scholarly journals Environmental and Molecular Modulation of Motor Individuality in Larval Zebrafish

2021 ◽  
Vol 15 ◽  
Author(s):  
John Hageter ◽  
Matthew Waalkes ◽  
Jacob Starkey ◽  
Haylee Copeland ◽  
Heather Price ◽  
...  
Keyword(s):  
Environmental Factors ◽  
Notch Signaling ◽  
Individual Variation ◽  
Environmental Enrichment ◽  
Notch Pathway ◽  
Dependent Manner ◽  
Behavioral Variation ◽  
Directional Bias ◽  
Larval Zebrafish ◽  
Genetic And Environmental Factors

Innate behavioral biases such as human handedness are a ubiquitous form of inter-individual variation that are not strictly hardwired into the genome and are influenced by diverse internal and external cues. Yet, genetic and environmental factors modulating behavioral variation remain poorly understood, especially in vertebrates. To identify genetic and environmental factors that influence behavioral variation, we take advantage of larval zebrafish light-search behavior. During light-search, individuals preferentially turn in leftward or rightward loops, in which directional bias is sustained and non-heritable. Our previous work has shown that bias is maintained by a habenula-rostral PT circuit and genes associated with Notch signaling. Here we use a medium-throughput recording strategy and unbiased analysis to show that significant individual to individual variation exists in wildtype larval zebrafish turning preference. We classify stable left, right, and unbiased turning types, with most individuals exhibiting a directional preference. We show unbiased behavior is not due to a loss of photo-responsiveness but reduced persistence in same-direction turning. Raising larvae at elevated temperature selectively reduces the leftward turning type and impacts rostral PT neurons, specifically. Exposure to conspecifics, variable salinity, environmental enrichment, and physical disturbance does not significantly impact inter-individual turning bias. Pharmacological manipulation of Notch signaling disrupts habenula development and turn bias individuality in a dose dependent manner, establishing a direct role of Notch signaling. Last, a mutant allele of a known Notch pathway affecter gene, gsx2, disrupts turn bias individuality, implicating that brain regions independent of the previously established habenula-rostral PT likely contribute to inter-individual variation. These results establish that larval zebrafish is a powerful vertebrate model for inter-individual variation with established neural targets showing sensitivity to specific environmental and gene signaling disruptions. Our results provide new insight into how variation is generated in the vertebrate nervous system.

scholarly journals Environmental and molecular modulation of motor individuality in larval zebrafish

2021 ◽  
Author(s):  
John Hageter ◽  
Matthew Waalkes ◽  
Jacob Starkey ◽  
Haylee Copeland ◽  
Heather Price ◽  
...  
Keyword(s):  
Environmental Factors ◽  
Individual Variation ◽  
Environmental Enrichment ◽  
Gene Dosage ◽  
Notch Pathway ◽  
Dependent Manner ◽  
Behavioral Variation ◽  
Directional Bias ◽  
Larval Zebrafish ◽  
Genetic And Environmental Factors

Innate behavioral biases such as human handedness are a ubiquitous form of inter-individual variation that are not strictly hardwired into the genome and are influenced by diverse internal and external cues. Yet, genetic and environmental factors modulating behavioral variation remain poorly understood, especially in vertebrates. To identify genetic and environmental factors that influence behavioral variation, we take advantage of larval zebrafish light-search behavior. During light-search, individuals preferentially turn in leftward or rightward loops, in which directional bias is sustained and non-heritable, and maintained by a habenula-rostral PT circuit. Here we use a medium-throughput recording strategy and unbiased analysis to show that significant individual to individual variation exists in wildtype larval zebrafish turning preference. We classify stable left, right, and unbiased turning types, with most individuals exhibiting a directional preference. Raising larvae at elevated temperature selectively reduces the leftward turning type and impacts rostral PT neurons, specifically. Exposure to conspecifics, variable salinity, environmental enrichment, and physical disturbance does not significantly impact inter-individual turning bias. Pharmacological manipulation of Notch signaling and carrying a mutant allele of a known Notch pathway affecter gene, gsx2, disrupted turn bias individuality in a dose-dependent manner. These results establish that larval zebrafish is a powerful vertebrate model for inter-individual variation with sensitivity to specific environmental perturbations and gene dosage.

Abstract 3380: Acetylation-dependent Regulation Of Notch1 Signaling In Endothelial Cells

Circulation ◽  
2008 ◽  
Vol 118 (suppl_18) ◽  
Author(s):  
Virginia Guarani ◽  
Franck Dequiedt ◽  
Andreas M Zeiher ◽  
Stefanie Dimmeler ◽  
Michael Potente
Keyword(s):  
Endothelial Cells ◽  
Notch Signaling ◽  
Cell Fate ◽  
Molecular Mechanisms ◽  
Trichostatin A ◽  
Notch Pathway ◽  
Dependent Manner ◽  
Class Iii ◽  
Cell Fate Decisions ◽  
Knock Down

The Notch signaling pathway is a versatile regulator of cell fate decisions and plays an essential role for embryonic and postnatal vascular development. As only modest differences in Notch pathway activity suffice to determine dramatic differences in blood vessel development, this pathway is tightly regulated by a variety of molecular mechanisms. Reversible acetylation has emerged as an important post-translational modification of several non-histone proteins, which are targeted by histone deacetylases (HDACs). Here, we report that specifically the Notch1 intracellular domain (NICD) is itself an acetylated protein and that its acetylation level is tightly regulated by the SIRT1 deacetylase, which we have previously identified as a key regulator of endothelial angiogenic functions during vascular growth. Coexpression of NICD with histone acetyltransferases such as p300 or PCAF induced a dose- and time-dependent acetylation of NICD. Blocking HDAC activity using the class III HDAC inhibitor nicotinamid (NAM), but not the class I/II HDAC inhibior trichostatin A, resulted in a significant increase of NICD acetylation suggesting that NICD is targetd by class III HDACs for deacetylation. Among the class III HDACs with deacetylase activity (SIRT1, 2, 3, 5), knock down of specifically SIRT1 resulted in enhanced acetylation of NICD. Moreover, wild type SIRT1, but not a catalytically inactive mutant catalyzed the deacetylation of NICD in a nicotinamid-dependent manner. SIRT1, but SIRT2, SIRT3 or SIRT5, associated with NICD through its catalytic domain demonstrating that SIRT1 is a direct NICD deacetylase. Enhancing NICD acetylation by either overexpression of p300 or inhibition of SIRT1 activity using NAM or RNAi-mediated knock down resulted in enhanced NICD protein stability by blocking its ubiquitin-mediated degradation. Consistent with these results, loss of SIRT1 amplified Notch target gene expression in endothelial cells in response to NICD overexpression or treatment with the Notch ligand Dll4. In summary, our results identify reversible acetylation of NICD as a novel molecular mechanism to control Notch signaling and suggest that deacetylation of NICD by SIRT1 plays a key role in the dynamic regulation of Notch signaling in endothelial cells.

Possible Selective Cytotoxicity of Vanadium Complex on Breast Cancer Cells Involving Pathophysiological Pathways

2020 ◽  
Vol 19 (17) ◽  
pp. 2130-2139 ◽  
Author(s):  
Eman S. El-Shafey ◽  
Eslam S. Elsherbiny
Keyword(s):  
Gene Expression ◽  
Cell Death ◽  
Notch Signaling ◽  
Cancer Cells ◽  
Signaling Pathways ◽  
Caspase 3 ◽  
Therapeutic Agent ◽  
Notch Pathway ◽  
Vanadium Complex ◽  
Dependent Manner

Background: Triple-Negative Breast Cancers (TNBC) are among the most aggressive and therapyresistant breast tumors. Development of new treatment strategies that target pathways involved in cancer cells resistance is an attractive candidate to overcome therapeutic resistance. Objective: To clarify the antitumor activity of [VO (bpy)2 Cl] Cl complex as a new therapeutic agent through studying the interplay between apoptosis, autophagy and notch signaling pathways. Methods: Proliferation of MDA-MB-231 cells and IC50 value of the vanadium complex were assessed by MTT assay. Flow cytometry was utilized to detect cell cycle distribution, apoptosis assay, LC3 levels and Acid Vascular Organelles (AVOs). Caspase 3 levels were detected by ELISA. Changes in Notch1 gene expression were assessed by real-time PCR. AVOs qualitative detection was assessed by a fluorescence microscope. Results: The growth of MDA-MB-231 cells was suppressed after treatment with [VO (bpy)2 Cl] Cl complex, in a dose-dependent manner. The affinity for apoptotic cell death induction was shown through the increase in the sub G0 peak, the percentage of early and late apoptotic phases, and the elevation in caspase 3 levels. The affinity for autophagic cell death induction was observed through the increase in the G0/G1 phase, G2/M arrest, the increase of AVOs red fluorescence and elevated LC3 levels. The affinity for notch pathway inhibition was shown through the suppression of Notch 1 gene expression. Conclusion: [VO (bpy)2 Cl] Cl complex could be a promising candidate as therapeutic agent targeting different therapeutic targets including apoptosis, autophagy and notch signaling pathways.

Notch1 Signaling Promotes Survival of Leukemic Cells in Close Contact with Bone Marrow-Derived Stromal Cells by Activating AKT and Stat-3 Pathways.

Blood ◽  
2006 ◽  
Vol 108 (11) ◽  
pp. 1398-1398
Author(s):  
Yoko Tabe ◽  
Linhua Jin ◽  
Teresa McQueen ◽  
Michael Andreeff ◽  
Patrick Zweidler-McKay ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Stem Cell ◽  
Notch Signaling ◽  
Stromal Cells ◽  
Notch Pathway ◽  
Leukemic Cells ◽  
Dependent Manner ◽  
Secretase Inhibitor ◽  
Hes1 Expression ◽  
Reh Cells

Abstract We have previously demonstrated that the bone marrow (BM) microenvironment plays a crucial role in the pathogenesis of acute myeloid leukemia (AML) by influencing tumor growth, survival, and drug resistance. Integrin-linked kinase (ILK) directly interacts with β integrins and phosphorylates AKT in a PI3-kinase(PI3K)-dependent manner. HES1 encodes a basic helix-loop-helix transcription factor downstream of the Notch receptor, and functions as a positive regulator of hematopoietic stem cell self-renewal. In this study, we investigated the functional role of Notch/HES1 signaling in leukemic cell survival stimulated by BM stromal interactions. Direct co-culture of human mesenchymal stem cell (MSC) and leukemic NB4 (AML) or REH (preB-ALL) cells activated ILK kinase activity and enhanced phosphorylation of AKT and GSK3β along with increased Notch1 and HES1 expression. Both, ILK inhibitor QLT0267 or PI3K inhibitor LY294002 inhibited MSC induced p-AKT, p-GSK3β and HES1 expression, while GSK3 inhibitor BIO induced HES1 expression, suggesting that activation of Notch signaling in stromal co-cultures is at least in part mediated via ILK/GSK3β pathway. Because the co-factor Mastermind-like (MAML) is required to transcribe downstream target genes of Notch pathway, we introduced a dominant-negative form of MAML to prevent Notch signaling. dnMAML blocked both basal and MSC-induced expression of cleaved intracellular Notch1 and HES1 and unexpectedly prevented MSC-induced phosphorylation of AktSer473, but not of GSK3βSer9, as documented by Western blot and confocal microscopy analyses (Figure 1A). Co-culture with MSC induced Stat3Tyr705 phosphorylation in NB4 cells, and this effect was abrogated by γ-secretase inhibitor (Fig. 1B). In contrast, Stat3 was still activated in leukemic cells harboring dnMAML (Fig. 1A), suggesting that Stat3Tyr705 phosphorylation is mediated via non-transcriptional effects of Notch or non-Notch effects of γ-secretase inhibition. Co-culture with MSC or with HS27A cells expressing Notch ligand Jagged1 stimulated REH cells proliferation under serum-limited conditions, which was partially blocked by γ-secretase inhibitor and completely abrogated in dnMAML REH cells. Interestingly, dnMAML NB4 cells acquired cell growth ability upon serum limitation. Altogether, these results suggest that interaction of leukemic and bone marrow stromal cells results in activation of PI3K/ILK/GSK3β signaling, the latter in turn activating Notch pathway. Notch activation enhances cell-type dependent leukemia cell survival upon interaction with BM-derived stromal cells. These effects of Notch signaling are at least in part mediated by a feedback activation of AKT pathway in a transcription-dependent manner, and via activation of Stat3 signaling independent of MAML. Our data show Notch-mediated regulation of AKT/Stat3 pathways and provide a novel role for activated Notch signaling in the context of bone marrow microenvironment. Figure 1A Figure 1A. Figure 1B Figure 1B.

scholarly journals The zebrafish reveals dependence of the mast cell lineage on Notch signaling in vivo

Blood ◽  
2012 ◽  
Vol 119 (15) ◽  
pp. 3585-3594 ◽  
Author(s):  
Sahar I. Da'as ◽  
Andrew J. Coombs ◽  
Tugce B. Balci ◽  
Chloe A. Grondin ◽  
Adolfo A. Ferrando ◽  
...  
Keyword(s):  
Mast Cell ◽  
Notch Signaling ◽  
Notch Pathway ◽  
Cell Development ◽  
Notch Receptor ◽  
Receptor Expression ◽  
Specific Marker ◽  
Dependent Manner ◽  
Hematopoietic Stem

We used the opportunities afforded by the zebrafish to determine upstream pathways regulating mast cell development in vivo and identify their cellular origin. Colocalization studies demonstrated zebrafish notch receptor expression in cells expressing carboxypeptidase A5 (cpa5), a zebrafish mast cell-specific marker. Inhibition of the Notch pathway resulted in decreased cpa5 expression in mindbomb mutants and wild-type embryos treated with the γ-secretase inhibitor, Compound E. A series of morpholino knockdown studies specifically identified notch1b and gata2 as the critical factors regulating mast cell fate. Moreover, hsp70::GAL4;UAS::nicd1a transgenic embryos overexpressing an activated form of notch1, nicd1a, displayed increased cpa5, gata2, and pu.1 expression. This increase in cpa5 expression could be reversed and reduced below baseline levels in a dose-dependent manner using Compound E. Finally, evidence that cpa5 expression colocalizes with lmo2 in the absence of hematopoietic stem cells revealed that definitive mast cells initially delineate from erythromyeloid progenitors. These studies identify a master role for Notch signaling in vertebrate mast cell development and establish developmental origins of this lineage. Moreover, these findings postulate targeting the Notch pathway as a therapeutic strategy in mast cell diseases.

scholarly journals Necroptotic astrocytes contribute to maintaining stemness of disseminated medulloblastoma through CCL2 secretion

2019 ◽  
Vol 22 (5) ◽  
pp. 625-638 ◽  
Author(s):  
Hailong Liu ◽  
Youliang Sun ◽  
Jenny A O’Brien ◽  
Janusz Franco-Barraza ◽  
Xueling Qi ◽  
...  
Keyword(s):  
Notch Signaling ◽  
Critical Role ◽  
Notch Pathway ◽  
Prognostic Indicator ◽  
Kinase Domain ◽  
Janus Kinase ◽  
Dependent Manner ◽  
Primary Therapy ◽  
Essential Components ◽  
Signaling Activation

Abstract Background Medulloblastoma (MB) with metastases at diagnosis and recurrence correlates with poor prognosis. Unfortunately, the molecular mechanism underlying metastases growth has received less attention than primary therapy-naïve MB. Though astrocytes have been frequently detected in brain tumors, their roles in regulating the stemness properties of MB stem-like cells (MBSCs) in disseminated lesions remain elusive. Methods Effects of tumor-associated astrocyte (TAA)–secreted chemokine C-C ligand 2 (CCL2) on MBSC self-renewal was determined by immunostaining analysis. Necroptosis of TAA was examined by measuring necrosome activity. Alterations in Notch signaling were examined after inhibition of CCL2. Progression of MBSC-derived tumors was evaluated after pharmaceutical blockage of necroptosis. Results TAA, as the essential components of disseminated tumor, produced high levels of CCL2 to shape the inflammation microenvironment, which stimulated the enrichment of MBSCs in disseminated MB. In particular, CCL2 played a pivotal role in maintaining stem-like properties via Janus kinase 2/signal transducer and activator of transcription 3 (JAK2/STAT3)–mediated activation of Notch signaling. Loss of CCL2/C-C chemokine receptor 2 (CCR2) function repressed the JAK2/STAT3-Notch pathway and impaired MBSC proliferation, leading to a dramatic reduction of stemness, tumorigenicity, and metastasizing capability. Furthermore, necroptosis-induced CCL2 release depended on activation of receptor-interacting protein 1 (RIP1)/RIP3/mixed lineage kinase domain-like pseudokinase (MLKL) in TAA, which promoted the oncogenic phenotype. Blockade of necroptosis resulted in CCL2 deprivation and compromised MBSC self-proliferation, indicating MBSCs outsourced CCL2 from necroptotic TAA. Finally, CCL2 was upregulated in high-risk stages of MB, further supporting its value as a prognostic indicator. Conclusion These findings highlighted the critical role of CCL2/CCR2 in Notch signaling activation in MBSCs and revealed a necroptosis-associated glial cytokine microenvironment driving stemness maintenance in disseminations. Key Points 1. TAA-derived CCL2 promoted stemness in disseminated MBSCs through Notch signaling activation via the JAK2/STAT3 pathway. 2. TAA released CCL2 in a RIP1/RIP3/MLKL-dependent manner leading to necroptosis.

scholarly journals Notch Signaling Molecules Activate TGF-βin Rat Mesangial Cells under High Glucose Conditions

2013 ◽  
Vol 2013 ◽  
pp. 1-8 ◽  
Author(s):  
Li Liu ◽  
Chenlin Gao ◽  
Guo Chen ◽  
Xia Li ◽  
Jia Li ◽  
...  
Keyword(s):  
Diabetic Nephropathy ◽  
Notch Signaling ◽  
Signaling Pathway ◽  
High Glucose ◽  
Transforming Growth Factor ◽  
Mesangial Cells ◽  
Notch Pathway ◽  
Notch Signaling Pathway ◽  
Signaling Molecules ◽  
Dependent Manner

The involvement of the Notch signaling pathway in the cellular differentiation of the mammalian kidney is established. Recently, the dysregulation of Notch signaling molecules has been identified in acute and chronic renal injuries, fibrosis models, and diabetic kidney biopsies. The canonical Notch ligand , Jagged1, is upregulated in a transforming growth factor-beta- (TGF-β-) dependent manner during chronic kidney disease. TGF-β, a central mediator of renal fibrosis, also is a major contributor to the development of diabetic nephropathy. To explore the roles and possible mechanisms of Notch signaling molecules in the pathogenesis of diabetic nephropathy, we exposed cultured rat mesangial cells to aγ-secretase inhibitor (DAPT) or high glucose and measured the expression of Notch signaling molecules and the fibrosis index. Notch pathway-related molecules, TGF-β, and fibronectin increased with exposure to high glucose and decreased with DAPT treatment. Our results suggest that the Notch signaling pathway may precipitate diabetic nephropathy via TGF-βactivation.

scholarly journals The roles and activation of endocardial Notch signaling in heart regeneration

2021 ◽  
Vol 10 (1) ◽  
Author(s):  
Huicong Li ◽  
Cheng Chang ◽  
Xueyu Li ◽  
Ruilin Zhang
Keyword(s):  
Notch Signaling ◽  
Tissue Regeneration ◽  
Primary Cilia ◽  
Cardiac Injury ◽  
Notch Pathway ◽  
Heart Regeneration ◽  
Larval Zebrafish ◽  
Flow Change ◽  
Mechanosensitive Ion Channel ◽  
Stress Models

AbstractAs a highly conserved signaling pathway in metazoans, the Notch pathway plays important roles in embryonic development and tissue regeneration. Recently, cardiac injury and regeneration have become an increasingly popular topic for biomedical research, and Notch signaling has been shown to exert crucial functions during heart regeneration as well. In this review, we briefly summarize the molecular functions of the endocardial Notch pathway in several cardiac injury and stress models. Although there is an increase in appreciating the importance of endocardial Notch signaling in heart regeneration, the mechanism of its activation is not fully understood. This review highlights recent findings on the activation of the endocardial Notch pathway by hemodynamic blood flow change in larval zebrafish ventricle after partial ablation, a process involving primary cilia, mechanosensitive ion channel Trpv4 and mechanosensitive transcription factor Klf2.

Using the Zebrafish As a Tool for Modeling Systemic Mastocytosis,

Blood ◽  
2011 ◽  
Vol 118 (21) ◽  
pp. 3208-3208
Author(s):  
Tugce B Balci ◽  
Andrew J Coombs ◽  
Chloe Grondin ◽  
Sahar I Da'as ◽  
Ian Chute ◽  
...  
Keyword(s):  
Mast Cells ◽  
Notch Signaling ◽  
Systemic Mastocytosis ◽  
Notch Pathway ◽  
Notch Signalling ◽  
Blue Staining ◽  
Transgenic Zebrafish ◽  
Dependent Manner ◽  
Transgenic Embryos

Abstract Abstract 3208 The zebrafish system provides many advantages in investigating intricate molecular pathways regulating vertebrate blood cell development and disease in vivo. We have been exploiting these assets to elucidate normal mast cell (MC) function and previously described the structural and functional characteristics of zebrafish MC equivalents (Dobson et al, Blood 2008, Da'as et al, Dev Comp Imm 2011). We have used this knowledge to develop transgenic zebrafish models of systemic mastocytosis (SM). SM is a pre-leukemic myeloproliferative disease that results from perturbed MC development and proliferation. Our recent studies have suggested that zebrafish MCs are uniquely dependent on Notch pathway signaling in contrast to that observed for other myeloid cell populations. Whole mount in situ hybridization (WISH) studies on the zebrafish Notch signaling mutant, mindbomb and notch1b “morphant” embryos both displayed decreased to absent carboxypeptidase A5 (cpa5) positive mast cells by WISH. Furthermore, wild type embryos treated with Compound E (CpdE), a γ-secretase inhibitor that inhibits Notch signaling, showed a similar phenotype. Given the role for Notch signalling in normal MC development, we wanted to see if driving the Notch pathway would result in a phenotype reminiscent of SM. Through a heat-shock inducible Gal4-UAS based system; we now demonstrate that over-expression of Notch signalling results in increased cpa5 positive mast cells in embryos as observed by WISH at 30, 36 and 48 hours post fertilization (hpf). Importantly, we were able to inhibit this increase and even reduce MC numbers below baseline levels in a dose-dependent manner using CpdE. Concurrently, we have established a transgenic zebrafish model of SM that ubiquitously expresses the human c-KIT D816V mutation under the zebrafish β-actin promoter. Beginning at 9 months of age, adult fish develop a number of skin and visceral lesions, many of which have been found to contain an abundance of MCs as identified by toluidine blue staining and tryptase immunohistochemistry. Transgenic embryos lack a developmental phenotype but demonstrate evidence of decreased phospho-histone H3 (pH3) signaling, suggesting additional mutations are required to progress to SM. In support of this G2/M arrest phenotype, microarray studies conducted on transgenic embryos revealed upregulation of p53 and cyclin G1. These studies have provided new insights into the role of Notch signaling in MC development and the opportunity to use the zebrafish as an in vivo model to identify and evaluate novel therapeutic strategies in MC diseases. Disclosures: No relevant conflicts of interest to declare.

Reductionist thinking and animal models in neuropsychiatric research

2019 ◽  
Vol 42 ◽  
Author(s):  
Nicole M. Baran
Keyword(s):  
Animal Models ◽  
Mental Disorders ◽  
Environmental Factors ◽  
Neuropsychiatric Disorders ◽  
Model Organisms ◽  
Developmental Genetic ◽  
Term Study ◽  
Genetic And Environmental Factors ◽  
Mechanisms Of Plasticity

AbstractReductionist thinking in neuroscience is manifest in the widespread use of animal models of neuropsychiatric disorders. Broader investigations of diverse behaviors in non-model organisms and longer-term study of the mechanisms of plasticity will yield fundamental insights into the neurobiological, developmental, genetic, and environmental factors contributing to the “massively multifactorial system networks” which go awry in mental disorders.

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