Name: adult mouse digit amputation and regeneration: a simple model to investigate mammalian blastema formation and intramembranous ossification
Uploaded: 2019-07-12T14:00:00
Description: Watch this Scientific Journal Video about Adult Mouse Digit Amputation and Regeneration: A Simple Model to Investigate Mammalian Blastema Formation and Intramembranous Ossification at JoVE.com

我们感谢穆尼奥实验室和得克萨斯基因组医学研究所(TIGM)的成员。这项工作得到了德克萨斯A&M大学的支持。

所有动物使用和技术都符合得克萨斯A&M大学机构动物护理和使用委员会的标准操作程序。
1. 成年小鼠下肢截肢 P3
<ol><li>麻醉8至12周大的CD-1小鼠(材料表),在氧气中使用非氧气体;最初在腔室中麻醉3%,随后在手术期间由诺塞酮提供2%的配苯。在眼睛上涂上眼药膏,防止麻醉时干燥。 注:在我们的实验室中标准化的成人P3截肢研究是在8至12周大的小鼠身上?...

<ol>
	<li>Douglas, B. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Conservative+management+of+guillotine+amputation+of+the+finger+in+children.">Conservative management of guillotine amputation of the finger in children.</a> Australian Paediatric Journal. 8, 86-89 (1972).</li><li>Illingworth, 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=Trapped+fingers+and+amputated+finger+tips+in+children.">Trapped fingers and amputated finger tips in children.</a> Journal of Pediatric Surgery. 9, 853-858 (1974).</li><li>Borgens, R. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mice+regrow+the+tips+of+their+foretoes.">Mice regrow the tips of their foretoes.</a> Science. 217, 747-750 (1982).</li><li>Neufeld, D. A., Zhao, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Phalangeal+regrowth+in+rodents:+postamputational+bone+regrowth+depends+upon+the+level+of+amputation.">Phalangeal regrowth in rodents: postamputational bone regrowth depends upon the level of amputation.</a> Progress in Clinical and Biological Research. 383a, 243-252 (1993).</li><li>Han, M., Yang, X., Lee, J., Allan, C. H., Muneoka, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Development+and+regeneration+of+the+neonatal+digit+tip+in+mice.">Development and regeneration of the neonatal digit tip in mice.</a> Developmental Biology. 315, 125-135 (2008).</li><li>Takeo, 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=Wnt+activation+in+nail+epithelium+couples+nail+growth+to+digit+regeneration.">Wnt activation in nail epithelium couples nail growth to digit regeneration.</a> Nature. 499, 228-232 (2013).</li><li>Chamberlain, C. 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=Level-specific+amputations+and+resulting+regenerative+outcomes+in+the+mouse+distal+phalanx.">Level-specific amputations and resulting regenerative outcomes in the mouse distal phalanx.</a> Wound repair and Regeneration: Official Publication of the Wound Healing Society [and] the European Tissue Repair Society. 25, 443-453 (2017).</li><li>Dawson, L. 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=Analogous+cellular+contribution+and+healing+mechanisms+following+digit+amputation+and+phalangeal+fracture+in+mice.">Analogous cellular contribution and healing mechanisms following digit amputation and phalangeal fracture in mice.</a> Regeneration. 3, 39-51 (2016).</li><li>Seifert, A. W., Muneoka, 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+blastema+and+epimorphic+regeneration+in+mammals.">The blastema and epimorphic regeneration in mammals.</a> Developmental Biology. 433, 190-199 (2018).</li><li>Carlson, B. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Principles of Regenerative Biology. , (2007).</li><li>Fernando, W. 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=Wound+healing+and+blastema+formation+in+regenerating+digit+tips+of+adult+mice.">Wound healing and blastema formation in regenerating digit tips of adult mice.</a> Developmental Biology. 350, 301-310 (2011).</li><li>Simkin, 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=Epidermal+closure+regulates+histolysis+during+mammalian+(Mus)+digit+regeneration.">Epidermal closure regulates histolysis during mammalian (Mus) digit regeneration.</a> Regeneration. 2, 106-119 (2015).</li><li>Yu, 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=Angiogenesis+is+inhibitory+for+mammalian+digit+regeneration.">Angiogenesis is inhibitory for mammalian digit regeneration.</a> Regeneration. 1, 33-46 (2014).</li><li>Dolan, 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=Axonal+regrowth+is+impaired+during+digit+tip+regeneration+in+mice.">Axonal regrowth is impaired during digit tip regeneration in mice.</a> Developmental Biology. 445, 237-244 (2018).</li><li>Dawson, L. 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=Blastema+formation+and+periosteal+ossification+in+the+regenerating+adult+mouse+digit.">Blastema formation and periosteal ossification in the regenerating adult mouse digit.</a> Wound Repair and Regeneration: Official Publication of the Wound Healing Society [and] the European Tissue Repair Society. 26, 263-273 (2018).</li><li>Sammarco, M. C., 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=Endogenous+bone+regeneration+is+dependent+upon+a+dynamic+oxygen+event.">Endogenous bone regeneration is dependent upon a dynamic oxygen event.</a> Journal of Bone and Mineral Research: The Official Journal of the American Society for Bone and Mineral Research. 29, 2336-2345 (2014).</li><li>Einhorn, T. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+science+of+fracture+healing.">The science of fracture healing.</a> Journal of Orthopaedic Trauma. 19, S4-S6 (2005).</li><li>Shapiro, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Bone+development+and+its+relation+to+fracture+repair.+The+role+of+mesenchymal+osteoblasts+and+surface+osteoblasts.">Bone development and its relation to fracture repair. The role of mesenchymal osteoblasts and surface osteoblasts.</a> European Cells &amp; Materials. 15, 53-76 (2008).</li><li>Ilizarov, G. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+tension-stress+effect+on+the+genesis+and+growth+of+tissues.+Part+I.+The+influence+of+stability+of+fixation+and+soft-tissue+preservation.">The tension-stress effect on the genesis and growth of tissues. Part I. The influence of stability of fixation and soft-tissue preservation.</a> Clinical Orthopaedics And Related Research. (238), 249-281 (1989).</li><li>Thompson, Z., Miclau, T., Hu, D., Helms, 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=A+model+for+intramembranous+ossification+during+fracture+healing.">A model for intramembranous ossification during fracture healing.</a> Journal of Orthopaedic Research: Official Publication of the Orthopaedic Research Society. 20, 1091-1098 (2002).</li><li>Dolan, C. P., Dawson, L. A., Muneoka, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Digit+Tip+Regeneration:+Merging+Regeneration+Biology+with+Regenerative+Medicine.">Digit Tip Regeneration: Merging Regeneration Biology with Regenerative Medicine.</a> Stem Cells Translational Medicine. 7, 262-270 (2018).</li><li>Lee, 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=SDF-1alpha/CXCR4+signaling+mediates+digit+tip+regeneration+promoted+by+BMP-2.">SDF-1alpha/CXCR4 signaling mediates digit tip regeneration promoted by BMP-2.</a> Developmental Biology. 382, 98-109 (2013).</li><li>Doube, 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=BoneJ:+Free+and+extensible+bone+image+analysis+in+ImageJ.">BoneJ: Free and extensible bone image analysis in ImageJ.</a> Bone. 47, 1076-1079 (2010).</li></ol>

该协议描述了成人小鼠远端P3截肢的标准化程序,荧光免疫组织化学染色,以可视化和调查气肿形成和膜内性渗透,并连续在体内进行_CT扫描到识别截肢后的骨形态、体积和长度变化。P3截肢是一种独特的,程序简单,可重复的模型,以分析一个促进再生的伤口环境,触发出血形成。此外,P3数字模型具有许多优点,比传统的骨损伤模型,以研究膜内骨化。
为确保此协议的成功,必须执行完整...

哺乳动物,包括人类和小鼠,在端法兰克斯(P3)1,2,3的远端截肢后,有能力再生其数字的尖端。在小鼠中,再生反应是截肢水平依赖;越来越近的数字截肢显示逐渐衰减的再生反应,直到完全再生失败在截肢横断和近端的P3指甲矩阵4,5,6,7,8.P3再生由形成大爆炸体介导,定义为通过形态形成来再生被截肢结构的增殖细胞群9。形成大爆炸体以再生截肢失去的结构,这个过程称为表皮再生,将多组织级P3再生反应与损伤后的传统组织修复区分开来。 10.P3再生是一个可重复和程序简单的模型,研究复杂的再生过程,包括伤口愈合11,12,骨质变11,12,再血管化13,外周神经再生14,并通过宫内骨化15向骨转化。
以前使用免疫性化学的研究表明,气肿是异质的,血管,缺氧,和高度增殖11,13,15,16。在远端P3截肢后,早期大发肿最初与P3型骨质和内膜有关,其特征是骨表面15附近有旺盛的增殖和新生的成骨。在骨降解和伤口闭合之后,异质性大爆发肿由围锁细胞和内窥镜相关细胞的合并形成,然后通过膜内骨化分化包括骨骼在内的囊肿成分分化15.
骨修复对损伤的反应通常发生内肠骨化,即通过初始的软骨,形成一个模板,为后续的骨形成17,18 。长骨内膜化,即骨形成无软骨中间体,通常诱导使用复杂的分心装置或手术固定19,20。数字再生反应是一种临床前模型,与传统的膜内沉降模型相比具有优势:1) 它不需要外部或内部固定损伤后刺激膜内沉降,2)使用每只动物的 4 位数字进行,从而在最大程度地减少动物使用的同时最大化样品,并且 3) 可轻松、快速地执行体内连续微计算断层扫描 (μCT) 分析。
在本研究中,我们展示了标准化的P3截肢平面,以实现可重复和强健的再生反应。此外,我们演示了优化的荧光免疫组织化学方案,使用石蜡部分来可视化气肿形成、再生环境中的再血管化,以及通过膜内向骨的气管转换骨化。我们还演示了在再生过程中使用连续体内-CT来识别同一数字的骨骼形态、体积和长度的变化。该协议的目的是研究截肢后的哺乳动物气肿形成,并演示2种技术,荧光免疫组织化学和连续体内-CT,用于研究宫内骨再生。

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>Protein Block Serum Free</td>
			<td>DAKO</td>
			<td>X0909</td>
			<td>Ready to use</td>
		</tr>
		<tr>
			<td>Mouse anti-PCNA antibody</td>
			<td>Abcam</td>
			<td>ab29</td>
			<td>1:2000 dilution</td>
		</tr>
		<tr>
			<td>Rat anti-CXCR4 antibody</td>
			<td>R&#38;D Systems</td>
			<td>MAB21651</td>
			<td>1:500 dilution</td>
		</tr>
		<tr>
			<td>Rabbit anti-human vWF XIII antibody</td>
			<td>DAKO</td>
			<td>A0082</td>
			<td>1:800 dilution</td>
		</tr>
		<tr>
			<td>Rabbit anti-osterix, SP7 antibody</td>
			<td>Abcam</td>
			<td>ab22552</td>
			<td>1:400 dilution</td>
		</tr>
		<tr>
			<td>Rabbit anti-Runx2 antibody</td>
			<td>Sigma-Aldrich Co.</td>
			<td>HPA022040</td>
			<td>1:250 dilution</td>
		</tr>
		<tr>
			<td>Alexa Fluor 647-conjugated goat anti-mouse IgG (H+L)</td>
			<td>Invitrogen</td>
			<td>A21235</td>
			<td>1:500 dilution</td>
		</tr>
		<tr>
			<td>Alexa Fluor 488-conjugated goat anti-rabbit IgG (H+L)</td>
			<td>Invitrogen</td>
			<td>A11008</td>
			<td>1:500 dilution</td>
		</tr>
		<tr>
			<td>Alexa Fluor 568-conjugated goat anti-rat IgG (H+L)</td>
			<td>Invitrogen</td>
			<td>A11077</td>
			<td>1:500 dilution</td>
		</tr>
		<tr>
			<td>Prolong Gold antifade reagent</td>
			<td>Invitrogen</td>
			<td>P36930</td>
			<td>Ready to use</td>
		</tr>
		<tr>
			<td>Surgipath Decalicifier 1</td>
			<td>Leica Biosystems</td>
			<td>3800400</td>
			<td>Ready to use</td>
		</tr>
		<tr>
			<td>Z-Fix, Aqueous buffered zinc formalin fixative</td>
			<td>Anatech LTD</td>
			<td>174</td>
			<td>Ready to use</td>
		</tr>
		<tr>
			<td>CD-1 Female Mouse</td>
			<td>Envigo</td>
			<td>ICR(CD-1)</td>
			<td>8-12-weeks-old</td>
		</tr>
		<tr>
			<td>vivaCT 40</td>
			<td>SCANCO Medical</td>
			<td></td>
			<td></td>
		</tr>
	</tbody>
</table>

adult mouse digit amputation and regeneration: a simple model to investigate mammalian blastema formation and intramembranous ossification

在这里,我们提出了成人小鼠远端咽喉(P3)截肢的协议,一个程序简单和可重复的哺乳动物表皮再生模型,其中涉及气肿形成和膜内性内性性荧光免疫组织化学和连续体内微计算断层扫描(μCT)。哺乳动物再生仅限于截肢,横断端法兰克斯(P3) 的远端区域;在更接近的水平上被截肢的数字不能再生,并经历纤维化愈合和疤痕形成。再生反应由增殖性爆发性气肿的调解,然后通过宫内骨质再生来恢复被截肢的骨骼长度。P3截肢是研究哺乳动物表皮再生的临床前模型,是设计治疗策略的有力工具,用成功的再生反应取代纤维化愈合。我们的方案使用荧光免疫组织化学 1) 识别早期和晚期的囊肿细胞群,2) 在再生环境中研究再血管化,3) 无需复杂骨骼即可研究膜内渗透稳定装置。我们还演示了使用连续体在体内μCT来创建高分辨率图像来检查截肢后的形态变化,以及量化再生过程中相同数字的体积和长度变化。我们相信,该协议为研究哺乳动物的表型和组织再生反应提供了巨大的效用。

成年小鼠在 6/7 DPA(图2A-D)、9 DPA(图 2E-H)和 10 DPA(图 2I-L)时重新生成 P3 位数字,免疫中带有 Runx2、OSX 和 PCNA 的抗体,以可视化膜内骨骼再生,免疫染色与抗体CXCR4和vWF可视化爆发性形成。在截肢前和再生过程中不同时间点扫描的数字的代表性CT渲染(图3A,B)和用于识?...

Watch this Scientific Journal Video about Adult Mouse Digit Amputation and Regeneration: A Simple Model to Investigate Mammalian Blastema Formation and Intramembranous Ossification at JoVE.com

Adult Mouse Digit Amputation and Regeneration: A Simple Model to Investigate Mammalian Blastema Formation and Intramembranous Ossification

成年小鼠数字截肢和再生:研究哺乳动物暴发体形成和内膜性细胞化的简单模型

在这里,我们提出了一个成人小鼠端程phalanx截肢的协议,以研究哺乳动物的气肿形成和膜内性渗透,通过荧光免疫组织化学和连续体内微计算断层扫描进行分析。

Here, we present a protocol of adult mouse distal terminal phalanx (P3) amputation, a procedurally simple and reproducible mammalian model of epimorphic ...

P3截肢是研究哺乳动物表观再生的临床前模型，也是识别关键细胞群和控制内源性再生机制的有力工具。P3模型允许识别早期和晚期的细胞群，研究再生期间的再血管化，以及研究无复杂骨稳定装置要求的膜内渗透。与我展示这个程序的将是奥萨马·库雷希、阿莉莎·法尔克和凯瑟琳·齐梅尔，技术员，以及来自我们实验室的研究助理教授雷吉娜·布鲁内尔。在确认8至12周大的CD-1小鼠对手趾捏没有反应后，在动物的眼睛上涂抹软膏，将鼠标置于10X解剖显微镜下。使用手术波维酮碘和70%乙醇消毒每个后肢的数字，并使用微切口修剪头发远离手术现场。局部涂抹一个微升局部局部的布皮瓦卡因麻醉手术场地，并轻轻涂抹一只后爪以暴露中位数字表面。然后拿着手术刀与脂肪垫平行的角度，使用无菌数字10手术刀截肢数字2和4的终端法兰克的端角。将局部的 bupivacaine 再涂抹一个微升，将鼠标返回到干净的笼子，使伤口愈合，无需伤口敷料和监控，直到完全卧位。为了确保P3截肢成功，在截肢过程中将手术刀与脂肪垫平行，并进行微CT扫描以确认正确的截肢长度。在适当的实验终点，使用手术刀切断数字中途通过中间法兰克斯骨在大约第二腹脂肪垫缩进。将组织转移到含有10毫升新鲜缓冲锌的20毫升闪烁小瓶中，固定24至48小时。并使用微原子获取每个数字的四到五微米厚的部分，将切片放入 38 至 41 摄氏度的水浴中，并辅以组织胶粘剂溶液。然后捕获胶粘剂幻灯片上的切片，将幻灯片放在 37 摄氏度的幻灯片加热器上晾干。当幻灯片脱去仿光后，将样品浸入适当抗原检索溶液的染色罐中，将幻灯片加热至 95 摄氏度 25 分钟，确保不发生沸腾。在孵化结束时，让罐子达到室温，将滑梯平放在一个一英寸深的塑料幻灯片盒中，底部铺有湿润的纸巾。在每个样品中加入100微升预预热蛋白酶K。用一小块副膜盖住每张幻灯片，以确保部分不会变干。在 37 摄氏度下 12 分钟后，小心地取出石蜡膜，轻轻将幻灯片排在纸巾上，以去除多余的阻塞溶液。接下来，在每张幻灯片上加入100至200微升的原抗体溶液。将场地返回到覆盖着石蜡膜的加湿室，在摄氏四度下进行过夜孵化。第二天早上，在TRIS缓冲盐水加TWEEN或TBST中清洗幻灯片后，用适当的二次抗体溶液为幻灯片贴上标签，在室温下进行45分钟的孵育，防止光线。在孵育结束时，在TBST中清洗幻灯片，让它们干燥。然后使用 100 微升防褪色安装介质，小心地将盖玻片放在干燥的幻灯片上，并在室温下平整，防止光线，使安装介质干燥。对于连续体内微计算断层扫描或微CT安装带管的微CT成像器，使氧气能够流入和流出微CT动物室，确保流出的氧气配有活性炭过滤器，以捕获异氟。将 Voxelsize 设置为 10.5 微米，在 55 峰值千伏和 145 微安，每 180 度投影 1，000 个，连续旋转，集成时间为 200 毫秒，每次扫描最多 213 片。当扫描仪准备就绪时，轻轻地将麻醉小鼠放在微型CT动物室中，后肢数字排列在扁平的、紧密的关联腹侧向上，左爪在左侧，右侧位于扫描平台的右侧。然后应用手术胶带轻轻地将数字固定到位。在开始扫描之前，将眼科软膏涂抹在动物的眼睛上。扫描后，将动物返回到干净的笼子，并监控直至完全恢复，并允许长达几个小时的微CT图像重建。为了分析重建的3D骨骼图像，使用微CT提供的软件，将图像文件转换为一系列DICOM文件。转换所有文件后，使用 Web 浏览器中的批处理文件下载器在单个文件夹中下载 DICOM 系列。将 BoneJ 插件加载到 ImageJ 插件文件夹中。将 DICOM 文件系列文件夹拖到 ImageJ 栏以打开文件。将打开一个显示所有灰色值的 16 位图像堆栈。要仅显示骨骼，请单击"图像调整"和"阈值"。将上阈值栏滑到最右侧，并调整较低的阈值栏，直到只有骨骼高亮为红色。在最大信号强度为32，767时，较低的阈值约为10，000至13，000。单击"应用"，并在新对话框中单击"黑色背景"。接下来，单击"确定"。将生成显示黑白值的八位图像。与白色对应骨骼。在 P3 骨骼周围绘制一个矩形以选择它并复制堆栈。要生成 3D 图像，请单击插件和 3D 查看器，并使用徒手选择工具对要删除的骨骼进行圈出。然后右键单击并选择"填充选择"以从图像中裁剪任何不必要的骨骼。要量化 3D 渲染的骨骼体积，请打开 BoneJ 插件并选择"体积分数"。将显示一个结果窗口，骨骼体积以毫米为单位显示。要量化 3D 渲染的骨骼长度，请使用 ImageJ 多点工具。要捕获 3D 渲染的图像，请单击"查看"并拍摄快照。然后将图像另存为 TIF 或 JPEG 文件。在这些图像中，成年小鼠再生P3数字的部分被免疫与抗体，以可视化在截肢后的第6天至7天、第9天和10天的米内骨再生和爆炸性形成。在这里，显示了截肢前扫描的数字的代表性微CT渲染，以及再生过程中不同时间点的尺寸测量，用于识别长度测量的地标。鼠标数字模型是一个强大的系统，用于分析一个支持再生的伤口环境，在深部截肢时触发爆炸形成。相反，数字的近位截肢是研究再生失败的模型系统，也是测试增强再生策略的站点。

Research

JoVE Journal

Developmental Biology

Kidney Regeneration in Adult Zebrafish by Gentamicin Induced Injury

肾再生的成年斑马鱼庆大霉素性损伤

The kidney is essential for fluid homeostasis, blood pressure regulation and filtration of waste from the body. The fundamental unit of kidney function is ...

科研

发育生物学

Loss- and Gain-of-function Approach to Investigate Early Cell Fate Determinants in Preimplantation Mouse Embryos

损失 - 和增益的功能角度进行调查研究早期细胞命运决定在植入前小鼠胚胎

 Gene silencing and overexpression techniques are instrumental for the identification of genes involved in embryonic development. Direct target gene ...

Epigenetic Conversion as a Safe and Simple Method to Obtain Insulin-secreting Cells from Adult Skin Fibroblasts

后生转换为安全和简单的方法来获得成人皮肤成纤维细胞胰岛素分泌细胞

Regenerative medicine requires new, fully functional cells that are delivered to patients in order to repair degenerated or damaged tissues. When such ...

Building Finite Element Models to Investigate Zebrafish Jaw Biomechanics

建立有限元模型，研究斑马鱼下颌生物力学

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

Culture of Murine Embryonic Metatarsals: A Physiological Model of Endochondral Ossification

小鼠胚胎跖骨文化：软骨内骨化的生理模型

The fundamental process of endochondral ossification is under tight regulation in the healthy individual so as to prevent disturbed development and/or ...

Establishment of Larval Zebrafish as an Animal Model to Investigate Trypanosoma cruzi Motility In Vivo

Establishment of Larval Zebrafish as an Animal Model to Investigate Trypanosoma cruzi Motility In Vivo

斑马鱼幼虫动物模型的建立研究锥虫运动性体内

Chagas disease is a parasitic infection caused by Trypanosoma cruzi, whose motility is not only important for localization, but also for cellular binding ...

A RANKL-based Osteoclast Culture Assay of Mouse Bone Marrow to Investigate the Role of mTORC1 in Osteoclast Formation

RANKL 小鼠骨髓破骨细胞培养法研究 mTORC1 在破骨细胞形成中的作用

Osteoclasts are unique bone-resorbing cells that differentiate from the monocyte/macrophage lineage of bone marrow. Dysfunction of osteoclasts may result ...

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

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

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 ...

Partial Lobular Hepatectomy: A Surgical Model for Morphologic Liver Regeneration

部分小叶切除术: 形态学肝再生的外科模型

Morphological organ regeneration following acute tissue loss is common among lower vertebrates, but is rarely observed in mammalian postnatal life. Adult ...

A Simplified and Efficient Method to Isolate Primary Human Keratinocytes from Adult Skin Tissue

一种简便有效的成人皮肤组织中人角质形成细胞的分离方法

Primary human keratinocytes isolated from fresh skin tissues and their expansion in vitro have been widely used for laboratory research and for clinical ...