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 i høj grad Direction des Usages du Numérique (DUN) fra La Réunion Universitet til redigering af videoen (især Jean-François Février, Eric Esnault og Sylvain Ducasse), Lynda-Rose Mottagan til voiceover, Mary Osborne-Pellegrin til korrekturlæsning Voice-over og CYROI-platformen. Dette arbejde blev støttet af tilskud fra La Réunion University (Bonus Qualité Recherche, Dispositifs incitatifs), Conseil Régional de La Réunion, Den Europæiske Union (CPER / FEDER) og Philancia-foreningen. ACD er modtager af stipendium fra Ministère de l&#39;Education Nationale de l&#39;Enseignement Supérieur et de la Recherche, La Réunion Universitet (Contrat Doctoral).

Voksne vildtype zebrafisk ( Danio rerio ) blev opretholdt under standardfotoperiode (14/10 h lys / mørk) og temperatur (28 ° C) betingelser. Alle forsøg blev udført i overensstemmelse med de franske og europæiske retningslinjer for anvendelse af dyr i forskning (86/609 / EØF og 2010/63 / EU) og blev godkendt af det lokale etiske udvalg til dyreforsøg. 1. Etablering af en model for akut hyperglykæmi hos zebrafisk <ol><li> Forbered en stamopløsning af tricain (MS-222) ved a...

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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. 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Dette arbejde beskriver forskellige metoder til at etablere akutte og kroniske modeller af hyperglykæmi hos zebrafisk. De vigtigste fordele ved disse procedurer er, at: 1) de giver mulighed for en reduktion i antallet af pattedyr, der anvendes til forskning, (2) de er enkle at etablere og hurtige at gennemføre, og (3) de er økonomiske. Derfor tillader sådanne modeller at undersøge virkningen af ​​hyperglykæmi på et stort antal dyr for at studere dens indvirkning på forskellige fysiologiske processer, herunde...

Hyperglykæmi er defineret som overdrevne blodglukoseniveauer. Selvom det kunne afspejle en akut stress situation, er hyperglykæmi også en tilstand, der ofte fører til diagnose af diabetes, en kronisk lidelse af insulinsekretion og / eller resistens. I 2016 er antallet af voksne, der lever med diabetes, nået 422 millioner over hele verden, og hvert år dør 1,5 millioner mennesker af denne sygdom, hvilket gør det til et stort sundhedsproblem 1 . Faktisk fører ukontrolleret diabetes til flere fysiologiske lidelser, som påvirker hjerte-kar-systemet, nyrerne og perifere og centrale nervesystemer. Interessant nok kan akut og kronisk hyperglykæmi ændre kognition og bidrage til både demens og depression 2 , 3 , 4 , 5 , 6 . Derudover er adgangen til patienter wIth hyperglykæmi har været forbundet med dårligere funktionelle, neurologiske og overlevelsesresultater efter iskæmisk slagtilfælde 7 , 8 , 9 , 10 , 11 . Det blev også vist, at hyperglykæmi / diabetes påvirker voksen neurogenese, en proces, der fører til dannelsen af ​​nye neuroner, ved at påvirke neural stamcelleaktivitet og neuronal differentiering, migration og overlevelse 2 , 12 . I modsætning til pattedyr viser teleostfisk som zebrafisk intensiv neurogen aktivitet i hele hjernen og udviser en fremragende kapacitet til hjernearbejde i voksenalderen 13 , 14 , 15 , 16 . Navnlig er sådanne kapaciteter mulige på grund af neu&#39;s vedholdenhedRal stam / progenitor celler, herunder radial glia og neuroblaster 17 , 18 , 19 . Desuden har zebrafisken for nylig fremkommet som en model til undersøgelse af stofskifteforstyrrelser, herunder fedme og hyperglykæmi / diabetes 20 , 21 , 22 . Selv om zebrafisken er en velkendt model for hyperglykæmi og neurogenese, har få undersøgelser undersøgt virkningen af ​​hyperglykæmi på hjernens homeostase og kognitive funktion 12 , 23 . For at bestemme virkningen af ​​hyperglykæmi på konstitutiv og skadefremkaldt hjernecelleproliferation blev der skabt en model af akut hyperglykæmi ved intraperitoneal injektion af D-glucose. Derudover blev en model af kronisk hyperglykæmi reproduceret ved nedsænkning af fisk i vand suppleret wMed D-glucose 12 . Zebrafish udviser mange fordele ved forskningen. De er billige, nemme at hæve og gennemsigtige i de første udviklingsstadier, og deres genom er blevet sekventeret. I forbindelse med dette arbejde viser de også flere yderligere fordele: (1) de deler lignende fysiologiske processer med mennesker, hvilket gør dem til et kritisk redskab til biomedicinsk forskning; (2) de giver mulighed for hurtig undersøgelse af virkningen af ​​hyperglykæmi på hjernens homeostase og neurogenese under hensyntagen til deres udbredt og stærke neurogene aktivitet; Og (3) de er en alternativ model, der muliggør reduktion af antallet af pattedyr, der anvendes i forskning. Endelig kan zebrafisk bruges som en model til testning af biodistribution af radioaktivt mærke molekyler og potentielle terapeutiske midler ved anvendelse af PET / CT. Det overordnede mål med følgende procedure er at visuelt dokumentere, hvordan man opstiller modeller af akut og kronisk hyperglykæmi i zebrafisk, brug zebRafish til at vurdere hjernemodellering i hyperglykæmiske tilstande og overvåge radioaktivt mærket molekyler (her [18F] -FDG) ved anvendelse af PET / CT.

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			<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>
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		<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>
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			<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>
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			<td height="17" style="height:17px;">D-glucose</td>
			<td>Sigma-Aldrich, Germany</td>
			<td>67021</td>
			<td></td>
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		<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>
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		<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>
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		<tr height="17">
			<td height="17" style="height:17px;">Embedding center</td>
			<td>Thermo Scientific, USA</td>
			<td>Shandon Histocentre 3</td>
			<td></td>
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		<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>
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		<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>
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		<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>
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		<tr height="17">
			<td height="17" style="height:17px;">Microtome</td>
			<td>Thermo Scientific, USA</td>
			<td>Microm HM 355 S</td>
			<td></td>
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		<tr height="17">
			<td height="17" style="height:17px;">Monoclonal mouse anti-PCNA</td>
			<td>DAKO, USA</td>
			<td>clone PC10</td>
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			<td height="17" style="height:17px;">Paraformaldehyde (PFA)</td>
			<td>Sigma-Aldrich, Germany</td>
			<td>P6148-500G</td>
			<td></td>
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		<tr height="17">
			<td height="17" style="height:17px;">Polyclonal rabbit anti-GFAP</td>
			<td>DAKO, USA</td>
			<td>Z033429</td>
			<td></td>
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		<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

Hyperglykæmi er et stort sundhedsproblem, der fører til hjerte-kar-og cerebral dysfunktion. For eksempel er det forbundet med øgede neurologiske problemer efter slagtilfælde og er vist at svække neurogene processer. Interessant nok har den voksne zebrafisk for nylig fremkommet som en relevant og nyttig model til at efterligne hyperglykæmi / diabetes og at undersøge konstitutiv og regenerativ neurogenese. Dette arbejde giver metoder til udvikling af zebrafiskmodeller af hyperglykæmi for at undersøge virkningen af ​​hyperglykæmi på hjernecelleproliferation under homeostatiske og hjernedannelsesbetingelser. Akut hyperglykæmi er etableret ved intraperitoneal injektion af D-glucose (2,5 g / kg legemsvægt) til voksen zebrafisk. Kronisk hyperglykæmi induceres ved nedsænkning af voksne zebrafisk i D-glucose (111 mM) indeholdende vand i 14 dage. Målinger af blodglukose er beskrevet for disse forskellige tilgange. Metoder til at undersøge virkningen af ​​hyperglykæmi på konstitutiv aNd regenerativ neurogenese ved at beskrive telencephalonens mekaniske skade, dissekere hjernen, paraffinindlejring og snitning med et mikrotom og udføre immunhistokemeprocedurer, påvises. Endelig beskrives også fremgangsmåden til anvendelse af zebrafisk som en relevant model til undersøgelse af biodistribution af radioaktivt mærkede molekyler (her [18F] -FDG) ved anvendelse af PET / CT.

Ved anvendelse af procedurerne beskrevet i denne artikel blev intraperitoneal injektion af D-glucose (2,5 g / kg legemsvægt) udført på voksne zebrafisk og ført til en signifikant stigning i blodglukoseniveauet 1,5 time efter injektion ( Figur 1A ). 24 timer efter injektion var blodglukoseniveauerne ens mellem D-glucose og PBS-injiceret fisk 12 . Til kronisk behandling blev zebrafisk nedsænket i D-glucosevand (111 mM) og blev hype...

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 værk beskriver metoder til etablering af akutte og kroniske hyperglykæmi modeller i zebrafisk. Målet er at undersøge virkningen af ​​hyperglykæmi på fysiologiske processer, såsom konstitutiv og skadefremkaldt neurogenese. Arbejdet fremhæver også brugen af ​​zebrafisk for at følge radiomærkede molekyler (her, [ 18 F] -FDG) ved anvendelse af PET / CT.

Akutte og kroniske modeller af hyperglykæmi i zebrafisk: En metode til vurdering af virkningen af ​​hyperglykæmi på neurogenese og biodistribution af radiomærkede 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 af zebrafisk Larver Skeletmuskel Integritet med Evans blå farvestof

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

Brug Touch-fremkaldt reaktion og Locomotion Analyser til Vurdere Muskel Ydeevne og funktion i zebrafisk

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 til at undersøge lymfeknuder kirtel og hæmocytter 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

Opbygning Finite Element Modeller til undersøgelse Zebrafisk Jaw Biomekanik

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 af akut alkoholisk leverskader 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

Dissektion og immunfluorescent farvning af champignon krop og fotoreceptor neuroner i voksen Drosophila melanogaster hjerner

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 zebrafisk skade modeller at studere virkningerne af Prednisolon i regenererende knoglevæv

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-formation assay til at evaluere virkningen af 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

Elektro porationsmetode til in vivo-levering af plasmid-DNA i den voksne Zebrafish 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 ...

Akutte og kroniske modeller af hyperglykæmi i zebrafisk: En metode til vurdering af virkningen af ​​hyperglykæmi på neurogenese og biodistribution af radiomærkede molekyler

Akutte og kroniske modeller af hyperglykæmi i zebrafisk: En metode til vurdering af virkningen af hyperglykæmi på neurogenese og biodistribution af radiomærkede molekyler