Abstract
The functional responses of different overnight-stored in vitro tissues are not clearly described in any animal model. The influence of overnight storage in an animal model may vary between tissue types. We employed Sprague–Dawley rat as our animal model and investigated the functional changes of rat aorta, trachea, bronchus and bladder that were used (i) immediately after surgical removal (denoted as fresh) and (ii) after storage in aerated (95 % O2, 5 % CO2) Krebs-Ringer bicarbonate solution at 4 °C for 24 h (denoted as stored). The aorta ring was pre-contracted with phenylephrine, and the functional response of the tissue was investigated using isoprenaline, forskolin and carbachol. Carbachol was also used to increase the tone in trachea, bronchus rings and bladder strips. A clear reduced function of endothelium, with a minor if any effect in the smooth muscle function in rat aorta was observed after overnight storage. The contractile response of overnight-stored rat airway (trachea and bronchus) and bladder smooth muscles remained unchanged. Among all tested tissues, only bronchus showed a reduced response rate (only 40 % responded) after storage. In vitro rat tissues that are stored in Krebs solution at 4 °C for 24 h can still be used to investigate smooth muscle responses, however, not endothelium-mediated responses for aorta. The influence of overnight storage on different tissues from an animal model (Sprague–Dawley rat in our study) also provides an insight in maximising the use of sacrificed animals.
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References
Ebner A, Poitz DM, Augstein A, Strasser RH, Deussen A (2012) Functional, morphologic, and molecular characterization of cold storage injury. J Vasc Surg 56(1):189–198
Furchgott RF, Zawadzki JV (1980) The obligatory role of endothelial cells in the relaxation of arterial smooth muscle by acetylcholine. Nature 288:373–376
Janssen LJ, Wattie J, Lu-Chao H, Tazzeo T (2001) Muscarinic excitation-contraction coupling mechanisms in tracheal and bronchial smooth muscles. J Appl Physiol 91:1142–1151
Jiang F, Li CG, Rand MJ (2000) Mechanisms of nitric oxide-independent relaxations induced by carbachol and acetylcholine in rat isolated renal arteries. Br J Pharmacol 130:1191–1200
Kawaguchi T, Satoh K, Kuji A, Joh S (2010) Features of distinct contractions induced with a high and a low concentration of KCl, noradrenaline, and histamine in swine lingual artery. Naunyn-Schmied’s Arch Pharmacol 381:107–120
Michel MC (2014a) Do β-adrenoceptor agonists induce homologous or heterologous desensitization in rat urinary bladder? Naunyn-Schmiedeberg’s Arch Pharmacol 387:215–224
Michel MC (2014b) How significant are your data? The need for a culture shift. Naunyn-Schmied’s Arch Pharmacol 387:1015–1016
Michel MC (2015) Therapeutic modulation of urinary bladder function: multiple targets at multiple levels. Annu Rev Pharmacol Toxicol 55:269–87
Motulsky HJ (2014) Common misconceptions about data analysis and statistics. Naunyn-Schmied’s Arch Pharmacol 387:1017–1023
Piepot HA, Pneumatikos IA, Johan Groeneveld AB, van Lambalgen AA, Sipkema P (2002) Cold storage sensitizes rat femoral artery to an endotoxin-induced decrease in endothelium-dependent relaxation. J Surg Res 105:189–194
Rascado RR, Bendhack LM (2005) Activation of a2-adrenoceptors is necessary to induce nitric oxide release in isoprenaline-induced relaxation. Vasc Pharmacol 42:63–68
Sadeghi-Hashjin G, Henricks PAJ, Folkerts G, Verheyen AKCP, van der Linde HJ, Nijkamp FP (1996) Bovine tracheal responsiveness in vitro: role of the epithelium and nitric oxide. Eur Respir J 9:2286–2293
Satake N, Shibata M, Shibata S (1996) The inhibitory effects of iberiotoxin and 4-aminopyridine on the relaxation induced by β1- and β2-adrenoceptor activation in rat aortic rings. Br J Pharmacol 119:505–510
Schneider T, Fetscher C, Michel MC (2011) Human urinary bladder strip relaxation by the β-adrenoceptor agonist isoprenaline: methodological considerations and effects of gender and age. Front Pharmacol 2:11. doi:10.3389/fphar.2011.00011
Shimokawa H, Flavahan NA, Lorenz RR, Vanhoutte PM (1988) Prostacyclin releases endothelium-derived relaxing factor and potentiates its action in coronary arteries of the pig. Br J Pharmacol 95:1197–1203
Taylor K, Gordon N, Langley G, Higgins W (2008) Estimates for worldwide laboratory animal use in 2005. Altern Lab Anim 36:327–342
Török J, Kristek F, Mokrášová M (1993) Endothelium-dependent relaxation in rabbit aorta after cold storage. Eur J Pharmacol: Environ Toxicol Pharm 228(5–6):313–319
Vanhoutte PM, Rubanyi GM, Miller VM, Houston DS (1986) Modulation of vascular smooth muscle contraction by the endothelium. Annu Rev Physiol 48:307–320
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Miss BJ Loong receives scholarship for her PhD studies from the University of Nottingham Malaysia Campus.
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Loong, B.J., Tan, J.H., Lim, K.H. et al. Contractile function of smooth muscle retained after overnight storage. Naunyn-Schmiedeberg's Arch Pharmacol 388, 1061–1067 (2015). https://doi.org/10.1007/s00210-015-1140-3
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DOI: https://doi.org/10.1007/s00210-015-1140-3