| Autor: |
Shen FY; Medical Scientist Training Program, University of Michigan, Ann Arbor, MI, USA.; Neuroscience Graduate Program, University of Michigan, Ann Arbor, MI, USA., Harrington MM; Department of Cell and Developmental Biology, University of Michigan, Ann Arbor, MI, USA., Walker LA; LS & A, Program in Biophysics, University of Michigan, Ann Arbor, MI, USA., Cheng HPJ; Department of Cell and Developmental Biology, University of Michigan, Ann Arbor, MI, USA., Boyden ES; McGovern Institute, Koch Institute, Department of Media Arts and Sciences, Department of Biological Engineering, and Department of Brain and Cognitive Sciences, Massachusetts Institute of Technology, Cambridge, MA, USA.; Howard Hughes Medical Institute, Cambridge, MA, USA., Cai D; Neuroscience Graduate Program, University of Michigan, Ann Arbor, MI, USA. dwcai@umich.edu.; Department of Cell and Developmental Biology, University of Michigan, Ann Arbor, MI, USA. dwcai@umich.edu.; LS & A, Program in Biophysics, University of Michigan, Ann Arbor, MI, USA. dwcai@umich.edu. |
| Abstrakt: |
Mapping neuroanatomy is a foundational goal towards understanding brain function. Electron microscopy (EM) has been the gold standard for connectivity analysis because nanoscale resolution is necessary to unambiguously resolve synapses. However, molecular information that specifies cell types is often lost in EM reconstructions. To address this, we devise a light microscopy approach for connectivity analysis of defined cell types called spectral connectomics. We combine multicolor labeling (Brainbow) of neurons with multi-round immunostaining Expansion Microscopy (miriEx) to simultaneously interrogate morphology, molecular markers, and connectivity in the same brain section. We apply this strategy to directly link inhibitory neuron cell types with their morphologies. Furthermore, we show that correlative Brainbow and endogenous synaptic machinery immunostaining can define putative synaptic connections between neurons, as well as map putative inhibitory and excitatory inputs. We envision that spectral connectomics can be applied routinely in neurobiology labs to gain insights into normal and pathophysiological neuroanatomy. |
