Journal Article

Quantitative neuroanatomy for connectomics in Drosophila

Midgley, Frank M, Fetter, Richard D, Saalfeld, Stephan, Longair, Mark, Zwart, Maarten F, Champion, Andrew, Li, Feng, Gerhard, Stephan, Cardona, Albert, Kazimiers, Tom, Schneider-Mizell, Casey M

2016

Overview

Neuronal circuit mapping using electron microscopy demands laborious proofreading or reconciliation of multiple independent reconstructions. Here, we describe new methods to apply quantitative arbor and network context to iteratively proofread and reconstruct circuits and create anatomically enriched wiring diagrams. We measured the morphological underpinnings of connectivity in new and existing reconstructions of Drosophila sensorimotor (larva) and visual (adult) systems. Synaptic inputs were preferentially located on numerous small, microtubule-free 'twigs' which branch off a single microtubule-containing 'backbone'. Omission of individual twigs accounted for 96% of errors. However, the synapses of highly connected neurons were distributed across multiple twigs. Thus, the robustness of a strong connection to detailed twig anatomy was associated with robustness to reconstruction error. By comparing iterative reconstruction to the consensus of multiple reconstructions, we show that our method overcomes the need for redundant effort through the discovery and application of relationships between cellular neuroanatomy and synaptic connectivity. The nervous system contains cells called neurons, which connect to each other to form circuits that send and process information. Each neuron receives and transmits signals to other neurons via very small junctions called synapses. Neurons are shaped a bit like trees, and most input synapses are located in the tiniest branches. Understanding the architecture of a neuron’s branches is important to understand the role that a particular neuron plays in processing information. Therefore, neuroscientists strive to reconstruct the architecture of these branches and how they connect to one another using imaging techniques. One imaging technique known as serial electron microscopy generates highly detailed images of neural circuits. However, reconstructing neural circuits from such images is notoriously time consuming and error prone. These errors could result in the reconstructed circuit being very different than the real-life circuit. For example, an error that leads to missing out a large branch could result in researchers failing to notice many important connections in the circuit. On the other hand, some errors may not matter much because the neurons share other synapses that are included in the reconstruction. To understand what effect errors have on the reconstructed circuits, neuroscientists need to have a more detailed understanding of the relationship between the shape of a neuron, its synaptic connections to other neurons, and where errors commonly occur. Here, Schneider-Mizell, Gerhard et al. study this relationship in detail and then devise a faster reconstruction method that uses the shape and other properties of neurons without sacrificing accuracy. The method includes a way to include data from the shape of neurons in the circuit wiring diagrams, revealing circuit patterns that would otherwise go unnoticed. The experiments use serial electron microscopy images of neurons from fruit flies and show that, from the tiniest larva to the adult fly, neurons form synapses with each other in a similar way. Most errors in the reconstruction only affect the tips of the smallest branches, which generally only host a single synapse. Such omissions do not have a big effect on the reconstructed circuit because strongly connected neurons make multiple synapses onto each other. Schneider-Mizell, Gerhard et al.'s approach will help researchers to reconstruct neural circuits and analyze them more effectively than was possible before. The algorithms and tools developed in this study are available in an open source software package so that they can be used by other researchers in the future.

• Publisher: eLife Science Publications, Ltd,eLife Sciences Publications Ltd,eLife Sciences Publications, Ltd
• Subject: Anatomy / Animals / Collaboration / Connectome - methods / connectomics / Drosophila / Drosophila - anatomy & histology / Drosophila - physiology / Electron microscopy / Insects / Larvae / Medical research / Microscopy / Nervous system / Nervous System - anatomy & histology / Nervous System Physiological Phenomena / Neural circuitry / Neural networks / neuroanatomy / Neurons / Neuroscience / Physiological aspects / Proofreading / proprioception / Sensorimotor system / Synapses / Visual pathways