Developmental NeurobiologyQuestions we are interested in:-How are ectodermal cells induced to become neuronal cells?- How is Dorsal-Ventral polarity established?-How is Anterior-Posterior polarity established?-How are neurons born?-How do neurons acquire their fates?-How are different neuronal types established??With thanks to Natalia Caporale, Outstanding GSI 2003MCB 160The determination of different cell types (cell fates)involves progressive restrictions in their developmentalpotentials. When a cell “chooses” a particular fate, it is saidto be determined, although it still "looks" just like itsundetermined neighbors.Differentiation follows determination, as the cell elaboratesa cell-specific developmental program.Competence is the ability of a cell to respond to a signal. Itdepends on the receptors, transcription factors, etcexpressed by the cellInitial Steps in the Development of the NervousSystem:FertilizationCleavageGastrulationNeurulationEctoderm Major tissues of the Nervous System,EpidermisMesoderm Connective Tissue, Muscle, Blood, BloodVesselsEndoderm  GutGastrulation can be defined as the process bywhich the embryo acquires 2-3 germ layers. Thereare extensive cell movements and rearrangements.Gastrulation :Blastula GastrulaNeurulation: Formation of theNeural TubeAfter Gastrulation comes Neurulation. The formation of the neural plate, the neural folds, andtheir closure to form the neural tube is called Neurulation.Blastula GastrulaNeurulation: Formation of theNeural TubeNote: This drawing doesn’t show the neural crest cells.Long After Neurulation:Neural TubeRostral CaudalBrain Spinal CordNeural CrestPNS (sensoyganglia, ANS),melanocytes,adrenal medulaXenopus laevis (South African clawed toad)Early Induction of the NervousSystem1920s-Spemann & MangoldInductive Signals  They affect/determine which genesare expressed in a cell.There is a 2nd neuraxis, that contains cells form the host and the donorembryo. Cells in the Dorsal Lip of the Blastopore (DLB) orOrganizer induce ectoderm of the host embryo to become neuraltissue. This is an INDUCTIVE EVENT.Before Gastrulation, they transplant the DLB from a pigmentedanimal to a non pigmented one and they place it in the ventralectoderm.What makes cells become neural?A cell’s fate will depend on the signals to which it is exposed as well as thedevelopmental history of the cell.The default is tobecome neuralExperiment 1:Background: early lateExperiment 2:Expression of a dominant negative mutant of one of thereceptors for one type of BMP in xenopus oocytes results inneural fate. The interpretation is that the BMP signal instructs thecells not to become neuralNote: Adding extra BMP to the animal cap cells results in alager number of cells going to epidermal fate. This suggeststhat BMP promotes epidermal fate.What makes cells become neural?Experiment 3:Richard Harland was looking for genes that would induceneural fate.Take theanimal capInject “Random”mRNAs (firstpools till youisolate the mRNAyou areinterested in)Neuralfate (?)(lookingatneuronalmarkers)In this way he identified noggin. Which is a secretedprotein, expressed in the organizer and later in thenotocord.Other people identified follistatin and chordin ashaving similar functions.What makes cells become neural?Model: The ‘double inhibition model’ for neural fateMembers of the BMP family of proteins inhibit the neural fate bybinding heterodimeric receptors in the ectodermic cells andinitiating a cascade of events. These BMP proteins are secretedby the ectodermal cells. Inhibition of this binding by moleculessuch as chordin, noggin and follistatin, allows the cells to adoptthe neural fate. These molecules are all expressed by cells in theorganizer region. The fate (neural vs epidermal) depends on athreshold.What makes cells become neural?Dorsal-Ventral PatterningDuring development, various structures are formed as the posteriorpart of the neural tube becomes the spinal cord:Dorsal•Roof Plate•Sensory InterneuronsVentral•Floor Plate•Motor NeuronsNeural FoldSpinal CordExperiment 1: Ventral Induction The Notochord induces ventral fate in the spinal cord.Let’s consider the chick spinal cord as our model system.Sonic Hedgehog (Shh) is a secreted protein expressed in the floorplate and notochord and it is the signal that induces the ventral fate.The particular fate of the different cells depends on the amount ofShh that they are exposed to.Low Concentration  Ventral InterneuronsMedium Concentration  Motor NeuronsHigh Concentration  Floor Plate CellsWhat is the signal released by the notochord?Threshold effect Dorsal-Ventral PatterningExperiments by Tom Jessell and colleagues…Thresholds, in general terms:Different cellular responses according to positionof cell in concentration gradient of morphogen=> results in discrete differences in cell fate alongthe gradient.Experiment 2: Dorsal InductionControlAdditionalnotochordWhat was observed:•Ventralization of the spinal cord•Extinguished BMP expressionThis suggests that BMP is neededfor dorsalization but doesn’t prove itThe control presented:•Normal dorsal and ventraldevelopment of the spinal cord•BMP expression in the roof plateDorsal-Ventral PatterningIntermediateplate (IP)FloorPlate(FP)IPFPMNFPFP- BMP+ BMPWhen culturing intermediate plate explants on floor plateexplants, the cells adopt ventral fates (MN). However, if there isBMP in the medium, then this is inhibited. BMP inhibits ventralization (as shown by the lack ofdifferentiation into motor neurons)Experiment 3: Dorsal InductionExperiment 4:Removal of the notochordresults in• the lack of ventralphenotype•BMP expressed everywhereDorsal-Ventral PatterningModel:•Shh signaling: induction of ventral cell fates.•BMP signaling: induction for dorsal cell fates.•BMP inhibits ventral fate.•Shh inhibits dorsal fate.BMPSHHdorsal ventralBMPSHHdorsal ventralDorsal-Ventral PatterningThe A/P patterning begins at the stageof the neural plate. The neural tissuesinduced by follistatin, noggin andchordin appear to express the genesthat are characteristic of the forebrain.Additional signaling is required for theinduction of tissue that will give rise tothe midbrain, hindbrain and spinalchord.Focus on the Hindbrain:The hindbrain is ‘segmented’ intorhombomeres. These expressdifferent genes during developmentand present the soma of the motorneurons of different cranial nervesin the adult.Anterior-Posterior