Name: two-step approach to explore early- and late-stages of organ formation in the avian model: the thymus and parathyroid glands organogenesis paradigm
Uploaded: 2018-06-17T14:00:00
Description: Watch this Scientific Journal Video about Two-step Approach to Explore Early- and Late-stages of Organ Formation in the Avian Model: The Thymus and Parathyroid Glands Organogenesis Paradigm at JoVE.co

作者感谢 António Cidadão、伊莎贝尔 Alcobia 和德里奥诺佩雷拉对手稿的批判性阅读, 对的视频叙事 Akkapeddi, 并体 Proa 从组织学服务 Histologia e Biologia 做Desenvolvimento, Faculdade de Medicina 里斯本, 里斯本 de 葡京, 为技术支持。我们特别感谢圣保罗 Caeiro 和雨果席尔瓦从 Unidade de audiovisuais (视听股), Faculdade de Medicina 德葡京, 里斯本 de 葡京, 他们对制作这部影片的杰出承诺。我们承认莱卡微系统的亲切提供了一个立体镜配备了视频体系和 Interaves-博彩农业 Pecuária, S. 为鹌鹑受精卵子贡献。这项工作得到葡京 Faculdade de Medicina、里斯本葡京 (FMUL) 的支持。

所有这些实验都遵循 Medicina Académico 中心的动物保育和道德准则。
1. 受精鹌鹑和鸡卵的孵化
<ol><li>孵化日本鹌鹑 (鹌鹑鹌鹑) 和鸡肉 (鸡鸡) 受精卵3和8天, 分别。<ol>
<li>放置鸡蛋与空气室 (鸡蛋钝端) 面对在一个湿润的孵化器在38摄氏度。</li>
		<li>用大约20只鹌鹑蛋和40只鸡蛋做这个实验。 注: 第一次建立此程序时, 这些数字应加倍。</li>
	</ol>
</li>
</ol><p class="jove_...

<ol>
	<li>Le Douarin, N. 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+Nogent+Institute-50+years+of+embryology.">The Nogent Institute-50 years of embryology.</a> Int J Dev Biol. 49 (2-3), 85-103 (2005).</li><li>Chuong, C. M., Wu, P., Plikus, M., Jiang, T. X., Bruce Widelitz, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Engineering+stem+cells+into+organs:+topobiological+transformations+demonstrated+by+beak,+feather,+and+other+ectodermal+organ+morphogenesis.">Engineering stem cells into organs: topobiological transformations demonstrated by beak, feather, and other ectodermal organ morphogenesis.</a> Curr Top Dev Biol. 72, 237-274 (2006).</li><li>Davey, M. G., Tickle, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+chicken+as+a+model+for+embryonic+development.">The chicken as a model for embryonic development.</a> Cytogenet Genome Res. 117 (1-4), 231-239 (2007).</li><li>Nowak-Sliwinska, P., Segura, T., Iruela-Arispe, 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=The+chicken+chorioallantoic+membrane+model+in+biology,+medicine+and+bioengineering.">The chicken chorioallantoic membrane model in biology, medicine and bioengineering.</a> Angiogenesis. 17 (4), 779-804 (2014).</li><li>Figueiredo, M., Silva, J. 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O., Correia-Pinto, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=FGF+signalling+pathway+in+the+developing+chick+lung:+expression+and+inhibition+studies.">FGF signalling pathway in the developing chick lung: expression and inhibition studies.</a> PLoS One. 6 (3), e17660 (2011).</li><li>Hamburger, V., Hamilton, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+series+of+normal+stages+in+the+development+of+the+chick+embryo.">A series of normal stages in the development of the chick embryo.</a> J. Morphol. 88, 49-92 (1951).</li><li>Takahashi, Y., Bontoux, M., Le Douarin, N. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Epithelio-mesenchymal+interactions+are+critical+for+Quox+7+expression+and+membrane+bone+differentiation+in+the+neural+crest+derived+mandibular+mesenchyme.">Epithelio-mesenchymal interactions are critical for Quox 7 expression and membrane bone differentiation in the neural crest derived mandibular mesenchyme.</a> EMBO J. 10 (9), 2387-2393 (1991).</li><li>Maeda, Y., Noda, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Coordinated+development+of+embryonic+long+bone+on+chorioallantoic+membrane+in+ovo+prevents+perichondrium-derived+suppressive+signals+against+cartilage+growth.">Coordinated development of embryonic long bone on chorioallantoic membrane in ovo prevents perichondrium-derived suppressive signals against cartilage growth.</a> Bone. 32 (1), 27-34 (2003).</li><li>Ishida, K., Mitsui, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Generation+of+bioengineered+feather+buds+on+a+reconstructed+chick+skin+from+dissociated+epithelial+and+mesenchymal+cells.">Generation of bioengineered feather buds on a reconstructed chick skin from dissociated epithelial and mesenchymal cells.</a> Dev Growth Differ. 58 (3), 303-314 (2016).</li><li>Le Douarin, N. M., Jotereau, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Tracing+of+cells+of+the+avian+thymus+trough+embryonic+life+in+interspecific+chimeras.">Tracing of cells of the avian thymus trough embryonic life in interspecific chimeras.</a> J. Exp. Med. 142, 17-40 (1975).</li><li>Farley, A. 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=Dynamics+of+thymus+organogenesis+and+colonization+in+early+human+development.">Dynamics of thymus organogenesis and colonization in early human development.</a> Development. 140, 2015-2026 (2013).</li><li>Gordon, J., Bennett, A. R., Blackburn, C. C., Manley, N. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Gcm2+and+Foxn1+mark+early+parathyroid-+and+thymus-specific+domains+in+the+developing+third+pharyngeal+pouch.">Gcm2 and Foxn1 mark early parathyroid- and thymus-specific domains in the developing third pharyngeal pouch.</a> Mech Dev. 103, 141-143 (2001).</li><li>Neves, H., Dupin, E., Parreira, L., Le Douarin, N. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Modulation+of+Bmp4+signalling+in+the+epithelial-mesenchymal+interactions+that+take+place+in+early+thymus+and+parathyroid+development+in+avian+embryos.">Modulation of Bmp4 signalling in the epithelial-mesenchymal interactions that take place in early thymus and parathyroid development in avian embryos.</a> Dev Biol. 361 (2), 208-219 (2012).</li><li>Jerome, L. A., Papaioannou, V. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=DiGeorge+syndrome+phenotype+in+mice+mutant+for+the+T-box+gene,+Tbx1.">DiGeorge syndrome phenotype in mice mutant for the T-box gene, Tbx1.</a> Nat Genet. 27 (3), 286-291 (2001).</li><li>Nie, X., Brown, C. B., Wang, Q., Jiao, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Inactivation+of+Bmp4+from+the+Tbx1+expression+domain+causes+abnormal+pharyngeal+arch+artery+and+cardiac+outflow+tract+remodeling.">Inactivation of Bmp4 from the Tbx1 expression domain causes abnormal pharyngeal arch artery and cardiac outflow tract remodeling.</a> Cells Tissues Organs. 193 (6), 393-403 (2011).</li><li>Zou, D., Silvius, D., Davenport, J., Grifone, R., Maire, P., Xu, P. X. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Patterning+of+the+third+pharyngeal+pouch+into+thymus/parathyroid+by+Six+and+Eya1.">Patterning of the third pharyngeal pouch into thymus/parathyroid by Six and Eya1.</a> Dev Biol. 293 (2), 499-512 (2006).</li><li>Uematsu, E., 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=Use+of+in+ovo.+chorioallantoic+membrane+engraftment+to+culture+testes+from+neonatal+mice.">Use of in ovo. chorioallantoic membrane engraftment to culture testes from neonatal mice.</a> Comp Med. 64 (4), 264-269 (2014).</li></ol>

这个方法的成功的一个关键方面是鸡和鹌鹑蛋的质量。考虑到长期的潜伏期, 特别是在蛋试验期间, 良好的鸡卵质量提高了生存率 (高达 90%) 的过程结束。要做到这一点, 测试鸡蛋从不同的供应商。长时间孵化 unmanipulated 卵 (最多16-17 天) 并检查其发育情况。要被认为是一个良好的质量批次, 超过80% 的胚胎应该呈现正常的发展。同样重要的是要确保每个孵化步骤提供可重现的同步发展阶段, 以?...

禽胚在精发育生物学研究中得到了广泛的应用。鸟类模型的主要优点包括打开卵子的可能性, 相对容易进入胚胎, 以及执行微操作系统的能力。一些例子包括经典的鹌鹑鸡嵌合体系统研究细胞命运1, 具体生长因子的应用到胚胎2, 以及异位细胞结构的增长在凸轮1,3, 4。
为了对器官形成的不同阶段有新的认识, 我们最近开发了一种将嫁接技术与胚胎组织体外操作相结合的方法5。这两个步骤的方法使对器官的早期和后期阶段的歧视和探索, 这往往是有限的, 因为高度动态和复杂的组织相互作用2。此外, 缺乏适当的组织特异标记经常限制使用基因改良动物模型6。这两步方法的新方法基本上克服了这种限制。
为了研究器官形成的早期阶段, 在第一步, 在 48 h 的体外脊髓系统中分离和培养出包括前瞻性器官雏形的鹌鹑胚胎区。在此期间, 特定信号通路的药理调控可以通过加入药物到培养基5,7来进行。此外, 培养组织可以收集在任何阶段的体外生长和探索基因表达 (使用方法, 如 qPCR, 微阵列等)。
第二步, 在胚胎日 (E) 8 (cE8) (汉堡和汉密尔顿 (HH)-阶段 33-35)8中, 将48个 h 培养组织移植到鸡 (c) 胚的凸轮上。该凸轮作为营养物质的血管供应商, 允许气体交换1,3,4到嫁接组织, 使其在蛋的发展更长一段时间。这一实验步骤特别适合研究器官的后期阶段, 因为完全形成的器官可以获得后10天的蛋发展5,9,10,11.形态学分析很容易通过传统的组织学来确认适当的器官形成和捐献者的细胞来源可以通过免疫组化使用特定物种的抗体 (即单克隆鹌鹑核周 (QCPN)) 鉴定。在凸轮潜伏期, 移植物也可以生长在药理剂的存在, 并收集在任何发展阶段, 以评估器官的进展。
这两步方法, 在这里深入描述, 已经在 Figueiredo等使用。5探讨禽甲状旁腺/胸腺共同普利摩顿的发展。因此, 在胸腺和甲状旁腺器官的形成过程中, 胚胎领土和发育阶段的固有特殊性将列于下文。
胸腺和甲状旁腺上皮, 虽然功能上不同, 从咽囊的内胚层 (PP)12。在禽类中, 这些器官的上皮来源于第三和第四 pp 内胚层 (3/4 pp)12, 而在哺乳动物中, 胸腺上皮来源于 3PP, 而甲状旁腺上皮则来源于老鼠和人类的3PP 和 3/4 PP,分别为13、14。
这些器官形成的最早阶段之一是在普通普利摩顿中出现离散胸腺和甲状旁腺领域。在鸡肉中, 这些领域可以通过原位杂交识别, 用特定的分子标记, 在 E4.515。随着发展的进行, 这些器官的雏形是个性化的, 并与咽分离, 而由神经嵴衍生细胞形成的薄间质胶囊 (在 E5;HH-阶段 27)。随后, 胸腺上皮由造血祖细胞 (E6.5) 所殖民;HH-阶段 30)12。
与经典鹌鹑鸡研究1,12, 两步方法是特别有用的研究造血/淋巴器官的形成, 即胸腺5。当鹌鹑外植体, 与器官雏形, 是嫁接在鸡胚前造血祖细胞殖民化, 一个嵌合胸腺形成与鸡血源祖细胞浸润鹌鹑胸腺上皮对应。因此, 该方法是探讨造血细胞在禽换血术/淋巴系统发展中的作用的有用工具。

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>Chicken fertilized eggs (Gallus gallus)</td>
			<td>Pintobar, Portugal</td>
			<td></td>
			<td>Poultry farm&#160;</td>
		</tr>
		<tr>
			<td>Quail fertilized eggs (Coturnix coturnix)</td>
			<td>Interaves, Portugal</td>
			<td></td>
			<td>Bird farm&#160;</td>
		</tr>
		<tr>
			<td>15 mL PP centrifuge tubes</td>
			<td>Corning</td>
			<td>430052</td>
			<td></td>
		</tr>
		<tr>
			<td>50 mL PP centrifuge tubes</td>
			<td>Corning</td>
			<td>430290</td>
			<td></td>
		</tr>
		<tr>
			<td>60 x 20 mm pyrex dishes</td>
			<td>Duran group</td>
			<td>21 755 41</td>
			<td></td>
		</tr>
		<tr>
			<td>100 x 20 mm pyrex dishes</td>
			<td>Duran group</td>
			<td>21 755 48</td>
			<td></td>
		</tr>
		<tr>
			<td>Polycarbonate Membrane Insert&#160;</td>
			<td>Corning</td>
			<td>3412</td>
			<td>24 mm transwell with 0.4 mm Pore Polycarbonate Membrane Insert&#160;</td>
		</tr>
		<tr>
			<td>Membrane filter</td>
			<td>Millipore</td>
			<td>DTTP01300</td>
			<td>0.6 mm Isopore membrane filter</td>
		</tr>
		<tr>
			<td>6-well culture plates</td>
			<td>Nunc, Thermo Fisher Scientific</td>
			<td>140675</td>
			<td></td>
		</tr>
		<tr>
			<td>Petri dish, 35 x 10 mm</td>
			<td>Sigma-Aldrich</td>
			<td>P5112&#160;</td>
			<td></td>
		</tr>
		<tr>
			<td>Pyrex bowls</td>
			<td></td>
			<td></td>
			<td>from supermarket&#160;</td>
		</tr>
		<tr>
			<td>Transfer pipettes</td>
			<td>Samco Scientific, Thermo Fisher Scientific</td>
			<td>2041S</td>
			<td>2 mL plastic pipet</td>
		</tr>
		<tr>
			<td>Glass pasteur pipette</td>
			<td>normax</td>
			<td>5426015</td>
			<td></td>
		</tr>
		<tr>
			<td>Whatman&#160;qualitative filter paper</td>
			<td>Sigma-Aldrich</td>
			<td>WHA1001090</td>
			<td>Filter paper</td>
		</tr>
		<tr>
			<td>Clear plastic tape</td>
			<td></td>
			<td></td>
			<td>from supermarket&#160;</td>
		</tr>
		<tr>
			<td>Cytokeratin (pan; acidic and basic, type I and II cytokeratins), clone Lu-5</td>
			<td>BMA Biomedicals</td>
			<td>T-1302</td>
			<td></td>
		</tr>
		<tr>
			<td>Cyclopamine hydrate</td>
			<td>Sigma-Aldrich</td>
			<td>C4116</td>
			<td>Pharmacological inhibitor of Hh signalling</td>
		</tr>
		<tr>
			<td>Fetal Bovine Serum</td>
			<td>Invitrogen, Thermo Fisher Scientific</td>
			<td></td>
			<td>Standart FBS</td>
		</tr>
		<tr>
			<td>Paraformaldehyde</td>
			<td>Sigma-Aldrich</td>
			<td>P6148</td>
			<td></td>
		</tr>
		<tr>
			<td>Penicillin-Streptomycin</td>
			<td>Invitrogen, Thermo Fisher Scientific</td>
			<td>15140-122</td>
			<td></td>
		</tr>
		<tr>
			<td>Phosphate-Buffered Saline (PBS)</td>
			<td>GIBCO, Thermo Fisher Scientific</td>
			<td>10010023</td>
			<td></td>
		</tr>
		<tr>
			<td>QCPN antibody</td>
			<td>Developmental Studies Hybridoma Bank</td>
			<td>QCPN</td>
			<td></td>
		</tr>
		<tr>
			<td>RPMI 1640 Medium, GlutaMAX Supplement&#160;</td>
			<td>GIBCO, Thermo Fisher Scientific</td>
			<td>61870010</td>
			<td></td>
		</tr>
		<tr>
			<td>Bluesil RTV141A/B Silicone Elastomer 1.1Kg Kit</td>
			<td>ELKEM/Silmid</td>
			<td>RH141001KG</td>
			<td>To prepare the back base for petri dish</td>
		</tr>
		<tr>
			<td>Stemolecule LY411575</td>
			<td>Stemgent</td>
			<td>04-0054</td>
			<td>Pharmacological inhibitor of Notch signalling</td>
		</tr>
		<tr>
			<td>TRIzol Reagent</td>
			<td>Invitrogen, Thermo Fisher Scientific</td>
			<td>15596026</td>
			<td>Reagent for total RNA isolation</td>
		</tr>
		<tr>
			<td>Dumont #5 Forceps</td>
			<td>Fine Science Tools</td>
			<td>11251-30&#160;</td>
			<td>Thin forceps</td>
		</tr>
		<tr>
			<td>Extra fine Bonn scissors, curved</td>
			<td>Fine Science Tools</td>
			<td>14085-08</td>
			<td>Curved scissors</td>
		</tr>
		<tr>
			<td>Insect pins&#160;</td>
			<td>Fine Science Tools</td>
			<td>26001-30</td>
			<td></td>
		</tr>
		<tr>
			<td>Micro spatula&#160;</td>
			<td>Fine Science Tools</td>
			<td>10087-12</td>
			<td>Transplantation spoon</td>
		</tr>
		<tr>
			<td>Minutien Pins</td>
			<td>Fine Science Tools</td>
			<td>26002-20</td>
			<td>Microscalpel</td>
		</tr>
		<tr>
			<td>Moria Nickel Plated Pin Holder</td>
			<td>Fine Science Tools</td>
			<td>26016-12</td>
			<td>Holder</td>
		</tr>
		<tr>
			<td>Moria Perforated Spoon</td>
			<td>Fine Science Tools</td>
			<td>10370-17</td>
			<td>Skimmer</td>
		</tr>
		<tr>
			<td>Wecker Eye Scissor</td>
			<td>Fine Science Tools</td>
			<td>15010-11</td>
			<td></td>
		</tr>
		<tr>
			<td>Camera</td>
			<td>Leica Microsystems&#160;</td>
			<td>MC170 HD</td>
			<td></td>
		</tr>
		<tr>
			<td>Stereoscope</td>
			<td>Leica Microsystems&#160;</td>
			<td>Leica M80</td>
			<td></td>
		</tr>
		<tr>
			<td>Microscope</td>
			<td>Leica Microsystems&#160;</td>
			<td>DM2500</td>
			<td></td>
		</tr>
	</tbody>
</table>

two-step approach to explore early- and late-stages of organ formation in the avian model: the thymus and parathyroid glands organogenesis paradigm

禽胚作为一种实验模型, 对发育生物学中的精发现具有极其重要的意义。在几种方法中, 鹌鹑鸡合体的形成和胚膜 (CAM) 用于维持异位组织发育的时间追溯到上个世纪。目前, 这些经典技术与最近的体外方法相结合, 为进一步探索器官形成提供了新的前景。
在这里, 我们描述了一个两个步骤的方法来研究早期和后期的器官。简言之, 含有该器官推定区域的胚胎区域与鹌鹑胚胎分离, 并在脊髓系统中体外生长 (最多48小时)。培养的组织随后被嫁接到鸡胚的凸轮上。经过10天的蛋发展, 完全形成的器官是从嫁接组织获得。这种方法还允许在整个体外和蛋发育步骤中, 通过药物的常规管理和组织遗传操作来调制信号通路。此外, 开发组织可以收集在任何时间窗口, 以分析其基因表达谱 (使用定量 PCR (qPCR), 微阵列等) 和形态学 (评估与常规组织学和免疫化学)。
所描述的实验程序可作为一种工具, 遵循鸟类胚胎外的器官形成, 从器官的早期阶段到完全形成和功能器官。

上述协议详细说明了一种方法, 它允许对器官的早期和后期阶段进行调查, 通常受复杂的细胞和分子相互作用的限制。
这种方法以前曾在 Figueiredo等地使用过。5解开缺口和 Hh 信号在禽甲状旁腺/胸腺共同普利摩顿发展中的作用。
在此,图 1</stro...

Watch this Scientific Journal Video about Two-step Approach to Explore Early- and Late-stages of Organ Formation in the Avian Model: The Thymus and Parathyroid Glands Organogenesis Paradigm at JoVE.co

Two-step Approach to Explore Early- and Late-stages of Organ Formation in the Avian Model: The Thymus and Parathyroid Glands Organogenesis Paradigm

鸟类模型中器官形成早期和晚期的两步探讨: 胸腺和甲状旁腺器官发生范式

本文通过结合新颖的体外和蛋实验程序, 为经典鹌鹑-鸡嵌合体系统的研究提供了一种最新的方法。

The avian embryo, as an experimental model, has been of utmost importance for seminal discoveries in developmental biology. Among several approaches, the ...

Research

JoVE Journal

Developmental Biology

Understanding Early Organogenesis Using a Simplified In Situ Hybridization Protocol in Xenopus

Understanding Early Organogenesis Using a Simplified In Situ Hybridization Protocol in Xenopus

了解早期器官使用简体<em&gt;原位</em&gt;杂交协议<em&gt;爪</em

Organogenesis is the study of how organs are specified and then acquire their specific shape and functions during development.  The Xenopuslaevis embryo ...

科研

发育生物学

Protocols for Obtaining Zygotic and Somatic Embryos for Studying the Regulation of Early Embryo Development in the Model Legume Medicago truncatula

Protocols for Obtaining Zygotic and Somatic Embryos for Studying the Regulation of Early Embryo Development in the Model Legume Medicago truncatula

协议获得合子和体细胞胚胎研究早期胚胎发育的调控中的示范豆科<em&gt;苜蓿</em

Early embryogenesis starting from a single cell zygote goes through rapid cell division and morphogenesis, and is morphologically characterized by ...

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 ...

Gene Transfer into the Chicken Auditory Organ by In Ovo Micro-electroporation

Gene Transfer into the Chicken Auditory Organ by In Ovo Micro-electroporation

基因转移到鸡听觉器官通过<em&gt;在卵</em&gt;微电

Chicken embryos are ideal model systems for studying embryonic development as manipulations of gene function can be conducted with relative ease in ovo. ...

Methods to Examine the Lymph Gland and Hemocytes in Drosophila Larvae

Methods to Examine the Lymph Gland and Hemocytes in Drosophila Larvae

方法来检查淋巴腺和血细胞中<em&gt;果蝇</em&gt;幼虫

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

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

成人果蝇脑内蘑菇体和感光神经元的解剖和荧光染色

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

Application of RNAi and Heat-shock-induced Transcription Factor Expression to Reprogram Germ Cells to Neurons in C. elegans

Application of RNAi and Heat-shock-induced Transcription Factor Expression to Reprogram Germ Cells to Neurons in C. elegans

rna 干扰和热休克诱导转录因子表达在重编程生殖细胞到神经元中的应用C. 线虫

Studying the cell biological processes during converting the identities of specific cell types provides important insights into mechanism that maintain ...

Isolation of Embryonic Tissues and Formation of Quail-Chicken Chimeric Organs Using The Thymus Example

胸腺标本分离和鸡刺激器官的形成

The capacity to isolate embryonic tissues was an essential step for establishing the quail-chicken chimera system, which in turn has provided undisputed ...

Microinjection-based System for In Vivo Implantation of Embryonic Cardiomyocytes in the Avian Embryo

基于微注射的免疫胚胎心肌细胞体内植入系统

Interpreting the relative impact of cell autonomous patterning versus extrinsic microenvironmental influence on cell lineage determination represents a ...

Whole-Mount In Situ Hybridization in Zebrafish Embryos and Tube Formation Assay in iPSC-ECs to Study the Role of Endoglin in Vascular Development

斑马鱼胚胎全山原位杂交和管状测定在iPSC-ECs中研究恩多林在血管发育中的作用

Vascular development is determined by the sequential expression of specific genes, which can be studied by performing in situ hybridization assays in ...