scholarly journals Study of the release of endogenous amines in Drosophila brain in vivo in response to stimuli linked to aversive olfactory conditioning

2020 ◽  
Author(s):  
Sergio Hidalgo ◽  
Nicolás Fuenzalida‐Uribe ◽  
Daniela Molina‐Mateo ◽  
Angélica P. Escobar ◽  
Carlos Oliva ◽  
...  
Keyword(s):  
Olfactory Conditioning ◽  
Drosophila Brain

scholarly journals The Disease-Associated Proteins Drosophila Nab2 and Ataxin-2 Interact with Shared RNAs and Coregulate Neuronal Morphology

Genetics ◽  
2021 ◽  
Author(s):  
J Christopher Rounds ◽  
Edwin B Corgiat ◽  
Changtian Ye ◽  
Joseph A Behnke ◽  
Seth M Kelly ◽  
...  
Keyword(s):  
Rna Binding ◽  
Rna Binding Proteins ◽  
Target Selection ◽  
High Specificity ◽  
Neuronal Morphology ◽  
Functional Protein ◽  
Specific Subset ◽  
Drosophila Brain ◽  
Ataxin 2

Abstract Nab2 encodes the Drosophila melanogaster member of a conserved family of zinc finger polyadenosine RNA-binding proteins (RBPs) linked to multiple steps in post-transcriptional regulation. Mutation of the Nab2 human ortholog ZC3H14 gives rise to an autosomal recessive intellectual disability but understanding of Nab2/ZC3H14 function in metazoan nervous systems is limited, in part because no comprehensive identification of metazoan Nab2/ZC3H14-associated RNA transcripts has yet been conducted. Moreover, many Nab2/ZC3H14 functional protein partnerships remain unidentified. Here, we present evidence that Nab2 genetically interacts with Ataxin-2 (Atx2), which encodes a neuronal translational regulator, and that these factors coordinately regulate neuronal morphology, circadian behavior, and adult viability. We then present the first high-throughput identifications of Nab2- and Atx2-associated RNAs in Drosophila brain neurons using RNA immunoprecipitation-sequencing (RIP-Seq). Critically, the RNA interactomes of each RBP overlap, and Nab2 exhibits high specificity in its RNA associations in neurons in vivo, associating with a small fraction of all polyadenylated RNAs. The identities of shared associated transcripts (e.g., drk, me31B, stai) and of transcripts specific to Nab2 or Atx2 (e.g., Arpc2 and tea) promise insight into neuronal functions of, and genetic interactions between, each RBP. Consistent with prior biochemical studies, Nab2-associated neuronal RNAs are overrepresented for internal A-rich motifs, suggesting these sequences may partially mediate Nab2 target selection. These data support a model where Nab2 functionally opposes Atx2 in neurons, demonstrate Nab2 shares associated neuronal RNAs with Atx2, and reveal Drosophila Nab2 associates with a more specific subset of polyadenylated mRNAs than its polyadenosine affinity alone may suggest.

scholarly journals Micromirror structured illumination microscope for high-speed in vivo drosophila brain imaging

2014 ◽  
Vol 22 (2) ◽  
pp. 1243 ◽  
Author(s):  
A. Masson ◽  
M. Pedrazzani ◽  
S. Benrezzak ◽  
P. Tchenio ◽  
T. Preat ◽  
...  
Keyword(s):  
Brain Imaging ◽  
High Speed ◽  
Structured Illumination ◽  
Drosophila Brain

scholarly journals FLARIM v2.0, an improved method to quantify transcript-ribosome interactions in vivo in the adult Drosophila brain

2021 ◽  
Author(s):  
Petra Richer ◽  
Sean D Speese ◽  
Mary A Logan
Keyword(s):  
Immune Response ◽  
Morphological Changes ◽  
Immune Gene ◽  
Promising Tool ◽  
Matrix Metalloproteinase 1 ◽  
New Variant ◽  
Wide Range ◽  
Drosophila Brain

Neural injury triggers striking immune reactions from glial cells, including significant transcriptional and morphological changes, but it is unclear how these events are coordinated to mount an effective immune response. Here, we present a new variant of the Fluorescence assay to detect ribosome interactions with mRNA (FLARIM), which we term FLARIM v2.0, to visualize single immune gene transcripts and association with ribosomes in glia responding to neurodegeneration. Specifically, using an in vivo axotomy assay in Drosophila, we show that matrix metalloproteinase-1 (Mmp-1) mRNAs and associated ribosomes are detected in distal processes of reactive glia where they are actively engulfing degenerating axonal material, suggesting that local translation is an important component of the glial immune response to axotomy. This work also validates our enhanced FLARIM assay as a promising tool to investigate mechanisms of mRNA transport and translation in a wide range of in vitro and in vivo paradigms.

scholarly journals PARIS, an optogenetic method for functionally mapping gap junctions

eLife ◽  
2019 ◽  
Vol 8 ◽  
Author(s):  
Ling Wu ◽  
Ao Dong ◽  
Liting Dong ◽  
Shi-Qiang Wang ◽  
Yulong Li
Keyword(s):  
Gap Junctions ◽  
Cell Communication ◽  
Cell Type Specificity ◽  
Spatiotemporal Resolution ◽  
Wide Range ◽  
Junctional Coupling ◽  
Chemical Coupling ◽  
Drosophila Brain ◽  
And Function

Cell-cell communication via gap junctions regulates a wide range of physiological processes by enabling the direct intercellular electrical and chemical coupling. However, the in vivo distribution and function of gap junctions remain poorly understood, partly due to the lack of non-invasive tools with both cell-type specificity and high spatiotemporal resolution. Here, we developed PARIS (pairing actuators and receivers to optically isolate gap junctions), a new fully genetically encoded tool for measuring the cell-specific gap junctional coupling (GJC). PARIS successfully enabled monitoring of GJC in several cultured cell lines under physiologically relevant conditions and in distinct genetically defined neurons in Drosophila brain, with ~10 s temporal resolution and sub-cellular spatial resolution. These results demonstrate that PARIS is a robust, highly sensitive tool for mapping functional gap junctions and study their regulation in both health and disease.

scholarly journals An original model of brain infection identifies the hijacking of host lipoprotein import as a bacterial strategy for blood-brain barrier crossing

2020 ◽  
Author(s):  
Billel Benmimoun ◽  
Florentia Papastefanaki ◽  
Bruno Périchon ◽  
Katerina Segklia ◽  
Nicolas Roby ◽  
...  
Keyword(s):  
Blood Brain Barrier ◽  
Group B Streptococcus ◽  
Ldl Receptor ◽  
Systemic Infection ◽  
Brain Barrier ◽  
Brain Infection ◽  
Blood Brain ◽  
Drosophila Brain ◽  
Group B

AbstractPathogens able to cross the blood-brain barrier (BBB) induce long-term neurological sequelae and death. Understanding how neurotropic pathogens bypass this strong physiological barrier is a prerequisite to devise therapeutic strategies. Here we propose an innovative model of infection in the developing Drosophila brain, combining whole brain explants with in vivo systemic infection. We identified several mammalian pathogens able to cross the Drosophila BBB, including Group B Streptococcus (GBS). Amongst GBS surface components, lipoproteins, and in particular the B leucin-rich Blr, were important for BBB crossing and virulence in Drosophila. Further, we identified (V)LDL receptor LpR2, expressed in the BBB, as a host receptor for Blr, allowing GBS translocation through endocytosis. Finally, we demonstrated that Blr is required for BBB crossing and pathogenicity in a murine model of infection. Our results support the relevance of Drosophila for studying host-pathogen interactions and identify a new mechanism by which pathogens exploit host barriers to generate brain infection.

Whole-Cell In Vivo Patch-Clamp Recordings in the Drosophila Brain

2013 ◽  
Vol 2013 (2) ◽  
pp. pdb.prot071704-pdb.prot071704 ◽  
Author(s):  
M. Murthy ◽  
G. Turner
Keyword(s):  
Patch Clamp ◽  
Whole Cell ◽  
Drosophila Brain

scholarly journals Neural stem cell temporal patterning and brain tumour growth rely on oxidative phosphorylation

eLife ◽  
2019 ◽  
Vol 8 ◽  
Author(s):  
Jelle van den Ameele ◽  
Andrea H Brand
Keyword(s):  
Stem Cells ◽  
Stem Cell ◽  
Oxidative Phosphorylation ◽  
Tumour Cells ◽  
Neuronal Diversity ◽  
Temporal Patterning ◽  
Cell Behaviour ◽  
Drosophila Brain ◽  
Insight Into

Translating advances in cancer research to clinical applications requires better insight into the metabolism of normal cells and tumour cells in vivo. Much effort has focused on understanding how glycolysis and oxidative phosphorylation (OxPhos) support proliferation, while their impact on other aspects of development and tumourigenesis remain largely unexplored. We found that inhibition of OxPhos in neural stem cells (NSCs) or tumours in the Drosophila brain not only decreases proliferation, but also affects many different aspects of stem cell behaviour. In NSCs, OxPhos dysfunction leads to a protracted G1/S-phase and results in delayed temporal patterning and reduced neuronal diversity. As a consequence, NSCs fail to undergo terminal differentiation, leading to prolonged neurogenesis into adulthood. Similarly, in brain tumours inhibition of OxPhos slows proliferation and prevents differentiation, resulting in reduced tumour heterogeneity. Thus, in vivo, highly proliferative stem cells and tumour cells require OxPhos for efficient growth and generation of diversity.

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