Metabolite localization in living drosophila using High Resolution Magic Angle Spinning NMR

Sarou‐Kanian, Vincent; Joudiou, Nicolas; Louat, Fanny; Yon, Maxime; Szeremeta, Frédéric; Même, Sandra; Massiot, Dominique; Decoville, Martine; Fayon, Franck; Belœil, Jean‐Claude

doi:10.1038/srep09872

Cited by 28 publications

(30 citation statements)

References 23 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Although it is debatable whether results obtained in this way qualify as in cell, the important factor is that the studied molecules are never taken out of their native environment. Sample spinning has also been criticized as being too violent for cells because of its centrifugal and dehydrating effects, but spinning rates and times can be adjusted to preserve cellular integrity and even cell survival (14,(18)(19)(20).…”

Section: Sample Preparationmentioning

confidence: 99%

“…Solid-state NMR has been applied to a variety of biomolecules that are naturally solid, ranging from human biopsies (10) to amyloid fibrils (11), mussel byssus (12), whole nematodes (13), and whole living flies (14), but it has also proved useful for studying membrane-bound molecules, which become solid because of the boundary imposed by the membrane. Such molecules include lipids, membrane proteins, and components of the cell wall and extracellular matrix that can also be studied separately or reconstituted into model systems (15).…”

mentioning

confidence: 99%

See 1 more Smart Citation

In-Cell Solid-State NMR: An Emerging Technique for the Study of Biological Membranes

Warnet

Arnold

Marcotte

et al. 2015

Biophysical Journal

View full text Add to dashboard Cite

Biological molecular processes are often studied in model systems, which simplifies their inherent complexity but may cause investigators to lose sight of the effects of the molecular environment. Information obtained in this way must therefore be validated by experiments in the cell. NMR has been used to study biological cells since the early days of its development. The first NMR structural studies of a protein inside a cell (by solution-state NMR) and of a membrane protein (by solid-state NMR) were published in 2001 and 2011, respectively. More recently, dynamic nuclear polarization, which has been used to enhance the signal in solid-state NMR, has also been applied to the study of frozen cells. Much progress has been made in the past 5 years, and in this review we take stock of this new technique, which is particularly appropriate for the study of biological membranes.

show abstract

Section: Sample Preparationmentioning

confidence: 99%

mentioning

confidence: 99%

In-Cell Solid-State NMR: An Emerging Technique for the Study of Biological Membranes

Warnet

Arnold

Marcotte

et al. 2015

Biophysical Journal

View full text Add to dashboard Cite

show abstract

“…Therefore, when applied to a living organism, swollen components, including solutions (metabolites) and gels (mobile membranes, parts of tissue), become easy to observe and information‐rich 1 H NMR profiles are obtained. As such, HR‐MAS has enjoyed wide applications to intact biological systems such as tissues or whole cells as well as a range of living organisms including Drosophila melanogaster, Caenorhabditis elegans, Aporrectodea caliginosa, Calanus finmarchicus, Daphnia magna, Saccharomyces cervisiae cells, and mice …”

Section: Introductionmentioning

confidence: 99%

In vivo comprehensive multiphase NMR

Mobarhan

Soong

Lane

et al. 2019

Magnetic Reson in Chemistry

View full text Add to dashboard Cite

Traditionally, due to different hardware requirements, nuclear magnetic resonance (NMR) has developed as two separate fields: one dealing with solids, and one with solutions. Comprehensive multiphase (CMP) NMR combines all electronics and hardware (magic angle spinning [MAS], gradients, high power Radio Frequency (RF) handling, lock, susceptibility matching) into a universal probe that permits a comprehensive study of all phases (i.e., liquid, gel‐like, semisolid, and solid), in intact samples. When applied in vivo, it provides unique insight into the wide array of bonds in a living system from the most mobile liquids (blood, fluids) through gels (muscle, tissues) to the most rigid (exoskeleton, shell). In this tutorial, the practical aspects of in vivo CMP NMR are discussed including: handling the organisms, rotor preparation, sample spinning, water suppression, editing experiments, and finishes with a brief look at the potential of other heteronuclei (2H, 15N, 19F, 31P) for in vivo research. The tutorial is aimed as a general resource for researchers interested in developing and applying MAS‐based approaches to living organisms. Although the focus here is CMP NMR, many of the approaches can be adapted (or directly applied) using conventional high‐resolution magic angle spinning, and in some cases, even standard solid‐state NMR probes.

show abstract

“…In Drosophila , beta-alanine is part of carcinine dipeptide (beta-alanine-histamine), is mainly localized in the head and thorax [25] and plays an important role in cuticule melanization [26] and signaling in photoreceptor synapse [27]. Similarly to tyrosine, decrease of beta-alanine could be explained by its consumption during the melanization process.…”

Section: Discussionmentioning

confidence: 99%

Metabolomics with Nuclear Magnetic Resonance Spectroscopy in a Drosophila melanogaster Model of Surviving Sepsis

et al. 2016

View full text Add to dashboard Cite

Abstract:Patients surviving sepsis demonstrate sustained inflammation, which has been associated with long-term complications. One of the main mechanisms behind sustained inflammation is a metabolic switch in parenchymal and immune cells, thus understanding metabolic alterations after sepsis may provide important insights to the pathophysiology of sepsis recovery. In this study, we explored metabolomics in a novel Drosophila melanogaster model of surviving sepsis using Nuclear Magnetic Resonance (NMR), to determine metabolite profiles. We used a model of percutaneous infection in Drosophila melanogaster to mimic sepsis. We had three experimental groups: sepsis survivors (infected with Staphylococcus aureus and treated with oral linezolid), sham (pricked with an aseptic needle), and unmanipulated (positive control). We performed metabolic measurements seven days after sepsis. We then implemented metabolites detected in NMR spectra into the MetExplore web server in order to identify the metabolic pathway alterations in sepsis surviving Drosophila. Our NMR metabolomic approach in a Drosophila model of recovery from sepsis clearly distinguished between all three groups and showed two different metabolomic signatures of inflammation. Sham flies had decreased levels of maltose, alanine, and glutamine, while their level of choline was increased. Sepsis survivors had a metabolic signature characterized by decreased glucose, maltose, tyrosine, beta-alanine, acetate, glutamine, and succinate.

show abstract

Metabolite localization in living drosophila using High Resolution Magic Angle Spinning NMR

Cited by 28 publications

References 23 publications

In-Cell Solid-State NMR: An Emerging Technique for the Study of Biological Membranes

In-Cell Solid-State NMR: An Emerging Technique for the Study of Biological Membranes

In vivo comprehensive multiphase NMR

Metabolomics with Nuclear Magnetic Resonance Spectroscopy in a Drosophila melanogaster Model of Surviving Sepsis

Contact Info

Product

Resources

About