Trabecular meshwork bradykinin receptors: MRNA levels, immunohistochemical visualization, signaling processes pharmacology, and linkage to IOP reduction

Najam A. Sharif, Parvaneh Katoli, Curtis R. Kelly, Linya Li, Shouxi Xu, Yu Wang, Laura Klekar, David Earnest, Shenouda Yacoub, Gwenette Hamilton, Nasreen Jacobson, Allan R. Shepard, Dorette Zoe Ellis

Research output: Contribution to journalArticle

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Abstract

Purpose: To localize mRNA and protein of bradykinin (BK) receptors, BK precursor polypeptide (kininogen) mRNA, and to study functional biochemical pharmacology of the signal transduction processes mediated by B 2-receptors in isolated human trabecular meshwork (h-TM) cells. Intraocular pressure (IOP) lowering effects of 2 kinins were also investigated. Methods: Previously documented procedures were utilized throughout these studies. Results: Kinninogen mRNA was most abundant in TM, ciliary body (CB), and optic nerve head and appeared elevated in glaucomatous h-TM tissue. High levels of B2-receptor mRNA were found in the sclera, iris, TM, and CB. B2-receptor subtype protein was localized in cells of the monkey and h-TM, and the treatment of isolated h-TM cells with transforming growth factor-β2 (5 ng/mL) caused significant (P<0.04) downregulation of B 2-receptor mRNA. In isolated primary h-TM cells, BK (EC 50=0.8±0.2 nM; n=19) and Met-Lys-BK (EC50=6. 5±1.5 nM) mobilized intracellular Ca2+ and induced the release of prostaglandins (PGs) that was blocked by 2 B2-receptor antagonists [HOE-140; (S)-WIN-64338]. The cyclooxygenase inhibitor, bromfenac, abolished BK-induced PGs production. BK concentration dependently increased cell impedance, and it significantly (P<0.05) decreased h-TM cell volume in vitro. Intravitreal (ivt) administration of BK (50 μg), but not a B 1-agonist (Sar-[D-Phe9]-Des-Arg9-BK; also at 50 μg), efficaciously lowered IOP (22.9% to 37% from baseline) of Dutch-Belted rabbits that naturally have high IOPs (27-28 mmHg). Conclusions: BK activates multiple signal transduction pathways in h-TM cells via B2-receptors that also mediate IOP reduction as observed in rabbits following ivt administration of BK. These ocular hypotensive effects of BK may be physiologically important and suggest a novel therapeutic potential of BK-related B2-agonists.

Original languageEnglish
Pages (from-to)21-34
Number of pages14
JournalJournal of Ocular Pharmacology and Therapeutics
Volume30
Issue number1
DOIs
StatePublished - 1 Feb 2014

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Bradykinin Receptors
Trabecular Meshwork
Bradykinin
Intraocular Pressure
Pharmacology
Messenger RNA
Ciliary Body
Prostaglandins
Signal Transduction
Kininogens
Rabbits
Kinins
Sclera
Cyclooxygenase Inhibitors
Optic Disk
Transforming Growth Factors
Iris
Electric Impedance
Cell Size
Haplorhini

Cite this

Sharif, Najam A. ; Katoli, Parvaneh ; Kelly, Curtis R. ; Li, Linya ; Xu, Shouxi ; Wang, Yu ; Klekar, Laura ; Earnest, David ; Yacoub, Shenouda ; Hamilton, Gwenette ; Jacobson, Nasreen ; Shepard, Allan R. ; Ellis, Dorette Zoe. / Trabecular meshwork bradykinin receptors : MRNA levels, immunohistochemical visualization, signaling processes pharmacology, and linkage to IOP reduction. In: Journal of Ocular Pharmacology and Therapeutics. 2014 ; Vol. 30, No. 1. pp. 21-34.
@article{d45fac4669f242718da1c3bb3b53d4a6,
title = "Trabecular meshwork bradykinin receptors: MRNA levels, immunohistochemical visualization, signaling processes pharmacology, and linkage to IOP reduction",
abstract = "Purpose: To localize mRNA and protein of bradykinin (BK) receptors, BK precursor polypeptide (kininogen) mRNA, and to study functional biochemical pharmacology of the signal transduction processes mediated by B 2-receptors in isolated human trabecular meshwork (h-TM) cells. Intraocular pressure (IOP) lowering effects of 2 kinins were also investigated. Methods: Previously documented procedures were utilized throughout these studies. Results: Kinninogen mRNA was most abundant in TM, ciliary body (CB), and optic nerve head and appeared elevated in glaucomatous h-TM tissue. High levels of B2-receptor mRNA were found in the sclera, iris, TM, and CB. B2-receptor subtype protein was localized in cells of the monkey and h-TM, and the treatment of isolated h-TM cells with transforming growth factor-β2 (5 ng/mL) caused significant (P<0.04) downregulation of B 2-receptor mRNA. In isolated primary h-TM cells, BK (EC 50=0.8±0.2 nM; n=19) and Met-Lys-BK (EC50=6. 5±1.5 nM) mobilized intracellular Ca2+ and induced the release of prostaglandins (PGs) that was blocked by 2 B2-receptor antagonists [HOE-140; (S)-WIN-64338]. The cyclooxygenase inhibitor, bromfenac, abolished BK-induced PGs production. BK concentration dependently increased cell impedance, and it significantly (P<0.05) decreased h-TM cell volume in vitro. Intravitreal (ivt) administration of BK (50 μg), but not a B 1-agonist (Sar-[D-Phe9]-Des-Arg9-BK; also at 50 μg), efficaciously lowered IOP (22.9{\%} to 37{\%} from baseline) of Dutch-Belted rabbits that naturally have high IOPs (27-28 mmHg). Conclusions: BK activates multiple signal transduction pathways in h-TM cells via B2-receptors that also mediate IOP reduction as observed in rabbits following ivt administration of BK. These ocular hypotensive effects of BK may be physiologically important and suggest a novel therapeutic potential of BK-related B2-agonists.",
author = "Sharif, {Najam A.} and Parvaneh Katoli and Kelly, {Curtis R.} and Linya Li and Shouxi Xu and Yu Wang and Laura Klekar and David Earnest and Shenouda Yacoub and Gwenette Hamilton and Nasreen Jacobson and Shepard, {Allan R.} and Ellis, {Dorette Zoe}",
year = "2014",
month = "2",
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doi = "10.1089/jop.2013.0105",
language = "English",
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pages = "21--34",
journal = "Journal of Ocular Pharmacology and Therapeutics",
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publisher = "Mary Ann Liebert Inc.",
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}

Sharif, NA, Katoli, P, Kelly, CR, Li, L, Xu, S, Wang, Y, Klekar, L, Earnest, D, Yacoub, S, Hamilton, G, Jacobson, N, Shepard, AR & Ellis, DZ 2014, 'Trabecular meshwork bradykinin receptors: MRNA levels, immunohistochemical visualization, signaling processes pharmacology, and linkage to IOP reduction', Journal of Ocular Pharmacology and Therapeutics, vol. 30, no. 1, pp. 21-34. https://doi.org/10.1089/jop.2013.0105

Trabecular meshwork bradykinin receptors : MRNA levels, immunohistochemical visualization, signaling processes pharmacology, and linkage to IOP reduction. / Sharif, Najam A.; Katoli, Parvaneh; Kelly, Curtis R.; Li, Linya; Xu, Shouxi; Wang, Yu; Klekar, Laura; Earnest, David; Yacoub, Shenouda; Hamilton, Gwenette; Jacobson, Nasreen; Shepard, Allan R.; Ellis, Dorette Zoe.

In: Journal of Ocular Pharmacology and Therapeutics, Vol. 30, No. 1, 01.02.2014, p. 21-34.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Trabecular meshwork bradykinin receptors

T2 - MRNA levels, immunohistochemical visualization, signaling processes pharmacology, and linkage to IOP reduction

AU - Sharif, Najam A.

AU - Katoli, Parvaneh

AU - Kelly, Curtis R.

AU - Li, Linya

AU - Xu, Shouxi

AU - Wang, Yu

AU - Klekar, Laura

AU - Earnest, David

AU - Yacoub, Shenouda

AU - Hamilton, Gwenette

AU - Jacobson, Nasreen

AU - Shepard, Allan R.

AU - Ellis, Dorette Zoe

PY - 2014/2/1

Y1 - 2014/2/1

N2 - Purpose: To localize mRNA and protein of bradykinin (BK) receptors, BK precursor polypeptide (kininogen) mRNA, and to study functional biochemical pharmacology of the signal transduction processes mediated by B 2-receptors in isolated human trabecular meshwork (h-TM) cells. Intraocular pressure (IOP) lowering effects of 2 kinins were also investigated. Methods: Previously documented procedures were utilized throughout these studies. Results: Kinninogen mRNA was most abundant in TM, ciliary body (CB), and optic nerve head and appeared elevated in glaucomatous h-TM tissue. High levels of B2-receptor mRNA were found in the sclera, iris, TM, and CB. B2-receptor subtype protein was localized in cells of the monkey and h-TM, and the treatment of isolated h-TM cells with transforming growth factor-β2 (5 ng/mL) caused significant (P<0.04) downregulation of B 2-receptor mRNA. In isolated primary h-TM cells, BK (EC 50=0.8±0.2 nM; n=19) and Met-Lys-BK (EC50=6. 5±1.5 nM) mobilized intracellular Ca2+ and induced the release of prostaglandins (PGs) that was blocked by 2 B2-receptor antagonists [HOE-140; (S)-WIN-64338]. The cyclooxygenase inhibitor, bromfenac, abolished BK-induced PGs production. BK concentration dependently increased cell impedance, and it significantly (P<0.05) decreased h-TM cell volume in vitro. Intravitreal (ivt) administration of BK (50 μg), but not a B 1-agonist (Sar-[D-Phe9]-Des-Arg9-BK; also at 50 μg), efficaciously lowered IOP (22.9% to 37% from baseline) of Dutch-Belted rabbits that naturally have high IOPs (27-28 mmHg). Conclusions: BK activates multiple signal transduction pathways in h-TM cells via B2-receptors that also mediate IOP reduction as observed in rabbits following ivt administration of BK. These ocular hypotensive effects of BK may be physiologically important and suggest a novel therapeutic potential of BK-related B2-agonists.

AB - Purpose: To localize mRNA and protein of bradykinin (BK) receptors, BK precursor polypeptide (kininogen) mRNA, and to study functional biochemical pharmacology of the signal transduction processes mediated by B 2-receptors in isolated human trabecular meshwork (h-TM) cells. Intraocular pressure (IOP) lowering effects of 2 kinins were also investigated. Methods: Previously documented procedures were utilized throughout these studies. Results: Kinninogen mRNA was most abundant in TM, ciliary body (CB), and optic nerve head and appeared elevated in glaucomatous h-TM tissue. High levels of B2-receptor mRNA were found in the sclera, iris, TM, and CB. B2-receptor subtype protein was localized in cells of the monkey and h-TM, and the treatment of isolated h-TM cells with transforming growth factor-β2 (5 ng/mL) caused significant (P<0.04) downregulation of B 2-receptor mRNA. In isolated primary h-TM cells, BK (EC 50=0.8±0.2 nM; n=19) and Met-Lys-BK (EC50=6. 5±1.5 nM) mobilized intracellular Ca2+ and induced the release of prostaglandins (PGs) that was blocked by 2 B2-receptor antagonists [HOE-140; (S)-WIN-64338]. The cyclooxygenase inhibitor, bromfenac, abolished BK-induced PGs production. BK concentration dependently increased cell impedance, and it significantly (P<0.05) decreased h-TM cell volume in vitro. Intravitreal (ivt) administration of BK (50 μg), but not a B 1-agonist (Sar-[D-Phe9]-Des-Arg9-BK; also at 50 μg), efficaciously lowered IOP (22.9% to 37% from baseline) of Dutch-Belted rabbits that naturally have high IOPs (27-28 mmHg). Conclusions: BK activates multiple signal transduction pathways in h-TM cells via B2-receptors that also mediate IOP reduction as observed in rabbits following ivt administration of BK. These ocular hypotensive effects of BK may be physiologically important and suggest a novel therapeutic potential of BK-related B2-agonists.

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