IN VITRO DENERVATION OF THE RAT VAS-DEFERENS THROUGH HYPOTHERMIC STORAGE
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1992-10-01
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Jurkiewicz, Neide Hyppolito [UNIFESP]
Garcia, Antonio G.
Jurkiewicz, Aron [UNIFESP]
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1 The rat vas deferens was excised, stored at 4-6-degrees-C and tested after 24, 48, 72 or 96 h for its contractile activity and for the presence of innervation.2 The maximal contractile capacity of the vas, tested through cumulative concentrations of barium chloride (3 x 10(-2) m) was progressively reduced from about 110 mm to about 63 mm after 72 h, without further decay after 96 h. Spontaneous rhythmic contractions were practically absent.3 A loss of endogenous pools of catecholamines was indicated by four parameters: (a) a decline of about 80% after 24 h and of more than 95% after 48 h of the contractile effect of the indirect sympathomimetic agonist tyramine; (b) a fall of about 20%, 50% and 85% on the concentration of noradrenaline, respectively after 24, 48 and 72 h; (c) a fall of about 25% and 90% after respectively 24 and 48 h, of the activity of dopamine-beta-hydroxylase (DBH); (d) a decline of noradrenaline-induced histofluorescence on cross sections of the vas.4 A loss of neuronal uptake capacity was indicated by: (a) a progressive variation of the apparent affinity for adrenaline, expressed as pD2 values, that increased by about 1.5 log units (corresponding to a 30 fold potentiation) after 72 h, and (b) a reduction of the ability of cocaine to potentiate the contractile effects of adrenaline.5 The pD2 values for barium chloride, 5-hydroxytryptamine (5-HT) and histamine were not significantly changed, while the corresponding value for acetylcholine was slightly but significantly reduced by about 0.8 log units.6 The maximal heights of concentration-response curves for noradrenaline, acetylcholine, histamine and 5-HT were reduced by 42-66% in relation to controls. However, when this reduction was measured in relation to the corresponding barium effect, by means of the relative responsiveness ratio (rho), a small though significant increase was observed for noradrenaline, and a fall for the other drugs.7 It is concluded that: (1) the values for the various biochemical and pharmacological parameters decline at different rates, though revealing altogether that denervation is completed by at least 85% after 72 h of hypothermic storage; (2) two of the results, i.e., the lack of spontaneous rhythmic contractions and the lack of increased contractile effects for acetylcholine, 5-HT and histamine, indicate that in these conditions the vas is devoid of the so-called nonspecific sips of denervation.
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British Journal Of Pharmacology. Basingstoke: Stockton Press, v. 107, n. 2, p. 610-615, 1992.