Multiphon imaging for understanding social brain function in tadpoles

项目摘要 母婴关系是为终生健康奠定基础的一种关键关系。社交 认知是这种关系的重要组成部分，因为婴儿会在母亲和婴儿的气味上留下印记 用嗅觉将它们的母亲与其他动物区分开来。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 幼崽很难对大脑的基本机制有一个精确的机械理解 母婴在幼年时的亲情与沟通。母婴认同感和亲和力也发生在 分类群，这表明还有其他物种可以研究亲子代的概括性神经原理 互动。最近，我们的实验室开发了(1)毒蛙蝌蚪作为研究社会大脑的模型 开发和(2)多光子成像方法，以便能够在体内记录有色水生幼虫。 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 社会毒蛙蝌蚪，可以区分它们的母亲和陌生人。我们假设嗅觉 随着小蝌蚪学习它们母亲的气味，整个发育过程中的反应景观都会发生变化。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不谋而合 通过嗅觉系统神经活动的多光子成像测量，这种火对母体气味具有特异性 在整个发展过程中反复出现。在这些实验之前，我们将评估各种类型的性能 多种毒蛙脑内光学可及深度的多光子成像技术 有不同程度色素沉着的蝌蚪。我们将通过严格的统计分析和 神经活动数据与脑片免疫组织学成像和活动依赖性的比较 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 同种身份将揭示编码基于嗅觉的识别的替代机制，或者 嗅觉编码模式是脊椎动物嗅觉加工的祖先或可概括的特征。 重要的是，我们的方法将导致开发用于全脑成像的多光子方法 色素水生动物，这是神经科学界广泛使用的宝贵工具包。 这个项目的成功完成将使我们能够获得提出活体成像的原则证明数据 在整个发育过程中，在比较的背景下，在行为学的框架内 相关行为，这对未来的R01应用至关重要。此外，制定记录协议 将允许两栖动物神经功能的其他方面，这一研究领域迄今一直是 由于技术上的限制，这一点很有限。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的印记原理，并揭示了这些原理是如何对嗅觉编码的 可以在脊椎动物类群中推广。

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.