Name: visualization of motor axon navigation and quantification of axon arborization in mouse embryos using light sheet fluorescence microscopy
Uploaded: 2018-05-11T15:00:00
Description: Watch this Scientific Journal Video about Visualization of Motor Axon Navigation and Quantification of Axon Arborization In Mouse Embryos Using Light Sheet Fluorescence Microscopy at JoVE.com

LSFM eksperimenter og dataanalyse blev udført delvist ved hjælp af de avancerede optiske mikroskoper Division Instrument tjeneste i Academia Sinica og med bistand fra Ms. Shu-Chen Shen. Vi takke Ms. Sue-Ping Lee fra den Imaging Core facilitet af IMB for betydelig teknisk bistand med Imaris billedanalyse. IMBS videnskabelige engelsk redigering Core gennemgik håndskriftet. Dette arbejde er finansieret af Academia Sinica karriere udvikling Award (CDA-107-L05), mest (104-2311-B-001-030-MY3) og NHRI (NHRI-EX106-10315NC).

Alle levende dyr var holdt i en specifik pathogen gratis (SPF) Dyrefacilitet, godkendt og overvåget af IACUC af Academia Sinica.
1. fiksation
<ol><li>Indsamle embryoner af embryonale dag 12,5 (E12.5) og derefter halshugge og rense dem.</li>
	<li>Fix embryonerne individuelt i 24-godt plader med 1 mL/godt af frisklavede 4% PARAFORMALDEHYD i 1 x fosfat buffer saltvand (PBS) natten over. Der inkuberes ved 4 ° C på en shaker.</li>
	<li>Vaske fast embryoner mindst 3 x, hver med 5-10 min, med 1 ...

<ol>
	<li>Davis-Dusenbery, B. N., Williams, L. A., Klim, J. R., Eggan, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=How+to+make+spinal+motor+neurons.">How to make spinal motor neurons.</a> Development. 141, 491-501 (2014).</li><li>Stifani, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Motor+neurons+and+the+generation+of+spinal+motor+neuron+diversity.">Motor neurons and the generation of spinal motor neuron diversity.</a> Front Cell Neurosci. 8, 293 (2014).</li><li>Kania, A., Jessell, T. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Topographic+motor+projections+in+the+limb+imposed+by+LIM+homeodomain+protein+regulation+of+ephrin-A:EphA+interactions.">Topographic motor projections in the limb imposed by LIM homeodomain protein regulation of ephrin-A:EphA interactions.</a> Neuron. 38, 581-596 (2003).</li><li>Livet, J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=ETS+gene+Pea3+controls+the+central+position+and+terminal+arborization+of+specific+motor+neuron+pools.">ETS gene Pea3 controls the central position and terminal arborization of specific motor neuron pools.</a> Neuron. 35, 877-892 (2002).</li><li>Kanning, K. C., Kaplan, A., Henderson, C. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Motor+neuron+diversity+in+development+and+disease.">Motor neuron diversity in development and disease.</a> Annu Rev Neurosci. 33, 409-440 (2010).</li><li>Matsumoto, S., Kiryu-Seo, S., Kiyama, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Motor+Nerve+Arborization+Requires+Proteolytic+Domain+of+Damage-Induced+Neuronal+Endopeptidase+(DINE)+during+Development.">Motor Nerve Arborization Requires Proteolytic Domain of Damage-Induced Neuronal Endopeptidase (DINE) during Development.</a> J Neurosci. 36, 4744-4757 (2016).</li><li>Nagata, K., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=ECEL1+mutation+implicates+impaired+axonal+arborization+of+motor+nerves+in+the+pathogenesis+of+distal+arthrogryposis.">ECEL1 mutation implicates impaired axonal arborization of motor nerves in the pathogenesis of distal arthrogryposis.</a> Acta Neuropathol. 132, 111-126 (2016).</li><li>Catela, C., Shin, M. M., Lee, D. H., Liu, J. P., Dasen, J. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hox+Proteins+Coordinate+Motor+Neuron+Differentiation+and+Connectivity+Programs+through+Ret/Gfralpha+Genes.">Hox Proteins Coordinate Motor Neuron Differentiation and Connectivity Programs through Ret/Gfralpha Genes.</a> Cell Rep. 14, 1901-1915 (2016).</li><li>Bonanomi, D., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ret+is+a+multifunctional+coreceptor+that+integrates+diffusible-+and+contact-axon+guidance+signals.">Ret is a multifunctional coreceptor that integrates diffusible- and contact-axon guidance signals.</a> Cell. 148, 568-582 (2012).</li><li>Tung, Y. T., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mir-17+approximately+92+Governs+Motor+Neuron+Subtype+Survival+by+Mediating+Nuclear+PTEN.">Mir-17 approximately 92 Governs Motor Neuron Subtype Survival by Mediating Nuclear PTEN.</a> Cell Rep. 11, 1305-1318 (2015).</li><li>Svoboda, K., Yasuda, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Principles+of+two-photon+excitation+microscopy+and+its+applications+to+neuroscience.">Principles of two-photon excitation microscopy and its applications to neuroscience.</a> Neuron. 50, 823-839 (2006).</li><li>Jonkman, J., Brown, C. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Any+Way+You+Slice+It-A+Comparison+of+Confocal+Microscopy+Techniques.">Any Way You Slice It-A Comparison of Confocal Microscopy Techniques.</a> J Biomol Tech. 26, 54-65 (2015).</li><li>Reynaud, E. G., Peychl, J., Huisken, J., Tomancak, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Guide+to+light-sheet+microscopy+for+adventurous+biologists.">Guide to light-sheet microscopy for adventurous biologists.</a> Nat Methods. 12, 30-34 (2015).</li><li>Santi, P. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Light+sheet+fluorescence+microscopy:+a+review.">Light sheet fluorescence microscopy: a review.</a> J Histochem Cytochem. 59, 129-138 (2011).</li><li>Hanley, O., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Parallel+Pbx-Dependent+Pathways+Govern+the+Coalescence+and+Fate+of+Motor+Columns.">Parallel Pbx-Dependent Pathways Govern the Coalescence and Fate of Motor Columns.</a> Neuron. 91, 1005-1020 (2016).</li><li>De Marco Garcia, N. V., Jessell, T. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Early+motor+neuron+pool+identity+and+muscle+nerve+trajectory+defined+by+postmitotic+restrictions+in+Nkx6.1+activity.">Early motor neuron pool identity and muscle nerve trajectory defined by postmitotic restrictions in Nkx6.1 activity.</a> Neuron. 57, 217-231 (2008).</li><li>Li, C. J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=MicroRNA+filters+Hox+temporal+transcription+noise+to+confer+boundary+formation+in+the+spinal+cord.">MicroRNA filters Hox temporal transcription noise to confer boundary formation in the spinal cord.</a> Nat Commun. 8, 14685 (2017).</li><li>Swoger, J., Pampaloni, F., Stelzer, E. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Light-sheet-based+fluorescence+microscopy+for+three-dimensional+imaging+of+biological+samples.">Light-sheet-based fluorescence microscopy for three-dimensional imaging of biological samples.</a> Cold Spring Harb Protoc. 2014, 1-8 (2014).</li><li>Pantazis, P., Supatto, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Advances+in+whole-embryo+imaging:+a+quantitative+transition+is+underway.">Advances in whole-embryo imaging: a quantitative transition is underway.</a> Nat Rev Mol Cell Biol. 15, 327-339 (2014).</li><li>Reynaud, E. G., Tomancak, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Meeting+report:+first+light+sheet+based+fluorescence+microscopy+workshop.">Meeting report: first light sheet based fluorescence microscopy workshop.</a> Biotechnol J. 5, 798-804 (2010).</li></ol>

Flere trin i protokollen kan blive udsat for ændringer under visse omstændigheder. For eksempel, afhænger varigheden af fiksering af alder af embryoner, varierende fra 2 h til 1 eller flere dage ved hjælp af frisklavede PARAFORMALDEHYD. Da fiksering er udført før hele mount immunfarvning for antistoffer, der er følsomme over for protein crosslinking, kan methanol bruges som en alternativ Fikseringsvæske agent. For en høj signal-støj forhold ved farvningen, er det nødvendigt at optimere vaskemiddel koncentratio...

Spinal MNs er del af det centrale nervesystem, men innerverer perifere muskler til at styre bevægelsen. I udvikle rygmarven, er MN stamfaderen (pMNs) udarbejdet i overensstemmelse med signalerne fra notochord og tilgrænsende somites. Alle opdelte post mitotiske MNs genereres derefter fra pMNs, i sidste ende giver anledning til en række MN undertyper langs rostrocaudal aksen af rygmarven1,2. Spinal MNs er Topografisk og anatomisk godt organiseret. Deres morfologiske arrangement korrelerer med placeringen af deres respektive mål i periferien3. Efter at nå deres mål, muskel, axoner modtage cellulære og eksogen neurotrope faktorer, at få dem til at udvide og filial yderligere ind i musklerne. Innervation og forgrening defekter kan bidrage til at danne neuromuskulære vejkryds (NMJ). For eksempel glial-afledte neurotrope faktorer (GDNF)-induceret Pea3 er uundværlig for axon arborization til kutane maximus (CM) og latissimus dorsi (LD) muskler4,5. Derudover vise Spis knockout mus embryoner defekte arborization af phrenic nerver i mellemgulvet, forårsager respirationssvigt og dødelighed umiddelbart efter fødslen6,7. Dette sidste trin i MN modning (dvs., cytoskeletale projektion og forgrening) er derfor afgørende at sikre kommunikationen mellem neuroner og target-cellerne.
For at få arborization mønstre, foretage forskere normalt Konfokal imaging eller to-foton fluorescens lysmikroskopi sektioneret eller hele-mount prøver8,9,10. Begge disse mikroskopi-teknikker generere acceptabel opløsning og dybde penetration. To-foton fluorescens lysmikroskopi indebærer excitation af fluorophores ved samtidige absorption af to lavere-energi fotoner11. Da to-foton excitation bruger nær-infrarød stråling, bidrager nedsat excitation frekvens til reduceret spredning og bedre væv penetration op til 1 mm i væv, således at billeddannelse med større dybde. Konfokal mikroskopi fjerner af filtre out-of-fokus-signaler og kun indsamler lys inden for brændplanet12. Med denne fremgangsmåde, kan billeder af prøver fra forskellige fokale planer kombineres for at producere et tre-dimensionelle (3D) billede via en Z-stakken funktion. Ikke desto mindre reduceres signal intensitet som de fleste af lys er blokeret og høj numeriske åbninger obskure dybde-af-felt. Endnu vigtigere, bidrage begge teknikker til svær solskader og fototoksicitet da hele prøven modtager belysning, selv når kun ét fly er afbildet på et givet tidspunkt.
For at omgå disse mangler, er LSFM blevet et yndet alternativ med fordelene ved at være hurtig, lys-effektiv, og mindre fototoksisk13,14. Desuden giver LSFM multi-view billeddannelse. Denne fremgangsmåde er især velegnet til at visualisere motor axoner og deres sprede terminaler, som de spredes gennem 3D-rum. LSFM udkonkurrerer de andre to muligheder, fordi prøver er monteret på en scene, der tillader rotation omkring en lodret akse og bevægelser langs x, y og z-akser. Dette set-up giver ikke kun mulighed for en minimalt blokerede visning af prøven men også valget af en ønskelig belysning sti, en mangel ved to-foton og konfokal mikroskopi, som begge kræver montering af prøver på en flad dias. LSFM er derfor den mest velegnede værktøj til 3D imaging af axon arborization og til kvantificering af motoriske nerve terminaler i mus embryoner.

<table>
	<tbody>
		<tr>
			<td>Hb9::GFP</td>
			<td>The Jackson labortory</td>
			<td>005029</td>
			<td>Collect embryos of embryonic day 13.5 (E13.5)</td>
		</tr>
		<tr>
			<td>4% Paraformaldehyde (PFA)</td>
			<td></td>
			<td></td>
			<td>For 200ml: Add 20ml 10X PBS, 8g PFA in ddH2O. Adjust pH to 7.4 with NaOH (10N). Filter sterilize and store at -20 &#176;C.</td>
		</tr>
		<tr>
			<td>Phosphate buffer saline 10X (PBS 10X)</td>
			<td></td>
			<td></td>
			<td>For 1L: Add 80 g NaCl, 2 g KCl,14.4g Na2HPO4, 2.4 g KH2PO4 and top up with ddH2O. Autoclave and store at RT.</td>
		</tr>
		<tr>
			<td>Triton X-100</td>
			<td>Sigma</td>
			<td>X100-500ML</td>
			<td></td>
		</tr>
		<tr>
			<td>Fetal Bovine Serum</td>
			<td>ThermoFisher</td>
			<td>26140079</td>
			<td></td>
		</tr>
		<tr>
			<td>Sheep polyclonal anti-GFP</td>
			<td>AbD Serotec</td>
			<td>4745-1051</td>
			<td>1:1000</td>
		</tr>
		<tr>
			<td>Alexa Fluor 488 donkey anti-sheep</td>
			<td>Invitrogen</td>
			<td>A-11015</td>
			<td>1:1000</td>
		</tr>
		<tr>
			<td>RapiClear 1.47 clearing reagent</td>
			<td>SunJin Lab</td>
			<td>RC147001</td>
			<td></td>
		</tr>
		<tr>
			<td>1.5ml micro tube</td>
			<td>Sarstedt</td>
			<td>72.690.001</td>
			<td></td>
		</tr>
		<tr>
			<td>24 wells plate</td>
			<td>ThermoFisher</td>
			<td>142475</td>
			<td></td>
		</tr>
		<tr>
			<td>5 SA Tweezer</td>
			<td>ideal-tek</td>
			<td>3480641</td>
			<td></td>
		</tr>
		<tr>
			<td>Iris Scissors striaght sharp/sharp</td>
			<td>Aesculap</td>
			<td>BC110R</td>
			<td></td>
		</tr>
		<tr>
			<td>Microsurgery Scalpels, single use</td>
			<td>Aesculap</td>
			<td>BA365</td>
			<td></td>
		</tr>
		<tr>
			<td>Dissecting microscope</td>
			<td>Nikon</td>
			<td>SMZ800</td>
			<td></td>
		</tr>
		<tr>
			<td>Shaker</td>
			<td>TKS</td>
			<td>RS-01</td>
			<td></td>
		</tr>
		<tr>
			<td>Lightsheet Z.1 microscope</td>
			<td>Carl Zeiss Microscopy</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Imaris 8.4.0 image analysis software</td>
			<td>Bitplane, Zurich, Switzerland</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>B6.Cg-Tg(Hlxb9-GFP)1Tmj/J (Hb9::GFP mice)</td>
			<td>The Jackson Laboratory</td>
			<td>005029</td>
			<td></td>
		</tr>
	</tbody>
</table>

visualization of motor axon navigation and quantification of axon arborization in mouse embryos using light sheet fluorescence microscopy

Spinal motor neuroner (MNs) udvide deres axoner til at kommunikere med deres innervating mål, derved kontrollere bevægelse og komplekse opgaver i hvirveldyr. Det er således afgørende for at afdække de molekylære mekanismer af hvordan motor axoner navigere til, arborize og innerverer deres perifere muskel mål i løbet af udvikling og degeneration. Selvom transgene Hb9::GFP mus linjer har længe tjente til at visualisere motor axon baner under fosterudviklingen, detaljerede beskrivelser af hele spektret af axon terminal arborization fortsat er ufuldstændige på grund af mønster kompleksiteten og begrænsninger af nuværende Optisk mikroskopi. Her, beskriver vi en forbedret protokol, der kombinerer lette ark Fluorescens mikroskopi (LSFM) og robust billedanalyse til kvalitativt og kvantitativt visualisere udvikle motoriske axoner. Dette system kan let vedtages for at krydse genetiske mutanter eller MN sygdomsmodeller med Hb9::GFP linier, afslørende roman molekylære mekanismer, der fører til fejl i motor axon navigation og arborization.

LSFM indeholder detaljerede 3D visualisering af MN bane og axon arborization i mus embryoner. Under lysfelt vises væv helt gennemsigtig efter at blive nedsænket i den kommercielle clearing reagens. Ingen svind eller hævelse af prøven blev bemærket efter op til en uge efter opbevaring i clearing reagens før billeddannelse. Under fluorescens kanal, er motoriske neuroner mærket med transgenically udtrykt normal god landbrugspraksis (film 1). I nogle tilfælde, er oply...

Watch this Scientific Journal Video about Visualization of Motor Axon Navigation and Quantification of Axon Arborization In Mouse Embryos Using Light Sheet Fluorescence Microscopy at JoVE.com

Visualization of Motor Axon Navigation and Quantification of Axon Arborization In Mouse Embryos Using Light Sheet Fluorescence Microscopy

Her, beskriver vi en protokol til at visualisere motorneuron projektion og axon arborization i transgene Hb9::GFP mus embryoner. Efter immunfarvning for motoriske neuroner brugte vi lys ark Fluorescens mikroskopi billede embryoner til efterfølgende kvantitativ analyse. Denne protokol finder anvendelse på andre neuron navigation processer i centralnervesystemet.

Visualisering af Motor Axon Navigation og kvantificering af Axon Arborization i mus embryoner ved hjælp af lys ark Fluorescens mikroskopi

Spinal motor neurons (MNs) extend their axons to communicate with their innervating targets, thereby controlling movement and complex tasks in ...

Research

JoVE Journal

Neuroscience

Study Glial Cell Heterogeneity Influence on Axon Growth Using a New Coculture Method

In the central nervous system of all mammals, severed axons after injury are unable to regenerate to their original targets and functional recovery is ...

Micron-scale Resolution Optical Tomography of Entire Mouse Brains with Confocal Light Sheet Microscopy

Micron-skala Opløsning Optical Tomography af hele Mouse Brains med konfokal Light Sheet Microscopy

Understanding the architecture of mammalian  brain at single-cell resolution is one of the key issues of neuroscience.  However, mapping neuronal soma and ...

Whole-mount Imaging of Mouse Embryo Sensory Axon Projections

Hele-mount Imaging af museembryo Sensory Axon Fremskrivninger

The visualization of full-length neuronal projections in embryos is essential to gain an understanding of how mammalian neuronal networks develop. Here we ...

The Neuromuscular Junction: Measuring Synapse Size, Fragmentation and Changes in Synaptic Protein Density Using Confocal Fluorescence Microscopy

Den neuromuskulære forbindelse: Måling Synapse Størrelse, fragmentering og ændringer i Synaptic Protein Density Brug Konfokal Fluorescens mikroskopi

The neuromuscular junction (NMJ) is the large, cholinergic relay synapse through which mammalian motor neurons control voluntary muscle contraction. ...

Imaging Ca2+ Dynamics in Cone Photoreceptor Axon Terminals of the Mouse Retina

Imaging Ca2+ Dynamics in Cone Photoreceptor Axon Terminals of the Mouse Retina

Imaging Ca<sup&gt; 2+</sup&gt; Dynamics i Cone Lysmodtageren Axon Terminals i Mouse Retina

Retinal cone photoreceptors (cones) serve daylight vision and are the basis of color discrimination. They are subject to degeneration, often leading to ...

Utilizing Combined Methodologies to Define the Role of Plasma Membrane Delivery During Axon Branching and Neuronal Morphogenesis

Udnytte Kombineret Metoder til Definer rolle plasmamembranen Delivery Under Axon Forgrening og neuronal Morfogenese

During neural development, growing axons extend to multiple synaptic partners by elaborating axonal branches. Axon branching is promoted by extracellular ...

Quadruple Immunostaining of the Olfactory Bulb for Visualization of Olfactory Sensory Axon Molecular Identity Codes

Quadruple Immunostaining af Olfactory Bulb til Visualisering af Olfactory Sensory Axon Molecular Identity Codes

The mouse olfactory system is often used to study mechanisms of neural circuit formation&#12288;because of its simple anatomical structure. An Olfactory ...

Visualization of Thalamocortical Axon Branching and Synapse Formation in Organotypic Cocultures

Visualisering af Thalamocortical Axon forgrening og Synapse dannelse i Organotypic Cocultures

Axon branching and synapse formation are crucial processes for establishing precise neuronal circuits. During development, sensory thalamocortical (TC) ...

Investigating Mammalian Axon Regeneration: In Vivo Electroporation of Adult Mouse Dorsal Root Ganglion

Investigating Mammalian Axon Regeneration: In Vivo Electroporation of Adult Mouse Dorsal Root Ganglion

Behandlende pattedyr Axon regenerering: In Vivo elektroporation af voksen mus dorsalrods Ganglion

Electroporation is an essential non-viral gene transfection approach to introduce DNA plasmids or small RNA molecules into cells. A sensory neuron in the ...

In Situ Visualization of Axon Growth and Growth Cone Dynamics in Acute Ex Vivo Embryonic Brain Slice Cultures

In Situ Visualization of Axon Growth and Growth Cone Dynamics in Acute Ex Vivo Embryonic Brain Slice Cultures

In Situ Visualisering af axonvækst og vækstkegledynamik i akutte ex vivo embryonale hjerneskivekulturer

During neuronal development, axons navigate the cortical environment to reach their final destinations and establish synaptic connections. Growth cones ...