Analysis of sigma receptor (σR1) expression in retinal ganglion cells cultured under hyperglycemic conditions and in diabetic mice

M. Shamsul Ola, Pamela Moore Martin, Dennis Maddox, Amira El-Sherbeny, Wei Huang, Penny Roon, Neeraj Agarwal, Vadivel Ganapathy, Sylvia B Smith

Research output: Contribution to journalArticle

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Abstract

The type 1 sigma receptor (σR1) is a nonopiate and nonphencyclidine binding site that has numerous pharmacological and physiological functions. In some studies, agonists for σR1 have been shown to afford neuroprotection against overstimulation of the NMDA receptor. σR1 expression has been demonstrated recently in retinal ganglion cells (RGC). RGCs undergo apoptosis early in diabetic retinopathy via NMDA receptor overstimulation. In the present study we asked whether RGCs cultured under hyperglycemic conditions and RGCs of diabetic mice continue to express σR1. RGCs were cultured 48 h in RPMI medium containing either 45 mM glucose or 11 mM glucose plus 34 mM mannitol (osmolar control). C57BL/6 mice were made diabetic using streptozotocin. The retina was dissected from normal and streptozotocin-induced diabetic mice 3, 6 and 12 weeks post-onset of diabetes. σR1 was analyzed in cells using semiquantitative RT-PCR and in tissues by semiquantitative RT-PCR, in situ hybridization, Western blot analysis and immunolocalization. The RT-PCR analysis of cultured RGCs showed that σR1 mRNA is expressed under hyperglycemic conditions at levels similar to control cells. Similarly, analysis of retinas of diabetic mice showed no difference in levels of mRNA encoding σR1 compared to retinas of control mice. In situ hybridization analysis showed that expression patterns of σR1 mRNA in the ganglion cell layer were similar between diabetic and control mice. Western blot analysis suggested that levels of σR1 in retina were similar between diabetic and control retinas. Immunohistochemical analysis of σR1 showed a similar pattern of σR1 protein expression between control and diabetic retina. These studies demonstrate that σR1 is expressed under hyperglycemic conditions in vitro and in vivo.

Original languageEnglish (US)
Pages (from-to)97-107
Number of pages11
JournalMolecular Brain Research
Volume107
Issue number2
DOIs
StatePublished - Nov 15 2002

Fingerprint

sigma Receptors
Retinal Ganglion Cells
Retina
Streptozocin
N-Methyl-D-Aspartate Receptors
Polymerase Chain Reaction
Messenger RNA
In Situ Hybridization
Western Blotting
Glucose
Mannitol
Diabetic Retinopathy
Inbred C57BL Mouse
Ganglia
Binding Sites
Pharmacology
Apoptosis

Keywords

  • Diabetes
  • Diabetic retinopathy
  • Retina
  • Retinal ganglion cells
  • Sigma receptor

ASJC Scopus subject areas

  • Molecular Biology
  • Cellular and Molecular Neuroscience

Cite this

Analysis of sigma receptor (σR1) expression in retinal ganglion cells cultured under hyperglycemic conditions and in diabetic mice. / Shamsul Ola, M.; Martin, Pamela Moore; Maddox, Dennis; El-Sherbeny, Amira; Huang, Wei; Roon, Penny; Agarwal, Neeraj; Ganapathy, Vadivel; Smith, Sylvia B.

In: Molecular Brain Research, Vol. 107, No. 2, 15.11.2002, p. 97-107.

Research output: Contribution to journalArticle

Shamsul Ola, M. ; Martin, Pamela Moore ; Maddox, Dennis ; El-Sherbeny, Amira ; Huang, Wei ; Roon, Penny ; Agarwal, Neeraj ; Ganapathy, Vadivel ; Smith, Sylvia B. / Analysis of sigma receptor (σR1) expression in retinal ganglion cells cultured under hyperglycemic conditions and in diabetic mice. In: Molecular Brain Research. 2002 ; Vol. 107, No. 2. pp. 97-107.
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