Name: methods for the modulation and analysis of nf-&#954;b-dependent adult neurogenesis
Uploaded: 2014-02-13T23:00:00
Description: Watch this Scientific Journal Video about Methods for the Modulation and Analysis of NF-ÃŽÂºB-dependent Adult Neurogenesis at JoVE.com

我们感谢安吉拉Kralemann - 科勒为优秀的技术支持。本文所述的实验工作是在我们的实验室进行，由授予德国研究理事会（DFG）的CK和BK和BK的资助研究和教育的德国卫生部部（BMBF）的支持。

伦理声明这项研究进行了严格按照政府的动物和保健使用委员会，国家北莱茵 - 威斯特法伦州，（德国杜塞尔多夫）的LANUV的规定。所有的动物实验已根据许可证号8.87-51.04.20.09.317（LANUV，NRW）批准LANUV，杜塞尔多夫。所做的一切努力，尽量减少痛苦和需要研究的动物的数量。 1。动物护理及房屋<ol><li>在本文描述的协议中使用的所有动物应保持无特定病原?...

<ol>
	<li>Gutierrez, H., Davies, 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=Regulation+of+neural+process+growth,+elaboration+and+structural+plasticity+by+NF-kappaB.">Regulation of neural process growth, elaboration and structural plasticity by NF-kappaB.</a> Trends Neurosci. 34, 316-325 (2011).</li><li>Imielski, Y., 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=Regrowing+the+adult+brain:+NF-kappaB+controls+functional+circuit+formation+and+tissue+homeostasis+in+the+dentate+gyrus.">Regrowing the adult brain: NF-kappaB controls functional circuit formation and tissue homeostasis in the dentate gyrus.</a> PLoS One. 7, (2012).</li><li>Zheng, 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=Intrahippocampal+injection+of+A+beta(1-42)+inhibits+neurogenesis+and+down-regulates+IFN-gamma+and+NF-kappaB+expression+in+hippocampus+of+adult+mouse+brain.">Intrahippocampal injection of A beta(1-42) inhibits neurogenesis and down-regulates IFN-gamma and NF-kappaB expression in hippocampus of adult mouse brain.</a> Amyloid. 20 (1-42), 13-20 (2013).</li><li>Denis-Donini, 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=Impaired+adult+neurogenesis+associated+with+short-term+memory+defects+in+NF-kappaB+p50-deficient+mice.">Impaired adult neurogenesis associated with short-term memory defects in NF-kappaB p50-deficient mice.</a> J. Neurosci. 28, 3911-3919 (2008).</li><li>Bracchi-Ricard, V., 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=Astroglial+nuclear+factor-kappaB+regulates+learning+and+memory+and+synaptic+plasticity+in+female+mice.">Astroglial nuclear factor-kappaB regulates learning and memory and synaptic plasticity in female mice.</a> J, Neurochem. 104, 611-623 (2008).</li><li>Koo, J. W., Russo, S. J., Ferguson, D., Nestler, E. J., Duman, R. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Nuclear+factor-kappaB+is+a+critical+mediator+of+stress-impaired+neurogenesis+and+depressive+behavior.">Nuclear factor-kappaB is a critical mediator of stress-impaired neurogenesis and depressive behavior.</a> Proc. Natl. Acad. Sci. U.S.A. 107, 2669-2674 (2010).</li><li>Fridmacher, V., 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=Forebrain-specific+neuronal+inhibition+of+nuclear+factor-kappaB+activity+leads+to+loss+of+neuroprotection.">Forebrain-specific neuronal inhibition of nuclear factor-kappaB activity leads to loss of neuroprotection.</a> J. Neurosci. 23, 9403-9408 (2003).</li><li>Whiteside, S. 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=C-terminal+sequences+control+degradation+of+MAD3/I+kappa+B+alpha+in+response+to+inducers+of+NF-kappa.">C-terminal sequences control degradation of MAD3/I kappa B alpha in response to inducers of NF-kappa.</a> B activity. Mol. Cell. Biol. 15, 5339-5345 (1995).</li><li>Mayford, 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=Control+of+memory+formation+through+regulated+expression+of+a+CaMKII+transgene.">Control of memory formation through regulated expression of a CaMKII transgene.</a> Science. 274, 1678-1683 (1996).</li><li>Rosenfeld, M. E., Prichard, L., Shiojiri, N., Fausto, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Prevention+of+hepatic+apoptosis+and+embryonic+lethality+in+RelA/TNFR-1+double+knockout+mice.">Prevention of hepatic apoptosis and embryonic lethality in RelA/TNFR-1 double knockout mice.</a> Am. J. Pathol. 156, 997-1007 (2000).</li><li>Morris, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Developments+of+a+water-maze+procedure+for+studying+spatial+learning+in+the+rat.">Developments of a water-maze procedure for studying spatial learning in the rat.</a> J Neurosci. Methods. 11, 47-60 (1984).</li><li>Barnes, 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=Memory+deficits+associated+with+senescence:+a+neurophysiological+and+behavioral+study+in+the+rat.">Memory deficits associated with senescence: a neurophysiological and behavioral study in the rat.</a> J. Compar. Physiol. Psychol. 93, 74-104 (1979).</li><li>Brun, V. 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=Place+cells+and+place+recognition+maintained+by+direct+entorhinal-hippocampal+circuitry.">Place cells and place recognition maintained by direct entorhinal-hippocampal circuitry.</a> Science. 296, 2243-2246 (2002).</li><li>Meshi, D., 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=Hippocampal+neurogenesis+is+not+required+for+behavioral+effects+of+environmental+enrichment.">Hippocampal neurogenesis is not required for behavioral effects of environmental enrichment.</a> Nat. Neurosci. 9, 729-731 (2006).</li><li>Bakker, A., Kirwan, C. B., Miller, M., Stark, C. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Pattern+separation+in+the+human+hippocampal+CA3+and+dentate+gyrus.">Pattern separation in the human hippocampal CA3 and dentate gyrus.</a> Science. 319, 1640-1642 (2008).</li><li>Dupret, D., 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=Spatial+relational+memory+requires+hippocampal+adult+neurogenesis.">Spatial relational memory requires hippocampal adult neurogenesis.</a> PLoS One. 3, (2008).</li><li>Leutgeb, J. K., Leutgeb, S., Moser, M. B., Moser, E. I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Pattern separation in the dentate gyrus and CA3 of the hippocampus. Science. 315. , 961-966 (2007).</li><li>Kaltschmidt, B., 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=NF-kappaB+regulates+spatial+memory+formation+and+synaptic+plasticity+through+protein+kinase+A/CREB+signaling.">NF-kappaB regulates spatial memory formation and synaptic plasticity through protein kinase A/CREB signaling.</a> Mol. Cell. Biol. 26, 2936-2946 (2006).</li><li>Clelland, C. D., 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=A+functional+role+for+adult+hippocampal+neurogenesis+in+spatial+pattern+separation.">A functional role for adult hippocampal neurogenesis in spatial pattern separation.</a> Science. 325, 210-213 (2009).</li><li>Kempermann, G., Jessberger, S., Steiner, B., Kronenberg, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Milestones+of+neuronal+development+in+the+adult+hippocampus.">Milestones of neuronal development in the adult hippocampus.</a> Trends Neurosci. 27, 447-452 (2004).</li></ol>

成年神经发生，并通过经多西环素在神经元抑制NF-κB的，其激活后其操纵的可能性，提供了调查，​​在成人大脑，以及为神经元去和重新生成的新生神经元引人入胜系统。该系统的优点在于，NF-κB信号通路抑制神经元不仅导致改变神经细胞死亡，祖细胞的增殖和迁移，以及严重的结构和解剖学上的变化，而且在学习的明显缺陷。重要的是，IκB/ tTA的动物的表型可逆转实验和多西环素给药后获救?...

本体论上，海马是已知的最古老的大脑解剖结构之一。它负责不同的复杂的任务，如在长期记忆，空间定位，并形成各自的存储器中，并固结的调节枢转功能。在解剖学上，海马由锥体细胞层（ 层锥体 ）包括大角Ammonis（CA1，CA2，CA3，CA4和）区和齿状回（ 齿状回 ），其中包含的颗粒细胞和一些神经祖细胞内的颗粒下层区。的颗粒细胞经所谓苔藓纤维（颗粒细胞的轴突）突出朝向CA3区。 直到最后世纪末，成年哺乳动物大脑被认为是静态的器官缺乏细胞可塑性和神经发生。然而，在过去的二十年中，越来越多的证据量清楚地表明了成年神经发生在至少2脑区，脑室下区（SVZ）和海马的颗粒下层区域。 我们以前的研究，并与其他团体，都表明，转录因子NF-κB的是成年神经发生的关键分子调节剂之一，其放松管制导致严重的结构缺陷，海马和认知障碍1-6。 NF-κB是五DNA结合亚基二聚体的不同组合组成的诱导转录因子的通用名称：P50，P52，C-REL，RelB基因和P65（的RelA），后三家的反式激活域。于大脑，在细胞质中发现的最丰富的形式是P50和P65，这是由卡巴B（IκB）蛋白的抑制剂保持在非活性形式的异源二聚体。 学习和直接操纵的NF-κB驱动的神经，我们用转基因小鼠模型，以使所有的NF-κBsubun的简单的抑制作用它的，特别是在脑7（ 见图1）。为了这个目的，我们杂交以下转基因小鼠系，IκB/ - 和-/tTA。采用NF-κB抑制剂IκBa（超阻遏IκBa-AA1）的反式显性负突变体产生的8号线-转基因IκB/。相反，野生型IκBα，IκBα-AA1具有两个丝氨酸残基突变为丙氨酸（V32和V36），这阻碍了该抑制剂的磷酸化和随后的蛋白酶体降解。对IκBa-AA1-转基因的前脑神经元特异性表达，IκB/ - 小鼠杂交小鼠窝藏钙 - 钙调蛋白依赖性蛋白激酶IIα（CAMKIIα）的启动子，可以通过四环素反式激活物（TTA）被驱动9。 P65基因敲除小鼠的胚胎致死表型，由于大量肝细胞凋亡10，所以这里显示的方法提供了一个优雅的方法为调查的NF-κB在产后和成年神经的作用。 经典的行为学测试，以空间学习和记忆研究20世纪80年代由理查德·莫里斯，测试被称为Morris水迷宫（MWM）11被描述。在这个开放的场水迷宫，动物学会从不透明水中逃脱到基于方向和额外迷宫的线索隐藏的平台。 MWM的干变体是所谓的巴恩斯迷宫（BM）12。本次测试采用了圆形板布置在板的边缘20圆孔，用一个定义孔作为转义框，视觉多余的迷宫线索方向。这两种实验范式依靠飞行行为引起的啮齿动物单曲厌恶水，或打开，明亮的照明空间。这两项测试让空间方位的调查，以及相关的内存性能。虽然海马起着空间记忆形成的一般和重要作用，海马Ř参与egions不同，根据不同的应用测试。在BM测试的内存源于enthorinal皮层（EC）和位于海马CA1区域无危险品13-16的贡献锥体神经元之间的神经元活动。特别是，经典的BM主要是通过突触颞ammonic途径从欧盟三CA1至欧盟五，重要的是，DG关键是参与所谓的空间模式识别17，这意味着不仅是处理依赖于导航视觉和空间信息，但也差不多陈述或记忆转化为不同的，不重叠的陈述。这个任务需要的功能的三突触回路从欧盟二DG可以CA3到CA1和EC六，不能在BM 15进行测试。 为应对这些挑战，我们设计了一套SPS-BM作为一个行为测试，具体测试齿状回依赖的认知表现在合作ntrol动物，并在IκB/ tTA的超阻抑物模型下的NF-κB的抑制。重要的是，与此相反的MWM或骨髓，在SPS-BM可以揭示从神经发生减值产生微妙的行为缺陷。因为空间图案分离是严格依赖于EC II和DG和CA3和CA1和EC VI之间的功能性电路，该测试是内中的神经发生电位变化，苔藓纤维通路的修改或组织稳态的变化高度敏感DG。 技术上，设定了我们的试验是基于研究Clelland 等，其中的空间间隔图案是用一个木制的8臂旋臂迷宫（RAM）19进行测试。在我们修改设置，八臂，7个相同的黄色食物房屋所取代。总之，这里显示的方法，包括doublecortin表达（DCX +）海马，苔藓纤维突起，神经元细胞内德细胞分析ATH特别是SPS-BM这里提出，可应用到的结合具有在成年神经发生的影响的转基因其他小鼠模型的研究。进一步的应用可能包括药物制剂的研究和测量它们对DG和空间格局分离的影响。

<table border="0" cellpadding="0" cellspacing="0" width="872">
	<colgroup>
		<col />
		<col />
		<col />
		<col />
	</colgroup>
	<tbody>
		<tr height="20">
			<td height="20" style="height:20px;width:447px;">Moria MC17 Perforated Spoon&#160;</td>
			<td style="width:127px;">FST</td>
			<td style="width:132px;">10370-18</td>
			<td style="width:167px;">removal of the brains</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Dissecting microscope</td>
			<td>Carl Zeiss</td>
			<td>Stemi SV8</td>
			<td>removal of the brains</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Surgical scissors&#160;</td>
			<td style="width:127px;">FST</td>
			<td>14084-08</td>
			<td>removal of the brains</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Surgical scissors&#160;</td>
			<td style="width:127px;">FST</td>
			<td>14381-43</td>
			<td>removal of the brains</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Dumont #5 forceps</td>
			<td style="width:127px;">FST</td>
			<td>11254-20</td>
			<td>removal of the brains</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:447px;">SuperFrost Slides</td>
			<td>Carl Roth&#160;</td>
			<td align="right">1879</td>
			<td>slides for immunohistochemistry</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:447px;">Paraformaldehyde</td>
			<td style="width:127px;">Sigma-Aldrich</td>
			<td>P6148</td>
			<td>fixative</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:447px;">TissueTek OCT compound</td>
			<td>Sakura Finetek</td>
			<td style="width:132px;">1004200018</td>
			<td>embedding of the brains</td>
		</tr>
		<tr height="35">
			<td height="35" style="height:35px;width:447px;">Normal Goat Serum</td>
			<td style="width:127px;">Jackson Immunolabs</td>
			<td style="width:132px;">005-000-001</td>
			<td>blocking in IHC</td>
		</tr>
		<tr height="35">
			<td height="35" style="height:35px;width:447px;">Normal Rabbit Serum</td>
			<td style="width:127px;">Jackson Immunolabs</td>
			<td style="width:132px;">011-000-001</td>
			<td>blocking in IHC</td>
		</tr>
		<tr height="35">
			<td height="35" style="height:35px;">Normal Donkey Serum</td>
			<td style="width:127px;">Jackson Immunolabs</td>
			<td style="width:132px;">017-000-001</td>
			<td>blocking in IHC</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">anti-Neurofilament-M antibody</td>
			<td>Developmental Studies Hybridoma Bank</td>
			<td>2H3</td>
			<td>IHC, Dilution 1:200</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">anti-doublecortin&#160;antibody</td>
			<td>sc-8066</td>
			<td>Santa Cruz</td>
			<td>IHC, Dilution 1:800</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:447px;">anti-GFP antibody</td>
			<td>ab290</td>
			<td>abcam</td>
			<td>IHC, Dilution 1:2000</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:447px;">anti-BrdU antibody</td>
			<td>OBT0030G</td>
			<td>Accurate Chemicals</td>
			<td>IHC, Dilution 1:2000&#160;</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:447px;">Fluoro Jade-C</td>
			<td>FJ-C</td>
			<td>HistoChem</td>
			<td>Determination of neuronal cell death</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:447px;">Betadine</td>
			<td style="width:127px;">&#160;MUNDIPHARMA</td>
			<td>D08AG02&#160;</td>
			<td>disinfectant</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">cryomicrotome</td>
			<td>Leica</td>
			<td>CM1900</td>
			<td>preparation of brain slices</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:447px;">Heparin sodium salt</td>
			<td style="width:127px;">Sigma-Aldrich</td>
			<td style="width:132px;">H3393</td>
			<td>perfusion</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">circular plate made from hard-plastic (diameter 120 cm)</td>
			<td style="width:127px;">lab made</td>
			<td style="width:132px;">none</td>
			<td>plate for SPS-BM, diameter 120cm</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Buraton rapid disinfectant&#160;</td>
			<td style="width:127px;">Sch&#252;lke &#38; Mayr</td>
			<td style="width:132px;">113 911</td>
			<td>disinfectant</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">video-tracking system TSE VideoMot 2 with Software Package VideoMot2</td>
			<td>tse systems</td>
			<td style="width:132px;">302050-SW-KIT</td>
			<td>tracking and analysis of SPS-BM</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Triton X-100&#160;</td>
			<td>Sigma Aldrich</td>
			<td>T8787</td>
			<td>permeabilisation/IHC</td>
		</tr>
		<tr height="35">
			<td height="35" style="height:35px;width:447px;">cryotome</td>
			<td style="width:127px;">Reichert&#160; Jung/Leica</td>
			<td style="width:132px;">Frigomobil 1206</td>
			<td>preparation of 40&#181;m brain slices</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:447px;">Mowiol 4-88</td>
			<td style="width:127px;">Carl Roth&#160;</td>
			<td>Art.-Nr. 0713</td>
			<td>embedding of the slides</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:447px;">SYTOX green</td>
			<td style="width:127px;">Invitrogen</td>
			<td>S7020</td>
			<td>Nuclear staining</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:447px;">Food pellets (Kellog`s Froot Loops)</td>
			<td>Kellog`s</td>
			<td style="width:132px;"></td>
			<td>SPS-BM</td>
		</tr>
		<tr height="52">
			<td height="52" style="height:52px;width:447px;">Prism, Version 3.0</td>
			<td style="width:127px;">Graph Pad Software, San Diego, USA&#160;</td>
			<td style="width:132px;"></td>
			<td>Statistical evaluation of SPS-BM</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:447px;">Zen 2008 or Zen 2011 Software</td>
			<td style="width:127px;">Carl Zeiss</td>
			<td style="width:132px;"></td>
			<td>Software (Confocal microscope)</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:447px;">D.P.X</td>
			<td style="width:127px;">Sigma-Aldrich</td>
			<td style="width:132px;">317616</td>
			<td>mounting medium for Fluoro Jade C staining</td>
		</tr>
	</tbody>
</table>

methods for the modulation and analysis of nf-&#954;b-dependent adult neurogenesis

海马起着举足轻重的作用，在情节记忆的形成和巩固，并在空间定位。从历史上看，成年海马一直被看作是哺乳动物大脑的一个非常静态的解剖区域。然而，最近的研究结果已经表明，在海马的齿状回是巨大的可塑性在成人的区域，不仅包括修改现有的神经元回路的，但也成年神经发生。此可塑性被复杂的转录调控网络，其中，所述转录因子NF-κB中起着突出的作用。研究和操纵成年神经发生，转基因小鼠模型的NF-κB活性的前脑特异性神经元的抑制可以使用。 在这项研究中，方法是用于对NF-κB依赖的神经发生的分析，包括其结构方面，神经元凋亡和祖细胞的增殖和认知意义，whic描述h的通过齿状回（DG）相关的行为测试，空间格局分离 - 巴恩斯迷宫（SPS-BM）的特别评估。在SPS-BM协议可以简单地适用于与设计，以评估特定基因对成年海马神经发生的影响等转基因动物模型的使用。此外，SPS-BM可以在其他实验设置旨在调查和处理DG-依赖的学习，例如，使用药剂使用。

杂交育种的IκB/ -以及tTA的转基因小鼠系导致NF-κB活性的海马条件抑制。 调查IκBα-AA1-转基因的双转基因小鼠（ 图1A）的表达，大脑分离，cryosectioned和使用抗体抗GFP（绿色荧光蛋白）染色。共聚焦激光扫描显微镜观察发现高表达在CA1和CA3区的转基因，并在危险品（ 图1B）。 该IκBα-AA1-转基因表达类型-2B祖细胞?...

Watch this Scientific Journal Video about Methods for the Modulation and Analysis of NF-ÃŽÂºB-dependent Adult Neurogenesis at JoVE.com

Methods for the Modulation and Analysis of NF-ÃƒÅ½Ã‚ÂºB-dependent Adult Neurogenesis

方法的NF-κB依赖型成年海马神经发生的操作和分析的描述。详细的协议提出了一个齿状回依赖性行为测试（称为空间格局分离巴恩斯迷宫）的认知结果小鼠的调查。这项技术也将有助于使在其他实验设置的调查。

方法对NF-κB依赖的成年神经发生的调制与分析

The hippocampus plays a pivotal role in the formation and consolidation of episodic memories, and in spatial orientation. Historically, the adult ...

Research

JoVE Journal

Neuroscience

Methods for the Modulation and Analysis of NF-&#954;B-dependent Adult Neurogenesis

Cerebrovascular Casting of the Adult Mouse for 3D Imaging and Morphological Analysis

成年鼠脑铸造三维成像和形态分析

Vascular imaging is crucial in the clinical diagnosis and management of cerebrovascular diseases, such as brain arteriovenous malformations (BAVMs). ...

科研

神经科学

Functional Interrogation of Adult Hypothalamic Neurogenesis with Focal Radiological Inhibition

成人下丘脑神经发生局灶性放射性抑制功能审讯

The functional characterization of adult-born neurons remains a significant challenge. Approaches to inhibit adult neurogenesis via invasive viral ...

Heat-Induced Antigen Retrieval: An Effective Method to Detect and Identify Progenitor Cell Types during Adult Hippocampal Neurogenesis

Traditional  methods of immunohistochemistry (IHC) following tissue fixation allow visualization  of various cell types. These typically proceed with the ...

Stab Wound Injury of the Zebrafish Adult Telencephalon: A Method to Investigate Vertebrate Brain Neurogenesis and Regeneration

斑马鱼成人端脑的一刀，受伤：一个方法调查脊椎动物脑部神经发​​生与再生

Adult zebrafish have an amazing capacity to regenerate their central nervous system after injury. To investigate the cellular response and the molecular ...

Immunohistochemistry and Multiple Labeling with Antibodies from the Same Host Species to Study Adult Hippocampal Neurogenesis

免疫组化和多标签与同一宿主物种抗体研究成人神经发生

Adult neurogenesis is a highly regulated, multi-stage process in which new neurons are generated from an activated neural stem cell via increasingly ...

Assessing Primary Neurogenesis in Xenopus Embryos Using Immunostaining

Assessing Primary Neurogenesis in Xenopus Embryos Using Immunostaining

评估主要神经再生<em&gt;爪</em&gt;胚胎使用免疫染色

Primary neurogenesis is a dynamic and complex process during embryonic development that sets up the initial layout of the central nervous system. During ...

Analyzing Synaptic Modulation of Drosophila melanogaster Photoreceptors after Exposure to Prolonged Light

Analyzing Synaptic Modulation of Drosophila melanogaster Photoreceptors after Exposure to Prolonged Light

分析突触调制<em&gt;果蝇</em&gt;曝光时间延长后，光光感受器

The nervous system has the remarkable ability to adapt and respond to various stimuli. This neural adjustment is largely achieved through plasticity at ...

The Mouse Hindbrain As a Model for Studying Embryonic Neurogenesis

小鼠后脑作为研究胚胎神经再生的模型

The mouse embryo forebrain is the most commonly employed system for studying mammalian neurogenesis during development. However, the highly folded ...

Metabolic Analysis of Drosophila melanogaster Larval and Adult Brains

Metabolic Analysis of Drosophila melanogaster Larval and Adult Brains

果蝇幼虫和成年脑的代谢分析

This protocol describes a method for measuring the metabolism in Drosophila melanogaster larval and adult brains. Quantifying metabolism in whole organs ...

Isolation of Adult Spinal Cord Nuclei for Massively Parallel Single-nucleus RNA Sequencing

Probing an individual cell&#39;s gene expression enables the identification of cell type and cell state. Single-cell RNA sequencing has emerged as a ...