Name: chemical amputation and regeneration of the pharynx in the planarian schmidtea mediterranea
Uploaded: 2018-03-26T13:30:00
Description: Watch this Scientific Journal Video about Chemical Amputation and Regeneration of the Pharynx in the Planarian Schmidtea mediterranea at JoVE.com

我们要感谢阿尔瓦拉多, 他支持这项技术的初步优化和发展。在卡罗琳·阿德勒的实验室工作是由康奈尔大学开办基金支持的。

1. 准备
<ol><li>
涡水的制备17<ol>
<li>在1X 蒙锥山盐溶液中保持三角涡虫。要准备真涡虫水, 请在超纯水中制作1米CaCl2、1米MgSO4、1米氯化镁2、1米氯化钾和5米氯化钠的个人库存解决方案。过滤-消毒与0.2 µm 瓶顶 filterfor 长期储存。 注意:只使用超纯去离子水 (电阻率为 18.2 MΩ在25摄氏度), 以制备蒙锥山盐。</li>
		<li>要准备 1 L 的5X ?...

<ol>
	<li>Tanaka, E. M., Reddien, P. W. <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+basis+for+animal+regeneration.">The cellular basis for animal regeneration.</a> Dev. Cell. 21, 172-185 (2011).</li><li>Tanaka, E. 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+Molecular+and+Cellular+Choreography+of+Appendage+Regeneration.">The Molecular and Cellular Choreography of Appendage Regeneration.</a> Cell. 165, 1598-1608 (2016).</li><li>Curado, S., Stainier, D. Y. R., Anderson, R. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Nitroreductase-mediated+cell/tissue+ablation+in+zebrafish:+a+spatially+and+temporally+controlled+ablation+method+with+applications+in+developmental+and+regeneration+studies.">Nitroreductase-mediated cell/tissue ablation in zebrafish: a spatially and temporally controlled ablation method with applications in developmental and regeneration studies.</a> Nat. Protoc. 3, 948-954 (2008).</li><li>White, D. T., Mumm, 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=The+nitroreductase+system+of+inducible+targeted+ablation+facilitates+cell-specific+regenerative+studies+in+zebrafish.">The nitroreductase system of inducible targeted ablation facilitates cell-specific regenerative studies in zebrafish.</a> Methods. 62, 232-240 (2013).</li><li>Smith-Bolton, R. K., Worley, M. I., Kanda, H., Hariharan, I. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Regenerative+growth+in+Drosophila+imaginal+discs+is+regulated+by+Wingless+and+Myc.">Regenerative growth in Drosophila imaginal discs is regulated by Wingless and Myc.</a> Dev. Cell. 16, 797-809 (2009).</li><li>Smith-Bolton, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Drosophila+Imaginal+Discs+as+a+Model+of+Epithelial+Wound+Repair+and+Regeneration.">Drosophila Imaginal Discs as a Model of Epithelial Wound Repair and Regeneration.</a> Adv. Wound Care. 5, 251-261 (2016).</li><li>Newmark, P. A., S&#225;nchez Alvarado, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Not+your+father's+planarian:+a+classic+model+enters+the+era+of+functional+genomics.">Not your father's planarian: a classic model enters the era of functional genomics.</a> Nat. Rev. Genet. 3, 210-219 (2002).</li><li>Reddien, P. W., S&#225;nchez Alvarado, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Fundamentals+of+planarian+regeneration.">Fundamentals of planarian regeneration.</a> Annu. Rev. Cell Dev. Biol. 20, 725-757 (2004).</li><li>Wagner, D. E., Wang, I. E., Reddien, P. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Clonogenic+neoblasts+are+pluripotent+adult+stem+cells+that+underlie+planarian+regeneration.">Clonogenic neoblasts are pluripotent adult stem cells that underlie planarian regeneration.</a> Science. 332, 811-816 (2011).</li><li>Scimone, M. L., Kravarik, K. M., Lapan, S. W., Reddien, P. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Neoblast+specialization+in+regeneration+of+the+planarian+Schmidtea+mediterranea.">Neoblast specialization in regeneration of the planarian Schmidtea mediterranea.</a> Stem Cell Reports. 3, 339-352 (2014).</li><li>van Wolfswinkel, J. C., Wagner, D. E., Reddien, P. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Single-cell+analysis+reveals+functionally+distinct+classes+within+the+planarian+stem+cell+compartment.">Single-cell analysis reveals functionally distinct classes within the planarian stem cell compartment.</a> Cell Stem Cell. 15, 326-339 (2014).</li><li>Adler, C. E., Seidel, C. W., McKinney, S. A., S&#225;nchez Alvarado, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Selective+amputation+of+the+pharynx+identifies+a+FoxA-dependent+regeneration+program+in+planaria.">Selective amputation of the pharynx identifies a FoxA-dependent regeneration program in planaria.</a> Elife. 3, e02238 (2014).</li><li>Adler, C. E., S&#225;nchez Alvarado, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=PHRED-1+is+a+divergent+neurexin-1+homolog+that+organizes+muscle+fibers+and+patterns+organs+during+regeneration.">PHRED-1 is a divergent neurexin-1 homolog that organizes muscle fibers and patterns organs during regeneration.</a> Dev. Biol. 427, 165-175 (2017).</li><li>Wong, M. C., Martynovsky, M., Schwarzbauer, J. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Analysis+of+cell+migration+using+Caenorhabditis+elegans+as+a+model+system.">Analysis of cell migration using Caenorhabditis elegans as a model system.</a> Methods Mol. Biol. 769, 233-247 (2011).</li><li>Margie, O., Palmer, C., Chin-Sang, I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=C+elegans+chemotaxis+assay.">C elegans chemotaxis assay.</a> J. Vis. Exp. , e50069 (2013).</li><li>Fang-Yen, C., Gabel, C. V., Samuel, A. D. T., Bargmann, C. I., Avery, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Laser+microsurgery+in+Caenorhabditis+elegans.">Laser microsurgery in Caenorhabditis elegans.</a> Methods Cell Biol. 107, 177 (2012).</li><li>Tasaki, J., Uchiyama-Tasaki, C., Rouhana, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Analysis+of+Stem+Cell+Motility+In+Vivo+Based+on+Immunodetection+of+Planarian+Neoblasts+and+Tracing+of+BrdU-Labeled+Cells+After+Partial+Irradiation.">Analysis of Stem Cell Motility In Vivo Based on Immunodetection of Planarian Neoblasts and Tracing of BrdU-Labeled Cells After Partial Irradiation.</a> Methods Mol. Biol. 1365, 323-338 (2016).</li><li>Roberts-Galbraith, R. H., Brubacher, J. L., Newmark, 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=A+functional+genomics+screen+in+planarians+reveals+regulators+of+whole-brain+regeneration.">A functional genomics screen in planarians reveals regulators of whole-brain regeneration.</a> Elife. 5, (2016).</li><li>Arnold, C. P., 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=Pathogenic+shifts+in+endogenous+microbiota+impede+tissue+regeneration+via+distinct+activation+of+TAK1/MKK/p38.">Pathogenic shifts in endogenous microbiota impede tissue regeneration via distinct activation of TAK1/MKK/p38.</a> Elife. 5, (2016).</li><li>Pearson, B. 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=Formaldehyde-based+whole-mount+in+situ+hybridization+method+for+planarians.">Formaldehyde-based whole-mount in situ hybridization method for planarians.</a> Dev. Dyn. 238, 443-450 (2009).</li><li>LoCascio, S. A., Lapan, S. W., Reddien, P. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Eye+Absence+Does+Not+Regulate+Planarian+Stem+Cells+during+Eye+Regeneration.">Eye Absence Does Not Regulate Planarian Stem Cells during Eye Regeneration.</a> Dev. Cell. 40, 381-391 (2017).</li><li>Nishimura, 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=Regeneration+of+dopaminergic+neurons+after+6-hydroxydopamine-induced+lesion+in+planarian+brain.">Regeneration of dopaminergic neurons after 6-hydroxydopamine-induced lesion in planarian brain.</a> J. Neurochem. 119, 1217-1231 (2011).</li><li>Wilson, D. F., Chance, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Azide+inhibition+of+mitochondrial+electron+transport.+I.+The+aerobic+steady+state+of+succinate+oxidation.">Azide inhibition of mitochondrial electron transport. I. The aerobic steady state of succinate oxidation.</a> Biochim. Biophys. Acta. 131, 421-430 (1967).</li><li>Duncan, H. M., Mackler, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Electron+Transport+Systems+of+Yeast:+III.+Preparation+and+properties+of+cytochrome+oxidase.">Electron Transport Systems of Yeast: III. Preparation and properties of cytochrome oxidase.</a> J. Biol. Chem. 241, 1694-1697 (1966).</li><li>Buchan, J. R., Yoon, J. H., Parker, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Stress-specific+composition,+assembly+and+kinetics+of+stress+granules+in+Saccharomyces+cerevisiae.">Stress-specific composition, assembly and kinetics of stress granules in Saccharomyces cerevisiae.</a> J. Cell Sci. 124, 228-239 (2011).</li><li>Cebri&#224;, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Organization+of+the+nervous+system+in+the+model+planarian+Schmidtea+mediterranea:+An+immunocytochemical+study.">Organization of the nervous system in the model planarian Schmidtea mediterranea: An immunocytochemical study.</a> Neurosci. Res. 61, 375-384 (2008).</li><li>Shimoyama, S., Inoue, T., Kashima, M., Agata, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Multiple+Neuropeptide-Coding+Genes+Involved+in+Planarian+Pharynx+Extension.">Multiple Neuropeptide-Coding Genes Involved in Planarian Pharynx Extension.</a> Zoolog. Sci. 33, 311-319 (2016).</li><li>Ito, H., Saito, Y., Watanabe, K., Orii, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Epimorphic+regeneration+of+the+distal+part+of+the+planarian+pharynx.">Epimorphic regeneration of the distal part of the planarian pharynx.</a> Dev. Genes Evol. 211, 2-9 (2001).</li><li>Bueno, D., Espinosa, L., Bagu&#241;&#224;, J., Romero, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Planarian+pharynx+regeneration+in++tail+fragments+monitored+with+cell-specific+monoclonal+antibodies.">Planarian pharynx regeneration in tail fragments monitored with cell-specific monoclonal antibodies.</a> Dev. Genes Evol. 206, 425-434 (1997).</li><li>Hyman, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> The invertebrates: Platyhelminthes and Rhynchocoela, the acoelomate Bilateria. , (1951).</li></ol>

本协议描述了用叠氮化钠选择性切除咽部的方法。三角涡虫的其他靶向消融研究使用改良手术去除感光细胞21 或药理治疗消融多巴胺能神经元22。化学截肢对现有方法的一个显著优点是它不需要手术。与真涡虫身体的其余部分相比, 咽部的刚性结构便于它完全从动物身上去除, 这是非常柔软的。此外, 咽与动物在一个小交界 (食管)30连接咽和肠道...

再生是整个动物王国发生的一种现象, 其再生能力从某些无脊椎动物的全身再生到脊椎动物的限制性能力为1。更换功能组织是一个复杂的过程, 往往需要同时恢复多种细胞类型。例如, 要再生蝾螈肢, 成骨细胞, 软骨细胞, 神经元, 肌肉和上皮细胞需要被替换为2。这些新产生的细胞类型也需要适当组织, 以促进新的肢体功能。了解这些复杂的过程需要的技术, 重点是再生的特定细胞类型和他们融入器官。
用于简化再生反应研究的策略之一是对某些细胞类型或较大的组织集合进行靶向消融。例如, 在斑马鱼中, nitroreductase 在特定细胞类型中的表达在应用甲硝唑3、4后导致其破坏。在果蝇幼虫中, 在组织特定的启动子下表达亲凋亡基因可以选择性地消融形象盘56的特定区域。这两种策略都造成了快速但受控的损伤, 并被用来解剖分子和细胞机制负责再生。
在这篇手稿中, 我们描述了一个方法, 有选择地消融整个器官称为咽在真涡虫Schmidtea mediterranea。三角涡虫是一种经典的再生模型, 以其多产的再生能力著称,其中即使是微小的碎片也能再生整个动物7, 8.它们有一个大的异构的干细胞群, 由多潜能细胞和受血统限制的祖细胞组成,9,10,11。这些细胞增殖和分化, 以取代所有缺失的组织, 包括咽, 神经, 消化系统和排泄系统, 肌肉和上皮细胞9,10,12。虽然我们知道这些干细胞开始再生, 但我们并不完全理解促使它们取代所有这些不同细胞类型的分子机制。定义的伤人方法, 使精确的干细胞反应可以帮助描绘这个复杂的过程。
咽是一个大的, 圆柱管需要喂养, 并包含神经元, 肌肉, 上皮和分泌细胞13,29。通常藏在动物腹侧的眼袋里, 在感觉到食物的存在时, 它会通过动物的单一身体开放。为了选择性地切除咽, 我们将三角涡虫浸泡在一种叫做叠氮化钠的化学物质中, 一种常用的麻醉剂,在线虫 14, 15, 16.它在三角涡虫中的使用首先由阿德勒et al 报告, 在2014年12。在接触叠氮化钠的几分钟内, 三角涡虫挤压他们的 pharynges, 并与温和的搅拌, 咽分离的动物。我们指的是这种完全和选择性的咽部 "化学截肢" 的损失。截肢后一周, 完全功能性咽部恢复为12。由于咽是需要喂养, 功能再生可以通过监测喂养行为来衡量。下面, 我们描述了化学截肢的协议, 以及评估咽部的再生和喂养行为的恢复。

<table border="0" cellpadding="0" cellspacing="0" width="1042">
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:447px;">Calcium Chloride Dihydrate (CaCl2)</td>
			<td style="width:204px;">Fisher Chemical</td>
			<td style="width:155px;">C79-3</td>
			<td style="width:236px;">Montju&#239;c salt solution</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Magnesium Sulfate Anhydrous (MgSO4)</td>
			<td>Fisher Chemical</td>
			<td>M65-3</td>
			<td>Montju&#239;c salt solution</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Magnesium Chloride Hexahydrate (MgCl2)</td>
			<td>Acros/VWR</td>
			<td>41341-5000</td>
			<td>Montju&#239;c salt solution</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Potassium Chloride (KCl)</td>
			<td>Acros Organics/VWR</td>
			<td>196770010</td>
			<td>Montju&#239;c salt solution</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Sodium Chloride (NaCl)</td>
			<td>Acros Organics/VWR</td>
			<td>207790050</td>
			<td>Montju&#239;c salt solution</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Sodium Bicarbonate (NaHCO3)</td>
			<td>Acros Organics/VWR</td>
			<td>123360010</td>
			<td>Montju&#239;c salt solution</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Nalgene autoclavable polypropylene copolymer lowboy with spigot</td>
			<td>ThermoFisher Scientific/VWR</td>
			<td>2324-0015</td>
			<td>Storing planaria water</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Instant Ocean Sea Salt</td>
			<td>Spectrum Brands</td>
			<td>Amazon</td>
			<td>Planaria water</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Gentamicin Sulfate</td>
			<td>Gemini Bio-products</td>
			<td>400-100P</td>
			<td>Planaria water</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Razor blades</td>
			<td>Electron Microscopy Sciences</td>
			<td>71970</td>
			<td>Mincing liver&#160;</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Disposable pastry bags-16&#8221;, 12 pack</td>
			<td>Wilton</td>
			<td>Amazon</td>
			<td>Liver aliquots</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">5 mL syringes</td>
			<td>BD/VWR</td>
			<td>309647</td>
			<td>Liver aliquots</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Petri dishes-35mm/60mm</td>
			<td>Greiner Bio-One/VWR</td>
			<td>82050-536/82050-544</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Plastic containers (various sizes)</td>
			<td>Ziploc</td>
			<td>Amazon</td>
			<td>Housing planarians in static culture</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Sodium Azide</td>
			<td>Sigma</td>
			<td>S2002</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Transfer pipette</td>
			<td>Globe Scientific</td>
			<td>138030</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Forceps - Dumont Tweezer, Style 5</td>
			<td>Electron Microscopy Sciences&#160;</td>
			<td>72700-D (0203-5-PO)</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Triton X-100</td>
			<td>Sigma</td>
			<td>T8787</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">DAPI&#160;</td>
			<td>ThermoFisher</td>
			<td>62247</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Streptavidin, Alexa Fluor 488 conjugate</td>
			<td>ThermoFisher</td>
			<td>S11223</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Glycerol</td>
			<td>Fisher BioReagents</td>
			<td>BP229-1</td>
			<td></td>
		</tr>
	</tbody>
</table>

chemical amputation and regeneration of the pharynx in the planarian schmidtea mediterranea

三角涡虫是虫非常有效的再生。他们把这种能力归功于大量的干细胞, 这些细胞能够迅速应对任何类型的损伤。在这些动物中常见的伤害模型去除大量的组织, 损害了多个器官。为了克服这种广泛的组织损伤, 我们这里描述了一种方法, 有选择地移除一个器官, 咽, 在真涡虫Schmidtea mediterranea。我们通过在含有细胞色素氧化酶抑制剂叠氮化钠的溶液中浸泡动物来实现这一目标。对叠氮化钠的短暂接触会导致咽部从动物身上挤出, 我们称之为 "化学截肢"。化学截肢除去整个咽部, 并产生一个小伤口, 咽附着在肠道。经过广泛的冲洗, 所有被截肢的动物在大约一周内再生一个完全功能的咽。身体其他部位的干细胞驱动新咽的再生。在这里, 我们提供了一个详细的化学截肢协议, 并描述了组织学和行为方法, 以评估成功的截肢和再生。

接触叠氮化钠扰乱了三角涡虫的正常运动, 导致动物伸展和扭动。这些运动迫使咽从动物腹侧出现, 在叠氮化钠溶液中大约6分钟后, 咽部的白色尖端可以看到(图 1B左面板)。几分钟后, 动物们积极地收缩, 并通过强行将咽从身体里推出来, 从而充分地伸展咽喉。(图 1B-中间面板)。在叠氮化物暴露后约11分钟,...

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Chemical Amputation and Regeneration of the Pharynx in the Planarian Schmidtea mediterranea

真涡虫的化学截肢和咽部再生Schmidtea mediterranea

真涡虫Schmidtea mediterranea 是研究干细胞和组织再生的优良模型。本刊物描述一种方法, 选择性地移除一个器官, 咽, 通过暴露动物的化学叠氮化钠。本议定书还概述了监测咽部再生的方法。

Planarians are flatworms that are extremely efficient at regeneration. They owe this ability to a large number of stem cells that can rapidly respond to ...

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Chemical Amputation and Regeneration of the Pharynx in the Planarian Schmidtea mediterranea

Unit 1

Regeneration and Repair

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