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Novel Insights Into Muscarinic and Purinergic Responses in Primary Cultures of Rat Lacrimal Gland Myoepithelial Cells
PURPOSE: The functional characteristics of receptors that regulate lacrimal gland myoepithelial cells are still somewhat unclear. To date, mainly muscarinic receptors have been of interest; however, further knowledge is needed regarding their expression and functional roles. For this purpose, primar...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
The Association for Research in Vision and Ophthalmology
2021
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8458779/ https://www.ncbi.nlm.nih.gov/pubmed/34546325 http://dx.doi.org/10.1167/iovs.62.12.19 |
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author | Dankis, Martin Carlsson, Thomas Aronsson, Patrik Tobin, Gunnar Winder, Michael |
author_facet | Dankis, Martin Carlsson, Thomas Aronsson, Patrik Tobin, Gunnar Winder, Michael |
author_sort | Dankis, Martin |
collection | PubMed |
description | PURPOSE: The functional characteristics of receptors that regulate lacrimal gland myoepithelial cells are still somewhat unclear. To date, mainly muscarinic receptors have been of interest; however, further knowledge is needed regarding their expression and functional roles. For this purpose, primary cultures of rat lacrimal gland myoepithelial cells were established and examined functionally. METHODS: Rat lacrimal glands were excised, minced, and further digested, yielding mixtures of cells that were seeded in culturing flasks. After 4-6 weeks, primary monocultures of myoepithelial cells were established, verified by immunocytochemistry. The cells were stained for all muscarinic receptor subtypes (M1–M5) and examined functionally regarding intracellular [Ca(2+)] responses upon activation of muscarinic receptors. For methodological verification, purinergic functional responses were also studied. RESULTS: Expression of muscarinic receptor subtypes M2-M5 was detected, whereas expression of muscarinic M1 receptors could not be shown. Activation of muscarinic receptors by the non-selective muscarinic agonist methacholine (3 × 10(−11)–10(−3) M) did not cause a significant increase in intracellular [Ca(2+)]. However, activation of purinergic receptors by the non-selective purinergic agonist ATP (10(−8)–10(−3) M) caused a concentration-dependent increase in intracellular [Ca(2+)] that could be blocked by the P2 antagonists PPADS and suramin. CONCLUSIONS: Primary cultures of rat lacrimal gland myoepithelial cells were established that displayed a heterogeneous expression of muscarinic receptors. Purinergic functional responses demonstrated a viable cell population. Upon treatment with methacholine, no significant increase in intracellular [Ca(2+)] could be detected, indicating that cholinergic activation of myoepithelial cells occurs via other intracellular messengers or is dependent on interaction with other cell types. |
format | Online Article Text |
id | pubmed-8458779 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | The Association for Research in Vision and Ophthalmology |
record_format | MEDLINE/PubMed |
spelling | pubmed-84587792021-10-05 Novel Insights Into Muscarinic and Purinergic Responses in Primary Cultures of Rat Lacrimal Gland Myoepithelial Cells Dankis, Martin Carlsson, Thomas Aronsson, Patrik Tobin, Gunnar Winder, Michael Invest Ophthalmol Vis Sci Biochemistry and Molecular Biology PURPOSE: The functional characteristics of receptors that regulate lacrimal gland myoepithelial cells are still somewhat unclear. To date, mainly muscarinic receptors have been of interest; however, further knowledge is needed regarding their expression and functional roles. For this purpose, primary cultures of rat lacrimal gland myoepithelial cells were established and examined functionally. METHODS: Rat lacrimal glands were excised, minced, and further digested, yielding mixtures of cells that were seeded in culturing flasks. After 4-6 weeks, primary monocultures of myoepithelial cells were established, verified by immunocytochemistry. The cells were stained for all muscarinic receptor subtypes (M1–M5) and examined functionally regarding intracellular [Ca(2+)] responses upon activation of muscarinic receptors. For methodological verification, purinergic functional responses were also studied. RESULTS: Expression of muscarinic receptor subtypes M2-M5 was detected, whereas expression of muscarinic M1 receptors could not be shown. Activation of muscarinic receptors by the non-selective muscarinic agonist methacholine (3 × 10(−11)–10(−3) M) did not cause a significant increase in intracellular [Ca(2+)]. However, activation of purinergic receptors by the non-selective purinergic agonist ATP (10(−8)–10(−3) M) caused a concentration-dependent increase in intracellular [Ca(2+)] that could be blocked by the P2 antagonists PPADS and suramin. CONCLUSIONS: Primary cultures of rat lacrimal gland myoepithelial cells were established that displayed a heterogeneous expression of muscarinic receptors. Purinergic functional responses demonstrated a viable cell population. Upon treatment with methacholine, no significant increase in intracellular [Ca(2+)] could be detected, indicating that cholinergic activation of myoepithelial cells occurs via other intracellular messengers or is dependent on interaction with other cell types. The Association for Research in Vision and Ophthalmology 2021-09-21 /pmc/articles/PMC8458779/ /pubmed/34546325 http://dx.doi.org/10.1167/iovs.62.12.19 Text en Copyright 2021 The Authors https://creativecommons.org/licenses/by/4.0/This work is licensed under a Creative Commons Attribution 4.0 International License. |
spellingShingle | Biochemistry and Molecular Biology Dankis, Martin Carlsson, Thomas Aronsson, Patrik Tobin, Gunnar Winder, Michael Novel Insights Into Muscarinic and Purinergic Responses in Primary Cultures of Rat Lacrimal Gland Myoepithelial Cells |
title | Novel Insights Into Muscarinic and Purinergic Responses in Primary Cultures of Rat Lacrimal Gland Myoepithelial Cells |
title_full | Novel Insights Into Muscarinic and Purinergic Responses in Primary Cultures of Rat Lacrimal Gland Myoepithelial Cells |
title_fullStr | Novel Insights Into Muscarinic and Purinergic Responses in Primary Cultures of Rat Lacrimal Gland Myoepithelial Cells |
title_full_unstemmed | Novel Insights Into Muscarinic and Purinergic Responses in Primary Cultures of Rat Lacrimal Gland Myoepithelial Cells |
title_short | Novel Insights Into Muscarinic and Purinergic Responses in Primary Cultures of Rat Lacrimal Gland Myoepithelial Cells |
title_sort | novel insights into muscarinic and purinergic responses in primary cultures of rat lacrimal gland myoepithelial cells |
topic | Biochemistry and Molecular Biology |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8458779/ https://www.ncbi.nlm.nih.gov/pubmed/34546325 http://dx.doi.org/10.1167/iovs.62.12.19 |
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