Name: micro-dissection of enamel organ from mandibular incisor of rats exposed to environmental toxicants
Uploaded: 2018-03-29T13:30:00
Description: Watch this Scientific Journal Video about Micro-dissection of Enamel Organ from Mandibular Incisor of Rats Exposed to Environmental Toxicants at JoVE.com

Dette arbejde blev finansieret ved universitetet Paris-Diderot, franske nationale Institut for sundhed og medicinsk forskning (INSERM) og det franske Institut for Odontological Research (IFRO).

Alle dyr, der anvendes i den foreliggende undersøgelse blev opretholdt i overensstemmelse med retningslinjer for pleje og anvendelse af forsøgsdyr fra det franske landbrugsministerium (A-75-06-12).
1. dyrs eksponering for giftige stoffer
<ol><li>Inden du udfører denne protokol, indhente nødvendige institutionelle godkendelse og sørg for at overholde alle dyrs pleje retningslinjer.</li>
	<li>Anvende design forskning protokollen tillader forfatningen af forskellige eksperimentelle grupper...

<ol>
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L., 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=Interaction+of+lifestyle,+behaviour+or+systemic+diseases+with+dental+caries+and+periodontal+diseases:+consensus+report+of+group+2+of+the+joint+EFP/ORCA+workshop+on+the+boundaries+between+caries+and+periodontal+diseases.">Interaction of lifestyle, behaviour or systemic diseases with dental caries and periodontal diseases: consensus report of group 2 of the joint EFP/ORCA workshop on the boundaries between caries and periodontal diseases.</a> J Clin Periodontol. 44, S39-S51 (2017).</li><li>Nanci, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Enamel:+Composition,+Formation,+and+Structure.">Enamel: Composition, Formation, and Structure.</a> Ten Cate's Oral Histology Development, Structure, and Function. , 122-164 (2012).</li><li>Leite, G. A., Sawan, R. M., Teofilo, J. M., Porto, I. M., Sousa, F. B., Gerlach, R. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Exposure+to+lead+exacerbates+dental+fluorosis.">Exposure to lead exacerbates dental fluorosis.</a> Arch Oral Biol. 56, 695-702 (2011).</li><li>Jedeon, 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=Enamel+hypomineralization+due+to+endocrine+disruptors.">Enamel hypomineralization due to endocrine disruptors.</a> Connect Tiss Res. 55, 1-5 (2014).</li><li>Jedeon, 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=Androgen+receptor+involvement+in+rat+amelogenesis:+an+additional+way+for+endocrine+disrupting+chemicals+to+affect+enamel+synthesis.">Androgen receptor involvement in rat amelogenesis: an additional way for endocrine disrupting chemicals to affect enamel synthesis.</a> Endocrinology. 157, 4287-4296 (2016).</li><li>Weerheijm, K. L., Jalevik, B., Alaluusua, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Molar-incisor+hypomineralisation.">Molar-incisor hypomineralisation.</a> Caries Res. 35, 390-391 (2001).</li><li>J&#228;levik, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Prevalence+and+Diagnosis+of+Molar-Incisor-+Hypomineralisation+(MIH):+A+systematic+review.">Prevalence and Diagnosis of Molar-Incisor- Hypomineralisation (MIH): A systematic review.</a> Eur Arch Paediatr Dent. 11, 59-64 (2010).</li><li>Alaluusua, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Aetiology+of+Molar-Incisor+Hypomineralisation:+A+systematic+review.">Aetiology of Molar-Incisor Hypomineralisation: A systematic review.</a> Eur Arch Paediatr Dent. 11, 53-58 (2010).</li><li>Jedeon, K., Berdal, A., Babajko, S., Gibert, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+tooth,+target+organ+of+Bisphenol+A,+could+be+used+as+a+biomarker+of+exposure+to+this+agent.">The tooth, target organ of Bisphenol A, could be used as a biomarker of exposure to this agent.</a> Bisphenol A: Sources, Risks of Environmental Exposure and Human Health Effects. , 205-225 (2015).</li><li>Fejerskov, O., Larsen, M. J., Richards, A., Baelum, V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Dental+tissue+effects+of+fluoride.">Dental tissue effects of fluoride.</a> Adv Dent Res. 8, 15-31 (1994).</li><li>Robinson, C., Connell, S., Kirkham, J., Brookes, S. J., Shore, R. C., Smith, A. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+effect+of+fluoride+on+the+developing+tooth.">The effect of fluoride on the developing tooth.</a> Caries Res. 38, 268-276 (2004).</li><li>Sahlberg, C., Pavlic, A., Ess, A., Lukinmaa, P. L., Salmela, E., Alaluusua, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Combined+effect+of+amoxicillin+and+sodium+fluoride+on+the+structure+of+developing+mouse+enamel+in+vitro.">Combined effect of amoxicillin and sodium fluoride on the structure of developing mouse enamel in vitro.</a> Arch Oral Biol. 58, 1155-1164 (2013).</li><li>Pritchett-Corning, K. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Euthanasia+of+neonatal+rats+with+carbon+dioxide.">Euthanasia of neonatal rats with carbon dioxide.</a> J Am Assoc Lab Anim Sci. 48, 23-27 (2009).</li><li>Hiller, C. R., Robinson, C., Weatherell, J. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Variations+in+the+composition+of+developing+rat+incisor+enamel.">Variations in the composition of developing rat incisor enamel.</a> Calcif Tissue Res. 18, 1-12 (1975).</li><li>Robinson, C., Kirkham, J., Nutman, C. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Relationship+between+enamel+formation+and+eruption+rate+in+rat+mandibular+incisors.">Relationship between enamel formation and eruption rate in rat mandibular incisors.</a> Cell Tissue Res. 254, 655-658 (1988).</li><li>Smith, C. E., Nanci, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+method+for+sampling+the+stages+of+amelogenesis+on+mandibular+rat+incisors+using+the+molars+as+a+reference+for+dissection.">A method for sampling the stages of amelogenesis on mandibular rat incisors using the molars as a reference for dissection.</a> Anat Rec. 225, 257-266 (1989).</li><li>Chavez, M. G., 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=Isolation+and+culture+of+dental+epithelial+stem+cells+from+the+adult+mouse+incisor.">Isolation and culture of dental epithelial stem cells from the adult mouse incisor.</a> J Vis Exp. (87), (2014).</li><li>Brookes, S. J., Kingswell, N. J., Barron, M. J., Dixon, M. J., Kirkham, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Is+the+32-kDa+fragment+the+functional+enamelin+unit+in+all+species?.">Is the 32-kDa fragment the functional enamelin unit in all species?.</a> Eur J Oral Sci. 119, 345-350 (2011).</li><li>Babajko, S., Jedeon, K., Houari, S., Loiodice, S., Berdal, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Disruption+of+Steroid+Axis,+a+New+Paradigm+for+Molar+Incisor+Hypomineralization+(MIH).">Disruption of Steroid Axis, a New Paradigm for Molar Incisor Hypomineralization (MIH).</a> Front Physiol. 8, 343 (2017).</li><li>Houari, S., 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=Asporin+and+the+mineralization+process+in+fluoride-treated+rats.">Asporin and the mineralization process in fluoride-treated rats.</a> J Bone Min Res. 29, 1446-1455 (2014).</li><li>Denbesten, P., Li, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Chronic+fluoride+toxicity:+dental+fluorosis.">Chronic fluoride toxicity: dental fluorosis.</a> Monographs in oral science. 22, 81-96 (2011).</li><li>Kirkham, J., Robinson, C., Phull, J. K., Shore, R. C., Moxham, B. J., Berkovitz, B. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+effect+of+rate+of+eruption+on+periodontal+ligament+glycosylaminoglycan+content+and+enamel+formation+in+the+rat+incisor.">The effect of rate of eruption on periodontal ligament glycosylaminoglycan content and enamel formation in the rat incisor.</a> Cell Tissue Res. 274, 413-419 (1993).</li><li>Lacruz, R. S., 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=Identification+of+novel+candidate+genes+involved+in+mineralization+of+dental+enamel+by+genome-wide+transcript+profiling.">Identification of novel candidate genes involved in mineralization of dental enamel by genome-wide transcript profiling.</a> J Cell Physiol. 227, 2264-2275 (2012).</li><li>Wen, X., Paine, M. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Iron+deposition+and+ferritin+heavy+chain+(Fth)+localization+in+rodent+teeth.">Iron deposition and ferritin heavy chain (Fth) localization in rodent teeth.</a> BMC research notes. 6, 1 (2013).</li><li>Houari, S., Loiodice, S., Jedeon, K., Berdal, A., Babajko, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Expression+of+Steroid+Receptors+in+Ameloblasts+during+Amelogenesis+in+Rat+Incisors.">Expression of Steroid Receptors in Ameloblasts during Amelogenesis in Rat Incisors.</a> Front Physiol. 7, 503 (2016).</li><li>Kawano, S., 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=Establishment+of+dental+epithelial+cell+line+(HAT-7)+and+the+cell+differentiation+dependent+on+Notch+signaling+pathway.">Establishment of dental epithelial cell line (HAT-7) and the cell differentiation dependent on Notch signaling pathway.</a> Connect Tissue Res. 43, 409-412 (2002).</li><li>Zhou, Y. L., Snead, M. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Identification+of+CCAAT/enhancer-binding+protein+alpha+as+a+transactivator+of+the+mouse+amelogenin+gene.">Identification of CCAAT/enhancer-binding protein alpha as a transactivator of the mouse amelogenin gene.</a> J Biol Chem. 275, 12273-12280 (2000).</li><li>Nakata, A., 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=Establishment+and+characterization+of+a+spontaneously+immortalized+mouse+ameloblast-lineage+cell+line.">Establishment and characterization of a spontaneously immortalized mouse ameloblast-lineage cell line.</a> Biochem Biophys Res Commun. 308, 834-839 (2003).</li><li>Harada, H., 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=Establishment+of+ameloblastoma+cell+line,+AM-1.">Establishment of ameloblastoma cell line, AM-1.</a> Journal of oral pathology &amp; medicine: official publication of the International Association of Oral Pathologists and the American Academy of Oral Pathology. 27, 207-212 (1998).</li><li>Jussila, M., Thesleff, I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Signaling+networks+regulating+tooth+organogenesis+and+regeneration,+and+the+specification+of+dental+mesenchymal+and+epithelial+cell+lineages.">Signaling networks regulating tooth organogenesis and regeneration, and the specification of dental mesenchymal and epithelial cell lineages.</a> Cold Spring Harb Perspect Biol. 4, a008425 (2012).</li><li>Tucker, A., Sharpe, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+cutting-edge+of+mammalian+development;+how+the+embryo+makes+teeth.">The cutting-edge of mammalian development; how the embryo makes teeth.</a> Nat Rev Genet. 5, 499-508 (2004).</li><li>Vos, 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=Years+lived+with+disability+(YLDs)+for+1160+sequelae+of+289+diseases+and+injuries+1990-2010:+a+systematic+analysis+for+the+Global+Burden+of+Disease+Study+2010.">Years lived with disability (YLDs) for 1160 sequelae of 289 diseases and injuries 1990-2010: a systematic analysis for the Global Burden of Disease Study 2010.</a> Lancet. 380, 2163-2196 (2012).</li><li>Marcenes, W., 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=Global+burden+of+oral+conditions+in+1990-2010:+a+systematic+analysis.">Global burden of oral conditions in 1990-2010: a systematic analysis.</a> J Dent Res. 92, 592-597 (2013).</li><li>. 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Ændrede ameloblast aktivitet og/eller forstyrret ameloblast spredning, differentiering og modning processer fører til irreversible emalje defekter, og til gengæld, karakterisering af emalje defekter kan hjælpe med at udvikle forståelse af den ændrede ameloblast aktivitet under amelogenesis. Undersøgelser på isolerede emalje orgel er således afgørende at belyse de patologiske begivenheder, der førte til emalje mangler uanset deres oprindelse, miljømæssige eller genetiske.
Denne tek...

Mange udviklingsmæssige emalje defekter kan skyldes eksponering for miljømæssige giftstoffer og/eller uhensigtsmæssig livsstil1,2,3,4. Karakterisering af forstyrrende begivenheder og molekyler af amelogenesis ved hjælp af de i øjeblikket beskrevne procedure vil fremme brugen af resulterende emalje defekter som tidlige markører for eksponering for flere giftige stoffer, og kan hjælpe til at rekonstruere historien om sundhed hver patient i den perinatale periode når emalje er synthetized1,2. Emalje syntese kan opdeles i fire hovedfaser afhængigt af ameloblast aktivitet5. Det første trin omfatter forløber celle og pre-ameloblast spredning. Under det andet trin udskiller differentierede ameloblasts emalje matrix proteiner (EMPs), hovedsageligt amelogenin, enamelin og ameloblastin, der bestemmer tykkelsen af den endelige emalje. Således fører enhver afbrydelse af EMP syntese til kvantitative defekter af emalje. Efter aflejring af fuld emalje tykkelse begynder modning fase. I denne fase tillader apatit crystallite vækst i bredde og tykkelse emalje at nå den højeste mineralisering forholdet findes i en biologisk væv, med op til 96% af vægten. Forstyrrende hændelser, der opstår under modning Stadium fører til kvalitative emalje defekter. Endelig ameloblasts ind i en fase af efter modning, også kaldet pigmentering i gnavere, og gennemgå apoptose i løbet af tand vulkanen gør emalje defekter (hvis nogen) uoprettelig og irreversibel, således defekter give potentielle retrospektiv optagelse af ameloblast understreger. I gnavere følger amelogenesis en lignende sekvens af begivenheder med den særegenhed, at deres fortænder vokser hele tiden, hvilket gør dem en passende model til at studere den generelle proces af amelogenesis. Således resulterer enhver afbrydelse af amelogenesis i ændringer af emalje kvalitet og/eller antal, afhængig af tidsvinduet af hændelsen forstyrrende. I forstand, eksponering for dioxin, bly og hormonforstyrrende kemikalier (EDC) som bisphenol A (BPA), genistein og vinclozolin, har vist sig at generere emalje hypomineralizations1,2,3 ,6,7,8. Asymmetrisk hvid uigennemsigtig steder blev identificeret på fortænderne rotter udsat for en lav-dosis BPA dosis i fostrets periode og den første måned efter fødslen1. Disse emalje defekter i rotter, og dem af menneskelige kindtand chopper hypomineralization (MIH), del lignende kliniske, strukturelle og biokemiske egenskaber. MIH er en nylig beskrevet dental emalje patologi, for som ætiologien stadig forbliver uklart9,10 trods mange årsagsfaktorer har været en hypotese9,10,11 ,12.
En anden vigtig emalje hypomineralization patologi på grund af miljømæssige faktorer er dental fluorose (DF), der er resultatet af overdreven fluor absorption (> 0,1 mg/kg/dag)13,14. Den vigtigste kilde til fluor er drikkevand, der er enten suppleres eller naturligt beriget med fluor. Fluor er også ofte ordineres for at forhindre karies, men profylaktisk dosis er kun 50% lavere end giftige one (≤0.05 mg/kg/dag). MIH og DF, to hyppige sygdomme som følge af eksponering for miljøfaktorer, kan fremlægge fælles træk, som skal karakteriseres på grund af potensering af hypomineralizing effekter af fluor kombineret med andre giftige stoffer såsom EDC2 eller amoxicillin15.
Mikro-dissektion af rotte emalje orgel indeholdende ameloblasts på forskellige differentiering faser vil bidrage til at forstå virkningsmekanismen af molekyler kan forstyrre ameloblast aktivitet og forårsage emalje mangler at blive diagnosticeret efter tand udbrud. Med andre ord, karakterisering af ændringer af emalje gen expression og emalje matrix sammensætning på grund af miljømæssige giftige stoffer tillader rekonstituering af historien om eksponering for giftige stoffer, og letter miljømæssige sikkerhed overvågning for offentlige sundhed.

<table>
	<tbody>
		<tr>
			<td>Bisphenol A</td>
			<td>Sigma Aldrich, Saint Louis MO</td>
			<td>239658</td>
			<td></td>
		</tr>
		<tr>
			<td>formalin 10%</td>
			<td>Sigma-Aldrich, Saint Louis, MO</td>
			<td>HT5012</td>
			<td></td>
		</tr>
		<tr>
			<td>Tri-Reagent</td>
			<td>Euromedex, France</td>
			<td>TR118</td>
			<td></td>
		</tr>
		<tr>
			<td>RLT buffer</td>
			<td>Qiagen, Les Ulis, France</td>
			<td>74126</td>
			<td>RNeasy Protect Mini Kit</td>
		</tr>
		<tr>
			<td>Androgen receptor antibody</td>
			<td>Santa Cruz Biotechnology, Santa Cruz, CA)</td>
			<td>sc-816</td>
			<td>rabbit polyclonal antibody</td>
		</tr>
		<tr>
			<td>PBS 10x</td>
			<td>EUOMEDEX</td>
			<td>ET330.A</td>
			<td></td>
		</tr>
		<tr>
			<td>Sodium fluoride (NaF)</td>
			<td>Sigma-Aldrich, Saint Louis, MO</td>
			<td>S-1504</td>
			<td></td>
		</tr>
		<tr>
			<td>paraplast regular</td>
			<td>Leica microsystems, Nanterre cedex,&#160;France</td>
			<td>39601006</td>
			<td>called was/parafin in the text</td>
		</tr>
		<tr>
			<td>tissue OCT</td>
			<td>VWR, Fontenay-sous-Bois, France</td>
			<td>411243</td>
			<td></td>
		</tr>
		<tr>
			<td>Extra Fine Bonn Scissors - Straight/8.5 cm</td>
			<td>PHYMEP , Paris, France</td>
			<td>14084-08</td>
			<td></td>
		</tr>
		<tr>
			<td>Handle for Scalpel Blades - 12.5 cm</td>
			<td>PHYMEP, Paris, France</td>
			<td>10035-12</td>
			<td></td>
		</tr>
		<tr>
			<td>Curved Scalpel Blade</td>
			<td>PHYMEP , Paris, France</td>
			<td>10035-20</td>
			<td></td>
		</tr>
		<tr>
			<td>Dissecting Knife - Fine/Straight Tip</td>
			<td>PHYMEP , Paris, France</td>
			<td>10055-12</td>
			<td></td>
		</tr>
		<tr>
			<td>Circle Knife</td>
			<td>PHYMEP, Paris, France</td>
			<td>10059-15</td>
			<td></td>
		</tr>
		<tr>
			<td>scalpel blades n&#176;11 Swann-Morton</td>
			<td>VWR, Fontenay-sous-Bois, France</td>
			<td>233-0024</td>
			<td></td>
		</tr>
		<tr>
			<td>binocular lens</td>
			<td>Leica biosystems, Nanterre cedex,&#160;France</td>
			<td>MZFLIII</td>
			<td></td>
		</tr>
	</tbody>
</table>

micro-dissection of enamel organ from mandibular incisor of rats exposed to environmental toxicants

Emalje defekter som følge af miljøforhold og livsformer er hensynet til folkesundheden på grund af deres høje prævalens. Disse mangler skyldes ændret aktivitet af celler der er ansvarlig for emalje syntese ved navn ameloblasts, som er til stede i emalje orgel. Under amelogenesis følge ameloblasts en konkret og præcis rækkefølge af hændelser for spredning, differentiering og død. En rotte konstant voksende fortænder er en egnet eksperimentel model til at studere ameloblast aktivitet og differentiering stadier i fysiologiske og patologiske betingelser. Her, beskriver vi en pålidelig og ensartet metode til mikro-dissekere emalje orgel af rotter udsat for miljømæssige giftige stoffer. De mikro-dissekeret dental epitheler indeholder sekretion og modning fase ameloblasts, der kan bruges til kvalitative eksperimenter, som Immunhistokemi assays og i situ hybridisering, samt for kvantitative analyser som RT-qPCR, RNA-FF. og Western blotting.

Mange emalje defekter, såsom dental fluorose12,13, kan skyldes miljømæssige forhold på grund af overdreven fluor absorption eller emalje hypomineralization ligner MIH som følge af eksponering for nogle EDC1, 7,22. Disse udviklingsmæssige emalje defekter kan gengives eksperimentelt på rotter (figur 1)<...

Watch this Scientific Journal Video about Micro-dissection of Enamel Organ from Mandibular Incisor of Rats Exposed to Environmental Toxicants at JoVE.com

Micro-dissection of Enamel Organ from Mandibular Incisor of Rats Exposed to Environmental Toxicants

Forståelse emalje dannelse og mulige ændringer kræver undersøgelse af ameloblast aktivitet. Her, beskriver vi en pålidelig og ensartet metode til mikro-dissekere emalje organer indeholdende sekretion og modning fase ameloblasts, der kan bruges til yderligere kvantitative og kvalitative eksperimentelle procedurer.

Mikro-dissektion af emalje orgel fra Mandibular chopper rotter udsat for miljømæssige giftige stoffer

Enamel defects resulting from environmental conditions and ways of life are public health concerns because of their high prevalence. These defects result ...

Research

JoVE Journal

Medicine

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