Campos Rafael, Mónica Fabíola Z, Rodrigues Renata Lopes, Rojas-Moscoso Julio Alejandro, Moreno Ronilson Agnaldo, Cogo José Carlos, de Oliveira Marco Antonio, Antunes Edson, De Nucci Gilberto
Faculty of Medical Sciences, Department of Pharmacology, University of Campinas (UNICAMP), Campinas, Brazil.
Faculty of Health Sciences, Department of Pharmacology, FACISB, Barretos, Brazil.
PLoS One. 2017 Aug 24;12(8):e0183766. doi: 10.1371/journal.pone.0183766. eCollection 2017.
Reptiles are the first amniotes to develop an intromitent penis, however until now the mechanisms involved in the electrical field stimulation-induced contraction on corpora cavernosa isolated from Crotalus durissus terrificus were not investigated. Crotalus and rabbit corpora cavernosa were mounted in 10 mL organ baths for isometric tension recording. Electrical field stimulation (EFS)-induced contractions were performed in presence/absence of phentolamine (10 μM), guanethidine (30 μM), tetrodotoxin (1 μM and 1mM), A-803467 (10 μM), 3-iodo-L-Tyrosine (1 mM), salsolinol (3 μM) and a modified Krebs solution (equimolar substitution of NaCl by N-methyl-D-glucamine). Immuno-histochemistry for tyrosine hydroxylase was also performed. Electrical field stimulation (EFS; 8 Hz and 16 Hz) caused contractions in both Crotalus and rabbit corpora cavernosa. The contractions were abolished by previous incubation with either phentolamine or guanethidine. Tetrodotoxin (1 μM) also abolished the EFS-induced contractions of rabbit CC, but did not affect EFS-induced contractions of Crotalus CC. Addition of A-803467 (10 μM) did not change the EFS-induced contractions of Crotalus CC but abolished rabbit CC contractions. 3-iodo-L-Tyrosine and salsolinol had no effect on EFS-induced contractions of Crotalus CC and Rabbit CC. Replacement of NaCl by N- Methyl-D-glucamine (NMDG) abolished EFS-induced contractions of rabbit CC, but did not affect Crotalus CC. The presence of tyrosine hydroxylase was identified in endothelial cells only of Crotalus CC. Since the EFS-induced contractions of Crotalus CC is dependent on catecholamine release, insensitive to TTX, insensitive to A803467 and to NaCl replacement, it indicates that the source of cathecolamine is unlikely to be from adrenergic terminals. The finding that tyrosine hydroxylase is present in endothelial cells suggests that these cells can modulate Crotalus CC tone.
爬行动物是最早发育出可插入阴茎的羊膜动物,然而到目前为止,尚未对电场刺激诱导的杜氏响尾蛇离体海绵体收缩所涉及的机制进行研究。将响尾蛇和兔子的海绵体置于10 mL器官浴槽中进行等长张力记录。在存在/不存在酚妥拉明(10 μM)、胍乙啶(30 μM)、河豚毒素(1 μM和1 mM)、A-803467(10 μM)、3-碘-L-酪氨酸(1 mM)、去甲猪毛菜碱(3 μM)以及一种改良的克雷布斯溶液(用N-甲基-D-葡糖胺等摩尔替代NaCl)的情况下进行电场刺激(EFS)诱导的收缩实验。还进行了酪氨酸羟化酶的免疫组织化学检测。电场刺激(EFS;8 Hz和16 Hz)在响尾蛇和兔子的海绵体中均引起收缩。预先用酚妥拉明或胍乙啶孵育可消除收缩。河豚毒素(1 μM)也消除了EFS诱导的兔子海绵体收缩,但不影响EFS诱导的响尾蛇海绵体收缩。添加A-803467(10 μM)不会改变EFS诱导的响尾蛇海绵体收缩,但消除了兔子海绵体收缩。3-碘-L-酪氨酸和去甲猪毛菜碱对EFS诱导的响尾蛇海绵体和兔子海绵体收缩均无影响。用N-甲基-D-葡糖胺(NMDG)替代NaCl消除了EFS诱导的兔子海绵体收缩,但不影响响尾蛇海绵体。仅在响尾蛇海绵体的内皮细胞中鉴定出酪氨酸羟化酶的存在。由于EFS诱导的响尾蛇海绵体收缩依赖于儿茶酚胺释放,对河豚毒素不敏感,对A803467不敏感且对NaCl替代不敏感,这表明儿茶酚胺的来源不太可能是肾上腺素能神经末梢。酪氨酸羟化酶存在于内皮细胞中的这一发现表明这些细胞可以调节响尾蛇海绵体的张力。