Retina-derived microglial cells induce photoreceptor cell death in vitro.
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 Müller 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 approximately 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 approximately 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.
Department of Anatomy and Cell Biology, University of North Texas Health Science Center, 3500 Camp Bowie Blvd., Fort Worth, TX 76107-2699, USA. rroque@molly.hsc.unt.edu
R S
RS
Roque
A A
AA
Rosales
L
L
Jingjing
N
N
Agarwal
M R
MR
Al-Ubaidi
D000818:Animals; D000952:Antigens, Polyomavirus Transforming / Q000032:analysis; D017209:Apoptosis / Q000502:physiology; D015415:Biomarkers; D016923:Cell Death / Q000502:physiology*; D002455:Cell Division / Q000502:physiology; D002460:Cell Line; D020287:In Situ Nick-End Labeling; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D051379:Mice; D008822:Mice, Transgenic; D017628:Microglia / Q000166:cytology*; D020419:Photoreceptor Cells, Vertebrate / Q000166:cytology*; D051381:Rats; D012160:Retina / Q000166:cytology*
D000952:Antigens, Polyomavirus Transforming; D015415:Biomarkers; D007770:L-Lactate Dehydrogenase