Name: acute and chronic models of hyperglycemia in zebrafish: a method to assess the impact of hyperglycemia on neurogenesis and the biodistribution of radiolabeled molecules
Uploaded: 2017-06-26T12:30:00
Description: Watch this Scientific Journal Video about Acute and Chronic Models of Hyperglycemia in Zebrafish: A Method to Assess the Impact of Hyperglycemia on Neurogenesis and the Biodistribution of Radiolabeled

Vi takker sterkt Direction des Usages du Numérique (DUN) fra La Réunion University for redigering av videoen (spesielt Jean-François Février, Eric Esnault og Sylvain Ducasse), Lynda-Rose Mottagan for voiceover, Mary Osborne-Pellegrin for korrekturlesing Voice-over, og CYROI-plattformen. Dette arbeidet ble støttet av tilskudd fra La Réunion University (Bonus Qualité Recherche, Dispositifs incitatifs), Conseil Regional de La Réunion, European Union (CPER / FEDER) og Philancia-foreningen. ACD er mottaker av stipend stipend fra Ministère de l&#39;Education Nationale, de l&#39;Enseignement Supérieur et de la Recherche, La Réunion Universitetet (Contrat Doctoral).

Voksne wildtype zebrafish ( Danio rerio ) ble opprettholdt under standard fotoperiod (14/10 h lys / mørk) og temperatur (28 ° C) forhold. Alle eksperimenter ble utført i samsvar med retningslinjene for fransk og europeisk fellesskap for bruk av dyr i forskning (86/609 / EEC og 2010/63 / EU) og ble godkjent av det lokale etikkutvalget for dyreforsøk. 1. Etablere en modell for akutt hyperglykemi hos sebrafisk <ol><li> Forbered en stamløsning av tricaine (MS-222) ved å oppløse...

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W., Tropepe, V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+comparative+framework+for+understanding+the+biological+principles+of+adult+neurogenesis.">A comparative framework for understanding the biological principles of adult neurogenesis.</a> Prog Neurobiol. 80 (6), 281-307 (2006).</li><li>M&#228;rz, M., 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=Heterogeneity+in+progenitor+cell+subtypes+in+the+ventricular+zone+of+the+zebrafish+adult+telencephalon.">Heterogeneity in progenitor cell subtypes in the ventricular zone of the zebrafish adult telencephalon.</a> Glia. 58 (7), 870-888 (2010).</li><li>Chapouton, P., Jagasia, R., Bally-Cuif, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Adult+neurogenesis+in+non-mammalian+vertebrates.">Adult neurogenesis in non-mammalian vertebrates.</a> Bioessays. 29 (8), 745-757 (2007).</li><li>Lindsey, B. W., Darabie, A., Tropepe, V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+cellular+composition+of+neurogenic+periventricular+zones+in+the+adult+zebrafish+forebrain.">The cellular composition of neurogenic periventricular zones in the adult zebrafish forebrain.</a> J Comp Neurol. 520 (10), 2275-2316 (2012).</li><li>Sarras, M. P., Intine, R. 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Dette arbeidet beskriver ulike metoder for å etablere akutte og kroniske modeller av hyperglykemi hos sebrafisk. De viktigste fordelene ved disse prosedyrene er at: (1) de tillater reduksjon i antall pattedyr som brukes til forskning, (2) de er enkle å sette opp og raskt å implementere, og (3) de er økonomiske. Derfor tillater slike modeller undersøkelsen av virkningen av hyperglykemi på et stort antall dyr for å studere dens innvirkning på forskjellige fysiologiske prosesser, inkludert aterotrombose, kardiovask...

Hyperglykemi er definert som for høyt blodsukkernivå. Selv om det kan gjenspeile en situasjon med akutt stress, er hyperglykemi også en tilstand som ofte fører til diagnose av diabetes, en kronisk lidelse av insulinsekresjon og / eller motstand. I 2016 har antall voksne som lever med diabetes nådd 422 millioner over hele verden, og hvert år dør 1,5 millioner mennesker av denne sykdommen, noe som gjør det til et stort helseproblem 1 . Faktisk fører ukontrollert diabetes til flere fysiologiske lidelser som påvirker kardiovaskulærsystemet, nyrene og perifere og sentrale nervesystemer. Interessant kan akutt og kronisk hyperglykemi endre kognisjon og bidra til både demens og depresjon 2 , 3 , 4 , 5 , 6 . I tillegg er inntak av pasienter wI hyperglykemi er det blitt forbundet med dårligere funksjonelle, neurologiske og overlevelsesutfall etter iskemisk berøring 7 , 8 , 9 , 10 , 11 . Det ble også vist at hyperglykemi / diabetes påvirker voksen neurogenese, en prosess som fører til generering av nye nevroner, ved å påvirke nevrale stamcelleaktivitet og neuronal differensiering, migrasjon og overlevelse 2 , 12 . I motsetning til pattedyr, viser teleostfisk, som sebrafisk, intensiv nevogenisk aktivitet gjennom hele hjernen og har en fremragende evne til hjernearbeid i voksen alder 13 , 14 , 15 , 16 . Spesielt er slike kapasiteter mulige på grunn av persistensen av neuRal stamme / stamceller, inkludert radial glia og neuroblaster 17 , 18 , 19 . I tillegg har zebrafisken nylig kommet fram som en modell for å studere metabolske forstyrrelser, inkludert fedme og hyperglykemi / diabetes 20 , 21 , 22 . Selv om zebrafisken er en velkjent modell for hyperglykemi og nevrogenese, har få studier undersøkt virkningen av hyperglykemi på hjernens homeostase og kognitive funksjon 12 , 23 . For å fastslå effekten av hyperglykemi på konstitutiv og skadefremkalt hjernecelleproliferasjon ble en modell av akutt hyperglykemi opprettet ved intraperitoneal injeksjon av D-glukose. I tillegg ble en modell av kronisk hyperglykemi reprodusert ved nedsenkning av fisk i vann tilsatt wMed D-glukose 12 . Sebrafisk har mange fordeler i forskning. De er billige, enkle å øke og gjennomsiktige i de første utviklingsstadiene, og deres genom er blitt sekvensert. I sammenheng med dette arbeidet viser de også flere flere fordeler: (1) de deler lignende fysiologiske prosesser med mennesker, noe som gjør dem til et kritisk verktøy for biomedisinsk forskning; (2) de tillater hurtig undersøkelse av virkningen av hyperglykemi på hjernens homeostase og neurogenese, gitt deres utbredt og sterk nevogenisk aktivitet; Og (3) de er en alternativ modell som gjør det mulig å redusere antallet pattedyr som brukes i forskning. Til slutt kan sebrafisk brukes som en modell for testing av biodistribusjon av radiomerkede molekyler og potensielle terapeutiske midler ved bruk av PET / CT. Det overordnede målet med følgende prosedyre er å visuelt dokumentere hvordan man oppretter modeller av akutt og kronisk hyperglykemi i sebrafisk, bruk zebElektroish for å vurdere hjernemodellering i hyperglykemiske forhold og overvåke radiomerkede molekyler (her, [ 18 F] -FDG) ved bruk av PET / CT.

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	<tbody>
		<tr height="17">
			<td height="17" style="height:17px;width:328px;">1mL Luer-Lok Syringe</td>
			<td style="width:151px;">BD, USA</td>
			<td style="width:93px;">309628</td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">4&#39;,6&#39;-diamidino-2-phenylindole (DAPI)</td>
			<td>Sigma-Aldrich, Germany</td>
			<td style="width:93px;">D8417</td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">7 mL bijou container plain lab</td>
			<td style="width:151px;">Dutscher, France</td>
			<td style="width:93px;">080171</td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">D-glucose</td>
			<td>Sigma-Aldrich, Germany</td>
			<td>67021</td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">Digital camera</td>
			<td>Life Sciences, Japan</td>
			<td>Hamamatsu ORCA-ER</td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">Disposable base molds&#160;</td>
			<td>Simport, Canada</td>
			<td>M475-2</td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">Donkey anti-rabbit Alexa fluor 488</td>
			<td>Life Technologies, USA</td>
			<td>A21206</td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">Embedding center</td>
			<td>Thermo Scientific, USA</td>
			<td>Shandon Histocentre 3</td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">Fluorescence microscope</td>
			<td>Nikon, Japan</td>
			<td>Eclipse 80i</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Fluorodeoxyglucose (18F-FDG)</td>
			<td>Cyclotron, France</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">Glucometer test strip</td>
			<td>LifeScan, France</td>
			<td>One-Touch 143 Ultra</td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">Goat anti-mouse Alexa fluor 594</td>
			<td>Life Technologies, USA</td>
			<td>A11005</td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">In-Vivo Imaging System</td>
			<td>TriFoil Imaging, Canada</td>
			<td>Triumph Trimodality&#160;</td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">Microtome</td>
			<td>Thermo Scientific, USA</td>
			<td>Microm HM 355 S</td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">Monoclonal mouse anti-PCNA</td>
			<td>DAKO, USA</td>
			<td>clone PC10</td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">Paraformaldehyde (PFA)</td>
			<td>Sigma-Aldrich, Germany</td>
			<td>P6148-500G</td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">Polyclonal rabbit anti-GFAP</td>
			<td>DAKO, USA</td>
			<td>Z033429</td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">Slide drying bench</td>
			<td>Electrothermal, USA</td>
			<td>MH6616</td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">Sodium chloride</td>
			<td>Sigma-Aldrich, Germany</td>
			<td>S9888</td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">Sodium citrate trisodium salt dehydrate&#160;</td>
			<td>Prolabo, France</td>
			<td>27833.294</td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">Sterile needle</td>
			<td>BD Microlance 3</td>
			<td>30 G 1/2 ; 0.3 mm x 13 mm</td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">Student Dumont #5 Forceps</td>
			<td>Fine Science Tools</td>
			<td>91150-20</td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">Student surgical scissors</td>
			<td>Fine Science Tools</td>
			<td>91400-14</td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">Superfros Plus Gold Slides</td>
			<td>Thermo Scientific, USA</td>
			<td>FT4981GLPLUS</td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">Surgical microscope</td>
			<td>Leica, France</td>
			<td>M320-F12</td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">Tissue embedding cassettes</td>
			<td>Simport, Canada</td>
			<td>M490-10</td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">Tissue embedding medium</td>
			<td>LeicaBiosystems, USA</td>
			<td>39602004</td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">Toluene</td>
			<td>Sigma-Aldrich, Germany</td>
			<td>244511</td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;width:328px;">Tricaine MS-222</td>
			<td>Sigma-Aldrich, Germany</td>
			<td>A5040</td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">Triton X100</td>
			<td>Sigma-Aldrich, Germany</td>
			<td>X100-500 mL</td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">Vectashield medium&#160;</td>
			<td>Vector Laboratories, USA</td>
			<td>H-1000</td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">Xylene</td>
			<td>Sigma-Aldrich, Germany</td>
			<td>534056</td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">Fish Strain</td>
			<td>AB</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">Saline phosphate buffer (10X PBS) pH 7.4 (for 1 liter)</td>
			<td></td>
			<td></td>
			<td>For preparing 10X PBS, add the following&#160; salts and complete to 1 liter with distilled water</td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">Potassium chloride (MM : 74.55 g/mol): 2.00 g</td>
			<td>Sigma-Aldrich, Germany</td>
			<td>746436</td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">Potassium phosphate monobasic (MM: 136,09 g/mol): 2.40g</td>
			<td>Sigma-Aldrich, Germany</td>
			<td>795488</td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">Sodium chloride (MM : 58.44 g/mol): 80.00 g&#160;</td>
			<td>Sigma-Aldrich, Germany</td>
			<td>S9888</td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">Sodium phosphate dibasic (MM: 141,96 g): 14,40 g</td>
			<td>Sigma-Aldrich, Germany</td>
			<td>795410</td>
			<td></td>
		</tr>
	</tbody>
</table>

acute and chronic models of hyperglycemia in zebrafish: a method to assess the impact of hyperglycemia on neurogenesis and the biodistribution of radiolabeled molecules

Hyperglykemi er et stort helseproblem som fører til kardiovaskulær og cerebral dysfunksjon. For eksempel er det forbundet med økte nevrologiske problemer etter slag og er vist å svekke nevogene prosesser. Interessant nok har den voksne zebrafisken nylig kommet fram som en relevant og nyttig modell for å etterligne hyperglykemi / diabetes og å undersøke konstitutiv og regenerativ neurogenese. Dette arbeidet gir metoder for å utvikle sebrafiskmodeller av hyperglykemi for å undersøke virkningen av hyperglykemi på hjernecelleproliferasjon under hjemmestatiske og hjernens reparasjonsbetingelser. Akutt hyperglykemi er etablert ved intraperitoneal injeksjon av D-glukose (2,5 g / kg kroppsvekt) til voksen sebrafisk. Kronisk hyperglykemi er indusert ved nedsenking av voksen sebrafisk i D-glukose (111 mM) som inneholder vann i 14 dager. Blodglukose-nivåmålinger er beskrevet for disse forskjellige tilnærmingene. Metoder for å undersøke effekten av hyperglykemi på konstitutiv aOg regenerativ neurogenese ved å beskrive telensfalonens mekaniske skade, dissekere hjernen, paraffininkonstruksjon og snitting med et mikrotom og utføre immunhistokjemeprosedyrer, blir demonstrert. Endelig beskrives også metoden for bruk av sebrafisk som en relevant modell for å studere biodistribusjon av radioaktive merkede molekyler (her, [ 18 F] -FDG) ved bruk av PET / CT.

Ved bruk av prosedyrene beskrevet i denne artikkelen ble intraperitoneal injeksjon av D-glukose (2,5 g / kg kroppsvekt) utført på voksen sebrafisk og ført til en signifikant økning i blodsukkernivået 1,5 time etter injeksjon ( figur 1A ). 24 timer etter injeksjon var blodglukosenivåene like mellom D-glukose og PBS-injisert fisk 12 . For kronisk behandling ble sebrafisk nedsenket i D-glukose-vann (111 mM) og ble hyperglykemisk ve...

Watch this Scientific Journal Video about Acute and Chronic Models of Hyperglycemia in Zebrafish: A Method to Assess the Impact of Hyperglycemia on Neurogenesis and the Biodistribution of Radiolabeled

Acute and Chronic Models of Hyperglycemia in Zebrafish: A Method to Assess the Impact of Hyperglycemia on Neurogenesis and the Biodistribution of Radiolabeled Molecules

Dette arbeidet beskriver metoder for å etablere akutte og kroniske hyperglykemodeller i sebrafisk. Målet er å undersøke effekten av hyperglykemi på fysiologiske prosesser, som konstitutiv og skadefremkalt neurogenese. Arbeidet fremhever også bruk av sebrafisk for å følge radiomerkede molekyler (her, [ 18 F] -FDG) ved hjelp av PET / CT.

Akutte og kroniske modeller av hyperglykemi i sebrafisk: En metode for å vurdere virkningen av hyperglykemi på neurogenese og biodistribusjon av radiomerkede molekyler

Hyperglycemia is a major health issue that leads to cardiovascular and cerebral dysfunction. For instance, it is associated with increased neurological ...

Research

JoVE Journal

Developmental Biology

Analysis of Zebrafish Larvae Skeletal Muscle Integrity with Evans Blue Dye

Analyse av Sebrafisk Larver Skeletal Muscle Integrity med Evans blå Dye

The zebrafish model is an emerging system for the study of neuromuscular disorders.  In the study of neuromuscular diseases, the integrity of the muscle ...

Using Touch-evoked Response and Locomotion Assays to Assess Muscle Performance and Function in Zebrafish

Ved hjelp av Touch-fremkalt respons og locomotion Analyser for å vurdere muskel ytelse og funksjon i Sebrafisk

Zebrafish muscle development is highly conserved with mammalian systems making them an excellent model to study muscle function and disease. Many ...

Methods to Examine the Lymph Gland and Hemocytes in Drosophila Larvae

Methods to Examine the Lymph Gland and Hemocytes in Drosophila Larvae

Metoder for å undersøke lymfekjertel og Hemocytes i<em&gt; Drosophila</em&gt; Larver

Many parallels exist between the Drosophila and mammalian hematopoietic systems, even though Drosophila lack the lymphoid lineage that characterize ...

Building Finite Element Models to Investigate Zebrafish Jaw Biomechanics

Bygge Finite Element Modeller for å etterforske sebrafisk Jaw Biomekanikk

Skeletal morphogenesis occurs through tightly regulated cell behaviors during development; many cell types alter their behavior in response to mechanical ...

Histological Analyses of Acute Alcoholic Liver Injury in Zebrafish

Histologiske analyser av akutt alkoholskader i sebrafisk

Alcoholic Liver Disease (ALD) refers to damage to the liver due to acute or chronic alcohol abuse. It is among the leading causes of alcohol-related ...

Live Imaging of Mouse Secondary Palate Fusion

The fusion of the secondary palatal shelves to form the intact secondary palate is a key process in mammalian development and its disruption can lead to ...

Dissection and Immunofluorescent Staining of Mushroom Body and Photoreceptor Neurons in Adult Drosophila melanogaster Brains

Dissection and Immunofluorescent Staining of Mushroom Body and Photoreceptor Neurons in Adult Drosophila melanogaster Brains

Disseksjon og Immunofluorescent flekker av sopp kropp og Photoreceptor Neurons i voksen Drosophila melanogaster hjernen

Nervous system development involves a sequential series of events that are coordinated by several signaling pathways and regulatory networks. Many of the ...

Adult Zebrafish Injury Models to Study the Effects of Prednisolone in Regenerating Bone Tissue

Voksen sebrafisk skade modeller for å studere virkningene av prednisolon i å regenerere benvev

Zebrafish are able to regenerate various organs, including appendages (fins) after amputation. This involves the regeneration of bone, which regrows ...

HUVEC Tube-formation Assay to Evaluate the Impact of Natural Products on Angiogenesis

HUVEC tube-formasjon analysen å evaluere virkningen av naturlige produkter på angiogenese

Angiogenesis is a phenomenon that includes different processes, such as endothelial cell proliferation, differentiation, and migration, that lead to the ...

Electroporation Method for In Vivo Delivery of Plasmid DNA in the Adult Zebrafish Telencephalon

Electroporation metode for in vivo levering av plasmider DNA i Adult sebrafisk Telencephalon

Electroporation is a transfection method in which an electrical field is applied to cells to create temporary pores in a cell membrane and increase its ...