Intraspinal circuits supporting synergy between the bladder and urethral sphincter in mice

支持小鼠膀胱和尿道括约肌协同作用的椎内回路

• 批准号: 10662280
• 负责人: SERGEI V KARNUP
• 金额: $ 34.98万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-07-01 至 2025-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10662280
• 关键词: 3-Dimensional; Anatomy; Anesthesia procedures; Animals; Area; Ataxia; Axon; Bladder; Brain; C57BL/10 Mouse; Cell Nucleus; Chronic; Development; Electric Stimulation; Electrophysiology (science); Exhibits; Feedback; Female; Functional disorder; Future; Gender; Generations; Genetic; Glutamates; Goals; Horns; In Vitro; Injections; Interneurons; Interruption; Knowledge; Label; Light; Lower urinary tract; Mammals; Maps; Mediating; Methods; Micturition Reflex; Modification; Motor Neurons; Mus; Muscle; Neurons; Neurotransmitters; Optics; Pathway interactions; Periodicity; Persons; Pontine structure; Population; Preparation; Rattus; Reaction; Recommendation; Recovery; Recovery of Function; Recurrence; Reflex action; Regulation; Relaxation; Research Project Grants; Residual state; Rhodopsin; Role; Sex Differences; Slice; Sphincter; Spinal; Spinal Cord; Spinal Cord Diseases; Spinal cord injury; Structure; Synapses; Synaptic Potentials; Techniques; Testing; Therapeutic; Therapeutic Intervention; Tracer; Transgenic Mice; Urethane; Urethral sphincter; Urinary Retention; Urination; Urine; Vertebral column; Viral; biocytin; cholinergic neuron; clinical practice; design; efficacy evaluation; excitatory neuron; experimental study; gender difference; improved; in vivo; inhibitory neuron; injury recovery; insight; intravesical; mad itch virus; male; mouse model; neural; neural circuit; neuronal circuitry; neuroregulation; novel therapeutics; optogenetics; pharmacologic; postsynaptic; pressure; response; restoration; sex; sexual dimorphism; side effect; spinal pathway; spinal tract; synergism; therapeutically effective; urologic

Project summary Detrusor-sphincter-dyssynergia (DSD) is a major urological problem inducing inefficient voiding, increased amount of post-void residual urine and high intravesical pressure after spinal cord injury (SCI). Until recently the majority of studies on spinal mechanisms of coordination between external urethral sphincter (EUS) and the bladder (BL) were conducted in female rats, although in clinical practice DSD develops more often in males. Furthermore, sexual dimorphism in anatomy and function of the lower urinary tract (LUT) suggests differences in the spinal neural circuits of male and female. With development of genetic modifications and optogenetic techniques the mouse model of LUT dysfunction is becoming more useful than the rat. Therefore, in the proposed project “Intraspinal circuits supporting synergy between the bladder and urethral sphincter in mice” we will use transgenic mice of both sexes for electrophysiological, optogenetic, immunohistochemical, pharmacological and anatomical studies of spinal cord circuits involved in interaction between the bladder (BL) and the external urethral sphincter (EUS) in spinal intact animals and after spinal cord injury. We will test several hypotheses in in vivo and in vitro experiments. In urethane anesthetized animals of both sexes, we will determine functional differences in spinal regulation of micturition and in control and SCI mice. Male and female animals will be also used to determine hypothesized sexual dimorphism in LUT-related spinal circuits and their plasticity after chronic SCI. Using electrical stimulation of the Lumbar Spinal Coordinating Center (LSCC) which we have recently discovered in L3/L4 spinal segments, we will test whether an external command can initiate voiding. Using trans-synaptic viral labeling, we will trace neuronal populations involved in coordination between EUS and BL to determine structural and functional differences in LUT spinal networks between males and females. In spinal cord slice preparations, we will use transgenic mice expressing channel rhodopsin (ChR2) in inhibitory or excitatory neurons to study reactions of EUS motoneurons to light-evoked activity of LUT-related spinal interneurons embedded in specific spinal pathways, and will identify and characterize the inhibitory circuit which is responsible for EUS relaxation. We will define the role of the L3/L4 LSCC in activity of EUS motoneurons and EUS relaxation and its mode of interaction with L6/S1 motor neuronal pool. We will evaluate the contribution of a hypothesized recurrent inhibitory circuit in L6/S1 to the generation of EUS bursting and relaxation as well as to bladder-sphincter coordination. The long-term objectives of the research project are to increase our understanding of the pathophysiological mechanisms inducing DSD and voiding problems after SCI and to develop new and effective therapeutic interventions for the treatment of DSD in spinal cord disorders. Considering gender related differences in neural control of the LUT different approaches to DSD treatment in males and females may be recommended.

项目摘要 逼尿肌括约肌协同失调（DSD）是一个主要的泌尿系统问题，导致排尿效率低下，增加 脊髓损伤后排尿后残余尿量和膀胱内压增高。直到最近 大多数关于外尿道括约肌（EUS）与脊髓之间协调机制的研究， 在雌性大鼠中进行了膀胱（BL）试验，尽管在临床实践中，DSD更常在雄性大鼠中发生。 此外，下尿路（LUT）的解剖和功能的性别二型性表明差异 在男性和女性的脊髓神经回路中。随着遗传修饰和光遗传学的发展， 技术，LUT功能障碍的小鼠模型变得比大鼠更有用。因此在 建议的项目“脊柱内电路支持膀胱和尿道括约肌之间的协同作用， 小鼠”我们将使用两种性别的转基因小鼠进行电生理学，光遗传学，免疫组织化学， 参与膀胱（BL）之间相互作用的脊髓回路的药理学和解剖学研究 以及脊髓完整动物和脊髓损伤后的尿道外括约肌（EUS）。 我们将在体内和体外实验中测试几个假设。在乌拉坦麻醉的动物中， 两种性别，我们将确定脊髓排尿调节功能的差异，并在控制和SCI 小鼠雄性和雌性动物也将用于确定LUT相关的假设性二态性。 慢性脊髓损伤后脊髓回路及其可塑性电刺激对腰椎协调功能的影响 中心（LSCC），我们最近发现在L3/L4脊柱节段，我们将测试是否外部 命令可以启动作废。使用跨突触病毒标记，我们将追踪参与神经元群体， EUS和BL之间的协调，以确定LUT脊髓网络的结构和功能差异 男性和女性之间。在脊髓切片制备中，我们将使用表达通道的转基因小鼠， 视紫红质（ChR 2）在抑制性或兴奋性神经元研究EUS运动神经元对光诱发的反应 活动的LUT相关的脊髓中间神经元嵌入特定的脊髓通路，并将确定和 表征负责EUS松弛的抑制回路。我们将定义L3/L4的角色 喉鳞状细胞癌EUS运动神经元的活动、EUS松弛及其与L 6/S1运动神经元的相互作用 池我们将评估L 6/S1中假设的复发性抑制回路对产生 EUS爆破和松弛以及膀胱括约肌协调。 该研究项目的长期目标是增加我们对 脊髓损伤后引起DSD和排尿问题的病理生理机制，并开发新的有效的 治疗脊髓疾病DSD的治疗干预措施。考虑性别相关 LUT神经控制的差异，男性和女性中DSD治疗的不同方法可能是 推荐

项目成果

Spinal interneurons of the lower urinary tract circuits.

• DOI: 10.1016/j.autneu.2021.102861
• 发表时间: 2021-11
• 期刊: Autonomic neuroscience : basic & clinical
• 作者: Karnup S

Current Knowledge and Novel Frontiers in Lower Urinary Tract Dysfunction after Spinal Cord Injury: Basic Research Perspectives.

脊髓损伤后尿路功能障碍的当前知识和新型边界：基础研究观点。

• DOI: 10.4103/uros.uros_31_22
• 发表时间: 2022-07
• 期刊: Urological science
• 影响因子: 0.5
• 作者: Wada N;Karnup S;Kadekawa K;Shimizu N;Kwon J;Shimizu T;Gotoh D;Kakizaki H;de Groat WC;Yoshimura N
• 通讯作者: Yoshimura N

Molecular Mechanisms of Neurogenic Lower Urinary Tract Dysfunction after Spinal Cord Injury.

• DOI: 10.3390/ijms24097885
• 发表时间: 2023-04-26
• 期刊: INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
• 影响因子: 5.6
• 作者: Shimizu, Nobutaka;Saito, Tetsuichi;Wada, Naoki;Hashimoto, Mamoru;Shimizu, Takahiro;Kwon, Joonbeom;Cho, Kang Jun;Saito, Motoaki;Karnup, Sergei;de Groat, William C.;Yoshimura, Naoki
• 通讯作者: Yoshimura, Naoki