Bhattacharya Anindya, Dang Hong, Zhu Quan-Ming, Schnegelsberg Birthe, Rozengurt Nora, Cain Gary, Prantil Rachelle, Vorp David A, Guy Nicholas, Julius David, Ford Anthony P D W, Lester Henry A, Cockayne Debra A
Roche Pharmaceuticals, Palo Alto, California 94304, USA.
J Neurosci. 2004 Jun 16;24(24):5537-48. doi: 10.1523/JNEUROSCI.5658-03.2004.
Mutant mice with a hypersensitive serotonin (5-HT)3A receptor were generated through targeted exon replacement. A valine to serine mutation (V13'S) in the channel-lining M2 domain of the 5-HT3A receptor subunit rendered the 5-HT3 receptor 70-fold more sensitive to serotonin and produced constitutive activity when combined with the 5-HT3B subunit. Mice homozygous for the mutant allele (5-HT3Avs/vs) had decreased levels of 5-HT3A mRNA. Measurements on sympathetic ganglion cells in these mice showed that whole-cell serotonin responses were reduced, and that the remaining 5-HT3 receptors were hypersensitive. Male 5-HT3Avs/vs mice died at 2-3 months of age, and heterozygous (5-HT3Avs/+) males and homozygous mutant females died at 4-6 months of age from an obstructive uropathy. Both male and female 5-HT3A mutant mice had urinary bladder mucosal and smooth muscle hyperplasia and hypertrophy, whereas male mutant mice had additional prostatic smooth muscle and urethral hyperplasia. 5-HT3A mutant mice had marked voiding dysfunction characterized by a loss of micturition contractions with overflow incontinence. Detrusor strips from 5-HT3Avs/vs mice failed to contract to neurogenic stimulation, despite overall normal responses to a cholinergic agonist, suggestive of altered neuronal signaling in mutant mouse bladders. Consistent with this hypothesis, decreased nerve fiber immunoreactivity was observed in the urinary bladders of 5-HT3Avs/vs compared with 5-HT3A wild-type (5-HT3A+/+) mice. These data suggest that persistent activation of the hypersensitive and constitutively active 5-HT3A receptor in vivo may lead to excitotoxic neuronal cell death and functional changes in the urinary bladder, resulting in bladder hyperdistension, urinary retention, and overflow incontinence.
通过靶向外显子替换产生了具有超敏5-羟色胺(5-HT)3A受体的突变小鼠。5-HT3A受体亚基的通道内衬M2结构域中的缬氨酸到丝氨酸突变(V13'S)使5-HT3受体对5-羟色胺的敏感性提高了70倍,并在与5-HT3B亚基结合时产生组成性活性。突变等位基因纯合的小鼠(5-HT3Avs/vs)5-HT3A mRNA水平降低。对这些小鼠交感神经节细胞的测量表明,全细胞5-羟色胺反应降低,并且剩余的5-HT3受体超敏。雄性5-HT3Avs/vs小鼠在2至3个月龄时死亡,杂合子(5-HT3Avs/+)雄性和纯合突变雌性在4至6个月龄时死于梗阻性尿路病。雄性和雌性5-HT3A突变小鼠均有膀胱黏膜和平滑肌增生及肥大,而雄性突变小鼠还有前列腺平滑肌和尿道增生。5-HT3A突变小鼠有明显的排尿功能障碍,其特征为排尿收缩丧失并伴有充溢性尿失禁。来自5-HT3Avs/vs小鼠的逼尿肌条对神经源性刺激无收缩反应,尽管对胆碱能激动剂的总体反应正常,这提示突变小鼠膀胱中的神经元信号传导发生了改变。与该假设一致,与5-HT3A野生型(5-HT3A+/+)小鼠相比,在5-HT3Avs/vs小鼠的膀胱中观察到神经纤维免疫反应性降低。这些数据表明,体内超敏且组成性激活的5-HT3A受体的持续激活可能导致兴奋性毒性神经元细胞死亡和膀胱功能改变,从而导致膀胱过度扩张、尿潴留和充溢性尿失禁。