Determining the Molecular Mechanisms of Vascular Sympathetic Innervation

确定血管交感神经支配的分子机制

• 批准号: 7600367
• 负责人: Jennifer B Long
• 金额: $ 0.72万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-03-05 至 2009-09-04
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7600367
• 关键词: 3-Dimensional; Actins; Adult; Age; Animals; Arteries; Automobile Driving; Axon; Bacteria; Biological Assay; Blocking Antibodies; Blood Vessels; Blood capillaries; Blood flow; CD31 Antigens; Candidate Disease Gene; Cardiac; Carotid Arteries; Cavia; Cells; Coculture Techniques; Collagen; Coronary Vessels; Cues; Culture Media; Denervation; Development; Disease; Distal; Ear; Embryo; Endothelial Cells; Environment; Etiology; Exercise; Gel; Gene Expression; Genes; Hypertension; Immunofluorescence Immunologic; In Vitro; Individual; Length; Life; Mediating; Mesentery; Messenger RNA; Microspheres; Modeling; Molecular; Molecular Profiling; Mus; Names; Nerve; Nerve Plexus; Neural Crest Cell; Neurites; Neurons; Organ; Oryctolagus cuniculus; Parkinson Disease; Pathway interactions; Patients; Pattern; Peripheral; Play; RNA Interference; Rattus; Role; Sampling; Sequence Homology; Series; Smooth Muscle; Stress; Structure of superior cervical ganglion; Sympathetic Ganglia; Sympathetic Nervous System; Synapses; Synaptophysin; Techniques; Temperature; Time; Tissue Sample; Tissues; Transcript; Transplantation; Travel; Tyrosine 3-Monooxygenase; Variant; Varicosity; Vascular resistance; Veins; Western Blotting; axon guidance; base; blood pressure regulation; capillary; cell type; cremaster muscle; density; design; ethylenevinylacetate copolymer; feeding; femoral artery; in vivo; knock-down; mRNA Expression; migration; nerve supply; neuroblast; neurogenesis; neuron development; peripheral blood; postnatal; protein expression; receptor; repaired; research study; response; spatiotemporal; synaptogenesis; venule

DESCRIPTION (provided by applicant): The sympathetic nervous system is integral to blood pressure regulation through changes in peripheral vascular resistance. Sympathetic nerves typically innervate major feed arteries and extend through the precapillary arteriolar network, but not into capillaries, venules or collecting veins. During development, axon extension closely follows arteries as neurons approach and innervate target organs. This close proximity to blood vessels suggests that vessel derived molecules may play a role in axon outgrowth and target innervation. Due to the selective sympathetic innervation of pre-capillary vessels, our hypothesis is that molecular cues that could promote chemoattraction between arteries and neurons or chemorepulsion between veins and neurons exist and may be expressed by vascular cells. Our specific aims are designed to allow us to identify candidate molecules that may drive selective vascular sympathetic innervation and evaluate their functions in vitro. We will (1) characterize sympathetic innervation patterns and synapse formation by immunofluorescence in several blood vessels in late embryonic and postnatal animals, (2) conduct a subtractive hybridization of innervated and non-innervated vessels to generate a list of differentially expressed candidate molecules and validate their expression through western blot and immunofluorescence and (3) evaluate the functional impacts of any identified candidate molecules using a three-dimensional (3-D) co-culture model using superior cervical ganglion explants with innervated and non-innervated vessel segments to assess directed neurite outgrowth in response to the application of identified candidate molecules to the culture medium. Relevance: Alterations in vascular sympathetic has been implicated in diseases such as hypertension (hyperinnervation) and the pathological sequalae following cardiac transplant (denervation). For example, loss of sympathetic neurons after transplant compromises the ability to regulate peripheral blood flow and to adequately tolerate exercise, temperature fluctuations and other physical stresses. By understanding the molecular mechanisms governing vascular sympathetic innervation, therapies may be developed that would allow for the repair of peripheral vascular sympathetic nerve patterning and therefore allow patients to regain normal function.

描述(申请人提供)：交感神经系统是通过改变外周血管阻力来调节血压的不可或缺的部分。交感神经通常支配主要的供血动脉，并通过毛细血管前小动脉网络延伸，但不进入毛细血管、小静脉或集合静脉。在发育过程中，随着神经元接近并支配靶器官，轴突延伸密切跟随动脉。这种接近血管的情况表明，血管衍生分子可能在轴突生长和靶神经支配中发挥作用。由于毛细血管前血管的选择性交感神经支配，我们的假设是，可能存在促进动脉与神经元之间的化学吸引或静脉与神经元之间的化学排斥的分子线索，并可能由血管细胞表达。 我们的具体目标是使我们能够识别可能驱动选择性血管交感神经支配的候选分子，并在体外评估它们的功能。我们将(1)通过免疫荧光鉴定胚胎晚期和出生后动物几个血管中的交感神经支配模式和突触形成，(2)对神经支配和非神经支配的血管进行消减杂交，以生成差异表达的候选分子列表，并通过蛋白质印迹和免疫荧光验证它们的表达，(3)使用具有神经支配和非神经支配的血管片段的颈上神经节外植体建立三维(3-D)共培养模型，评估任何已确定的候选分子的功能影响，以评估响应于将已确定的候选分子应用到培养液中而定向突起生长。 相关性：血管交感神经的改变与高血压(过度神经支配)和心脏移植后的病理后遗症(去神经支配)等疾病有关。例如，移植后交感神经元的丧失损害了调节外周血流量和充分耐受运动、温度波动和其他身体压力的能力。通过了解支配血管交感神经的分子机制，可以开发出允许修复周围血管交感神经模式的治疗方法，从而使患者恢复正常功能。