Expression, subcellular localization, and regulation of sigma receptor in retinal Müller cells

Guoliang Jiang, Barbara Mysona, Ying Dun, Jaya P. Gnana-Prakasam, Navjotsin Pabla, Weiguo Li, Zheng Dong, Vadivel Ganapathy, Sylvia B. Smith

Research output: Contribution to journalArticle

47 Citations (Scopus)

Abstract

PURPOSE. Sigma receptors (σrRs) are nonopioid, nonphencyclidine binding sites with robust neuroprotective properties. Type 1 σrR1 (σTR1) is expressed in brain oligodendrocytes, but its expression and binding capacity have not been analyzed in retinal glial cells. This study examined the expression, subcellular localization, binding activity, and regulation of σR1 in retinal Müller cells. METHODS. Primary mouse Müller cells (MCs) were analyzed by RT-PCR, immunoblotting, and immunocytochemistry for the expression of σ-R1, and data were compared with those of the rat Müller cell line (rMC-1) and the rat ganglion cell line (RGC-5). Confocal microscopy was used to determine the subcellular σR1 location in primary mouse MCs. Membranes prepared from these cells were used for binding assays with [3H]-pentazocine (PTZ). The kinetics of binding, the ability of various σR1 ligands to compete with σR1 binding, and the effects of donated nitric oxide (NO) and reactive oxygen species (ROS) on binding were examined. RESULTS. σR1 is expressed in primary mouse MCs and is localized to the nuclear and endoplasmic reticulum membranes. Binding assays showed that in primary mouse MCs, rMC-1, and RGC-5, the binding of PTZ was saturable. [3H]-PTZ bound with high affinity in RGC-5 and rMC-1 cells, and the binding was similarly robust in primary mouse MCs. Competition studies showed marked inhibition of [3H]-PTZ binding in the presence of σR1-specific ligands. Incubation of cells with NO and ROS donors markedly increased σR1 binding activity. CONCLUSIONS. MCS express σR1 and demonstrate robust σR1 binding activity, which is inhibited by σR1 ligands and is stimulated during oxidative stress. The potential of Müller cells to bind σR1 ligands may prove beneficial in retinal degenerative diseases such as diabetic retinopathy.

Original languageEnglish (US)
Pages (from-to)5576-5582
Number of pages7
JournalInvestigative Ophthalmology and Visual Science
Volume47
Issue number12
DOIs
StatePublished - Dec 1 2006

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sigma Receptors
Pentazocine
Ligands
Reactive Oxygen Species
Nitric Oxide
Cell Line
Retinal Diseases
Oligodendroglia
Diabetic Retinopathy
Immunoblotting
Confocal Microscopy
Neuroglia
Ganglia
Endoplasmic Reticulum
Oxidative Stress
Immunohistochemistry
Binding Sites
Cell Membrane
Polymerase Chain Reaction

ASJC Scopus subject areas

  • Ophthalmology
  • Sensory Systems
  • Cellular and Molecular Neuroscience

Cite this

Expression, subcellular localization, and regulation of sigma receptor in retinal Müller cells. / Jiang, Guoliang; Mysona, Barbara; Dun, Ying; Gnana-Prakasam, Jaya P.; Pabla, Navjotsin; Li, Weiguo; Dong, Zheng; Ganapathy, Vadivel; Smith, Sylvia B.

In: Investigative Ophthalmology and Visual Science, Vol. 47, No. 12, 01.12.2006, p. 5576-5582.

Research output: Contribution to journalArticle

Jiang, Guoliang ; Mysona, Barbara ; Dun, Ying ; Gnana-Prakasam, Jaya P. ; Pabla, Navjotsin ; Li, Weiguo ; Dong, Zheng ; Ganapathy, Vadivel ; Smith, Sylvia B. / Expression, subcellular localization, and regulation of sigma receptor in retinal Müller cells. In: Investigative Ophthalmology and Visual Science. 2006 ; Vol. 47, No. 12. pp. 5576-5582.
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abstract = "PURPOSE. Sigma receptors (σrRs) are nonopioid, nonphencyclidine binding sites with robust neuroprotective properties. Type 1 σrR1 (σTR1) is expressed in brain oligodendrocytes, but its expression and binding capacity have not been analyzed in retinal glial cells. This study examined the expression, subcellular localization, binding activity, and regulation of σR1 in retinal M{\"u}ller cells. METHODS. Primary mouse M{\"u}ller cells (MCs) were analyzed by RT-PCR, immunoblotting, and immunocytochemistry for the expression of σ-R1, and data were compared with those of the rat M{\"u}ller cell line (rMC-1) and the rat ganglion cell line (RGC-5). Confocal microscopy was used to determine the subcellular σR1 location in primary mouse MCs. Membranes prepared from these cells were used for binding assays with [3H]-pentazocine (PTZ). The kinetics of binding, the ability of various σR1 ligands to compete with σR1 binding, and the effects of donated nitric oxide (NO) and reactive oxygen species (ROS) on binding were examined. RESULTS. σR1 is expressed in primary mouse MCs and is localized to the nuclear and endoplasmic reticulum membranes. Binding assays showed that in primary mouse MCs, rMC-1, and RGC-5, the binding of PTZ was saturable. [3H]-PTZ bound with high affinity in RGC-5 and rMC-1 cells, and the binding was similarly robust in primary mouse MCs. Competition studies showed marked inhibition of [3H]-PTZ binding in the presence of σR1-specific ligands. Incubation of cells with NO and ROS donors markedly increased σR1 binding activity. CONCLUSIONS. MCS express σR1 and demonstrate robust σR1 binding activity, which is inhibited by σR1 ligands and is stimulated during oxidative stress. The potential of M{\"u}ller cells to bind σR1 ligands may prove beneficial in retinal degenerative diseases such as diabetic retinopathy.",
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T1 - Expression, subcellular localization, and regulation of sigma receptor in retinal Müller cells

AU - Jiang, Guoliang

AU - Mysona, Barbara

AU - Dun, Ying

AU - Gnana-Prakasam, Jaya P.

AU - Pabla, Navjotsin

AU - Li, Weiguo

AU - Dong, Zheng

AU - Ganapathy, Vadivel

AU - Smith, Sylvia B.

PY - 2006/12/1

Y1 - 2006/12/1

N2 - PURPOSE. Sigma receptors (σrRs) are nonopioid, nonphencyclidine binding sites with robust neuroprotective properties. Type 1 σrR1 (σTR1) is expressed in brain oligodendrocytes, but its expression and binding capacity have not been analyzed in retinal glial cells. This study examined the expression, subcellular localization, binding activity, and regulation of σR1 in retinal Müller cells. METHODS. Primary mouse Müller cells (MCs) were analyzed by RT-PCR, immunoblotting, and immunocytochemistry for the expression of σ-R1, and data were compared with those of the rat Müller cell line (rMC-1) and the rat ganglion cell line (RGC-5). Confocal microscopy was used to determine the subcellular σR1 location in primary mouse MCs. Membranes prepared from these cells were used for binding assays with [3H]-pentazocine (PTZ). The kinetics of binding, the ability of various σR1 ligands to compete with σR1 binding, and the effects of donated nitric oxide (NO) and reactive oxygen species (ROS) on binding were examined. RESULTS. σR1 is expressed in primary mouse MCs and is localized to the nuclear and endoplasmic reticulum membranes. Binding assays showed that in primary mouse MCs, rMC-1, and RGC-5, the binding of PTZ was saturable. [3H]-PTZ bound with high affinity in RGC-5 and rMC-1 cells, and the binding was similarly robust in primary mouse MCs. Competition studies showed marked inhibition of [3H]-PTZ binding in the presence of σR1-specific ligands. Incubation of cells with NO and ROS donors markedly increased σR1 binding activity. CONCLUSIONS. MCS express σR1 and demonstrate robust σR1 binding activity, which is inhibited by σR1 ligands and is stimulated during oxidative stress. The potential of Müller cells to bind σR1 ligands may prove beneficial in retinal degenerative diseases such as diabetic retinopathy.

AB - PURPOSE. Sigma receptors (σrRs) are nonopioid, nonphencyclidine binding sites with robust neuroprotective properties. Type 1 σrR1 (σTR1) is expressed in brain oligodendrocytes, but its expression and binding capacity have not been analyzed in retinal glial cells. This study examined the expression, subcellular localization, binding activity, and regulation of σR1 in retinal Müller cells. METHODS. Primary mouse Müller cells (MCs) were analyzed by RT-PCR, immunoblotting, and immunocytochemistry for the expression of σ-R1, and data were compared with those of the rat Müller cell line (rMC-1) and the rat ganglion cell line (RGC-5). Confocal microscopy was used to determine the subcellular σR1 location in primary mouse MCs. Membranes prepared from these cells were used for binding assays with [3H]-pentazocine (PTZ). The kinetics of binding, the ability of various σR1 ligands to compete with σR1 binding, and the effects of donated nitric oxide (NO) and reactive oxygen species (ROS) on binding were examined. RESULTS. σR1 is expressed in primary mouse MCs and is localized to the nuclear and endoplasmic reticulum membranes. Binding assays showed that in primary mouse MCs, rMC-1, and RGC-5, the binding of PTZ was saturable. [3H]-PTZ bound with high affinity in RGC-5 and rMC-1 cells, and the binding was similarly robust in primary mouse MCs. Competition studies showed marked inhibition of [3H]-PTZ binding in the presence of σR1-specific ligands. Incubation of cells with NO and ROS donors markedly increased σR1 binding activity. CONCLUSIONS. MCS express σR1 and demonstrate robust σR1 binding activity, which is inhibited by σR1 ligands and is stimulated during oxidative stress. The potential of Müller cells to bind σR1 ligands may prove beneficial in retinal degenerative diseases such as diabetic retinopathy.

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