图书简介
This volume provides experimeTal approaches aimedTo charaTerizeThe Cerebral Cavernous MalformTions (CCM) disease andTo defineThe cellular and molecular mechanisms underlyingThis pThology. ChaTers are divided iTo four seTions providing a general overview ofThe nTural hiTory, epidemiology, and pThogenTic mechanisms of CCM disease, describing mThods curreTly used for diagnosis andTreTmeT, produTion and analysis of diTinT cellular and animal models in which CCM can be Tudied, and differeT mThodological approachesTo TudyThe mechanisms underlyingThe CCM onsT and progression. WrTten inThe highly successful MThods in Molecular Biology series formT, chaTers include iTroduTionsToTheir respeTiveTopics, liTs ofThe necessary mTerials and reageTs, Tep-by-Tep, readily reproducible laborTory prTocols, andTips onTroubleshoTing and avoiding known pTfalls.AThorTTive and cTting-edge,Cerebral Cavernous MalformTions (CCM): MThods and PrTocols aimsTo ensure successful resuTs inThe fuTher Tudy ofThis complex disease and Ts pThophysiologic correlTes.
From Genes and MChanisms to MolCular-targeted Therapies: the LongClimb to theCure ofCerebralCavernous Malformation CCM) Disease.- ICideCe, PrevaleCe, andClinCal Presentation ofCerebralCavernous Malformations.- Natural History,ClinCal, and SurgCal Management ofCavernous Malformations.- MolCular GenetCs Creening ofCCM patients, An Overview.- Next Generation SequCing (NGS) Strategies for GenetC Testing and Diagnosis ofCerebralCavernous Malformation CCM) Disease.- Genome-wide Genotyping ofCerebralCavernous Malformation Type 1 Individuals to Identify GenetC Modifiers of Disease Severity.-ClinCal Imaging ofCerebralCavernous Malformations:Computed Tomography and MagnetC ResonaCe Imaging.- Neuroradiology: Differential Diagnosis, Follow-Up, and Reporting.- SurgCal Management of BrainCavernous Malformations.- Generation ofCCM Phenotype by a Human MCrovaCular Endothelial Model.- Isolation and PurifCation of Mouse Brain EndothelialCells to StudyCerebralCavernous Malformation Disease.- ProdCtion of KRIT1-knCkout and KRIT1-knCkin Mouse EmbryonC Fibroblasts asCellular Models ofCCM Disease.CRISPRCas9-mediated Generation of Human EndothelialCell KnCkout Models ofCCM Disease.- DissCtion of the Role ofCCM Genes in Tubulogenesis using theDrosophilaTrCheal System as a Model.- Generation and Analysis ofCCM Phenotypes iC. elegans.-Generation of TransgenC Lines of Zebrafish Expressing FluoreCently-taggedCCM Proteins to Study Their FuCtion and SuCellular LCalization within the VaCulature.- Vertebrate Models to InvestigateCCM Pathogenesis: The Zebrafish and Mouse Model.- Generation ofCerebralCavernous Malformation in Neonatal Mouse Models using IndCibleCre-LoxP Strategy.- Isolation ofCerebral EndothelialCells fromCCM1/KRIT1 Null Mouse Brain.- IdentifCation of the KRIT1 Protein by LexA-based Yeast Two-Hybrid System.-CrystallographC Studies of theCerebralCavernous Malformations Proteins.- MCroCopy TChniques to InvestigateCCM Pathogenesis.- Preparation and Analysis of Protein ExtrCts to InvestigateCCM Pathogenesis.- Systems Wide Analysis ofCCM SignalingComplex CC) Alterations inCCM DefCient Models using OmCs ApproChes.- Study of MolCular InterCtions ofCCM Proteins by using a GAL4-based Yeast Two-hybrid Creening.- Study ofCCM MCrovaCular Endothelial Phenotype by anin vitroTubule Differentiation Model.- Bidimentionalin vitroAngiogenC Assays to StudyCCM Pathogenesis: EndothelialCell Proliferation and Migration.- Measurement of Endothelial Barrier FuCtion in Mouse Models oCerebralCavernous Malformations using Intravital MCroCopy.- ImmunofluoreCeCe ofCellCell andCell-ExtrCellular Matrix Adhesive DefCtsinin vitroEndothelialCCM Model. JuxtCrine Role of Mutant ExtrCellular Matrix on Wild-Type EndothelialCells.- DetCtion of p62/SQSTM1 Aggregates inCellular Models ofCCM Disease by ImmunofluoreCeCe.- NoCh Signaling in FamilialCerebralCavernous Malformations and ImmunohistChemCal DetCtion ofCleaved NoCh1 IntrCellular Domain.- Measuring the Kinase Ctivity of CKIII Proteinsin vitro.-SpCtrophotometrC Method for Determining Glyoxalase 1 Ctivity inCerebralCavernous Malformation CCM) Disease.- FluoreCeCe Analysis of ReCtive Oxygen SpCies (ROS) inCellular Models ofCerebralCavernous Malformation Disease.- Small RNA Library Preparation for Profiling Small NonCoding RNAs in Patients AffCted byCavernous Malformations.- Affinity PurifCation and Preparation of Peptides for Mass SpCtrometry froC. elegans.
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