■基因编码传感器和双光子成像的进展在单个树突棘水平上解锁了功能成像。可以在清醒的动物中实时测量突触活动。然而,需要工具来促进对该方法获得的大型数据集的分析。通常可用的用于对细胞体中的钙瞬变进行成像的软件套件不适合于脊柱成像,因为树突棘具有与细胞体不同的结构特征。我们提出了一个自动调整分析工具(AUTOTUNE),它提供了专门为从亚细胞区室提取和分析信号而开发的分析程序,包括树突亚区域和棘。
■尽管体内功能性突触成像数据的获取越来越容易获得,在对采集数据进行大量计算的分析中仍然存在障碍。这项研究的目的是通过提供一个全面的软件套件来克服这一障碍,该套件具有用户友好的界面,便于非程序员访问。
■我们通过体内从小鼠V1中的2/3层锥体神经元获得的树突成像数据的演示分析来证明我们的软件的实用性和有效性。还提供了用户手册和演示数据集。
■AUTOTUNE提供了一个强大的工作流程,用于分析来自神经元树突的功能成像数据。功能包括源图像配准,感兴趣区域的分割和结构周转的检测,荧光瞬态提取和平滑,从假定的反向传播动作电位中减去信号,以及刺激和行为参数反应调整分析。
■AUTOTUNE是开源的,可扩展用于多种功能性突触成像实验。由我们的软件提供的树突脊柱活动的功能表征的简易性可以加速新的功能研究,补充树突的形态学研究的几十年,并进一步扩大我们对健康和疾病中神经回路的理解。
UNASSIGNED: Advances in genetically encoded sensors and two-photon imaging have unlocked functional imaging at the level of single dendritic
spines. Synaptic activity can be measured in real time in awake animals. However, tools are needed to facilitate the analysis of the large datasets acquired by the approach. Commonly available software suites for imaging calcium transients in cell bodies are ill-suited for spine imaging as dendritic
spines have structural characteristics distinct from those of the cell bodies. We present an automated tuning analysis tool (AUTOTUNE), which provides analysis routines specifically developed for the extraction and analysis of signals from subcellular compartments, including dendritic subregions and
spines.
UNASSIGNED: Although the acquisition of in vivo functional synaptic imaging data is increasingly accessible, a hurdle remains in the computation-heavy analyses of the acquired data. The aim of this study is to overcome this barrier by offering a comprehensive software suite with a user-friendly interface for easy access to nonprogrammers.
UNASSIGNED: We demonstrate the utility and effectiveness of our software with demo analyses of dendritic imaging data acquired from layer 2/3 pyramidal neurons in mouse V1 in vivo. A user manual and demo datasets are also provided.
UNASSIGNED: AUTOTUNE provides a robust workflow for analyzing functional imaging data from neuronal dendrites. Features include source image registration, segmentation of regions-of-interest and detection of structural turnover, fluorescence transient extraction and smoothing, subtraction of signals from putative backpropagating action potentials, and stimulus and behavioral parameter response tuning analyses.
UNASSIGNED: AUTOTUNE is open-source and extendable for diverse functional synaptic imaging experiments. The ease of functional characterization of dendritic spine activity provided by our software can accelerate new functional studies that complement decades of morphological studies of dendrites, and further expand our understanding of neural circuits in health and in disease.