Identification of Neuronal Lineages in the Drosophila Peripheral Nervous System with a “Digital” Multi-spectral Lineage Tracing System

Veling, Macy W.; Li, Ye; Veling, Mike T.; Litts, Christopher; Michki, Nigel S; Liu, Hao; Yan, Bing; Cai, Dawen

doi:10.1016/j.celrep.2019.10.124

Cited by 20 publications

(20 citation statements)

References 68 publications

(87 reference statements)

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“…We can already observe tens of easily distinguishable color barcodes, which can be exponentially increased in the future by probing more FPs. For example, five FPs were used in the recently developed Bitbow, a "digital" format of Brainbow 40 . Expansion of the sample is important for increasing imaging resolution, but comes at the cost of diluting antibody signal.…”

Section: Discussionmentioning

confidence: 99%

Light microscopy based approach for mapping connectivity with molecular specificity

et al. 2020

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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.

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Section: Discussionmentioning

confidence: 99%

Light microscopy based approach for mapping connectivity with molecular specificity

et al. 2020

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“…We can already observe tens of easily distinguishable color barcodes, which can be exponentially increased in the future by probing more FPs. For example, 5 FPs were used in the recently developed Bitbow, a "digital" format of Brainbow 39 . Expansion of the sample is important for increasing imaging resolution, but comes at the cost of diluting antibody signal.…”

Section: Discussionmentioning

confidence: 99%

Light microscopy based approach for mapping connectivity with molecular specificity

Shen

Harrington

Walker

et al. 2020

Preprint

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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 chemical and electrical synapses. However, molecular information that specifies cell types is often lost in EM reconstructions. To address this, we devised a light microscopy approach for connectivity analysis of defined cell types called spectral connectomics. We combined multicolor genetic labeling (Brainbow) of neurons with a m ult i-r ound i mmunostaining Ex pansion Microscopy (miriEx) strategy to simultaneously interrogate morphology, molecular markers, and connectivity in the same brain section. We applied our multimodal profiling strategy to directly link inhibitory neuron cell types with their network morphologies. Furthermore, we showed that correlative Brainbow and endogenous synaptic machinery immunostaining can be used to define putative synaptic connections between spectrally unique 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 across multiple animals and time points.

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“…Genetic constructs based around CRISPR-Cas9 (Raj et al, 2017;Spanjaard et al, 2018) and the Cre/Lox system (Kalhor et al, 2018;Pei et al, 2017;Weber et al, 2016) have been developed for this purpose, although which exact lineage was labeled by a particular barcode was still unknown. The introduction of a spectrally unique barcode for each neuroblast lineage, in a similar vein to the recently developed Bitbow lineage tracking strategy (Li et al, 2020;Veling et al, 2019), would be advantageous as they can provide direct in situ evidence for neuroblast lineage identity.…”

Section: Challenges and Opportunitiesmentioning

confidence: 99%

The molecular landscape of neural differentiation in the developing Drosophila brain revealed by targeted scRNA-seq and multi-informatic analysis

Michki

Sanjasaz

et al. 2021

Cell Reports

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The Drosophila type II neuroblast lineages present an attractive model to investigate the neurogenesis and differentiation process as they adapt to a process similar to that in the human outer subventricular zone. We perform targeted single-cell mRNA sequencing in third instar larval brains to study this process of the type II NB lineage. Combining prior knowledge, in silico analyses, and in situ validation, our multi-informatic investigation describes the molecular landscape from a single developmental snapshot. 17 markers are identified to differentiate distinct maturation stages. 30 markers are identified to specify the stem cell origin and/or cell division numbers of INPs, and at least 12 neuronal subtypes are identified. To foster future discoveries, we provide annotated tables of pairwise gene-gene correlation in single cells and MiCV, a web tool for interactively analyzing scRNA-seq datasets. Taken together, these resources advance our understanding of the neural differentiation process at the molecular level.

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Identification of Neuronal Lineages in the Drosophila Peripheral Nervous System with a “Digital” Multi-spectral Lineage Tracing System

Cited by 20 publications

References 68 publications

Light microscopy based approach for mapping connectivity with molecular specificity

Light microscopy based approach for mapping connectivity with molecular specificity

Light microscopy based approach for mapping connectivity with molecular specificity

The molecular landscape of neural differentiation in the developing Drosophila brain revealed by targeted scRNA-seq and multi-informatic analysis

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