Retina-derived microglial cells induce photoreceptor cell death in vitro

Rouel S. Roque, Antonio Armando Aviles Rosales, Liu Jingjing, Neeraj Agarwal, Muayyad R. Al-Ubaidi

Research output: Contribution to journalArticle

85 Citations (Scopus)

Abstract

In animals with retinal degeneration, the presence of activated microglial cells in the outer retina during the early stages of injury suggests that they may be involved in the ensuing photoreceptor cell death. In the following study, we investigated the effects of rat retina-derived microglial cells on a photoreceptor cell line (661w) using cell culture techniques. The difficulty of obtaining pure populations of photoreceptor cells necessitated our use of the 661w photoreceptor cells generated from retinas of transgenic mice. 661w cells were incubated for 24-48 h in basal medium or basal medium conditioned by activated microglial cells (MGCM) or Muller cells (MCCM), and tested for cell death using lactate dehydrogenase (LDH) assay. The induction of apoptosis in the 661w cells by MGCM was investigated using Terminal deoxynucleotidyl Transferase (TdT)-mediated dUTP nick-end labeling (TUNEL) and DNA laddering. Treatment of 661w cells with MGCM for 48 h resulted in ~ 73% of cells dead as compared with 19-20% of cells grown in either basal medium or MCCM. Serum supplementation or pretreatment with heat did not abolish the cytotoxicity of MGCM. More TUNEL- positive cells were observed in MGCM-treated cultures as compared with those in basal medium. Bands in multiples of ~ 180 bp formed DNA ladders in MGCM- treated but not in basal medium-treated samples. Our study shows that microglial cells release soluble product(s) that induce degeneration of cultured photoreceptor cells. Moreover, the mechanism of microglia-induced photoreceptor cell death may involve apoptosis similar to that observed in animals with retinal degeneration.

Original languageEnglish
Pages (from-to)110-119
Number of pages10
JournalBrain Research
Volume836
Issue number1-2
DOIs
StatePublished - 31 Jul 1999

Fingerprint

Photoreceptor Cells
Retina
Cell Death
Retinal Degeneration
In Vitro Techniques
Apoptosis
Ependymoglial Cells
DNA Nucleotidylexotransferase
DNA
Microglia
Conditioned Culture Medium
L-Lactate Dehydrogenase
Transgenic Mice
Cultured Cells
Cell Culture Techniques
Hot Temperature
Cell Line

Keywords

  • Apoptosis
  • Lactate dehydrogenase
  • Microglia
  • Muller cell
  • Necrosis
  • Nick-end labeling
  • Photoreceptor cell
  • Retinal degeneration

Cite this

Roque, Rouel S. ; Rosales, Antonio Armando Aviles ; Jingjing, Liu ; Agarwal, Neeraj ; Al-Ubaidi, Muayyad R. / Retina-derived microglial cells induce photoreceptor cell death in vitro. In: Brain Research. 1999 ; Vol. 836, No. 1-2. pp. 110-119.
@article{9fcb504455c34644a8904d29975e593d,
title = "Retina-derived microglial cells induce photoreceptor cell death in vitro",
abstract = "In animals with retinal degeneration, the presence of activated microglial cells in the outer retina during the early stages of injury suggests that they may be involved in the ensuing photoreceptor cell death. In the following study, we investigated the effects of rat retina-derived microglial cells on a photoreceptor cell line (661w) using cell culture techniques. The difficulty of obtaining pure populations of photoreceptor cells necessitated our use of the 661w photoreceptor cells generated from retinas of transgenic mice. 661w cells were incubated for 24-48 h in basal medium or basal medium conditioned by activated microglial cells (MGCM) or Muller cells (MCCM), and tested for cell death using lactate dehydrogenase (LDH) assay. The induction of apoptosis in the 661w cells by MGCM was investigated using Terminal deoxynucleotidyl Transferase (TdT)-mediated dUTP nick-end labeling (TUNEL) and DNA laddering. Treatment of 661w cells with MGCM for 48 h resulted in ~ 73{\%} of cells dead as compared with 19-20{\%} of cells grown in either basal medium or MCCM. Serum supplementation or pretreatment with heat did not abolish the cytotoxicity of MGCM. More TUNEL- positive cells were observed in MGCM-treated cultures as compared with those in basal medium. Bands in multiples of ~ 180 bp formed DNA ladders in MGCM- treated but not in basal medium-treated samples. Our study shows that microglial cells release soluble product(s) that induce degeneration of cultured photoreceptor cells. Moreover, the mechanism of microglia-induced photoreceptor cell death may involve apoptosis similar to that observed in animals with retinal degeneration.",
keywords = "Apoptosis, Lactate dehydrogenase, Microglia, Muller cell, Necrosis, Nick-end labeling, Photoreceptor cell, Retinal degeneration",
author = "Roque, {Rouel S.} and Rosales, {Antonio Armando Aviles} and Liu Jingjing and Neeraj Agarwal and Al-Ubaidi, {Muayyad R.}",
year = "1999",
month = "7",
day = "31",
doi = "10.1016/S0006-8993(99)01625-X",
language = "English",
volume = "836",
pages = "110--119",
journal = "Brain Research",
issn = "0006-8993",
publisher = "Elsevier",
number = "1-2",

}

Retina-derived microglial cells induce photoreceptor cell death in vitro. / Roque, Rouel S.; Rosales, Antonio Armando Aviles; Jingjing, Liu; Agarwal, Neeraj; Al-Ubaidi, Muayyad R.

In: Brain Research, Vol. 836, No. 1-2, 31.07.1999, p. 110-119.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Retina-derived microglial cells induce photoreceptor cell death in vitro

AU - Roque, Rouel S.

AU - Rosales, Antonio Armando Aviles

AU - Jingjing, Liu

AU - Agarwal, Neeraj

AU - Al-Ubaidi, Muayyad R.

PY - 1999/7/31

Y1 - 1999/7/31

N2 - In animals with retinal degeneration, the presence of activated microglial cells in the outer retina during the early stages of injury suggests that they may be involved in the ensuing photoreceptor cell death. In the following study, we investigated the effects of rat retina-derived microglial cells on a photoreceptor cell line (661w) using cell culture techniques. The difficulty of obtaining pure populations of photoreceptor cells necessitated our use of the 661w photoreceptor cells generated from retinas of transgenic mice. 661w cells were incubated for 24-48 h in basal medium or basal medium conditioned by activated microglial cells (MGCM) or Muller cells (MCCM), and tested for cell death using lactate dehydrogenase (LDH) assay. The induction of apoptosis in the 661w cells by MGCM was investigated using Terminal deoxynucleotidyl Transferase (TdT)-mediated dUTP nick-end labeling (TUNEL) and DNA laddering. Treatment of 661w cells with MGCM for 48 h resulted in ~ 73% of cells dead as compared with 19-20% of cells grown in either basal medium or MCCM. Serum supplementation or pretreatment with heat did not abolish the cytotoxicity of MGCM. More TUNEL- positive cells were observed in MGCM-treated cultures as compared with those in basal medium. Bands in multiples of ~ 180 bp formed DNA ladders in MGCM- treated but not in basal medium-treated samples. Our study shows that microglial cells release soluble product(s) that induce degeneration of cultured photoreceptor cells. Moreover, the mechanism of microglia-induced photoreceptor cell death may involve apoptosis similar to that observed in animals with retinal degeneration.

AB - In animals with retinal degeneration, the presence of activated microglial cells in the outer retina during the early stages of injury suggests that they may be involved in the ensuing photoreceptor cell death. In the following study, we investigated the effects of rat retina-derived microglial cells on a photoreceptor cell line (661w) using cell culture techniques. The difficulty of obtaining pure populations of photoreceptor cells necessitated our use of the 661w photoreceptor cells generated from retinas of transgenic mice. 661w cells were incubated for 24-48 h in basal medium or basal medium conditioned by activated microglial cells (MGCM) or Muller cells (MCCM), and tested for cell death using lactate dehydrogenase (LDH) assay. The induction of apoptosis in the 661w cells by MGCM was investigated using Terminal deoxynucleotidyl Transferase (TdT)-mediated dUTP nick-end labeling (TUNEL) and DNA laddering. Treatment of 661w cells with MGCM for 48 h resulted in ~ 73% of cells dead as compared with 19-20% of cells grown in either basal medium or MCCM. Serum supplementation or pretreatment with heat did not abolish the cytotoxicity of MGCM. More TUNEL- positive cells were observed in MGCM-treated cultures as compared with those in basal medium. Bands in multiples of ~ 180 bp formed DNA ladders in MGCM- treated but not in basal medium-treated samples. Our study shows that microglial cells release soluble product(s) that induce degeneration of cultured photoreceptor cells. Moreover, the mechanism of microglia-induced photoreceptor cell death may involve apoptosis similar to that observed in animals with retinal degeneration.

KW - Apoptosis

KW - Lactate dehydrogenase

KW - Microglia

KW - Muller cell

KW - Necrosis

KW - Nick-end labeling

KW - Photoreceptor cell

KW - Retinal degeneration

UR - http://www.scopus.com/inward/record.url?scp=0033620899&partnerID=8YFLogxK

U2 - 10.1016/S0006-8993(99)01625-X

DO - 10.1016/S0006-8993(99)01625-X

M3 - Article

C2 - 10415410

AN - SCOPUS:0033620899

VL - 836

SP - 110

EP - 119

JO - Brain Research

JF - Brain Research

SN - 0006-8993

IS - 1-2

ER -