Opponent vesicular transporters regulate the strength of glutamatergic neurotransmission in a C. elegans sensory circuit

Choi, Jung-Hwan; Horowitz, Lauren Bayer; Ringstad, Niels

doi:10.1038/s41467-021-26575-3

Cited by 11 publications

(11 citation statements)

References 70 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…B) Inhibiting neuron-neuron synaptic transmission in VC causes increased activity (membrane depolarization) in HSN, suggesting inhibition of HSN dependent on synaptic transmission from VC (Kopchock et al 2021) (Table 4H, this manuscript). C) Model of the mechanisms involved in RIA activation based on data from Choi et al (2021) (Table 4J-4K). Blue boxes represent a GO cellular component term.…”

Section: Resultsmentioning

confidence: 99%

Semantic Representation of Neural Circuit Knowledge inCaenorhabditis elegans

Prakash

Auken

Hill

et al. 2023

Preprint

View full text Add to dashboard Cite

In modern biology, new knowledge is generated quickly, making it challenging for researchers to efficiently acquire and synthesise new information from the large volume of primary publications. To address this problem, computational approaches that generate machine-readable representations of scientific findings in the form of knowledge graphs have been developed. These representations can integrate different types of experimental data from multiple papers and biological knowledge bases in a unifying data model, providing a complementary method to manual review for interacting with published knowledge. The Gene Ontology Consortium (GOC) has created a semantic modelling framework that extends individual functional gene annotations to structured descriptions of causal networks representing biological processes (Gene Ontology Causal Activity Modelling, or GO-CAM). In this study, we explored whether the GO-CAM framework could represent knowledge of the causal relationships between environmental inputs, neural circuits and behavior in the model nematode C. elegans (C. elegans Neural Circuit Causal Activity Modelling (CeN-CAM)). We found that, given extensions to several relevant ontologies, a wide variety of author statements from the literature about the neural circuit basis of egg-laying and carbon dioxide (CO2) avoidance behaviors could be faithfully represented with CeN-CAM. Through this process, we were able to generate generic data models for several categories of experimental results. We also generated representations of multisensory integration and sensory adaptation, and we discuss how semantic modelling may be used to functionally annotate the C. elegans connectome. Thus, Gene Ontology-based semantic modelling has the potential to support various machine-readable representations of neurobiological knowledge.

show abstract

Section: Resultsmentioning

confidence: 99%

Semantic Representation of Neural Circuit Knowledge inCaenorhabditis elegans

Prakash

Auken

Hill

et al. 2023

Preprint

View full text Add to dashboard Cite

show abstract

“…For this, the maximum background corrected fluorescence during stimulation was calculated (as a moving average of 1 s). If this was higher than the average background corrected fluorescence before the stimulation +3 × standard deviations (of background corrected fluorescence before stimulation; minimum of two arbitrary units of fluorescence increase), the animal was counted as a strong responder (adapted from Choi et al, 2021); see Supplementary Figure 6A for example traces that fit or do not fit these parameters; animals not fitting the cut-off showed no discernable light-evoked effect on DNC fluorescence. Also, animals that showed an increase of the fluorescence after the end of the stimulus, or animals showing spontaneous events, were excluded.…”

Section: Strainmentioning

confidence: 99%

“…Alternatively, exposure to stimuli such as odors can be used. However, this is difficult to control, and limited to applications in sensory neurons (Choi et al, 2021). Thus, an all-optical solution that is not limited to certain cell types would be ideal, e.g., a combination of genetically encoded non-invasive tools for in vivo stimulation of neurons, and pH-sensitive fluorescent proteins.…”

Section: Introductionmentioning

confidence: 99%

pOpsicle: An all-optical reporter system for synaptic vesicle recycling combining pH-sensitive fluorescent proteins with optogenetic manipulation of neuronal activity

Seidenthal

Jánosi

Rosenkranz

et al. 2023

Front. Cell. Neurosci.

View full text Add to dashboard Cite

pH-sensitive fluorescent proteins are widely used to study synaptic vesicle (SV) fusion and recycling. When targeted to the lumen of SVs, fluorescence of these proteins is quenched by the acidic pH. Following SV fusion, they are exposed to extracellular neutral pH, resulting in a fluorescence increase. SV fusion, recycling and acidification can thus be tracked by tagging integral SV proteins with pH-sensitive proteins. Neurotransmission is generally activated by electrical stimulation, which is not feasible in small, intact animals. Previous in vivo approaches depended on distinct (sensory) stimuli, thus limiting the addressable neuron types. To overcome these limitations, we established an all-optical approach to stimulate and visualize SV fusion and recycling. We combined distinct pH-sensitive fluorescent proteins (inserted into the SV protein synaptogyrin) and light-gated channelrhodopsins (ChRs) for optical stimulation, overcoming optical crosstalk and thus enabling an all-optical approach. We generated two different variants of the pH-sensitive optogenetic reporter of vesicle recycling (pOpsicle) and tested them in cholinergic neurons of intact Caenorhabditis elegans nematodes. First, we combined the red fluorescent protein pHuji with the blue-light gated ChR2(H134R), and second, the green fluorescent pHluorin combined with the novel red-shifted ChR ChrimsonSA. In both cases, fluorescence increases were observed after optical stimulation. Increase and subsequent decline of fluorescence was affected by mutations of proteins involved in SV fusion and endocytosis. These results establish pOpsicle as a non-invasive, all-optical approach to investigate different steps of the SV cycle.

show abstract

“…For this, the maximum background corrected fluorescence during stimulation was calculated (as a moving average of 1 s). If this was higher than the average background corrected fluorescence before the stimulation + 3 * standard deviation (of background corrected fluorescence before stimulation), the animal was counted as a strong responder (adapted from Choi et al, 2021); see Fig. S3A for example traces that fit or do not fit these parameters; animals not fitting the cut-off showed no discernable light-evoked effect on DNC fluorescence.…”

Section: Quantification Of Fluorescencementioning

confidence: 99%

“…Alternatively, exposure to stimuli such as odors can be used. Yet, this is difficult to control, and limited to applications in sensory neurons (Choi et al, 2021). Thus, an all-optical solution that is not limited to certain cell types would be ideal, e.g., involving a combination of genetically encoded non-invasive tools for in vivo stimulation of neurons, with pH-sensitive fluorescent proteins.…”

Section: Introductionmentioning

confidence: 99%

pOpsicle: An all-optical reporter system for synaptic vesicle recycling combining pH-sensitive fluorescent proteins with optogenetic manipulation of neuronal activity

Seidenthal

Jánosi

Rosenkranz

et al. 2022

Preprint

View full text Add to dashboard Cite

pH-sensitive fluorescent proteins are widely used to study synaptic vesicle (SV) fusion and recycling. When targeted to the lumen of SVs, fluorescence of these proteins is quenched by the acidic pH. Following SV fusion, they are exposed to extracellular neutral pH, resulting in a fluorescence increase. SV fusion, recycling and acidification can thus be tracked by tagging integral SV proteins with pHsensitive proteins. Neurotransmission is generally stimulated by electrophysiology, which is not feasible in small, intact animals, thus limiting the approach to cell culture regimes. Previous in vivo approaches depended on distinct (sensory) stimuli, thus limiting the addressable neuron types. To overcome these limitations, we established an all-optical approach to stimulate and visualize SV fusion and recycling. We combined distinct pH-sensitive fluorescent proteins (inserted into the SV protein synaptogyrin) and light-gated channelrhodopsins (ChRs) for optical stimulation, overcoming optical crosstalk and thus enabling an all-optical approach. We generated two different variants of the pHsensitive optogenetic reporter of vesicle recycling (pOpsicle) and tested them in cholinergic neurons of intact Caenorhabditis elegans nematodes. First, we combined the red fluorescent protein pHuji with the blue-light gated ChR2(H134R), and second, the green fluorescent pHluorin combined with the novel red-shifted ChR ChrimsonSA. In both cases, fluorescence increases were observed after optical stimulation. Increase and subsequent decline of fluorescence was affected by mutations of proteins involved in SV fusion and endocytosis. These results establish pOpsicle as a non-invasive, all-optical approach to investigate different steps of the SV cycle.

show abstract

Opponent vesicular transporters regulate the strength of glutamatergic neurotransmission in a C. elegans sensory circuit

Cited by 11 publications

References 70 publications

Semantic Representation of Neural Circuit Knowledge inCaenorhabditis elegans

Semantic Representation of Neural Circuit Knowledge inCaenorhabditis elegans

pOpsicle: An all-optical reporter system for synaptic vesicle recycling combining pH-sensitive fluorescent proteins with optogenetic manipulation of neuronal activity

pOpsicle: An all-optical reporter system for synaptic vesicle recycling combining pH-sensitive fluorescent proteins with optogenetic manipulation of neuronal activity

Contact Info

Product

Resources

About