Multiphon imaging for understanding social brain function in tadpoles

项目概要 母婴关系是一种重要的关系，为一生的健康奠定基础。社交 识别是这种关系的重要组成部分，因为婴儿会在母亲的气味上留下印记， 使用嗅觉将母亲与其他人区分开来。这种行为在哺乳动物中很普遍，尽管 哺乳动物大脑的复杂性以及研究新生儿的工具和重复成像能力的缺乏 幼崽很难获得对基本大脑机制的精确理解。 年轻时的母婴联系和沟通。母婴认知和联系也发生在其他方面 类群，表明还有其他物种可以研究亲子的普遍神经原理 互动。最近我们的实验室开发了（1）毒蛙蝌蚪作为研究社交大脑的模型 开发和（2）多光子成像方法，以实现色素水生幼虫的体内记录。 我们建议开发多光子体内成像方法来研究母体气味的编码 社会性毒蛙蝌蚪可以区分母亲和陌生人。我们假设嗅觉 当蝌蚪学习母亲的气味时，反应景观在整个发育过程中发生变化。我们预测 蝌蚪区分母亲和陌生人的能力与嗅觉细胞的增加相一致 通过嗅觉系统神经活动的多光子成像测量，对母体气味具有特异性 反复跨越发展。在这些实验之前，我们将评估各种性能 多光子成像技术可实现多种毒蛙大脑内部光学深度 具有不同程度色素沉着的蝌蚪。我们将通过严格的统计分析来验证结果 神经活动数据与脑切片免疫组织学成像的比较以及活动依赖性 嗅觉感觉神经元的测序。了解两栖动物如何学习和编码个体 同种身份要么揭示编码基于嗅觉的识别的替代机制，要么揭示 嗅觉编码模式是脊椎动物嗅觉处理的祖先或普遍特征。 重要的是，我们的方法将导致全脑成像多光子方法的发展 有色素的水生动物，这是神经科学界广泛使用的宝贵工具包。 该项目的成功完成将使我们能够获得提出体内成像的原理验证数据 在比较的背景下，在行为学的框架内，蝌蚪大脑在整个发育过程中的变化 相关行为，这对于未来的 R01 应用至关重要。此外，建立一个记录协议 蝌蚪的研究将允许两栖动物神经功能的其他方面，这是迄今为止已被研究的一个研究领域 由于技术限制而受到限制。总之，我们提出的研究将有助于阐明核心嗅觉 在相对简单的蝌蚪大脑中印记原理，并揭示这些原理如何进行嗅觉编码 可以推广到脊椎动物类群。

Project Summary Mother-infant bonding is a key relationship that lays a foundation for wellness throughout life. Social recognition is an important component of this relationship, as infants imprint on the smell of their mothers and use olfaction to distinguish their mother from others. This behavior is widespread in mammals, although the complexity of the mammalian brain and paucity of tools and repeated imaging capabilities to study neonate pups make it difficult to obtain a precise mechanistic understanding of the basic brain mechanisms for mother-infant bonding and communication in young. Mother-infant recognition and bonding also occur in other taxa, suggesting there are other species in which to study generalizable neural principles of parent-offspring interactions. Recently our labs have developed (1) poison frog tadpoles as a model to study social brain development and (2) multiphoton imaging approaches to enable in vivo recording of pigmented aquatic larvae. We propose to develop multiphoton in vivo imaging approaches to study the encoding of maternal odors in a social poison frog tadpole that can distinguish their mother from strangers. We hypothesize that the olfactory response landscape changes throughout development as tadpoles learn the smell of their mothers. We predict that a tadpole’s ability to distinguish their mother from strangers coincides with an increase in olfactory cells that fire with specificity to maternal odors, measured by multiphoton imaging of olfactory system neural activity repeatedly across development. Prior to these experiments, we will evaluate the performance of various multiphoton imaging techniques for optically accessible depth inside the brain of multiple species of poison frog tadpoles with varying levels of pigmentation. We will validate the results with rigorous statistical analyses and comparison of neural activity data with immunohistological imaging of brain slices and activity-dependent sequencing of olfactory sensory neurons. Understanding how amphibians learn and encode individual conspecific identity will either reveal alternative mechanisms of encoding olfactory-based recognition or which patterns of olfactory encoding are ancestral or generalizable features of vertebrate olfactory processing. Importantly, our approach will result in the development of multiphoton approaches for whole brain imaging of pigmented aquatic animals, which is a valuable toolkit of broad use for the neuroscience community. Successful completion of this project will allow us to obtain proof-of-principle data for proposing in vivo imaging of tadpole brains throughout development, in a comparative context, and within the framework of ethologically relevant behaviors, which is crucial for future R01 applications. Furthermore, establishing a recording protocol in tadpoles will allow for other aspects of neural function in amphibians, a research area that has thus far been limited due to technological constraints. In summary, our proposed research will help elucidate core olfactory imprinting principles in a relatively simple tadpole brain and reveal how these principles for olfactory encoding may generalize across vertebrate taxa.