PACAP Modulation of Ca2+ Activity in Neonatal Mouse OB

PACAP 对新生小鼠 OB 中 Ca2 活性的调节

• 批准号: 8603235
• 负责人: Mavis Amity Irwin
• 金额: $ 1.53万
• 依托单位: UNIVERSITY OF UTAH
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-12-01 至 2014-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8603235
• 关键词: Acute; Adult; Affect; Auditory; Autistic Disorder; Behavior; Birth; Body Temperature; Brain region; Cells; Cerebellum; Chemical Stimulation; Cytoplasmic Granules; Data; Development; Disease; Foundations; Future; G-Protein-Coupled Receptors; Gene Mutation; Hippocampus (Brain); Image; Immigration; Injury; Interneurons; Knock-out; Learning; Life; Maps; Mediating; Migration Assay; Monitor; Mus; Neonatal; Neurodegenerative Disorders; Neurons; Neuropeptides; Neurotransmitters; Newborn Infant; Nipples; Odors; Olfactory Epithelium; PACAPR-1 protein; Pathway interactions; Peptides; Physiological; Play; Proteins; Role; Schizophrenia; Sensory; Slice; System; Temperature; Testing; Therapeutic; Therapeutic Uses; Time; Transgenic Mice; Visual; Weaning; adult neurogenesis; base; cell type; central nervous system injury; density; dentate gyrus; design; feeding; gamma-Aminobutyric Acid; granule cell; insight; knockout animal; migration; natural hypothermia; neonatal death; nervous system disorder; neuroblast; neuron development; neuronal circuitry; neuronal survival; neurotransmitter release; new therapeutic target; novel; olfactory bulb; pituitary adenylate cyclase activating polypeptide; postnatal; prevent; pup; receptor; receptor expression; response; sensory system

Project Summary Pituitary Adenylate Cyclase-Activating Polypeptide (PACAP) is a pleiotropic peptide with functions in development and throughout life [4]. PACAP knockout (KO) animals will die shortly after birth due to an inability to maintain core body temperature [5]. Our observation that preventing hypothermia in PACAP KO pups does not eliminate poor survival suggests that other functions such as olfactory mediated feeding may be disturbed. Both PACAP and its G-protein coupled receptor (PAC1) are highly expressed in the olfactory bulb (OB) throughout life suggesting PACAP plays an important role in the development, survival and replacement of neurons within the bulb. Surprisingly, there have been no studies on the role of PACAP in modulating the Ca[2+] activity, migration, and maturation of developing OB neurons. In other CNS regions, Ca[2+] activity in the form of spontaneous oscillations is critical for the establishment and survival of developing neuronal circuitry, especially prior to sensory input [6]. GABA is excitatory in immature neurons and has been implicated in this early developmental activity but the control of maturation in olfactory interneurons is poorly understood. However, our preliminary data show that PACAP induces transient and sustained Ca[2+] oscillations in multiple cell types of the neonatal OB. Our focus in the first aim will be on the developing GABAergic granule cells (GCs) because they are thought to express PAC1Rs and can be identified using GAD2- and Dlx2- tdTomato transgenic mice. The developing GCs will be evaluated for PACAP responses and their maturity tested with GABA, which will be an excitatory neurotransmitter for the immature cells which maintain high intracellular Cl- levels and depolarize with GABA stimulation. The second aim will be determining the role of PACAP in the migration and maturation of immature OB interneurons. It will be directly accomplished using migration assays on cultured GAD2 or Dlx2 tdTomato mice OB cells. Morphological changes associated with maturation will also be examined. This proposal will examine the role of PACAP during early postnatal development of OB circuitry and is based on studies of PACAP and PAC1 receptor expression in the OB and other brain regions. However, my studies are the first dynamic imaging studies of PACAP in identified cells of neonatal OB. Because little is known about the function of PACAP in the OB, the emphasis of my proposal will be to lay the foundation for future understanding of how PACAP affects the development of OB circuitry. A number of neurological diseases such as autism, bipolar disease, and schizophrenia are thought to originate from problems in early development of neuronal circuits. Understanding the role of PACAP in early circuitry development may provide a novel therapeutic target for these devastating diseases. Furthermore, understanding how PACAP affects developing interneurons may aid in designing strategies for adult neurogenesis following injury or disease.

项目摘要 垂体腺苷酸环化酶激活多肽（PACAP）是一种多效肽，具有发育和整个生命的功能[4]。 PACAP敲除（KO）动物由于无法维持核心体温而出生后不久[5]。我们的观察结果是，预防PACAP KO PUP中的体温过低并不能消除较差的生存表明其他功能，例如嗅觉介导的喂养可能会受到干扰。 PACAP及其G蛋白偶联受体（PAC1）均在嗅球（OB）中高度表达，这表明PACAP在灯泡内神经元的发育，存活和替代中起重要作用。令人惊讶的是，没有关于PACAP在调节CA [2+]活性，迁移和发展OB神经元成熟的作用的研究。在其他中枢神经系统区域中，以自发振荡形式的Ca [2+]活性对于发展神经元电路的建立和存活至关重要，尤其是在感觉输入之前[6]。 GABA在未成熟的神经元中是兴奋性的，并且已经参与了这种早期的发育活动，但是嗅觉中间神经元的成熟度的控制知之甚少。但是，我们的初步数据表明，PACAP在新生儿OB的多种细胞类型中诱导瞬时和持续的Ca [2+]振荡。 我们在第一个目标上的重点是发展的GABA能细胞（GC），因为它们被认为是表达PAC1RS的，并且可以使用GAD2-和DLX2- TDTOMATO转基因小鼠进行鉴定。开发的GC将进行PACAP反应的评估及其与GABA测试的成熟度，这将是未成熟细胞的兴奋性神经递质，可维持高细胞内CL水平并随GABA刺激而去极化。 第二个目的是确定PACAP在未成熟OB中间神经元迁移和成熟中的作用。它将使用培养的GAD2或DLX2 TDTOMATO小鼠OB细胞上的迁移测定直接完成。还将检查与成熟相关的形态变化。 该提案将研究PACAP在OB回路产后早期发育中的作用，并基于对OB和其他大脑区域中PACAP和PAC1受体表达的研究。但是，我的研究是新生儿OB细胞中PACAP的首次动态成像研究。由于对PACAP在OB中的功能知之甚少，因此我的提议的重点是为将来了解PACAP如何影响OB电路的发展奠定基础。人们认为许多神经系统疾病，例如自闭症，躁郁症和精神分裂症，源于神经元回路的早期发展中的问题。了解PACAP在早期电路发展中的作用可能为这些毁灭性疾病提供了新的治疗靶点。此外，了解PACAP如何影响发展中间神经元可能有助于设计受伤或疾病后成人神经发生的策略。