Gene expression of muscarinic, tachykinin and purinergic receptors in porcine bladder: comparison with cultured cells
Urothelial cells, myofibroblasts, and smooth muscle cells are important cell types contributing to bladder function. Multiple receptors including muscarinic (M3/M5), tachykinin (NK1/NK2) and purinergic (P2X1/P2Y6) receptors are involved in bladder motor and sensory actions. Using female pig bladder,...
| Main Authors: | , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Frontiers Media S.A.
2013-11-01
|
| Series: | Frontiers in Pharmacology |
| Subjects: | |
| Online Access: | http://journal.frontiersin.org/Journal/10.3389/fphar.2013.00148/full |
| _version_ | 1828223207450083328 |
|---|---|
| author | Forough eBahadory Kate Hilda Moore Lu eLiu Elizabeth eBurcher |
| author_facet | Forough eBahadory Kate Hilda Moore Lu eLiu Elizabeth eBurcher |
| author_sort | Forough eBahadory |
| collection | DOAJ |
| description | Urothelial cells, myofibroblasts, and smooth muscle cells are important cell types contributing to bladder function. Multiple receptors including muscarinic (M3/M5), tachykinin (NK1/NK2) and purinergic (P2X1/P2Y6) receptors are involved in bladder motor and sensory actions. Using female pig bladder, our aim was to differentiate between various cell types in bladder by genetic markers. We compared the molecular expression pattern between the fresh tissue layers and their cultured cell counterparts. We also examined responses to agonists for these receptors in cultured cells. Urothelial, suburothelial (myofibroblasts) and smooth muscle cells isolated from pig bladder were cultured (10-14 days) and identified by marker antibodies. Gene (mRNA) expression level was demonstrated by real-time PCR. The receptor expression pattern was very similar between suburothelium and detrusor, and higher than urothelium. The gene expression of all receptors decreased in culture compared with the fresh tissue, although the reduction in cultured urothelial cells appeared less significant compared to suburothelial and detrusor cells. Cultured myofibroblasts and detrusor cells did not contract in response to the agonists acetylcholine, neurokinin A and β,γ-MeATP, up to concentrations of 0.1 and 1 mM. The significant reduction of M3, NK2 and P2X1 receptors under culture conditions may be associated with the unresponsiveness of cultured suburothelial and detrusor cells to their respective agonists. These results suggest that under culture conditions, bladder cells lose the receptors that are involved in contraction, as this function is no longer required. The study provides further evidence that cultured cells do not necessarily mimic the actions exerted by intact tissues. |
| first_indexed | 2024-04-12T17:04:13Z |
| format | Article |
| id | doaj.art-45d88bf636ff4594b890b7ad1c411e4a |
| institution | Directory Open Access Journal |
| issn | 1663-9812 |
| language | English |
| last_indexed | 2024-04-12T17:04:13Z |
| publishDate | 2013-11-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Pharmacology |
| spelling | doaj.art-45d88bf636ff4594b890b7ad1c411e4a2022-12-22T03:23:59ZengFrontiers Media S.A.Frontiers in Pharmacology1663-98122013-11-01410.3389/fphar.2013.0014864199Gene expression of muscarinic, tachykinin and purinergic receptors in porcine bladder: comparison with cultured cellsForough eBahadory0Kate Hilda Moore1Lu eLiu2Elizabeth eBurcher3University of New South WalesSt George HospitalUniversity of New South WalesUniversity of New South WalesUrothelial cells, myofibroblasts, and smooth muscle cells are important cell types contributing to bladder function. Multiple receptors including muscarinic (M3/M5), tachykinin (NK1/NK2) and purinergic (P2X1/P2Y6) receptors are involved in bladder motor and sensory actions. Using female pig bladder, our aim was to differentiate between various cell types in bladder by genetic markers. We compared the molecular expression pattern between the fresh tissue layers and their cultured cell counterparts. We also examined responses to agonists for these receptors in cultured cells. Urothelial, suburothelial (myofibroblasts) and smooth muscle cells isolated from pig bladder were cultured (10-14 days) and identified by marker antibodies. Gene (mRNA) expression level was demonstrated by real-time PCR. The receptor expression pattern was very similar between suburothelium and detrusor, and higher than urothelium. The gene expression of all receptors decreased in culture compared with the fresh tissue, although the reduction in cultured urothelial cells appeared less significant compared to suburothelial and detrusor cells. Cultured myofibroblasts and detrusor cells did not contract in response to the agonists acetylcholine, neurokinin A and β,γ-MeATP, up to concentrations of 0.1 and 1 mM. The significant reduction of M3, NK2 and P2X1 receptors under culture conditions may be associated with the unresponsiveness of cultured suburothelial and detrusor cells to their respective agonists. These results suggest that under culture conditions, bladder cells lose the receptors that are involved in contraction, as this function is no longer required. The study provides further evidence that cultured cells do not necessarily mimic the actions exerted by intact tissues.http://journal.frontiersin.org/Journal/10.3389/fphar.2013.00148/fullMyofibroblastsUrotheliumcell cultureimmunostainingSmooth musclemuscarinic receptors |
| spellingShingle | Forough eBahadory Kate Hilda Moore Lu eLiu Elizabeth eBurcher Gene expression of muscarinic, tachykinin and purinergic receptors in porcine bladder: comparison with cultured cells Frontiers in Pharmacology Myofibroblasts Urothelium cell culture immunostaining Smooth muscle muscarinic receptors |
| title | Gene expression of muscarinic, tachykinin and purinergic receptors in porcine bladder: comparison with cultured cells |
| title_full | Gene expression of muscarinic, tachykinin and purinergic receptors in porcine bladder: comparison with cultured cells |
| title_fullStr | Gene expression of muscarinic, tachykinin and purinergic receptors in porcine bladder: comparison with cultured cells |
| title_full_unstemmed | Gene expression of muscarinic, tachykinin and purinergic receptors in porcine bladder: comparison with cultured cells |
| title_short | Gene expression of muscarinic, tachykinin and purinergic receptors in porcine bladder: comparison with cultured cells |
| title_sort | gene expression of muscarinic tachykinin and purinergic receptors in porcine bladder comparison with cultured cells |
| topic | Myofibroblasts Urothelium cell culture immunostaining Smooth muscle muscarinic receptors |
| url | http://journal.frontiersin.org/Journal/10.3389/fphar.2013.00148/full |
| work_keys_str_mv | AT foroughebahadory geneexpressionofmuscarinictachykininandpurinergicreceptorsinporcinebladdercomparisonwithculturedcells AT katehildamoore geneexpressionofmuscarinictachykininandpurinergicreceptorsinporcinebladdercomparisonwithculturedcells AT lueliu geneexpressionofmuscarinictachykininandpurinergicreceptorsinporcinebladdercomparisonwithculturedcells AT elizabetheburcher geneexpressionofmuscarinictachykininandpurinergicreceptorsinporcinebladdercomparisonwithculturedcells |