Activation of the EGF Receptor by Histamine Receptor Subtypes Stimulates Mucin Secretion in Conjunctival Goblet Cells

Investigative Ophthalmology & Visual Science
Min HeDarlene A Dartt

Abstract

The purpose of this study was to determine if histamine receptors interact with the epidermal growth factor receptor (EGFR) in cultured rat conjunctival goblet cells. Goblet cells from rat conjunctiva were grown in organ culture. First-passage goblet cells were used in all experiments. Phosphorylated (active) and total EGFR, AKT, and extracellular signal-regulated kinase (ERK)1/2 were measured by Western blot analysis. Cells were preincubated with the EGFR antagonist AG1478 for 30 minutes or small interfering RNA specific to the EGFR for 3 days prior to stimulation with histamine or agonists specific for histamine receptor subtypes for 2 hours. Goblet cell secretion was measured using an enzyme-linked lectin assay. Goblet cells were incubated for 1 hour with the calcium indicator molecule fura-2/AM, and intracellular [Ca2+] ([Ca2+]i) was determined. Data were collected in real time and presented as the actual [Ca2+]i with time and as the change in peak [Ca2+]i. Histamine increased the phosphorylation of the EGFR. Mucin secretion and increase in [Ca2+]i stimulated by histamine, and agonists specific for each histamine receptor subtype were blocked by inhibition of the EGFR. Increase in [Ca2+]i stimulated by histamine and specifi...Continue Reading

References

Mar 7, 2003·American Journal of Physiology. Cell Physiology·Harumi KannoDarlene A Dartt
Jun 20, 2003·Annals of the New York Academy of Sciences·David C LeeLeslie F Jackson
Mar 3, 2004·The Journal of Cell Biology·Umut SahinCarl P Blobel
Apr 19, 2008·Investigative Ophthalmology & Visual Science·Marie A ShatosDarlene A Dartt
Mar 2, 2011·The Journal of Immunology : Official Journal of the American Association of Immunologists·Darlene A DarttCharles N Serhan
Jan 17, 2012·FASEB Journal : Official Publication of the Federation of American Societies for Experimental Biology·Nobuaki HirotaJames G Martin
Apr 3, 2012·Investigative Ophthalmology & Visual Science·Densen HayashiDarlene A Dartt
Sep 8, 2012·Investigative Ophthalmology & Visual Science·Dayu LiDarlene A Dartt
Sep 15, 2012·Experimental Eye Research·Robin R HodgesDarlene A Dartt
Jun 11, 2014·The Journal of Cell Biology·David J Erle, Dean Sheppard
Jul 26, 2014·Current Opinion in Allergy and Clinical Immunology·Darlene A Dartt, Sharmila Masli
Aug 11, 2015·Current Opinion in Allergy and Clinical Immunology·Andrea LeonardiDaniela Lazzarini
Nov 20, 2015·Neuropharmacology·Bassem Sadek, Holger Stark
May 8, 2016·The Ocular Surface·Anna F Ablamowicz, Jason J Nichols
Sep 12, 2017·Investigative Ophthalmology & Visual Science·Marit LippestadDarlene A Dartt
Nov 4, 2017·BMJ : British Medical Journal·Darshak S PatelRomesh Angunawela

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Citations

Mar 19, 2019·Current Opinion in Supportive and Palliative Care·Andrea M Stringer, Nicole Blijlevens
Jun 27, 2019·Current Medicinal Chemistry·Chen-Chen Chu, Shao-Zhen Zhao
Oct 29, 2021·Bulletin of Experimental Biology and Medicine·M HeX Wang
Jul 7, 2021·Ocular Immunology and Inflammation·Maurizio Rolando, Stefano Barabino

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Methods Mentioned

BETA
transfection
ELISA
protein assay
GTPase

Software Mentioned

ImageJ

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