Mass spectrometric measurement of neuropeptide secretion in the crab, Cancer borealis, by in vivo microdialysis

Liang, Zhidan; Schmerberg, Claire M.

doi:10.1039/c4an02016b

Cited by 12 publications

(20 citation statements)

References 47 publications

(129 reference statements)

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“…Other hemolymph functions regard the circulation of oxygen, nutrients, hormone‐like, and neuropeptides and xenobiotic detoxification . Despite these accomplishments, our understanding of the mechanisms underlying the functions of hemolymph and the immunomodulation processes in marine molluscs is still scarce and restricted to the study of individual proteins and applying principles of comparative physiology.…”

Section: Introductionmentioning

confidence: 99%

“…Despite these accomplishments, our understanding of the mechanisms underlying the functions of hemolymph and the immunomodulation processes in marine molluscs is still scarce and restricted to the study of individual proteins and applying principles of comparative physiology. Proteomics is helping to cover this gap enabling the identification of: (i) key elements of the immune response of Octopus vulgaris to the parasite Aggregata octopiana , (ii) markers of immune response to metal contamination , (iii) novel elements of the innate immune response of the ascidian Halocynthia roretzi showing soft tunic syndrome , (iv) 160 gene products from hemolymph of the ancient marine arthropod Limulus polyphemus , (v) novel signaling mechanisms and hormone‐mediated regulatory processes in crustaceans and molluscs . Furthermore, the use of combined proteomic and transcriptomic approaches have uncovered patterns of expression of constitutive haemoglobin isoforms in model crustacean Daphnia pulex that are related to acute temperature‐induced tissue hypoxia and to a mechanism of acclimation to moderately elevated temperatures .…”

Section: Introductionmentioning

confidence: 99%

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Shotgun analysis of the marine mussel Mytilus edulis hemolymph proteome and mapping the innate immunity elements

et al. 2015

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Shotgun analysis of the marine mussel Mytilus edulis hemolymph proteome and mapping the innate immunity elements. Campos A, Apraiz I, da Fonseca RR, Cristobal S. Proteomics 2015, 15, 4021-4029 DOI 10.1002 1 AbstractThe marine mussel innate immunity provides protection to pathogen invasion and inflammation. In this regard the mussel haemolymph takes a main role in the animal innate response. Despite the importance of this body fluid in determining the physiological condition of the animal, little is known about the molecular mechanisms underlying the cellular and humoral responses. In this work we have applied a mass spectrometry (nanoLC-MS/MS) strategy integrating genomic and transcriptomic data with the aim to: i) identify the main protein functional groups that characterize haemolymph and ii) to map the elements of innate immunity in the marine mussel Mytilus edulis haemolymph proteome.After sample analysis and first protein identification based on MS/MS data comparison, proteins with unknown functions were annotated with blast using public database (nrNCBI) information. Overall 595 haemolymph proteins were identified with high confidence and annotated. These proteins encompass primary cellular metabolic processes: energy production and metabolism of biomolecules, as well as processes related to oxidative stress defence, xenobiotic detoxification, drug metabolism, and immune response. A group of proteins was identified with putative immune effector, receptor and signalling functions in M. edulis. Data are available via ProteomeXchange with identifier PXD001951. Significance of the Study:Proteomic investigations have been hampered in Mytilus sp. by the scarce genomic data available.Here a high-throughput methodology was successfully employed to describe the overlooked haemolymph proteome from the marine mussel M. edulis, revealing a conjunct of molecular functions contributing to the immune defence. The information available contributes to enhance the Shotgun analysis of the marine mussel Mytilus edulis hemolymph proteome and mapping the innate immunity elements. Campos A, Apraiz I, da Fonseca RR, Cristobal S.

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Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Shotgun analysis of the marine mussel Mytilus edulis hemolymph proteome and mapping the innate immunity elements

et al. 2015

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“…Previously, our group has employed the MD sampling technique on crabs to study neuropeptide content in the circulating hemolymph . However, the study of NTs in the model crustacean system, which is an essential component of neuromodulators in the nervous system, has been lacking.…”

Section: Resultsmentioning

confidence: 99%

“…Animals were housed, treated, and sacrificed following the animal care protocol in accordance with the University of Wisconsin‐Madison's animal care guidelines. The surgery of MD was adapted from previous publication . After the probe was surgically implanted in the crab, the animal was allowed to recover for at least 24 h before dialysate was collected for MS analysis.…”

Section: Methodsmentioning

confidence: 99%

Investigation of signaling molecules and metabolites found in crustacean hemolymph via in vivo microdialysis using a multifaceted mass spectrometric platform

2016

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Neurotransmitters (NTs) are endogenous signaling molecules which play an important role in regulating various physiological processes in animals. Detection of these chemical messengers is often challenging due to their low concentration levels and fast degradation rate in vitro. In order to address these challenges, herein we employed in vivo microdialysis (MD) sampling to study neurotransmitters in the crustacean model Cancer borealis. Multi-faceted separation tools, such as capillary electrophoresis (CE) and ion mobility mass spectrometry were utilized in this work. Small molecules were separated by different mechanisms and detected by matrix-assisted laser desorption/ionization mass spectrometric imaging (MALDI-MSI). Performance of this separation-based MSI platform was also compared to liquid chromatography-electrospray ionization mass spectrometry (LC-ESI-MS). By utilizing both MALDI and ESI MS, a total of 208 small molecule neurotransmitters and metabolites were identified, of which 39 were identified as signaling molecules secreted in vivo. In addition, the inherent property of sub micro-scale sample consumption using CE enables shorter time of MD sample collection. Temporal resolution of in vivo MD was improved by approximately 10-fold compared to LC-ESI-MS, indicating the significant advantage of applying separation-assisted MALDI MS imaging platform.

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“…Whilst not experimentally demonstrated in nematodes to date, the existence and significance of additional forms of non-synaptic neuronal communication have been recognized among invertebrates [5, 6]. For example, in crustaceans, extrasynaptic volume transmission (EVT) operates beyond the synaptic connectome, mediating long-range hormonal communication in the absence of neuronal synapses [7, 8]. Notably, EVT has been implicated in nematode neuronal signaling [9-14].…”

Section: Introductionmentioning

confidence: 99%

Extrasynaptic volume transmission: A novel route for neuropeptide signaling in nematodes

Atkinson

Liu

McKay

et al. 2020

Preprint

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Neural circuit synaptic connectivities (the connectome) provide the anatomical foundation for our understanding of nematode nervous system function. However, other non-synaptic routes of communication are known in invertebrates including extrasynaptic volume transmission (EVT), which enables short- and/or long-range communication in the absence of synaptic connections. Although EVT has been highlighted as a facet of Caenorhabditis elegans neurosignaling, no experimental evidence identifies body cavity fluid (pseudocoelomic fluid; PCF) as a vehicle for either neuropeptide or biogenic amine transmission. In the parasitic nematode Ascaris suum FMRFamide-like peptides encoded on flp-18 potently stimulate female reproductive organs but are only expressed in cells that are anatomically distant from the reproductive organ, with no known synaptic connections to this tissue. Here we report a new non-synaptic mode of signaling in nematodes mediated by neuropeptides within the PCF. Our data show that: (i) A. suum PCF (As-PCF) contains a catalogue of neuropeptides including FMRFamide-like peptides and neuropeptide-like proteins; (ii) the A. suum FMRFamide-like peptide As-FLP-18A dominates the As-PCF peptidome; (iii) As-PCF potently modulates nematode reproductive muscle function ex vivo, mirroring the effects of synthetic FLP-18 peptides; (iv) As-PCF activates the C. elegans FLP-18 receptors NPR-4 and -5; (v) As-PCF alters C. elegans behavior and, (vi) FLP-18 and FLP-18 receptors display pan-phylum distribution in nematodes. Here we provide the first direct experimental evidence that supports an extrasynaptic volume route for neuropeptide transmission in nematodes. These data demonstrate non-synaptic signaling within the nematode functional connectome and are pertinent to receptor deorphanisation approaches underpinning drug discovery programs for nematode pathogens.

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Mass spectrometric measurement of neuropeptide secretion in the crab, Cancer borealis, by in vivo microdialysis

Cited by 12 publications

References 47 publications

Shotgun analysis of the marine mussel Mytilus edulis hemolymph proteome and mapping the innate immunity elements

Shotgun analysis of the marine mussel Mytilus edulis hemolymph proteome and mapping the innate immunity elements

Investigation of signaling molecules and metabolites found in crustacean hemolymph via in vivo microdialysis using a multifaceted mass spectrometric platform

Extrasynaptic volume transmission: A novel route for neuropeptide signaling in nematodes

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