Cajal间质细胞上M受体介导的胆碱能信号在输尿管自主收缩中的作用研究
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摘要
背景与目的:
Cajal间质细胞原来被认为是仅存在于胃肠道的一种特殊类型细胞,起着起搏胃肠道慢波、传递电信号、介导肠神经与平滑肌之间调节信号的作用,与病理状态下的多种胃肠动力紊乱疾病(如便秘、假性肠道梗阻等)密切相关,是胃肠道平滑肌节律性收缩功能调节的重要因素。后来,在胃肠道外其他组织中发现了与胃肠道Cajal间质细胞免疫学特性相同、形态学特点相似的细胞,既往曾称之为ICC样细胞(ICC-like cells),目前已统称为Cajal间质细胞(ICCs)。
生理情况下,输尿管平滑肌具有与胃肠道十分相似的特点,即存在不依赖于神经的自发性蠕动,但确切机制目前尚未阐明。目前已有的研究发现,在豚鼠和人的全尿路均存在ICCs。有文献报道,先天性肾盂输尿管梗阻的患儿肾盂和输尿管ICCs较正常儿童明显减少,提示ICCs与输尿管兴奋起源和调控有密切关系。进一步研究ICCs在输尿管自主收缩中的作用及其对神经递质的反应,有助于阐明ICCs在输尿管平滑肌功能调控中的作用及其机制,为上尿路功能障碍性疾病的治疗提供新的切入点。
本课题通过肌条实验观察离体的豚鼠输尿管肌条,在使用非特异性M型胆碱能受体激动剂卡巴可前后自主收缩的变化,并在特异性阻断ICCs功能后,观察肌条的自主收缩及对卡巴可的反应是否发生改变,此外还应用免疫荧光双标染色法检测输尿管ICCs上M受体亚型的表达情况,从而为阐明ICCs上M受体介导的胆碱能信号在输尿管平滑肌自主收缩中的作用提供新的实验依据。
方法:
实验动物均采用清洁级健康Hartley豚鼠。(1)卡巴可对输尿管平滑肌自主收缩的影响及ICCs在此影响中的作用:将20只豚鼠随机分为正常对照组、阻断ICCs组,每组10只。阻断ICCs组的肌条检测前平衡及检测中均置于含终浓度10-4mol/L Glivec的Kreb’s液中,正常对照组置于不含Glivec的Kreb’s液中。取豚鼠左侧输尿管上段1.0cm组织制成离体平滑肌肌条,检测其在终浓度为10-4mol/L卡巴可作用前后的自主收缩频率和幅度。(2)输尿管ICCs上M受体亚型的表达情况:通过c-kit+M1～M5免疫荧光双标染色法检测豚鼠输尿管ICCs上M受体亚型的表达情况。
结果:
(1)在一定前负荷(2.0g)下,加入1mol/L KCl溶液200μL可诱发正常对照组豚鼠的离体输尿管平滑肌肌条出现自主收缩,频率为(3.4±0.84)次/分,波幅为(0.105±0.0228)g。
(2)在肌条出现稳定的自主收缩后,加入0.01mol/L卡巴可100μL,肌条自主收缩频率为(4.9±0.88)次/分,较加药前明显增快(p<0.01);而收缩幅度为(0.106±0.0210)g,较加药前无明显变化(p>0.05)。
(3)用10-4mol/L Glivec平衡30分钟、特异性阻断ICCs后,同法进行检测发现,阻断ICCs组的输尿管肌条无自主收缩出现,加入0.01mol/L卡巴可100μL仍无反应。
(4)免疫荧光双标染色发现,豚鼠输尿管ICCs上表达M2、M3两种M受体亚型。
结论:
ICCs很可能是输尿管平滑肌自主收缩兴奋的起源,并通过M2、M3受体亚型接受胆碱能神经的调控。
Background and Objective:
Interstitial cells of Cajal are the special cells of gastrointestinal tract, which pacing the slow waves of gastrointestinal tract, transmit electrical signals, and mediate signals conduction and integration between nerve cells and smooth muscle cells. As an important factor of adjusting the rhythmic contraction of gastrointestinal smooth muscle, interstitial cells of Cajal have close correlation with some gastrointestinal motility disorders, such as constipation, intestinal pseudo-obstruction. Initially, interstitial cells of Cajal were supposed to exist only in gastrointestinal tract. Recently, interstitial cells of Cajal had been reported existed in other organizations outside gastrointestinal tract. Recent researches showed that morphological and immunological features of interstitial cells of Cajal outside gastrointestinal tract resemble those of gastrointestinal interstitial cells of Cajal. So, interstitial cells of Cajal outside gastrointestinal tract that had been called ICC-like cells were collectively referred to as interstitial cells of Cajal (ICCs) now.
Under normal circumstances, ureteral smooth muscle similar to gastrointestinal smooth muscle, has spontaneous myogenic contraction uncontrolled by nerve. However, the mechanism of spontaneous myogenic stimulation had not yet been fully illuminated. There was a study demonstrated ICCs been located in the entire urinary tract of human and guinea pig. Solari et al had compared the density of ICCs of renal pelvis and ureter between children with congenital ureteropelvic junction obstruction and normal children, and found that the distribution of ICCs of renal pelvis and ureter of normal children was scattered. The number of ICCs of renal pelvis and ureter of normal children was significantly decreased compared with that of children with congenital ureteropelvic junction obstruction. The study suggests that ICCs have close correlation with origin and regulation of ureter’s excitation. Therefore, to further research the role and neuromodulation of ICCs in smooth muscle spontaneous contraction of ureter is helpful to elucidate the role and mechanism of ICCs in the regulation of function of ureteral smooth muscle, and to treat the functional disorders of upper urinary tract.
In this study, we compared the smooth muscle spontaneous contraction of isolated guinea pig ureter by muscle strips experiment before and after the effect of carbachol, a nonspecific agonist of muscarinic receptor. Furthermore, we observed if the smooth muscle spontaneous contraction of isolated guinea pig ureter and its reaction to carbachol had been changed after blocking ICCs. Lastly, immunofluorescence (double-labeled) were used to study the expressions of muscarinic receptor subtypes on ICCs of guinea pig ureter. These results will provide new experimental evidences for elucidating the role of cholinergic signal mediated by muscarinic receptors on ICCs in smooth muscle spontaneous contraction of guinea pig ureter.
Materials and Methods:
The cleaning grade healthy Hartley guinea pigs were sacrificed in this study. Firstly, we investigated the effect of carbachol to smooth muscle spontaneous contraction of guinea pig ureter and the role of ICCs in this effect. 20 guinea pigs were randomly divided into two groups: normal group (n=10) and ICCs inhibited group (n=10). The smooth muscle strips obtained from guinea pigs of ICCs inhibited group were incubated with and measured in Kreb’s solution containing 10-4mol/L Glivec, but those of normal group were incubated with and measured in Kreb’s solution uncontaining Glivec. All of smooth muscle strips were about 1.0 centimeter obtained from superior segment of left guinea pig ureter. Frequency and amplitude of smooth muscle spontaneous contraction were measured in Kreb’s solution containing or uncontaining 10-4mol/L Glivec before and after the effect of 10-4mol/L carbachol. Then, we detected the expressions of muscarinic receptor subtypes on ICCs of guinea pig ureter. Both c-kit receptor and muscarinic receptor subtypes (M1~M5) on ICCs of guinea pig ureter were been observed with laser scanning confocal microscope after been stained by immunofluorescence.
Results:
(1) Spontaneous contractions were recorded on isolated smooth muscle strips of guinea pig ureter under certain preload (2.0g) and when 200μL 1.0mol/L KCl solution was added. The frequency and amplitude of spontaneous contraction were (3.4±0.84)/min and (0.105±0.0228)g respectively.
(2) When 100μL 0.01mol/L carbachol solution was added, the frequency of spontaneous contraction of ureteral smooth muscle strips increased to (4.9±0.88)/min (p<0.01). While the amplitude was (0.106±0.0210)g, and had no statistical significant difference compared with that before using carbachol (p>0.05).
(3) Incubated with Kreb’s solution containing 10-4mol/L Glivec 30 minutes for blocking ICCs, then repeat the test in advance, the spontaneous contraction of ureteral smooth muscle strips could not bring out, even in Kreb’s solution containing 10-4mol/L carbachol.
(4) M2 and M3 of muscarinic receptor subtypes were detected on ICCs of guinea pig ureter by immunofluorescence (double-labeled).
Conclusion:
Our results indicate that ICCs act as the origin of spontaneous contraction of ureteral smooth muscle and accept regulation of cholinergic nerve through M2 and M3 muscarinic receptor subtypes.
Cajal间质细胞原来被认为是仅存在于胃肠道的一种特殊类型细胞,起着起搏胃肠道慢波、传递电信号、介导肠神经与平滑肌之间调节信号的作用,与病理状态下的多种胃肠动力紊乱疾病(如便秘、假性肠道梗阻等)密切相关,是胃肠道平滑肌节律性收缩功能调节的重要因素。后来,在胃肠道外其他组织中发现了与胃肠道Cajal间质细胞免疫学特性相同、形态学特点相似的细胞,既往曾称之为ICC样细胞(ICC-like cells),目前已统称为Cajal间质细胞(ICCs)。
生理情况下,输尿管平滑肌具有与胃肠道十分相似的特点,即存在不依赖于神经的自发性蠕动,但确切机制目前尚未阐明。目前已有的研究发现,在豚鼠和人的全尿路均存在ICCs。有文献报道,先天性肾盂输尿管梗阻的患儿肾盂和输尿管ICCs较正常儿童明显减少,提示ICCs与输尿管兴奋起源和调控有密切关系。进一步研究ICCs在输尿管自主收缩中的作用及其对神经递质的反应,有助于阐明ICCs在输尿管平滑肌功能调控中的作用及其机制,为上尿路功能障碍性疾病的治疗提供新的切入点。
本课题通过肌条实验观察离体的豚鼠输尿管肌条,在使用非特异性M型胆碱能受体激动剂卡巴可前后自主收缩的变化,并在特异性阻断ICCs功能后,观察肌条的自主收缩及对卡巴可的反应是否发生改变,此外还应用免疫荧光双标染色法检测输尿管ICCs上M受体亚型的表达情况,从而为阐明ICCs上M受体介导的胆碱能信号在输尿管平滑肌自主收缩中的作用提供新的实验依据。
方法:
实验动物均采用清洁级健康Hartley豚鼠。(1)卡巴可对输尿管平滑肌自主收缩的影响及ICCs在此影响中的作用:将20只豚鼠随机分为正常对照组、阻断ICCs组,每组10只。阻断ICCs组的肌条检测前平衡及检测中均置于含终浓度10-4mol/L Glivec的Kreb’s液中,正常对照组置于不含Glivec的Kreb’s液中。取豚鼠左侧输尿管上段1.0cm组织制成离体平滑肌肌条,检测其在终浓度为10-4mol/L卡巴可作用前后的自主收缩频率和幅度。(2)输尿管ICCs上M受体亚型的表达情况:通过c-kit+M1～M5免疫荧光双标染色法检测豚鼠输尿管ICCs上M受体亚型的表达情况。
结果:
(1)在一定前负荷(2.0g)下,加入1mol/L KCl溶液200μL可诱发正常对照组豚鼠的离体输尿管平滑肌肌条出现自主收缩,频率为(3.4±0.84)次/分,波幅为(0.105±0.0228)g。
(2)在肌条出现稳定的自主收缩后,加入0.01mol/L卡巴可100μL,肌条自主收缩频率为(4.9±0.88)次/分,较加药前明显增快(p<0.01);而收缩幅度为(0.106±0.0210)g,较加药前无明显变化(p>0.05)。
(3)用10-4mol/L Glivec平衡30分钟、特异性阻断ICCs后,同法进行检测发现,阻断ICCs组的输尿管肌条无自主收缩出现,加入0.01mol/L卡巴可100μL仍无反应。
(4)免疫荧光双标染色发现,豚鼠输尿管ICCs上表达M2、M3两种M受体亚型。
结论:
ICCs很可能是输尿管平滑肌自主收缩兴奋的起源,并通过M2、M3受体亚型接受胆碱能神经的调控。
Background and Objective:
Interstitial cells of Cajal are the special cells of gastrointestinal tract, which pacing the slow waves of gastrointestinal tract, transmit electrical signals, and mediate signals conduction and integration between nerve cells and smooth muscle cells. As an important factor of adjusting the rhythmic contraction of gastrointestinal smooth muscle, interstitial cells of Cajal have close correlation with some gastrointestinal motility disorders, such as constipation, intestinal pseudo-obstruction. Initially, interstitial cells of Cajal were supposed to exist only in gastrointestinal tract. Recently, interstitial cells of Cajal had been reported existed in other organizations outside gastrointestinal tract. Recent researches showed that morphological and immunological features of interstitial cells of Cajal outside gastrointestinal tract resemble those of gastrointestinal interstitial cells of Cajal. So, interstitial cells of Cajal outside gastrointestinal tract that had been called ICC-like cells were collectively referred to as interstitial cells of Cajal (ICCs) now.
Under normal circumstances, ureteral smooth muscle similar to gastrointestinal smooth muscle, has spontaneous myogenic contraction uncontrolled by nerve. However, the mechanism of spontaneous myogenic stimulation had not yet been fully illuminated. There was a study demonstrated ICCs been located in the entire urinary tract of human and guinea pig. Solari et al had compared the density of ICCs of renal pelvis and ureter between children with congenital ureteropelvic junction obstruction and normal children, and found that the distribution of ICCs of renal pelvis and ureter of normal children was scattered. The number of ICCs of renal pelvis and ureter of normal children was significantly decreased compared with that of children with congenital ureteropelvic junction obstruction. The study suggests that ICCs have close correlation with origin and regulation of ureter’s excitation. Therefore, to further research the role and neuromodulation of ICCs in smooth muscle spontaneous contraction of ureter is helpful to elucidate the role and mechanism of ICCs in the regulation of function of ureteral smooth muscle, and to treat the functional disorders of upper urinary tract.
In this study, we compared the smooth muscle spontaneous contraction of isolated guinea pig ureter by muscle strips experiment before and after the effect of carbachol, a nonspecific agonist of muscarinic receptor. Furthermore, we observed if the smooth muscle spontaneous contraction of isolated guinea pig ureter and its reaction to carbachol had been changed after blocking ICCs. Lastly, immunofluorescence (double-labeled) were used to study the expressions of muscarinic receptor subtypes on ICCs of guinea pig ureter. These results will provide new experimental evidences for elucidating the role of cholinergic signal mediated by muscarinic receptors on ICCs in smooth muscle spontaneous contraction of guinea pig ureter.
Materials and Methods:
The cleaning grade healthy Hartley guinea pigs were sacrificed in this study. Firstly, we investigated the effect of carbachol to smooth muscle spontaneous contraction of guinea pig ureter and the role of ICCs in this effect. 20 guinea pigs were randomly divided into two groups: normal group (n=10) and ICCs inhibited group (n=10). The smooth muscle strips obtained from guinea pigs of ICCs inhibited group were incubated with and measured in Kreb’s solution containing 10-4mol/L Glivec, but those of normal group were incubated with and measured in Kreb’s solution uncontaining Glivec. All of smooth muscle strips were about 1.0 centimeter obtained from superior segment of left guinea pig ureter. Frequency and amplitude of smooth muscle spontaneous contraction were measured in Kreb’s solution containing or uncontaining 10-4mol/L Glivec before and after the effect of 10-4mol/L carbachol. Then, we detected the expressions of muscarinic receptor subtypes on ICCs of guinea pig ureter. Both c-kit receptor and muscarinic receptor subtypes (M1~M5) on ICCs of guinea pig ureter were been observed with laser scanning confocal microscope after been stained by immunofluorescence.
Results:
(1) Spontaneous contractions were recorded on isolated smooth muscle strips of guinea pig ureter under certain preload (2.0g) and when 200μL 1.0mol/L KCl solution was added. The frequency and amplitude of spontaneous contraction were (3.4±0.84)/min and (0.105±0.0228)g respectively.
(2) When 100μL 0.01mol/L carbachol solution was added, the frequency of spontaneous contraction of ureteral smooth muscle strips increased to (4.9±0.88)/min (p<0.01). While the amplitude was (0.106±0.0210)g, and had no statistical significant difference compared with that before using carbachol (p>0.05).
(3) Incubated with Kreb’s solution containing 10-4mol/L Glivec 30 minutes for blocking ICCs, then repeat the test in advance, the spontaneous contraction of ureteral smooth muscle strips could not bring out, even in Kreb’s solution containing 10-4mol/L carbachol.
(4) M2 and M3 of muscarinic receptor subtypes were detected on ICCs of guinea pig ureter by immunofluorescence (double-labeled).
Conclusion:
Our results indicate that ICCs act as the origin of spontaneous contraction of ureteral smooth muscle and accept regulation of cholinergic nerve through M2 and M3 muscarinic receptor subtypes.
引文
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1. Daniel EE. Communication between interstitial cells of Cajal and gastrointestinal muscle. Neurogastroenterol Motil, 2004, 16 Suppl 1: 118-122.
2. Smet PJ, Jonavicius J, Marshall VR, et al. Distribution of nitric oxide synthase-immunoreactive nerves and identification of the cellular targets of nitric oxide in guinea-pig and human urinary bladder by cGMP immunohistochemistry. Neuroscience, 1996, 71(2): 337-348.
3. Klemm MF, Exintaris B, Lang RJ. Identification of the cells underlying pacemaker activity in the guinea-pig upper urinary tract. J Physiol, 1999, 519 pt 3: 867-884.
4. McCloskey KD, Gurney AM. Kit positive cells in the guinea pig bladder. J Urol, 2002, 168(2): 832-836.
5. Ward SM, Sanders KM, Hirst GD. Role of interstitial cells of Cajal in neural control of gastrointestinal smooth muscles. Neurogastroenterol Motil, 2004, 16 Suppl 1: 112-117.
6.彭丽华,杨云生.肥大细胞与功能性胃肠疾病.世界华人消化杂志, 2003, 11(5): 651-653.
7.孙金山,王宝西. Cajal间质细胞与胃肠动力疾病.中国当代儿科杂志, 2006, 8(2): 164-168.
8.李毅,齐清会.胃肠道神经-Cajal间质细胞-平滑肌网络研究进展.国际消化病杂志, 2007, 27(3): 225-227.
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10. Epperson A, Hatton WJ, Callaghan B, et al. Molecular markers expressed in cultured and freshly isolated interstitial cells of Cajal. Am J Physiol Cell Physiol, 2000, 279(2): C529-C539.
11. Van-Nassauw L, Costagliola A, Van-Op-den-Bosch J, et al. Region-specific distribution of the P2Y4 receptor in enteric glial cells and interstitial cells of Cajal within the guinea-pig gastrointestinal tract. Auton Neurosci, 2006, 126-127: 299-306.
12. Ghanem E, Robaye B, Leal T, et al. The role of epithelial P2Y2 and P2Y4 receptors inthe regulation of intestinal chloride secretion. Br J Pharmacol, 2005, 146(3): 364-369.
13. Matos JE, Robaye B, Boeynaems JM, et al. K+ secretion activated by luminal P2Y2 and P2Y4 receptors in mouse colon. J Physiol, 2005, 564 (Pt 1): 269-279.
14. Suarez-Huerta N, Pouillon V, Boeynaems J, et al. Molecular cloning and characterization of the mouse P2Y4 nucleotide receptor. Eur J Pharmacol, 2001, 416(3): 197-202.
15. Burnstock G, Lavin S. Interstitial cells of Cajal and purinergic signalling. Auton Neurosci, 2002, 97(1): 68-72.
16. Wouters MM, Gibbons SJ, Roeder JL, et al. Exogenous serotonin regulates proliferation of interstitial cells of Cajal in mouse jejunum through 5-HT2B receptors. Gastroenterology, 2007, 133(3): 897-906.
17. Lang RJ, Takano H, Davidson ME, et al. Characterization of the spontaneous electrical and contractile activity of smooth muscle cells in the rat upper urinary tract. J Urol, 2001, 166(1): 329-334.
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28. Hashitani H, Suzuki H. Identification of interstitial cells of Cajal in corporal tissues of the guinea-pig penis. Br J Pharmacol, 2004, 141(2): 199-204.
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1. Daniel EE. Communication between interstitial cells of Cajal and gastrointestinal muscle. Neurogastroenterol Motil, 2004, 16 Suppl 1: 118-122.
2. Smet PJ, Jonavicius J, Marshall VR, et al. Distribution of nitric oxide synthase-immunoreactive nerves and identification of the cellular targets of nitric oxide in guinea-pig and human urinary bladder by cGMP immunohistochemistry. Neuroscience, 1996, 71(2): 337-348.
3. Klemm MF, Exintaris B, Lang RJ. Identification of the cells underlying pacemaker activity in the guinea-pig upper urinary tract. J Physiol, 1999, 519 pt 3: 867-884.
4. McCloskey KD, Gurney AM. Kit positive cells in the guinea pig bladder. J Urol, 2002, 168(2): 832-836.
5. Ward SM, Sanders KM, Hirst GD. Role of interstitial cells of Cajal in neural control of gastrointestinal smooth muscles. Neurogastroenterol Motil, 2004, 16 Suppl 1: 112-117.
6.彭丽华,杨云生.肥大细胞与功能性胃肠疾病.世界华人消化杂志, 2003, 11(5): 651-653.
7.孙金山,王宝西. Cajal间质细胞与胃肠动力疾病.中国当代儿科杂志, 2006, 8(2): 164-168.
8.李毅,齐清会.胃肠道神经-Cajal间质细胞-平滑肌网络研究进展.国际消化病杂志, 2007, 27(3): 225-227.
9. Orellana JA, Palacios-Prado N, Saez JC. Chlorpromazine reduces the intercellular communication via gap junctions in mammalian cells. Toxicol Appl Pharmacol, 2006, 213(3): 187-197.
10. Epperson A, Hatton WJ, Callaghan B, et al. Molecular markers expressed in cultured and freshly isolated interstitial cells of Cajal. Am J Physiol Cell Physiol, 2000, 279(2): C529-C539.
11. Van-Nassauw L, Costagliola A, Van-Op-den-Bosch J, et al. Region-specific distribution of the P2Y4 receptor in enteric glial cells and interstitial cells of Cajal within the guinea-pig gastrointestinal tract. Auton Neurosci, 2006, 126-127: 299-306.
12. Ghanem E, Robaye B, Leal T, et al. The role of epithelial P2Y2 and P2Y4 receptors inthe regulation of intestinal chloride secretion. Br J Pharmacol, 2005, 146(3): 364-369.
13. Matos JE, Robaye B, Boeynaems JM, et al. K+ secretion activated by luminal P2Y2 and P2Y4 receptors in mouse colon. J Physiol, 2005, 564 (Pt 1): 269-279.
14. Suarez-Huerta N, Pouillon V, Boeynaems J, et al. Molecular cloning and characterization of the mouse P2Y4 nucleotide receptor. Eur J Pharmacol, 2001, 416(3): 197-202.
15. Burnstock G, Lavin S. Interstitial cells of Cajal and purinergic signalling. Auton Neurosci, 2002, 97(1): 68-72.
16. Wouters MM, Gibbons SJ, Roeder JL, et al. Exogenous serotonin regulates proliferation of interstitial cells of Cajal in mouse jejunum through 5-HT2B receptors. Gastroenterology, 2007, 133(3): 897-906.
17. Lang RJ, Takano H, Davidson ME, et al. Characterization of the spontaneous electrical and contractile activity of smooth muscle cells in the rat upper urinary tract. J Urol, 2001, 166(1): 329-334.
18. Lang RJ, Hashitani H, Keller S, et al. Modulators of internal Ca2+ stores and the spontaneous electrical and contractile activity of the guinea-pig renal pelvis. Br J Pharmacol, 2002, 135(6): 1363-1374.
19. Pezzone MA, Watkins SC, Alber SM, et al. Identification of c-kit-positive cells in the mouse ureter: the interstitial cells of Cajal of the urinary tract. Am J Physiol Renal Physiol, 2003, 284(5): F925-F929.
20. Davidson RA, McCloskey KD. Morphology and localization of interstitial cells in the guinea pig bladder: structural relationships with smooth muscle and neurons. J Urol, 2005, 173(4): 1385-1390.
21. Piaseczna-Piotrowska A, Rolle U, Solari V, et al. Interstitial cells of Cajal in the human normal urinary bladder and in the bladder of patients with megacystis-microcolon intestinal hypoperistalsis syndrome. BJU Int, 2004, 94(1): 143-146.
22. Hashitani H, Fukuta H, Takano H, et al. Origin and propagation of spontaneous excitation in smooth muscle of the guinea-pig urinary bladder. J Physiol, 2001, 530(Pt 2): 273-286.
23. Kubota Y, Hashitani H, Fukuta H, et al. Role of mitochondria in the generation ofspontaneous activity in detrusor smooth muscles of the Guinea pig bladder. J Urol, 2003, 170(2 Pt 1): 628-633.
24. Exintaris B, Klemm MF, Lang RJ. Spontaneous slow wave and contractile activity of the guinea pig prostate. J Urol, 2002, 168(1): 315-322.
25. Sergeant GP, Hollywood MA, McCloskey KD, et al. Specialised pacemaking cells in the rabbit urethra. J Physiol, 2000, 526 Pt 2: 359-366.
26. Ciontea SM, Radu E, Regalia T, et al. C-kit immunopositive interstitial cells (Cajal-type) in human myometrium. J Cell Mol Med, 2005, 9(2): 407-420.
27. Popescu LM, Ciontea SM, Cretoiu D, et al. Novel type of interstitial cell (Cajal-like) in human fallopian tube. J Cell Mol Med, 2005, 9(2): 479-523.
28. Hashitani H, Suzuki H. Identification of interstitial cells of Cajal in corporal tissues of the guinea-pig penis. Br J Pharmacol, 2004, 141(2): 199-204.