11/5/2021 0 Comments Merlin Project Predictions 2017
The Hippo pathway is emerging as a key evolutionarily conserved signaling mechanism that controls organ size. Actually for feedforward networks I found it to be a bit more flexible to directly implement the prediction using a linear algebra library - mainly just a few matrix. 10:46 GMT+01:00 Markus Toman : Regarding C++: I did it by exporting the weights and then constructing the same network with another toolkit.Genetic perturbation of Hippo pathway members can lead to tumorigenesis in mice ( Dong, et al., 2007) and the Neurofibromatosis 2 (NF2) tumor syndrome in humans ( Rouleau, et al., 1993). Parallel studies in mammals have demonstrated that the components and overall functions of the Hippo pathway are evolutionarily conserved ( Yu, et al., 2015). The Hippo pathway is composed of upstream activators consisting of Merlin (Mer), Expanded (Ex), Kibra (Kib) and Salvador (Sav), a core kinase cassette consisting of Tao1, Hippo (Hpo) and Warts (Wts), and a downstream transcriptional co-activator, Yorkie (Yki) ( Boggiano and Fehon, 2012). Pioneering work in Drosophila has identified the Hippo pathway as having a key role in regulating organ growth ( Huang, et al., 2005 Harvey, et al., 2003 Wu, et al., 2003 Tapon, et al., 2002 Xu, et al., 1995). Here are five tips for gaining buy-in for projects.With 2.5+ years of experience in Software development, Machine learning and Data Analytics, I have worked majorly on Fintech product to detect Fraud and also on multiple projects like Health application, Careem Food Delivery product, Customer Revenue Prediction and Analytics of Food and Beverage System.How multicellular tissues reach and maintain appropriate size is a fundamental and fascinating question in developmental biology.
Mer, Ex and Kib also are known to form a complex, colocalize at the junctions, and function upstream of the core kinases, suggesting that these three proteins function together to regulate pathway activity ( Enderle and McNeill, 2013 Yu, et al., 2010). Hippo pathway components including Crumbs (Crb), Ex, Mer, Echinoid (Ed), Ajuba (Jub), Sav, Hpo and Wts have been shown to localize junctionally ( Enderle and McNeill, 2013 Boggiano and Fehon, 2012), and recent studies indicate that the junctional domain is a primary site of pathway activation ( Chung, et al., 2016 Sun, et al., 2015). In polarized epithelia such as the Drosophila imaginal discs, the cortex and associated plasma membrane are organized into apical, junctional and basolateral domains ( Tepass, 2012) ( Figure 1A). 29 September 2017 / Accepted: 2 October 2017 / Published: 16 October 2017.A striking aspect of Hippo pathway function and regulation is its close relationship with the cell cortex and intercellular junctions ( Enderle and McNeill, 2013 Boggiano and Fehon, 2012). We show that Mer and Kib function at the medial apical cell cortex where they can recruit Sav, Hpo and Wts to promote Hippo pathway activation, all independently of Ex. Mer and Kib show extensive co-localization junctionally (yellow arrows) and medially (white arrows).(FâFâ³) Co-staining of Mer and Sav shows co-localization at the junctions and medially.(G) Cartoons summarizing the observed localizations of proteins examined in this figure.All panels are taken from the blade region of third-instar wing imaginal discs.Here we use endogenously expressed, fluorescently-tagged proteins in intact tissues to explore the functional relationships between upstream Hippo pathway components and the core kinases. Junctional accumulation is indicated by yellow arrows and medial accumulation by white arrows.(EâEâ³) Co-staining of Mer and Kib in the wing epithelium. The junctional domain is composed of the adherens junction (AJ) and marginal zone (MZ).(BâD) Tangential sections through the apical domain of wing epithelial cells expressing Mer-YFP, Kib:GFP and Ex-YFP, respectively. Thus, despite considerable progress in understanding how Hippo pathway activity is regulated, we do not yet understand how these proteins are organized and function at the cell cortex, particularly in the context of living tissues.Mer and Kib accumulate at both the junctional and medial domains of wing imaginal epithelial cells, while Ex only localizes to junctions(A) Cartoons showing side (cross section) and top (tangential section) views of a typical imaginal disc cell. Furthermore, both Ex and Mer can recruit Wts to the plasma membrane ( Sun, et al., 2015 Yin, et al., 2013). All tagged, endogenously expressed proteins used in this study are fully functional, as judged by the ability to rescue null mutations in the corresponding genes (YFP-tagged transgenes) or homozygous viability ( kib:GFP data not shown).In living wing imaginal discs, Ex-YFP appeared exclusively localized to the junctions, while both Mer-YFP and Kib:GFP localized both junctionally and to non-junctional punctae localized within the medial apical cortex ( Figures 1Bâ1D). To assess the subcellular localization of endogenously expressed Mer and other Hippo pathway components in living tissues, we generated endogenously expressed, fluorescently tagged forms of Mer, Kib and Ex either using CRISPR/Cas-9 genome editing ( kib:GFP) or transgenes made by recombineering tags into genomic sequences contained within bacterial artificial chromosome (BAC) clones ( Mer-YFP and ex-YFP). However, close examination of Mer antibody staining shows significant non-junctional staining at the medial apical cell cortex ( McCartney, et al., 2000 McCartney and Fehon, 1996). Together, these findings reveal a previously unrecognized second cortical domain for pathway activation, elucidate the cellular basis underlying observed genetic redundancies between Mer and Ex, and provide a mechanistic better understanding for how Mer, the product of the NF2 tumor suppressor, regulates Hippo signaling.Mer, Kib and Sav colocalize at a non-junctional sitePrevious studies have shown that the upstream Hippo pathway regulators Mer, Kib and Ex primarily localize to the junctional domain in imaginal epithelia cells ( Sun, et al., 2015 Enderle and McNeill, 2013 Genevet, et al., 2010). Strikingly, although Crb synergizes with Ex at junctions to promote HSW pathway activation, Crb antagonizes Kibâs ability to restrict growth by recruiting it to the junctions. When we co-stained Mer and Sav in the wing imaginal disc, we found that these proteins also co-localized both junctionally and medially at the apical ends of imaginal epithelial cells ( Figure 1Fââ1Fâ³).To further define the specific apical domain at which these components localize, we co-stained for Merlin and three apically-localized markers - Cad99C, a marker for apical microvilli ( Glowinski, et al. We have not yet tagged Sav for imaging in live tissues, but published antisera recognize endogenous Sav ( Yue, et al., 2012). A previous study has reported that Sav localizes junctionally, where it interacts with the transmembrane protocadherin Ed ( Yue, et al., 2012). Silver from sonic coloring pageHigh magnification insets of anterior ( wild-type ROI 1) in Aâ³ and posterior ( Mer-RNAi, kib-RNAi ROI 2) in Aâ´ cells show loss of medial Sav in Kib/Mer depleted cells.(BâC) Kib expression enhances Mer and Sav abundance apically. Note that while total Sav staining is not strongly affected by reduction of Mer and Kib, Sav medial is dramatically reduced. Anti-E-cad staining (not shown) was used to mark the junctions and generate a binary mask to remove junctional Sav from the image (compare Sav total in A to Sav medial in Aâ²). Thus, our analysis reveals that three important Hippo pathway regulators, Kib, Mer and Sav, localize to the medial apical cortex, a previously unrecognized subcellular localization for pathway components ( Figure 1G).Kib and Mer work together to promote accumulation of Sav at the medial apical domain(AâAâ´) Sav localization in the wing epithelium expressing both UAS-Mer-RNAi and UAS-kib-RNAi in the posterior compartment under hh-Gal4 (dashed line indicates anterior-posterior compartment boundary). These results indicate that Merlin, and by association Kib and Sav, localize to the apical cortex, immediately adjacent to the apical membrane. Serial optical sectioning of co-stained tissues revealed that the plane of maximal Merlin staining intensity coincides well with that of Cad99C ( Figure S1A) and Squash ( Figure S1C), but is apical to the maximal plane of Rab5-CFP ( Figure S1B). The composite images indicate co-localization of Kib and Mer (Bâ²âBâ´) or Mer and Sav (Câ²âCâ´) at the junctions (yellow arrows) and medially (white arrows).
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