Human optic nerve head astrocytes as a target for endothelin-1

Ganesh Prasanna, Raghu Krishnamoorthy, Abbot Clark, Robert J. Wordinger, Thomas Yorio

Research output: Contribution to journalArticleResearchpeer-review

101 Citations (Scopus)

Abstract

PURPOSE. To determine whether human optic nerve head astrocytes (hONAs) are target cells for the actions of endothelin (ET)-I, a potent vasoactive peptide, by causing astrocyte proliferation, as occurs in glaucomatous optic nerve heads. ET-1 levels are elevated in glaucomatous eyes, and administration of ET-1 to the retina causes glial activation and optic nerve damage in animal models in a manner similar to that observed in glaucoma. METHODS. Well-characterized hONAs were used in this study. Cell proliferation of hONAs was assessed, after ET-1 treatment under serum-free culture conditions, with both a formazan assay and [3H] thymidine uptake. ET receptor involvement for cell proliferation was determined with BQ788 (an ETB antagonist), BQ610 (an ETA antagonist), PD142893 (an ETA/B mixed antagonist), and sarafotoxin 6C (S6C; an ETB agonist). ET-1-induced intracellular calcium ([Ca2+]i) in hONAs was measured by fura-2 imaging. RT-PCR was used to determine whether hONAs express mRNA for preproET-1, ETA, and ETB receptors. RESULTS. ET-1 (10 and 100 nM) caused a time-dependent proliferation of hONAs, which was completely blocked by PD142893, as detected by two different cell proliferation assays. The effects of ET-1 were blocked by BQ788 and were also mimicked by S6C, indicative of the involvement of ETB receptor activation. ET-1-induced elevation in [Ca2+]i, and cell proliferation were both blocked completely by the ETA antagonist BQ610, suggesting ETA receptor involvement. The hONAs expressed mRNA for ETA and ETB, receptors as well as preproET-1, suggesting that these cells may also be a source for ET-1 in the optic nerve head. CONCLUSIONS. ET-1 induces astroglial proliferation in cultured human optic nerve head astrocytes through ETA/B receptor activation. This is similar to the proliferation of ET-1 in brain astrocytes. These findings suggest that ET-1, which is elevated in glaucoma, could cause proliferation of ONAs in the optic nerve head.

Original languageEnglish
Pages (from-to)2704-2713
Number of pages10
JournalInvestigative Ophthalmology and Visual Science
Volume43
Issue number8
StatePublished - 5 Aug 2002

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Optic Disk
Endothelin-1
Astrocytes
Cell Proliferation
Glaucoma
Formazans
Endothelin Receptors
Messenger RNA
Endothelins
Optic Nerve
Neuroglia
Thymidine
Retina
Animal Models
Calcium
Polymerase Chain Reaction
Peptides

Cite this

@article{e962a3931cf242f0864957cc3b2623f5,
title = "Human optic nerve head astrocytes as a target for endothelin-1",
abstract = "PURPOSE. To determine whether human optic nerve head astrocytes (hONAs) are target cells for the actions of endothelin (ET)-I, a potent vasoactive peptide, by causing astrocyte proliferation, as occurs in glaucomatous optic nerve heads. ET-1 levels are elevated in glaucomatous eyes, and administration of ET-1 to the retina causes glial activation and optic nerve damage in animal models in a manner similar to that observed in glaucoma. METHODS. Well-characterized hONAs were used in this study. Cell proliferation of hONAs was assessed, after ET-1 treatment under serum-free culture conditions, with both a formazan assay and [3H] thymidine uptake. ET receptor involvement for cell proliferation was determined with BQ788 (an ETB antagonist), BQ610 (an ETA antagonist), PD142893 (an ETA/B mixed antagonist), and sarafotoxin 6C (S6C; an ETB agonist). ET-1-induced intracellular calcium ([Ca2+]i) in hONAs was measured by fura-2 imaging. RT-PCR was used to determine whether hONAs express mRNA for preproET-1, ETA, and ETB receptors. RESULTS. ET-1 (10 and 100 nM) caused a time-dependent proliferation of hONAs, which was completely blocked by PD142893, as detected by two different cell proliferation assays. The effects of ET-1 were blocked by BQ788 and were also mimicked by S6C, indicative of the involvement of ETB receptor activation. ET-1-induced elevation in [Ca2+]i, and cell proliferation were both blocked completely by the ETA antagonist BQ610, suggesting ETA receptor involvement. The hONAs expressed mRNA for ETA and ETB, receptors as well as preproET-1, suggesting that these cells may also be a source for ET-1 in the optic nerve head. CONCLUSIONS. ET-1 induces astroglial proliferation in cultured human optic nerve head astrocytes through ETA/B receptor activation. This is similar to the proliferation of ET-1 in brain astrocytes. These findings suggest that ET-1, which is elevated in glaucoma, could cause proliferation of ONAs in the optic nerve head.",
author = "Ganesh Prasanna and Raghu Krishnamoorthy and Abbot Clark and Wordinger, {Robert J.} and Thomas Yorio",
year = "2002",
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volume = "43",
pages = "2704--2713",
journal = "Investigative Ophthalmology and Visual Science",
issn = "0146-0404",
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number = "8",

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Human optic nerve head astrocytes as a target for endothelin-1. / Prasanna, Ganesh; Krishnamoorthy, Raghu; Clark, Abbot; Wordinger, Robert J.; Yorio, Thomas.

In: Investigative Ophthalmology and Visual Science, Vol. 43, No. 8, 05.08.2002, p. 2704-2713.

Research output: Contribution to journalArticleResearchpeer-review

TY - JOUR

T1 - Human optic nerve head astrocytes as a target for endothelin-1

AU - Prasanna, Ganesh

AU - Krishnamoorthy, Raghu

AU - Clark, Abbot

AU - Wordinger, Robert J.

AU - Yorio, Thomas

PY - 2002/8/5

Y1 - 2002/8/5

N2 - PURPOSE. To determine whether human optic nerve head astrocytes (hONAs) are target cells for the actions of endothelin (ET)-I, a potent vasoactive peptide, by causing astrocyte proliferation, as occurs in glaucomatous optic nerve heads. ET-1 levels are elevated in glaucomatous eyes, and administration of ET-1 to the retina causes glial activation and optic nerve damage in animal models in a manner similar to that observed in glaucoma. METHODS. Well-characterized hONAs were used in this study. Cell proliferation of hONAs was assessed, after ET-1 treatment under serum-free culture conditions, with both a formazan assay and [3H] thymidine uptake. ET receptor involvement for cell proliferation was determined with BQ788 (an ETB antagonist), BQ610 (an ETA antagonist), PD142893 (an ETA/B mixed antagonist), and sarafotoxin 6C (S6C; an ETB agonist). ET-1-induced intracellular calcium ([Ca2+]i) in hONAs was measured by fura-2 imaging. RT-PCR was used to determine whether hONAs express mRNA for preproET-1, ETA, and ETB receptors. RESULTS. ET-1 (10 and 100 nM) caused a time-dependent proliferation of hONAs, which was completely blocked by PD142893, as detected by two different cell proliferation assays. The effects of ET-1 were blocked by BQ788 and were also mimicked by S6C, indicative of the involvement of ETB receptor activation. ET-1-induced elevation in [Ca2+]i, and cell proliferation were both blocked completely by the ETA antagonist BQ610, suggesting ETA receptor involvement. The hONAs expressed mRNA for ETA and ETB, receptors as well as preproET-1, suggesting that these cells may also be a source for ET-1 in the optic nerve head. CONCLUSIONS. ET-1 induces astroglial proliferation in cultured human optic nerve head astrocytes through ETA/B receptor activation. This is similar to the proliferation of ET-1 in brain astrocytes. These findings suggest that ET-1, which is elevated in glaucoma, could cause proliferation of ONAs in the optic nerve head.

AB - PURPOSE. To determine whether human optic nerve head astrocytes (hONAs) are target cells for the actions of endothelin (ET)-I, a potent vasoactive peptide, by causing astrocyte proliferation, as occurs in glaucomatous optic nerve heads. ET-1 levels are elevated in glaucomatous eyes, and administration of ET-1 to the retina causes glial activation and optic nerve damage in animal models in a manner similar to that observed in glaucoma. METHODS. Well-characterized hONAs were used in this study. Cell proliferation of hONAs was assessed, after ET-1 treatment under serum-free culture conditions, with both a formazan assay and [3H] thymidine uptake. ET receptor involvement for cell proliferation was determined with BQ788 (an ETB antagonist), BQ610 (an ETA antagonist), PD142893 (an ETA/B mixed antagonist), and sarafotoxin 6C (S6C; an ETB agonist). ET-1-induced intracellular calcium ([Ca2+]i) in hONAs was measured by fura-2 imaging. RT-PCR was used to determine whether hONAs express mRNA for preproET-1, ETA, and ETB receptors. RESULTS. ET-1 (10 and 100 nM) caused a time-dependent proliferation of hONAs, which was completely blocked by PD142893, as detected by two different cell proliferation assays. The effects of ET-1 were blocked by BQ788 and were also mimicked by S6C, indicative of the involvement of ETB receptor activation. ET-1-induced elevation in [Ca2+]i, and cell proliferation were both blocked completely by the ETA antagonist BQ610, suggesting ETA receptor involvement. The hONAs expressed mRNA for ETA and ETB, receptors as well as preproET-1, suggesting that these cells may also be a source for ET-1 in the optic nerve head. CONCLUSIONS. ET-1 induces astroglial proliferation in cultured human optic nerve head astrocytes through ETA/B receptor activation. This is similar to the proliferation of ET-1 in brain astrocytes. These findings suggest that ET-1, which is elevated in glaucoma, could cause proliferation of ONAs in the optic nerve head.

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M3 - Article

VL - 43

SP - 2704

EP - 2713

JO - Investigative Ophthalmology and Visual Science

JF - Investigative Ophthalmology and Visual Science

SN - 0146-0404

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