In Vivo Ca2+ and Voltage Imaging on The Urinary Bladder

膀胱体内 Ca2 和电压成像

基本信息

批准号：
7197712
负责人：
Michael I. Kotlikoff
金额：
$ 32.16万
依托单位：
CORNELL UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2007
资助国家：
美国
起止时间：
2007-02-01 至 2011-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7197712
关键词：
Affect Bacterial Artificial Chromosomes Bladder Cardiac Cells Cholinergic Agents Complex Condition Coupling Data Development Disease Frequencies Functional disorder Gastrointestinal tract structure Gene Transfer Techniques Generations Genetic Green Fluorescent Proteins Heart Hormonal Hyperactive behavior Hypertrophy Image Imagery Imaging technology In Vitro Incontinence Individual Interstitial Cell of Cajal Life Maps Measurement Measures Methods Molecular Monitor Morbidity - disease rate Mus Muscle Muscle Cells Nature Nerve Noise Obstruction Organ Overactive Bladder Pacemakers Pattern Periodicity Physiological Play Property Quality of life Regulation Resolution Role Ryanodine Receptor Calcium Release Channel Sarcoplasmic Reticulum Secondary to Signal Transduction Smooth Muscle Smooth Muscle Myocytes Source Surface Symptoms Syndrome Techniques Temperature Testing Transgenic Mice Urge Incontinence Urinary tract Urination base cholinergic cholinergic neuron in vivo micturition urgency molecular scale novel psychologic relating to nervous system selective expression social tool voltage

项目摘要

DESCRIPTION (provided by applicant): Postobstructive urinary tract dysfunction is a disorder with extremely high morbidity and devastating social and psychological consequence, resulting in a complex of symptoms including incontinence, increased frequency of urination, and an inappropriate sense of urinary urgency, all of which appear to result from abnormal contractile activity of the smooth muscle lining the bladder. In this proposal we seek to understand the generation of rhythmic or phasic contractile activity in the normal and postobstructed urinary bladder in vivo. Although the study of the cellular signals underlying phasic activity is extraordinarily difficult in vivo, it is important to understand this activity in the context of neural and hormonal inputs that play an important role in the regulation of contractile activity. We have recently developed methods to monitor intracellular free Ca2+ in vivo through the development of transgenic mice expressing a novel high signal-noise, genetically encoded Ca2+ indicator. GCaMP2, the Ca2+ indicator, is stable at physiological temperatures and approaches the brightness of GFP, enabling sustained, high resolution recording of cellular Ca2+ transients in the living mouse. Mice in which this molecule is highly expressed in urinary bladder smooth muscle have been created and will be used to visualize cellular Ca2+ signals in vivo. We will use these mice to more fully understand the dysfunction associated with urinary tract outlet obstruction. The basis of abnormal contractile activity in postobstructive mice that develop spontaneous bladder overactivity will be studied to understand the cellular and molecular basis for abnormal bladder contractions. We hypothesize that bladder hyperactivity occurs secondary to the development of abnormal pacemaker activity and anomalous Ca2+ waves associated with dysregulated Ca2+ release from the sarcoplasmic reticulum (SR) of smooth muscle or Interstitial Cells of Cajal (ICC). To test this hypothesis, we will image phasic Ca2+ signals in smooth muscle cells under normal conditions, seek to understand the cellular and molecular basis for spontaneous activity, determine the extent to which this pattern is disturbed by sustained outflow obstruction, and test whether abnormal activity arises due to alterations in SR Ca2+ release in muscle or changes in the activity of pacemaker cells in the urinary bladder. These findings should provide a clearer understanding of smooth muscle dysfunction associated with urinary tract outflow obstruction.

描述（由申请人提供）：射击后尿路功能障碍是一种疾病，具有极高的发病率和毁灭性的社会和心理后果，导致症状复杂，包括尿失禁，排尿频率的增加以及尿液的不适当感，所有这些症状似乎都是由于平稳性lards linders ladder linder linder linders lind linder linder lind linders ladder ladder ladder ladder ladder ladder ladder ladder ladder ladder ladder ladder ladder ladder ladder的均匀效果。在该提案中，我们试图了解体内正常和后杀后膀胱中有节奏或阶段性收缩活动的产生。尽管体内对基础阶段活性的细胞信号的研究非常困难，但在神经和荷尔蒙输入的背景下，重要的是要了解这种活性，这在调节收缩活性中起着重要作用。我们最近开发了通过表达新型高信号噪声，遗传编码的Ca2+指标的转基因小鼠的发展来监测细胞内游离Ca2+体内的方法。 Ca2+指示剂GCAMP2在生理温度下是稳定的，并接近GFP的亮度，从而实现了活小鼠中细胞Ca2+瞬变的持续，高分辨率记录。已经创建了该分子高度表达该分子的小鼠，并将用于可视化体内细胞Ca2+信号。我们将使用这些小鼠更充分地了解与尿路出口阻塞相关的功能障碍。将研究发生自发性膀胱过度活动的后杀性小鼠收缩活性异常的基础，以了解异常膀胱收缩的细胞和分子基础。我们假设膀胱多动症发生在异常起搏器活性的发展以及与Cajal（ICC）平滑肌或间质细胞的肌肉网状肌肉或间质细胞中的肌肉网状（SR）相关的异常Ca2+波。为了检验这一假设，我们将在正常条件下在平滑肌细胞中成像阶段性CA2+信号，试图了解自发活性的细胞和分子基础，确定该模式在持续流出的流出阻塞受到干扰的程度，并测试由于SR CA2+在SR CA2+释放中是否会在Pacem pacemaker of Pacemaker in urearional Blad的肌肉中发生变化而导致的异常活性。这些发现应更清楚地了解与尿路流出阻塞相关的平滑肌功能障碍。