May 1995
Volume 36, Issue 6
Free
Articles  |   May 1995
IP3 generation increases rod outer segment phagocytosis by cultured Royal College of Surgeons retinal pigment epithelium.
Author Affiliations
  • C A Heth
    Berman-Gund Laboratory for the Study of Retinal Degenerations, Harvard Medical School, Massachusetts Eye and Ear Infirmary, Boston 02114, USA.
  • P A Marescalchi
    Berman-Gund Laboratory for the Study of Retinal Degenerations, Harvard Medical School, Massachusetts Eye and Ear Infirmary, Boston 02114, USA.
  • L Ye
    Berman-Gund Laboratory for the Study of Retinal Degenerations, Harvard Medical School, Massachusetts Eye and Ear Infirmary, Boston 02114, USA.
Investigative Ophthalmology & Visual Science May 1995, Vol.36, 984-989. doi:
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    • Get Citation

      C A Heth, P A Marescalchi, L Ye; IP3 generation increases rod outer segment phagocytosis by cultured Royal College of Surgeons retinal pigment epithelium.. Invest. Ophthalmol. Vis. Sci. 1995;36(6):984-989.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

PURPOSE: To measure outer segment phagocytosis in cultures of Royal College of Surgeons (RCS) rat retinal pigment epithelium (RPE) that have been treated with carbachol. Carbachol treatment of RCS RPE results in an increase in the second messenger inositol triphosphate, which mimics that observed in normal RPE after interaction with rod outer segments (ROS). METHODS: Cultures of RCS RPE were phagocytically challenged with isolated rat ROS for 2 hours. Carbachol (1 mM) was added to some cultures to stimulate inositol triphosphate synthesis, and incubation continued for 30 minutes at 37 degrees C. Inositol triphosphate concentration was measured by radioreceptor assay. Bound and ingested outer segments were quantified by double immunofluorescent staining. Ingestion of outer segment membranes was confirmed by electron microscopy and immunogold staining. RESULTS: Carbachol treatment was associated with a rapid and temporary increase in inositol triphosphate levels. Royal College of Surgeons rat RPE phagocytically challenged with outer segments and treated with carbachol showed significantly higher ingestion (34%) compared to untreated RCS RPE (9%) (P < 0.05). CONCLUSIONS: Exposure of cultured RCS RPE to carbachol increases the intracellular concentration of inositol triphosphate and enhances phagocytosis of bound ROS. These results support the hypothesis that the phagocytic defect in RCS RPE is related to an abnormality in the generation of inositol triphosphate as a second messenger after outer segment recognition and binding.

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