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Serum deprivation can suppress receptor-mediated calcium signaling in pterygial-derived fibroblasts.
Invest Ophthalmol Vis Sci. 2013 Jul 10; 54(7):4563-70.IO

Abstract

PURPOSE

Pterygium is characterized as invasive, proliferative fibrovascular altered conjunctival tissue. The extensive vascular network is likely to significantly contribute to the progression of the disease. In the present study, we investigated the effects of reduced serum (to mimic a suppressed blood supply) on cell signaling events and the functional role of the calcium store in cultured, pterygial-derived fibroblasts.

METHODS

Pure fibroblast cultures were established from cell outgrowths of pterygial tissue. Growth and migration of pterygial-derived fibroblast was evaluated using a patch growth assay, MTS assay, and a scratch wound assay. Intracellular calcium levels were determined using Fura-2 detection in response to ligand stimulation using a 96-well plate format.

RESULTS

A progressive increase in serum concentration resulted in promotion of pterygial cell growth detected using the MTS assay. A significant increase in intracellular calcium level was observed in response to histamine (10 and 100 μM), ATP (10 and 100 μM), acetylcholine (10 and 100 μM) and epidermal growth factor (10 ng/mL) in serum-maintained cells. However, no significant changes were observed when cells were maintained in serum-free medium. Thapsigargin (1 μM), a Ca-ATPase inhibitor, induced a significantly greater increase in intracellular calcium level in the serum-maintained group relative to serum-starved cells. In both cases, elevation of intracellular calcium was reduced when calcium-free bathing medium was used. Preincubation of cells with 1 μM thapsigargin ablated ligand-induced calcium responses. Disruption of calcium signaling through thapsigargin treatment significantly perturbed cell growth and migration.

CONCLUSIONS

Receptor-induced calcium signaling activity is suppressed in pterygial-derived fibroblasts in response to serum deprivation. This correlates with reduced growth rates and a depleted endoplasmic reticulum calcium store. The store plays a key role in cell growth and migration of pterygial-derived fibroblasts. Therefore, the strategic reduction of the vascular network in pterygium will affect the calcium store level and in turn affect functional responses associated with pterygia.

Authors+Show Affiliations

School of Biological Sciences, University of East Anglia, Norwich, United Kingdom.No affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

23761089

Citation

Fang, Chunlai, et al. "Serum Deprivation Can Suppress Receptor-mediated Calcium Signaling in Pterygial-derived Fibroblasts." Investigative Ophthalmology & Visual Science, vol. 54, no. 7, 2013, pp. 4563-70.
Fang C, Illingworth CD, Qian L, et al. Serum deprivation can suppress receptor-mediated calcium signaling in pterygial-derived fibroblasts. Invest Ophthalmol Vis Sci. 2013;54(7):4563-70.
Fang, C., Illingworth, C. D., Qian, L., & Wormstone, I. M. (2013). Serum deprivation can suppress receptor-mediated calcium signaling in pterygial-derived fibroblasts. Investigative Ophthalmology & Visual Science, 54(7), 4563-70. https://doi.org/10.1167/iovs.13-11604
Fang C, et al. Serum Deprivation Can Suppress Receptor-mediated Calcium Signaling in Pterygial-derived Fibroblasts. Invest Ophthalmol Vis Sci. 2013 Jul 10;54(7):4563-70. PubMed PMID: 23761089.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Serum deprivation can suppress receptor-mediated calcium signaling in pterygial-derived fibroblasts. AU - Fang,Chunlai, AU - Illingworth,Christopher D, AU - Qian,Limin, AU - Wormstone,I Michael, Y1 - 2013/07/10/ PY - 2013/6/14/entrez PY - 2013/6/14/pubmed PY - 2013/9/6/medline KW - angiogenesis KW - calcium KW - fibroblasts KW - pterygia KW - serum deprivation KW - signaling SP - 4563 EP - 70 JF - Investigative ophthalmology & visual science JO - Invest Ophthalmol Vis Sci VL - 54 IS - 7 N2 - PURPOSE: Pterygium is characterized as invasive, proliferative fibrovascular altered conjunctival tissue. The extensive vascular network is likely to significantly contribute to the progression of the disease. In the present study, we investigated the effects of reduced serum (to mimic a suppressed blood supply) on cell signaling events and the functional role of the calcium store in cultured, pterygial-derived fibroblasts. METHODS: Pure fibroblast cultures were established from cell outgrowths of pterygial tissue. Growth and migration of pterygial-derived fibroblast was evaluated using a patch growth assay, MTS assay, and a scratch wound assay. Intracellular calcium levels were determined using Fura-2 detection in response to ligand stimulation using a 96-well plate format. RESULTS: A progressive increase in serum concentration resulted in promotion of pterygial cell growth detected using the MTS assay. A significant increase in intracellular calcium level was observed in response to histamine (10 and 100 μM), ATP (10 and 100 μM), acetylcholine (10 and 100 μM) and epidermal growth factor (10 ng/mL) in serum-maintained cells. However, no significant changes were observed when cells were maintained in serum-free medium. Thapsigargin (1 μM), a Ca-ATPase inhibitor, induced a significantly greater increase in intracellular calcium level in the serum-maintained group relative to serum-starved cells. In both cases, elevation of intracellular calcium was reduced when calcium-free bathing medium was used. Preincubation of cells with 1 μM thapsigargin ablated ligand-induced calcium responses. Disruption of calcium signaling through thapsigargin treatment significantly perturbed cell growth and migration. CONCLUSIONS: Receptor-induced calcium signaling activity is suppressed in pterygial-derived fibroblasts in response to serum deprivation. This correlates with reduced growth rates and a depleted endoplasmic reticulum calcium store. The store plays a key role in cell growth and migration of pterygial-derived fibroblasts. Therefore, the strategic reduction of the vascular network in pterygium will affect the calcium store level and in turn affect functional responses associated with pterygia. SN - 1552-5783 UR - https://www.unboundmedicine.com/medline/citation/23761089/Serum_deprivation_can_suppress_receptor-mediated_calcium_signaling_in_pterygial-derived_fibroblasts L2 - https://iovs.arvojournals.org/article.aspx?doi=10.1167/iovs.13-11604 DB - PRIME DP - Unbound Medicine ER -