue, kainic acid, induces cell loss through apoptosis and/or through necrosis. The vulnerability of rabbit retinal cells was evaluated by routine histopathology. The DNA fragmentation was examined using an in situ method (TUNEL: TdT-mediated biotin-dUTP nick-end labelling) and agarose gel electrophoresis of extracted retinal DNA. Retinas were examined at 30 min, and 4, 16, 24 and 36 h, and 2-5 days following the intraocular administration of 140 nmol kainic acid. Although pyknotic cells could be seen already at 30 min post-injection, TUNEL-labelled nuclei were first observed 4 h after the injection. A relatively large number of pyknotic cells and of TUNEL-labelled nuclei were still seen at 5 days post-injection. Pyknotic cells were seen throughout the inner nuclear layer (mostly in the proximal half of the layer) and in the ganglion cell layer. The TUNEL-labelled nuclei were almost only seen in the proximal inner nuclear layer. Analysis of DNA by electrophoresis revealed the presence of large molecular weight fragments 4 h after the injection, and of oligonucleosome-size fragments between 16 h and 2 days after the injection. The present study thus presents evidence that, in our model, the retinal cell loss induced by kainic acid is preceded, probably in most cells, by a fragmentation of DNA characteristic of apoptotic cell death. The process of cell loss following kainic acid administration was found to be relatively slow, further suggesting that a programmed type of cell death, which eventually induces apoptosis, is involved. No indication that cells were lost also through necrosis was obtained.

(Less)
Please use this url to cite or link to this publication:
author
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
Animals, Apoptosis, DNA Fragmentation, Electrophoresis, Agar Gel, Female, Genetic Techniques, Kainic Acid, Male, Rabbits, Retina
in
Neurochemistry International
volume
31
issue
2
pages
10 pages
publisher
Elsevier
external identifiers
  • Scopus:0030971574
ISSN
0197-0186
language
English
LU publication?
yes
id
b2a37a66-8186-4817-bfe8-5de4082194d2
date added to LUP
2016-05-21 13:58:05
date last changed
2016-06-13 15:03:18
@misc{b2a37a66-8186-4817-bfe8-5de4082194d2,
  abstract     = {<p>We have examined whether in vivo exposure to the glutamate analogue, kainic acid, induces cell loss through apoptosis and/or through necrosis. The vulnerability of rabbit retinal cells was evaluated by routine histopathology. The DNA fragmentation was examined using an in situ method (TUNEL: TdT-mediated biotin-dUTP nick-end labelling) and agarose gel electrophoresis of extracted retinal DNA. Retinas were examined at 30 min, and 4, 16, 24 and 36 h, and 2-5 days following the intraocular administration of 140 nmol kainic acid. Although pyknotic cells could be seen already at 30 min post-injection, TUNEL-labelled nuclei were first observed 4 h after the injection. A relatively large number of pyknotic cells and of TUNEL-labelled nuclei were still seen at 5 days post-injection. Pyknotic cells were seen throughout the inner nuclear layer (mostly in the proximal half of the layer) and in the ganglion cell layer. The TUNEL-labelled nuclei were almost only seen in the proximal inner nuclear layer. Analysis of DNA by electrophoresis revealed the presence of large molecular weight fragments 4 h after the injection, and of oligonucleosome-size fragments between 16 h and 2 days after the injection. The present study thus presents evidence that, in our model, the retinal cell loss induced by kainic acid is preceded, probably in most cells, by a fragmentation of DNA characteristic of apoptotic cell death. The process of cell loss following kainic acid administration was found to be relatively slow, further suggesting that a programmed type of cell death, which eventually induces apoptosis, is involved. No indication that cells were lost also through necrosis was obtained.</p>},
  author       = {Perez, M T and Arnér, K and Håkansson, Anders P},
  issn         = {0197-0186},
  keyword      = {Animals,Apoptosis,DNA Fragmentation,Electrophoresis, Agar Gel,Female,Genetic Techniques,Kainic Acid,Male,Rabbits,Retina},
  language     = {eng},
  number       = {2},
  pages        = {251--60},
  publisher    = {ARRAY(0x9db15f0)},
  series       = {Neurochemistry International},
  title        = {DNA fragmentation characteristic of apoptosis and cell loss induced by kainic acid in rabbit retinas},
  volume       = {31},
  year         = {1997},
}