Analysis of the contractions evoked by sympathetic nerve stimulation, and thermal effect on the guinea-pig vas deferens--study as a model for the thermotherapy of benign prostatic hyperplasia

BACKGROUND: The present study was designed to investigate the mechanism of thermotherapy on benign prostatic hyperplasia (BPH), using the guinea-pig vas deferens as a model for BPH. The components of contractions elicited by electrical field stimulation and nicotine were analyzed, and the thermal effect on the vas deferens was examined. METHODS: The vas deferens was dissected, suspended vertically through two silver ring electrodes, and attached to an isometric transducer. The electrical stimulation of 10 constant current pulses (10 mA) with 0.3 msec in duration of 5, 10, and 40 Hz was achieved under air-gap condition. Drugs were added directly to a 5 ml Magnus tube containing Tyrode solution (36 degrees C) gassed with a 95% O2-5% CO2 mixture. The components of contractions evoked by electrical stimulation and nicotine were investigated by tetrodotoxin (TTX), and blocking agents of alpha 1-adrenoceptors and/or purinoceptors. Thermal effect on electrically evoked contractions was examined at incubation temperature of 25 degrees C (control), 43 degrees C, 45 degrees C, 46 degrees C and 47 degrees C for 1 hour. RESULTS: Nicotine (200 microM) elicited biphasic contractions, which were triggered by corelease of noradrenaline (NA) and ATP (N-ATP) from sympathetic nerve terminals by activation of prejunctional nicotine receptors. NA and N-ATP caused the corresponding contractions, alpha 1 and N-ATP components, respectively. Combined application of prazosin (1 microM) and suramin (50 microM) abolished these contractions. Activation of post-junctional alpha 1-adrenoceptors by NA caused release of ATP from muscle cells to produce the contraction (alpha 1-ATP component), which was sensitive to both suramin and prazosin. N-ATP and alpha 1 components attributed to fast and slow part of the contraction, respectively. Electrical field stimulation caused biphasic contractions which consisted of both neurogenic (TTX-sensitive) and non-neurogenic (TTX-insensitive) components. An increase in stimulation frequency (5 to 40 Hz) increased the neurogenic components, which contained alpha 1 and N-ATP components, as well as the case of nicotine. The non-neurogenic components consisted of alpha 1-ATP, muscle-derived ATP (m-ATP) and unknown substance 'X' components. Nifedipine (10 microM). L-type Ca2+ channel blocker, markedly reduced the contractions induced by bath applied phenylephrine (alpha 1-agonist, 100 microM) but only partially blocked the contractions produced by bath applied ATP (500 microM). The contractile force in amplitude and neurogenic components induced by electrical field stimulation did not change at 43 degrees C, but both declined significantly above 45 degrees C. The neurogenic components at 45 degrees C and 46 degrees C were suppressed to 22 +/- 6% and 14 +/- 3% (mean +/- SD) of control, respectively. All the contractile responses were abolished at 47 degrees C. CONCLUSION: The contractions of the guinea-pig vas deferens evoked by electrical field stimulation consisted of alpha 1, N-ATP, alpha 1-ATP, m-ATP and X components. Sympathetic nerve fibers in the muscles were completely inactivated by thermal exposure at 47 degrees C for 1 hour. The results suggest that the minimal temperature for thermotherapy of BPH should be 47 degrees C.     

Effects of the antidepressant St. John's wort (Hypericum perforatum) on rat and human vas deferens contractility

PURPOSE: Since sexual dysfunction related to vas deferens smooth muscle contractility is a possible side effect of St. John's wort (SJW) (Hypericum perforatum) we evaluated the effect of this herbal antidepressant on rat and human vas deferens contractility. MATERIALS AND METHODS: The effect of SJW was evaluated on contractions induced by electrical field stimulation or exogenous agonists (alpha,beta-methylene adenosine triphosphate and phenylephrine) in isolated rat and human vas deferens. RESULTS: SJW (1 to 300 microM) decreased in a concentration dependent manner the amplitude of electrical field stimulation and agonist induced contractions with the same potency, suggesting direct inhibition of rat vas deferens smooth muscle. Of the chemical constituents of SJW tested hyperforin but not hypericin or the flavonoids quercitrin, rutin and kaempferol inhibited phenylephrine induced contractions. SJW and hyperforin also inhibited phenylephrine induced contractions in human vas deferens CONCLUSIONS: The results of our study demonstrate that SJW directly inhibits rat and human vas deferens contractility. If confirmed in vivo, these results suggest that SJW might affect sexual function in humans. These results might explain delayed ejaculation described in patients receiving SJW.     

Saw palmetto is an indirectly acting sympathomimetic in the rat-isolated prostate gland

BACKGROUND: To investigate whether saw palmetto that inhibits alpha1-adrenoceptor binding in vitro affects contractility of the rat prostate gland. METHODS: The effects of a commercially available saw palmetto extract were examined on the contractility of rat-isolated prostate glands. The extract was tested in the presence and absence of phentolamine, prazosin, yohimbine, propranolol, hexamethonium, cocaine, desipramine, nifedipine, guanethidine, atropine, and alpha,beta-methylene ATP to evaluate the mechanism of action. Isolated preparations of rat vas deferens and bladder were used for comparison. RESULTS: Unexpectedly, saw palmetto extract caused contractions of the rat prostate gland that could be attenuated by prazosin, phentolamine, nifedipine, guanethidine, cocaine, and desipramine but not by any of the other pharmacological tools. Similar contractile effects were observed in rat-isolated vas deferens preparations but not in rat-isolated bladder preparations. CONCLUSIONS: In the rat prostate gland, saw palmetto extract causes indirect alpha1-adrenoceptor-mediated contractions via the release of noradrenaline from sympathetic neurons.     

L-NAME诱发的高血压大鼠输精管的胆碱能收缩：西地那非的作用

高血压是引起勃起功能障碍的一个危险因素,但其对输精管收缩和射精反应的作用还未有详尽的描述。本研究用氮氧合酶抑制剂-NG-硝基-L-精氨酸甲酯（L-NAME）,诱导一氧化氮（NO）缺乏的高血压。研究目的是评估L-NAME诱导的高血压对大鼠输精管收缩性的影响,检测西地那非是否影响高血压大鼠输精管的收缩性。将36R雄性大鼠分为三组：（1）空白对照组,（2）L-NAME诱导的高血压组,（3）用西地那非处理经L—NAME诱导的高血压组。第二组给予L-NAME（每只老鼠每天40mg）,处理4周。第三组大鼠同时给予西地那非（每只老鼠每天5mg,经口腔灌胃）和L—NAME。输精管的前列腺段给予电刺激（EFS,1—20赫兹）,P2X2激动剂—α,β亚甲基ATP（α,β-meATP,100μmolL-1μmolL－1）和α1－肾上腺素受体激动剂苯福林（Phe,100μmolL^-1－1mmolL－1）用于构建浓度反应曲线。用P2X受体拮抗剂、pyridoxalphosphate-6－azophenyl-2,4＇-disulfonicacid（PPADS,30μmolL。）重复实验。L-NAME可以明显加强输精管对EFS,α,β—meATP和Phe的收缩反应。L—NAME组经西地那非治疗后,输精管对EFS的收缩反应（20Hz）明显加强。高血压大鼠再经PPADS处理后,被EFS和α,β-meATP加强的输精管收缩反应得以恢复。在慢性NO缺乏的大鼠模型中,胆碱能、肾上腺素能组分及EFS可以影响输精管的收缩性。胆碱能系统对输精管收缩反应的调节作用比。肾上腺素能更强,而且西地那非可能可以调节PE患者的射精反应。     

Protein kinase c regulates purinergic component of neurogenic contractions in mouse bladder

PURPOSE: We evaluate the role of protein kinase C in excitatory purinergic neurotransmission in the mouse bladder. MATERIALS AND METHODS: In isolated mouse detrusor strips contractile responses to electrical field stimulation were mostly mediated by neural released acetylcholine and adenosine triphosphate (ATP). The changes in neurotransmission were measured indirectly by recording the contraction of detrusor strips in response to repetitive electrical field stimulation by trains of electrical pulses at 8 Hz. 1 second in duration. RESULTS: A protein kinase C activator, 1 to 2.5 nM. (beta-phorbol-12,13-dibutyrate (beta-PDBu), but not the inactive form alpha-phorbol-12,13-dibutyrate, significantly enhanced neurogenic detrusor contractions. The purinergic component of the evoked detrusor contractions in the presence of atropine was specifically sensitive to this enhancing effect by beta-PDBu but the cholinergic component in the alpha,beta-methylene ATP treated detrusors remained unaffected. This enhancing effect of beta-PDBu was dependent on the extracellular calcium (Ca2+) concentration. A P and/or Q type Ca2+ channel blocker, 0.1 and 0.3 microM. omega-conotoxin-MVIIC, and protein kinase C inhibitors, 0.3 and 1 microM. staurosporine and 0.3 and 1 microM. bisindolylmaleimide I but not 0.1 and 0.3 microM. omega-conotoxin-GVIA, an N type Ca2+ channel blocker, abolished the effect of beta-PDBu. Moreover, beta-PDBu did not affect the muscle responses induced by the exogenous agonists carbachol or alpha,beta-methylene ATP and potassium chloride. CONCLUSIONS: These results suggest that the activation of Ca2+ channel, especially the P and/or Q type, may be involved in the enhancing effect of protein kinase C activator beta-PDBu on muscle contractions elicited by excitatory purinergic neurotransmission in the mouse detrusor strips.     

Purinergic sensory neurotransmission in the urinary bladder: an in vitro study in the rat.

OBJECTIVES: To determine the response of mechanosensitive pelvic nerve afferents, arising from the rat urinary bladder, to the purinergic agonist alpha,beta-methylene ATP and to the purinergic antagonist suramin. MATERIALS AND METHODS: Using a newly developed in vitro bladder-pelvic nerve afferent model, multiunit recordings were taken from mechanosensitive pelvic nerve afferents arising from the rat urinary bladder, in response to bladder distension. Control experiments were performed by distending the bladder with saline at 0.04 mL/min, and recording the total afferent nerve activity and the bladder pressure response to the distension. Bladder distensions were then repeated using a solution of the stable purinergic agonist alpha,beta-methylene ATP (10 micromol/L), which is known to desensitize P2X-purinoceptors after prolonged exposure, and the total afferent activity and bladder pressure response were again measured. In a separate series of experiments the afferent nerve activity and bladder pressure response to bladder distension with saline was determined in the presence of the purinergic antagonist suramin (10 micromol/L) and repeated after washout of the drug. In both series of experiments, afferent nerve responses were compared with control using the paired t-test, whilst the bladder pressure responses were compared using one-way analysis of variance. RESULTS: Bladder distension with alpha,beta-methylene-ATP produced a statistically significant reduction in afferent nerve activity, by up to 75% compared with the control, whilst having no significant effect on the bladder pressure response. Bladder distension with saline in the presence of suramin (10 micromol/L) produced a significant reduction in the resultant afferent nerve activity, by 50%, which returned to normal after washout of the drug. CONCLUSION: These findings are consistent with the notion that ATP is released endogenously during bladder distension in the rat and is involved significantly in the activation of pelvic nerve afferents arising from the rat urinary bladder.     

DIFFERENTIAL EFFECTS OF SEX HORMONES AND PHYTOESTROGENS ON PEAK AND STEADY STATE CONTRACTIONS IN ISOLATED RABBIT DETRUSOR

PURPOSE: Recent evidence suggests that sex steroids may produce rapid inhibition of voltage operated Ca2+ channels (VOCCs). Detrusor smooth muscle is highly dependent upon Ca2+ influx for receptor-activated contractions. Thus, we examined the relative effectiveness of a select group of sex steroids and dietary phytoestrogens to relax detrusor contracted with the muscarinic receptor agonist, bethanechol (BE) and the purinergic P2X receptor agonist, alpha,beta-methylene ATP (alpha,beta-MeATP). MATERIALS AND METHODS: Isolated strips of rabbit detrusor were secured to isometric force transducers in a tissue bath and length-adjusted until maximum contractions were achieved. Peak (P) contractile responses were recorded for alpha,beta-MeATP (P(ATP)) and BE (P(BE)) and steady-state (SS) responses were recorded for BE (SS(BE)) in the presence and absence of selected sex steroids and phytoestrogens (10 microM, unless indicated). RESULTS: The L-type VOCC inhibitor, nifedipine (1 to 10 microM), completely inhibited P(ATP) but reduced SS(BE) by approximately 50%, whereas the VOCC and non-VOCC inhibitor, SKF 96365, inhibited SS(BE) by approximately 95%, suggesting that P(ATP) was entirely dependent on L-type VOCCs, but (BE)-induced contractions depended also on activation of non-VOCCs. 17Beta-estradiol (estradiol) and progesterone inhibited P(ATP) by approximately 60% and 20%, respectively, and 32 microM estradiol and ethinyl estradiol inhibited SS(BE) by approximately 80 and 95%, respectively. Inhibition by estradiol was potentiated, rather than blocked, by the nuclear estrogen receptor antagonist, tamoxifen. Moreover, tamoxifen alone nearly completely relaxed SS(BE). The inactive metabolite of estradiol, 17alpha-estradiol, inhibited both P(ATP) and P(BE) by approximately 40%. Testosterone had no effect on P(ATP) and P(BE). The phytoestrogen and tyrosine kinase inhibitor, genistein, inhibited SS(BE) by 44%, whereas daidzein, a phytoestrogen without tyrosine kinase inhibitory activity, produced only a 7% inhibition. None of the phytoestrogens examined inhibited P(BE), whereas all inhibited P(ATP) by approximately 20 to 35%. A comparison of inhibition of (BE) and alpha,beta-MeATP-induced contractions by selected estrogen isomers showed some distinct differences. For example, estrone did not inhibit P(BE) or SS(BE), but inhibited P(ATP) by approximately 20%, whereas DES inhibited SS(BE) by nearly 90%, but P(ATP) by a lesser degree (approximately 70%). CONCLUSIONS: Our data support the hypothesis that 17beta-estradiol, ethinyl estradiol, DES, tamoxifen and genistein may relax detrusor contractions by inhibition of both VOCCs and non-VOCCs. Moreover, our data show that genistein, a dietary phytoestrogen with tyrosine kinase inhibitory activity, selectively reduced alpha,beta-MeATP-induced peak and BE-induced steady-state contractions, sparing the maximum response to BE. Lastly, the inactive isomer, 17alpha-estradiol, inhibited both BE- and alpha,beta-MeATP-induced contractions. These data suggest that certain dietary phytoestrogens (for example, genistein) or sex steroids, especially those with weak activity at the nuclear steroid site (for example, 17alpha-estradiol), or tamoxifen may prove therapeutically useful in treating overactive bladder caused by elevated muscarinic and purinergic receptor activation.     

The evaluation of sympathetic system-related contractile activity of the rat vas deferens after ligation and intra-abdominal placement of the testis.

OBJECTIVE: To evaluate the contractile response of the vas deferens in a model of stress, to determine any changes in sympathetic activity as a result of stress in the ipsilateral testis, which decreases blood flow to the contralateral testis. MATERIALS AND METHODS: The study comprised two groups of six rats each; group 1 underwent a sham operation, and in group 2 the right testis was placed into the abdominal cavity and the vas deferens ligated. After 30 days, the vasa deferentia were resected bilaterally and their isometric contractions recorded. Electrical-field stimulation (EFS) was applied through a pair of platinum electrodes and concentration-response curves constructed for noradrenaline at 37 degrees C and to a solution containing 80 mmol/L K+. RESULTS: The vasa deferentia in both groups showed similar contractile responses to EFS, which were frequency-dependent and maximal at 80 Hz. Noradrenaline-induced contractile activity was lower in amplitude in the vasa deferentia of group 2 than in the contralateral and ipsilateral vasa deferentia of group 1, which were not significantly different from each other. All groups responded similarly to high K+. CONCLUSION: Intra-abdominal placement of the testes with vas deferens ligation decreased the contractile response to noradrenaline in the ipsilateral vas deferens without altering the contractile response to EFS and high K+. This difference could be caused by a reduction in the number of postjunctional alpha-adrenergic receptors or decreased receptor sensitivity. Both possibilities suggest that the vas deferens may initiate sympathetic activity, which may be responsible for contralateral testicular deterioration.     

Effects of mexiletine on electrical field stimulation-induced contractile responses in the ipsilateral and contralateral vasa deferentia after unilateral testicular torsion/detorsion

AIM: To investigate testicular torsion-induced changes on the electrical field stimulation (EFS)-induced contractions in rabbit vasa deferentia and to evaluate the effect of mexiletine. METHODS: 18 male New Zealand albino rabbits were used in this experiment. Rabbits were divided into three groups: (1) control group (n = 6); (2) torsion group (n = 6), and (3) mexiletine group (n = 6). In the control group, vasa deferentia on both sides were harvested. In the torsion and mexiletine groups, the left testes of the rabbits were subjected to 720 degrees of clockwise torsion for 2 h and then detorsion was performed. In the mexiletine group, 50 mg/kg i.p. mexiletine was administered 1 h before detorsion. Following 24 h of the torsion, vasa deferentia on both sides were harvested and 2-cm strips including both the prostatic and epididymal portions were prepared to record EFS-induced contractions. RESULTS: Testicular torsion caused a significant inhibition in both phases of EFS-induced biphasic contractions of the ipsi- and contralateral vasa deferentia. Mexiletine treatment did not affect these inhibitory responses. Torsion/detorsion of the spermatic cord did not alter exogenously applied noradrenaline-induced contractions in both vasa deferentia. However, KCl-induced contractions diminished significantly in ipsilateral vas deferens of the torsion group and mexiletine restored this inhibition. CONCLUSIONS: Unilateral testicular torsion/detorsion leads to inhibition in both phases of EFS-induced biphasic contractions of the ipsi- and contralateral vasa deferentia by causing a defect in presynaptic nerve transmission. However, mexiletine has no effect on this inhibition. Inhibition of the KCl-induced contractions in the ipsilateral vas deferens, which indicates postsynaptic tissue damage, is restored by administering mexiletine 1 h prior to detorsion.     

Effect of age on the responses of rat bladder detrusor strips to adenosine triphosphate

OBJECTIVE: To assess age-related changes in bladder function using the contractile responses to ATP of detrusor strips from rats of various ages. Materials and methods Urinary bladders were obtained from male Wistar rats aged 9 weeks (young), 24 weeks (adult) and 24 months (aged). Contractions of urinary bladder muscle strips to ATP were measured isometrically. The size of the initial phasic response and the secondary contractile response that developed after washing out ATP (postwashout contraction) were measured. The magnitudes of the ATP-induced phasic and postwashout contraction were compared among the age groups. During the contractions, prostanoid concentrations in the organ-bath medium were measured using an enzyme immunoassay. RESULTS: The ATP-induced postwashout contraction did not occur after stimulation with KCl or acetylcholine, but was induced by alpha,beta-methylene ATP. Both the phasic and postwashout contractions were concentration-dependent. Although the phasic contraction did not change progressively with age, the magnitude and duration of the postwashout contraction increased substantially with age. Nicardipine (a calcium antagonist) slightly inhibited both contractions. Suramin (a nonselective P2-receptor antagonist) did not significantly inhibit the phasic contraction, but reduced the postwashout contraction. PPADS (a selective P2X receptor antagonist) did not inhibit either contraction. Indomethacin (a prostaglandin synthesis inhibitor) had no effect on the phasic contraction but almost completely blocked the postwashout contraction when added before ATP stimulation, but was less effective when added after ATP. The prostaglandin E2 concentration in the organ bath increased during the postwashout contraction. CONCLUSIONS: These findings suggest that the ATP-induced postwashout contraction is not directly mediated by P2x purinoceptors, but results from the synthesis of prostaglandins, especially E2, which is a sensory autacoid. The age-linked increase in postwashout contraction may be involved in the changes in sensory and voiding mechanisms seen in the aged urinary bladder.     

Unusual absence of endothelium-dependent or -independent vasodilatation to purines or pyrimidines in the rat renal artery

BACKGROUND: Adenosine triphosphate (ATP) is a cotransmitter with noradrenaline (NA) in sympathetic perivascular nerves. It has a dual role in the maintenance of vascular tone as ATP, released from endothelial cells during shear stress or hypoxia, induces vasodilatation via endothelial P2Y receptors or by direct action on smooth muscle. The role and distribution of P2 receptors is well characterized for many blood vessels but not for the rat renal artery. This study aims to determine whether ATP is a vasoconstrictor cotransmitter with NA and whether ATP induces vasodilatation via the endothelium or smooth muscle. METHODS: On isolated rat renal arteries, electrical field stimulation (EFS) in the absence and presence of antagonists to P2X receptors and alpha1-adrenoceptors was examined. Concentration-response curves were constructed to NA, ATP, alpha,beta-methylene ATP (alpha,beta-meATP), uridine triphosphate (UTP), and 2-methylthio ADP (2-MeSADP) on low tone. Curves to acetylcholine (ACh), 2-MeSADP, and UTP were constructed on raised tone. Immunofluorescent localization of P2X and P2Y receptor subtypes was performed. RESULTS: Electrical field stimulation induced vasoconstriction, partially inhibited by the P2X receptor antagonist, pyridoxalphosphate-6-azophenyl-2',4'-disulfonic acid, and predominantly by prazosin. Exogenous NA and ATP mimicked EFS; immunostaining for P2X1 and P2X2 receptors was expressed on vascular smooth muscle. Unusually, ATP, 2-MeSADP, and UTP failed to induce vasodilatation. Acetylcholine induced vasodilatation. alpha,beta-meATP, 2-MeSADP, and UTP induced vasoconstriction via P2X1, P2Y1, and P2Y2 receptors, respectively. Immunostaining for P2X1, P2Y1, and P2Y2 receptors was expressed on the vascular smooth muscle. CONCLUSION: Adenosine triphosphate and NA are cotransmitters in sympathetic nerves supplying the rat renal artery, NA being the dominant partner. The novel feature of this vessel is that purines and pyrimidines do not produce either endothelium-dependent or -independent vasodilatation; P2X1, P2Y1, and P2Y2 receptors on the smooth muscle all mediate vasoconstriction.     

Stimulating and potentiating effects of sodium nitroprusside on the guinea-pig vas deferens and their comparison with the effects on the portal vein and the taenia coli

The effects of sodium nitroprusside on the electrical and mechanical properties of the guinea-pig vas deferens were studied and compared with those on the portal vein and the taenia coli. Sodium nitroprusside at concentrations higher than 0.5 mM caused depolarization of the membrane of the vas deferens and initiated spontaneous contractions, while spontaneous contractions of the portal vein were blocked by similar concentration of the drug. Noradrenaline- and carbachol-induced contractions of the vas deferens were markedly potentiated by sodium nitroprusside, whereas the noradrenaline-induced contraction of the portal vein was suppressed by the same concentration of the drug. Increasing the K+ concentration by 15 to 30 mM caused a similar potentiation of the contraction by noradrenaline or carbachol in the vas deferens. When sodium nitroprusside was applied during the course of noradrenaline- or carbachol-induced contracture, contraction was observed in the vas deferens, while relaxation was induced in the portal vein and taenia coli. In either case, however, the addition of Ca caused a relaxation of the preparations. These results suggest that the membrane depolarization may be involved in the stimulating effects sodium nitroprusside in the guinea-pig vas deferens.     

A QUANTITATIVE STUDY OF ATROPINE-RESISTANT CONTRACTILE RESPONSES IN HUMAN DETRUSOR SMOOTH MUSCLE, FROM STABLE, UNSTABLE AND OBSTRUCTED BLADDERS

PURPOSE: The objective of the study was to quantify in vitro the magnitude of atropine-resistant contractions using human detrusor samples and to determine the cellular processes underlying these contractions. MATERIALS AND METHODS: Isometric contractile responses were measured in isolated strips of human detrusor muscle obtained from patients with i) stable, ii) unstable or iii) obstructed bladders. Preparations were electrically stimulated or exposed to carbachol and ATP in the superfusate. RESULTS: Force-frequency curves were shifted to the right in samples from unstable and obstructed bladders. These same tissue groups also showed significant atropine-resistant contractions which were abolished by the neurotoxin TTX, or the non-hydrolysable ATP analog, alpha,beta-methylene ATP, suggesting that these contractions were mediated by neurally released ATP. Sub-division of the patient group with unstable bladders demonstrated that those with neuropathic instability did not show atropine-resistance, whereas those with idiopathic instability or secondary instability after obstruction did show atropine-resistant contractions. The potency of carbachol in generating a contracture was significantly greater than ATP (mean EC50 0.65 microM and 151 microM respectively) however, for each agonist there was no difference in potency between the three patient groups. Direct muscle excitability was similar in all three patient groups. CONCLUSIONS: It is concluded that purinergic, atropine-resistant contractions are present in some types of dysfunctional bladder, and these are not caused by a differential sensitivity of the muscle to ATP and cholinergic agonists.     

Hormonal influence on the neurogenic response of the hamster vas deferens

The effect of castration on in vitro contractility of smooth muscle of the vas deferens and body of the bladder has been studied in the hamster. Castration produced supersensitivity to in vitro electrical stimulation and norepinephrine in the vas deferens, but had no effect on the body of the bladder. Castration also increased the maximum contractile response of the vas deferens to electrical stimulation, norepinephrine, ATP, acetylcholine and histamine. The changes in contractility of smooth muscle of the vas deferens developed slowly and may be explained by specific effects upon adrenergic and purinergic neurotransmission and/or non-specific effects upon smooth muscle cell membranes.     

Potentiation by Bradykinin and Substance P of Purinergic Neurotransmission in Urinary Bladder

Purpose

The higher than normal levels of substance P (SP) and the kinins in patients suffering from interstitial cystitis suggest that they may contribute to the complex symptoms of the condition. The purpose of our experiments was to determine whether SP and bradykinin (BK) influence the excitatory motor innervation of the urinary bladder.

Materials and Methods

Strips of guinea pig urinary bladder were placed in isolated tissue baths, and the influence of SP and BK on contractions induced by transmural electrical stimulation and cholinergic and purinergic agonists was evaluated.

Results

Substance P and BK potentiated responses to the purinergic component of the neurogenic stimulation (that part of the contractile response that remains after treatment with atropine) and potentiated responses to exogenously applied adenosine triphosphate (ATP). The peptides did not potentiate the response to the cholinergic component of the nerve-induced contraction (that part of the neurogenic response that remains after desensitization of purinoceptors with alpha, beta-methylene ATP) nor responses to carbachol. The potentiating actions of SP and BK were reduced but not abolished by treatment with meclofenamic acid.

Conclusions

Substance P and BK potentiate the neurogenic response of the bladder by influencing the purinergic component of the excitatory motor innervation, apparently at a postjunctional site. Prostaglandins may be involved in mediating some of the actions of these peptides.     

Cholinergic and purinergic contribution to the micturition reflex in conscious rats with long-term bladder outlet obstruction

The urethra of female Wistar rats was partially obstructed for 15 weeks. The effects of atropine (1 mg/kg i.v.), suramin (100 mg/kg i.v.), and a combination of atropine and suramin on the peak micturition pressure (MP) were compared during cystometry in conscious rats controls or subjected to outlet obstruction. On the isolated bladder dome, we studied the inhibitory effect of 1 micromol/L atropine, 1 mmol/L suramin, and the combination of the two drugs on contractions induced by electrical field stimulation (EFS). We studied also the contractile response to 80 mmol/L KCl and the concentration-response curves to noradrenaline, phenylephrine, and carbachol on the bladder dome and bladder neck and alpha, beta-methylene adenosine triphosphate on the bladder dome. In conscious rats, the MP, bladder capacity, and micturition volume were significantly higher in obstructed rats than in controls. Suramin induced the same inhibition in the two groups of animals (-30.7 +/- 13.3% in controls and -29.2 +/- 8.5% in obstructed rats). Atropine decreased the MP, but this effect was twofold greater in obstructed animals (-28.1 +/- 3.1% and -65.1 +/- 6.9% in control and obstructed animals, respectively). However, the combined effect of atropine and suramin was additive in controls but not in obstructed (-56.7 +/- 5.4% and -55.9 +/- 9.4%, respectively). Similar results were obtained in vitro using 1 micromol/L atropine and 1 mmol/L suramin. In the obstructed bladder dome and bladder neck, we found a great reduction in KCl- and carbachol-induced contractility but no difference in the response to EFS. Responses to noradrenaline and phenylephrine were moderately reduced in the bladder neck only, whereas responses to alpha, beta-methylene adenosine triphosphate in the bladder dome were not reduced except at the concentration of 300 micromol/L. We conclude that long-term obstruction in rats could induce cholinergic nerve fiber proliferation as suggested by the decrease in M(3) muscarinic receptor contractility (desensitization) and by a greater sensitivity of the MP to atropine.     

Regulation of rat caput epididymidis contractility by prostaglandins

Mechanical activity of the rat caput epididymidis in vitro was recorded using a videomicrography system. The effects of prostaglandin (PG)F2 alpha, PGE2, and aspirin on caput epididymidis contractility were determined by measuring the frequency of contraction, luminal diameter, and amplitude of contraction at various concentrations of each test compound in vitro. PGF2 alpha stimulated contractility of the tubules at physiological concentrations, while PGE2 reduced contractility. Aspirin strongly inhibited contractility at concentrations of 10(-3) and 10(-2)M. Endogenous levels of PGF2 alpha and PGE were determined for rat testes, caput, corpus, and cauda epididymidis and vas deferens. While the concentrations of PGE were consistently higher than those of PGF2 alpha, both compounds were relatively low in the testes, high in the vas deferens, and intermediate throughout the epididymis. Results from these experiments strongly suggest that PGs are important regulators of proximal epididymidis contractions and thus may regulate sperm transport through that organ.     

Effects of intravenous and infravesical administration of suramin,terazosin and BMY 7378 on bladder instability in conscious rats with bladder outlet obstruction

OBJECTIVE: To evaluate the effect of the nonselective purinergic antagonist suramin and the alpha1-adrenergic antagonists, terazosin and BMY 7378, given intravenously or infused directly into the bladder during cystometry in conscious rats with bladder outlet obstruction induced by urethral ligation. MATERIALS AND METHODS: Cystometry was performed in conscious female rats recording bladder volume capacity (BVC), evaluated as the amount of saline infused between two voiding cycles, and micturition volume (MV). Changes in frequency and amplitude of spontaneous non-voiding bladder contractions (NVC) were also recorded. The effects of the intravenous administration of suramin (100 mg/kg), BMY 7378 (1 mg/kg), and terazosin (0.3 mg/kg) on NVC, BVC and MV were evaluated in obstructed rats with bladder infusion of saline. The effects of infravesical infusion of suramin (3-10 micromol/L), terazosin (1 micromol/L) and BMY 7378 (10 micromol/L) were also evaluated and compared with values observed in control rats during saline infusion into the bladder. RESULTS: Intravenous injection with suramin had no effects on NVC, BVC and MV, but suramin infused into the bladder induced a consistent reduction in the amplitude of NVC (significantly different from matched control animals) with a tendency to reduce their frequency. BVC and MV were slightly but significantly decreased by infravesical infusion of suramin. In contrast, BMY 7378 and terazosin, given intravenously, were extremely potent at inhibiting the frequency and amplitude of the NVC, but were inactive on NVC when infused into bladder. CONCLUSIONS: These findings confirm a role for alpha1-adrenergic receptors in bladder instability caused by bladder outlet obstruction. In addition, a purinergic neurotransmitter, presumably ATP, is shown to be involved.     

The contractile potency of adenosine triphosphate and ecto-adenosine triphosphatase activity in guinea pig detrusor and detrusor from patients with a stable,unstable or obstructed bladder

PURPOSE: We compared the potency of adenosine triphosphate (ATP) and its nonhydrolyzable analogue alpha,beta-methylene ATP for generating contractions in human detrusor smooth muscle from patients with a stable, unstable and obstructed bladders. The different ATP potencies were compared with the ecto-adenosine triphosphatase (ATPase) of these samples. MATERIALS AND METHODS: Contractile experiments were done in vitro by superfusing samples with purines and dose-response curves were generated. Ecto-ATPase activity was measured from the rate of ATP hydrolysis sensitive to the ecto-ATPase inhibitor ARL 67156 with a luciferin-luciferase assay. RESULTS: ATP generated contractions with a mean EC50 of 933 microM. in tissue from stable bladders and was significantly more potent in tissue from unstable and obstructed bladders (EC50 141 and 172 microM., respectively). alpha,beta-methylene ATP was more potent in tissue from stable and unstable bladders (mean combined EC50 3 microM.). In guinea pig detrusor the mean EC50 for ATP and alpha,beta-methylene ATP was 138 and 5.5 microM., respectively. Mean total ATPase activity in unstable bladder biopsies plus or minus standard deviation was about 50% of that in stable bladder biopsies (2.54 +/- 1.50 versus 1.37 +/- 0.46 nmol. per second per mg. protein ). The ARL 67156 sensitive fraction was also significantly less in samples from unstable compared with stable bladders (mean 0.94 +/- 0.41 versus 0.36 +/- 0.26 nmol. per second mg. protein ). CONCLUSIONS: The greater potency of ATP for generating contractions in detrusor from unstable bladders may be due to reduced extracellular hydrolysis, allowing purine greater access to detrusor smooth muscle. This finding may explain atropine resistant purine based contractions in detrusor from unstable bladders.     

Effect of extracellular Mg concentration on electrically induced contractions of rat vas deferens in vitro

Contraction of smooth muscles of the vas deferens plays an important role in the propulsion of sperm into the pelvic urethra. This study examined the influence of external Mg2+ concentration on reactivity of the rat vas deferens to electrical stimulation in vitro. Vasa deferentia isolated from adult male rats were set up in tissue baths containing physiological salt solution at 37 degrees C and were stimulated electrically. Thereafter, increasing concentrations of Mg2+ were added to the bath and their effects on electrically evoked contractions were recorded. The effect of external Mg2+ depletion on evoked contractions was also examined. External Mg2+ depletion enhanced the contractile response to electrical stimulation while increasing external Mg2+ concentration inhibited the contractions. The inhibitory effect of Mg2+ was partially reversed by increasing extracellular Ca2+ concentration and was not additive with nifedipine. The results indicate that reactivity of the vas deferens to electrical stimulation is modulated by extracellular Mg2+ concentration. The possible relevance of these data to sperm transport through the vas deferens is discussed.