Name: a lateralized odor learning model in neonatal rats for dissecting neural circuitry underpinning memory formation
Uploaded: 2014-08-18T16:00:00.000+00:00
Duration: 10 min 42 s
Description: Watch this Scientific Journal Video about A Lateralized Odor Learning Model in Neonatal Rats for Dissecting Neural Circuitry Underpinning Memory Formation at JoVE.com

This work was supported by a CIHR operating grant (MOP-102624) to Q. Y. We thank Dr Carolyn Harley for helpful discussions throughout the study, Dr. Qinlong Hou, Amin Shakhawat, and Andrea Darby-King for technical support.

无论男女，只SD大鼠（查尔斯河）的幼崽被使用。窝被扑杀，以12 PD1（出生是PD0）。水坝维持12小时的光/暗周期， 随意获取食物和水。实验步骤已通过纪念大学实验动物管理委员会。 1，鼻封堵施工注:此程序改编和卡明斯等人修改（1997）35。 <ol><li> AQUIRE聚乙烯20管和3-0丝线缝合线。 </li><li>剪一小块聚乙烯管材20到约0.8mm。 </li><li>螺纹丝缝线穿过准备管，使得有螺纹的管的部分的任一侧。 </li><li>在插头外螺纹的一端，打结的线。 </li><li>管拉下来了，在螺纹结的部分。结应提出管内。</li><li>修剪的螺纹，使得〜2毫米螺纹的从管的一端突出的两端（ 见图1A）。 </li></ol>训练前2娜丽丝闭塞<ol><li>在一个安全的菜铺上被褥定期删除大坝和地点的小狗。 </li><li>用棉签应用轻拍局部麻醉果冻，2％利多卡因，在鼻孔被遮挡。 </li><li>让小狗休息的菜〜3分钟。 </li><li>轻轻但稳固地握住小狗在非优势手。 </li><li>使用优势手，拿起一个鼻子插头并轻轻从该线程不突出的尖端周围的相同的局部麻醉果冻。这将作为这两种麻醉与插头插入相关的任何轻​​微疼痛，并会担任鼻孔内的润滑剂。注:利多卡因的效果开始不到两分钟，持续20-30分钟。幼仔一般表现出良好的耐受性的插头插入后利多卡因凝胶的应用（M挣扎的最小值和发声）。 </li><li>轻轻地紧紧抱着小狗，慢慢地旋转插头很温柔推，直到插头完全插入，只有2毫米螺纹从鼻孔插入伸出鼻子塞（ 见图1B）。应该有从任一鼻孔在此过程中没有出血。鼻子插头插入时有出血幼崽被排除在外，回到水坝</li><li>让动物休息的这盘5分钟才能到，观察者插头。 </li><li>取下习惯菜的小狗，并开始调节模式24。 </li></ol><img alt="图1" fo:content-width="5in" src="/files/ftp_upload/51808/51808fig1highres.jpg" width="500" /> 图1。施工鼻子塞。 A）的示意图显示使鼻子塞的步骤。线程是通过聚乙烯管拉;一个结是由和PUL领进油管，以阻止它的中间;两端切有2mm残留在一端从管接头。 的B）前，用在一个鼻孔鼻插头大鼠的侧视图。 3，香味床上准备<ol><li>戴新手套，在通风橱，以防止异味污染，将500毫升木屑铺垫成一个塑料袋。 </li><li>用注射器吸取了0.3毫升薄荷提取物，并喷上这种在塑料袋中的床上用品。 </li><li>扎紧袋口封住，摇晃剧烈袋，并允许被褥休息收入囊中5分钟。 </li><li>将香味寝具清晰，浅，丙烯酸培训发现在通风橱中使用的前5分钟盒（20×20×5厘米3， 图2A）。一旦被褥准备，放弃这些手套，不要让这些手套来与动物接触。 </li><li>将无迹寝具相同的透明塑料盒，以及连接确保不接触到香味的被褥或使用的手套接触。 </li></ol> 4，气味调节范式（参见图图2A） 幼崽接受任何一个单一的空调会议上，PD 6或多个审判庭（每天一次会议，PD 3-6）。 <ol><li>广场上香气四溢的床上用品习惯于小狗。对于控制气味只有（O / S - ）的幼崽，离开被褥10分钟这些幼崽，然后跳到步骤4.5。对于实验异味+行程（O / S +）的幼崽，继续在本节下面的步骤。 </li><li>行程小狗30秒使用小画笔。使用快速打圈主要围绕着小狗的后腿区域。 </li><li>让小狗休息30秒。 </li><li>重复步骤4.2和4.3共10分钟（ 即 10配对行程+异味）。 </li><li>从调理盒中取出的小狗，取出鼻子塞和小狗回到大坝。 </li></ol> &lt;P舱="jove_title"&gt; 5。单侧气味偏好测试（参见图图2B） 测试发生在不同时间点（ 例如 ，24或48小时）之后的最后的训练会话。试验是在一个不锈钢测试腔室（30×20×18 毫升 ），它被放置在两个训练拖曳（训练框在3.4中描述），以2厘米的中性区域分开顶部。一个培训箱含有薄荷香味的床上用品，而另一盒包含清洁，无味的床上用品。测试腔室的底板是金属网格，然后将其覆盖的塑料网状物的可移动片（ 图2B）。 <ol><li>制备1薄荷油和一种无味寝具按照第3章，并将其放置在测试室中，相距2厘米的相对侧上的每个方块。放在试验室的金属网格地板的塑料网。 </li><li>从大坝拆除的小狗和地点无味硅脂坚定民建联的鼻孔在训练中被遮挡。重新申请需要整个第一测试过程中润滑脂。注意:在训练和测试随机鼻孔闭塞，可以考虑，以避免偏差。 </li><li>放置小狗在测试室的中性区。 </li><li>让小狗探索室1分钟，记录多久的小狗花了室的两侧（ 即超过薄荷或中性香味的床上用品）。 </li><li>让小狗在一个带盖的塑料保持室休息1分钟。 </li><li>共10分钟，重复步骤5.2和5.3（ 即由5其余试验分离5试验试验）交换腔室中的幼犬的初始方向，以控制对方向的偏好。 </li><li>紧随其后的测试，擦去从鼻孔油脂。 </li><li>将聚乙烯noseplug到相反鼻孔按照第2节，让动物休息10分钟。 </li><li>再次测试的小狗在<html> 5.3 -5.6，拔下电源插头，并小狗回到大坝。除去和清洗试验室用95％乙醇的塑料网，并允许液体蒸发。测试下一小狗之前，将网格回来。 注意:在测试过程中的应用硅脂在第一鼻孔闭塞防止出血和鼻插头插入有关的应力的机会。 </li></ol><img alt="图2" fo:content-width="5in" src="/files/ftp_upload/51808/51808fig2highres.jpg" width="500" /> 图2:早期气味偏好的培训和测试。一）早期气味偏好使用气味+抚摸着范式乙训练。）的一个侧面薄荷寝具两个选择气味偏好测试，控制无味的床上用品在另一侧。 2厘米中立区被放置在中间。 6，测试单娜丽丝闭塞的有效性NT"&gt;本实验的目的是确定单个鼻孔闭塞是否导致嗅觉系统的单侧激活。 <ol><li>在第2节中描述到PD 6或7的幼崽进行单侧鼻孔闭塞。 </li><li>后〜5分钟习惯，将幼鼠在有盖的塑料容器中，并暴露于30微升纯薄荷油浸泡在一块组织10分钟。 </li><li>紧接着的薄荷气味曝光，腹腔（IP）与水合氯醛（400毫克/千克）为全身麻醉，或戊巴比妥（150毫克/千克）注入小狗。 </li><li>一旦完全麻醉（示于尾部或脚捏无响应），transcardially灌注小狗用生理盐水（0.9％）为〜1分钟的冰冷溶液，随后的多聚甲醛（4％，溶于0.1M磷酸盐缓冲溶液，PBS ）。 </li><li>后的多聚甲醛灌注的10分钟，收集大脑，并放置在多聚甲醛中过夜，在4℃，然后反FER大脑到蔗糖溶液（在PBS中的20％），另外24小时。 </li><li>冠状切开大脑切片在30微米厚度与低温恒温器。收集转播和PC片和安装到明胶包被的载玻片，其次是标准的免疫组化染色区pCREB抗体21,25,30。 </li></ol> 7，测试单娜丽丝闭塞的可逆性该实验测试的阻挡效果是否是可逆的，在24小时之后除去鼻子塞。 <ol><li>在第2节中描述到PD 6或7的幼崽进行单侧鼻孔闭塞。 </li><li> 15分钟后（相当于鼻孔闭塞的持续时间期间训练 - 5分钟驯化+ 10分钟训练），除去鼻插头和小狗返回到坝。 </li><li> 24小时后，在6.2中描述揭露小狗的薄荷气味在有盖的塑料容器10分钟。 </li><li>按照教派相同的步骤离子6.3-6.6。 </li></ol>

<ol>
	<li>Gregory, E. H., Pfaff, D. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Development+of+olfactory-guided+behavior+in+infant+rats.">Development of olfactory-guided behavior in infant rats.</a> Physiol Behav. 6, 573-576 (1971).</li><li>Alberts, J. R., May, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Nonnutritive,+thermotactile+induction+of+filial+huddling+in+rat+pups.">Nonnutritive, thermotactile induction of filial huddling in rat pups.</a> Dev Psychobiol. 17, 161-181 (1984).</li><li>Galef, B. G., Kaner, H. C. <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+maintenance+of+preference+for+natural+and+artificial+olfactory+stimuli+in+juvenile+rats.">Establishment and maintenance of preference for natural and artificial olfactory stimuli in juvenile rats.</a> J Comp Physiol Psychol. 94, 588-595 (1980).</li><li>Johanson, I. B., Hall, W. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Appetitive+learning+in+1-day-old+rat+pups.">Appetitive learning in 1-day-old rat pups.</a> Science. 205, 419-421 (1979).</li><li>Johanson, I. B., Hall, W. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Appetitive+conditioning+in+neonatal+rats:+conditioned+orientation+to+a+novel+odor.">Appetitive conditioning in neonatal rats: conditioned orientation to a novel odor.</a> Dev Psychobiol. 15, 379-397 (1982).</li><li>Johanson, I. B., Teicher, M. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Classical+conditioning+of+an+odor+preference+in+3-day-old+rats.">Classical conditioning of an odor preference in 3-day-old rats.</a> Behav Neural Biol. 29, 132-136 (1980).</li><li>McLean, J. H., Darby-King, A., Sullivan, R. M., King, S. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Serotonergic+influence+on+olfactory+learning+in+the+neonate+rat.">Serotonergic influence on olfactory learning in the neonate rat.</a> Behav Neural Biol. 60, 152-162 (1993).</li><li>Moore, C. L., Power, K. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Variation+in+maternal+care+and+individual+differences+in+play,+exploration,+and+grooming+of+juvenile+Norway+rat+offspring.">Variation in maternal care and individual differences in play, exploration, and grooming of juvenile Norway rat offspring.</a> Dev Psychobiol. 25, 165-182 (1992).</li><li>Pedersen, P. E., Williams, C. L., Blass, 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=Activation+and+odor+conditioning+of+suckling+behavior+in+3-day-old+albino+rats.">Activation and odor conditioning of suckling behavior in 3-day-old albino rats.</a> J Exp Psychol Anim Behav Process. 8, 329-341 (1982).</li><li>Sullivan, R. M., Hall, W. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Reinforcers+in+infancy:+classical+conditioning+using+stroking+or+intra-oral+infusions+of+milk+as+UCS.">Reinforcers in infancy: classical conditioning using stroking or intra-oral infusions of milk as UCS.</a> Dev Psychobiol. 21, 215-223 (1988).</li><li>Sullivan, R. M., Leon, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Early+olfactory+learning+induces+an+enhanced+olfactory+bulb+response+in+young+rats.">Early olfactory learning induces an enhanced olfactory bulb response in young rats.</a> Brain Res. 392, 278-282 (1986).</li><li>Weldon, D. A., Travis, M. L., Kennedy, D. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Posttraining+D1+receptor+blockade+impairs+odor+conditioning+in+neonatal+rats.">Posttraining D1 receptor blockade impairs odor conditioning in neonatal rats.</a> Behav Neurosci. 105, 450-458 (1991).</li><li>Sullivan, R. M., Hofer, M. A., Brake, S. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Olfactory-guided+orientation+in+neonatal+rats+is+enhanced+by+a+conditioned+change+in+behavioral+state.">Olfactory-guided orientation in neonatal rats is enhanced by a conditioned change in behavioral state.</a> Dev Psychobiol. 19, 615-623 (1986).</li><li>Camp, L. L., Rudy, J. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Changes+in+the+categorization+of+appetitive+and+aversive+events+during+postnatal+development+of+the+rat.">Changes in the categorization of appetitive and aversive events during postnatal development of the rat.</a> Dev Psychobiol. 21, 25-42 (1988).</li><li>Moriceau, S., Wilson, D. A., Levine, S., Sullivan, 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=Dual+circuitry+for+odor-shock+conditioning+during+infancy:+corticosterone+switches+between+fear+and+attraction+via+amygdala.">Dual circuitry for odor-shock conditioning during infancy: corticosterone switches between fear and attraction via amygdala.</a> J Neurosci. 26, 6737-6748 (2006).</li><li>Roth, T. L., Sullivan, 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=Endogenous+opioids+and+their+role+in+odor+preference+acquisition+and+consolidation+following+odor-shock+conditioning+in+infant+rats.">Endogenous opioids and their role in odor preference acquisition and consolidation following odor-shock conditioning in infant rats.</a> Dev Psychobiol. 39, 188-198 (2001).</li><li>Roth, T. L., Sullivan, 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=Consolidation+and+expression+of+a+shock-induced+odor+preference+in+rat+pups+is+facilitated+by+opioids.">Consolidation and expression of a shock-induced odor preference in rat pups is facilitated by opioids.</a> Physiol Behav. 78, 135-142 (2003).</li><li>Sullivan, 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=Developing+a+sense+of+safety:+the+neurobiology+of+neonatal+attachment.">Developing a sense of safety: the neurobiology of neonatal attachment.</a> Ann N Y Acad Sci. 1008, 122-131 (2003).</li><li>Wilson, D. A., Sullivan, 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=Olfactory+associative+conditioning+in+infant+rats+with+brain+stimulation+as+reward.+I.+Neurobehavioral+consequences.">Olfactory associative conditioning in infant rats with brain stimulation as reward. I. Neurobehavioral consequences.</a> Brain Res Dev Brain Res. 53, 215-221 (1990).</li><li>Sullivan, R. M., Wilson, D. A., Leon, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Associative+Processes+in+Early+Olfactory+Preference+Acquisition:+Neural+and+Behavioral+Consequences.">Associative Processes in Early Olfactory Preference Acquisition: Neural and Behavioral Consequences.</a> Psychobiology. , 29-33 (1989).</li><li>McLean, J. H., Harley, C. W., Darby-King, A., Yuan, Q. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=pCREB+in+the+neonate+rat+olfactory+bulb+is+selectively+and+transiently+increased+by+odor+preference-conditioned+training.">pCREB in the neonate rat olfactory bulb is selectively and transiently increased by odor preference-conditioned training.</a> Learn Mem. 6, 608-618 (1999).</li><li>Sullivan, R. M., Stackenwalt, G., Nasr, F., Lemon, C., Wilson, D. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Association+of+an+odor+with+activation+of+olfactory+bulb+noradrenergic+beta-receptors+or+locus+coeruleus+stimulation+is+sufficient+to+produce+learned+approach+responses+to+that+odor+in+neonatal+rats.">Association of an odor with activation of olfactory bulb noradrenergic beta-receptors or locus coeruleus stimulation is sufficient to produce learned approach responses to that odor in neonatal rats.</a> Behav Neurosci. 114, 957-962 (2000).</li><li>Yuan, Q., Harley, C. W., McLean, J. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mitral+cell+beta1+and+5-HT2A+receptor+colocalization+and+cAMP+coregulation:+a+new+model+of+norepinephrine-induced+learning+in+the+olfactory+bulb.">Mitral cell beta1 and 5-HT2A receptor colocalization and cAMP coregulation: a new model of norepinephrine-induced learning in the olfactory bulb.</a> Learn Mem. 10, 5-15 (2003).</li><li>Fontaine, C. J., Harley, C. W., Yuan, Q. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Lateralized+odor+preference+training+in+rat+pups+reveals+an+enhanced+network+response+in+anterior+piriform+cortex+to+olfactory+input+that+parallels+extended+memory.">Lateralized odor preference training in rat pups reveals an enhanced network response in anterior piriform cortex to olfactory input that parallels extended memory.</a> J Neurosci. 33, 15126-15131 (2013).</li><li>Morrison, G. L., Fontaine, C. J., Harley, C. W., Yuan, Q. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+role+for+the+anterior+piriform+cortex+in+early+odor+preference+learning:+evidence+for+multiple+olfactory+learning+structures+in+the+rat+pup.">A role for the anterior piriform cortex in early odor preference learning: evidence for multiple olfactory learning structures in the rat pup.</a> J Neurophysiol. 110, 141-152 (2013).</li><li>Nakamura, S., Kimura, F., Sakaguchi, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Postnatal+development+of+electrical+activity+in+the+locus+ceruleus.">Postnatal development of electrical activity in the locus ceruleus.</a> J Neurophysiol. 58, 510-524 (1987).</li><li>Harley, C. W., Darby-King, A., McCann, J., McLean, J. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Beta1-adrenoceptor+or+alpha1-adrenoceptor+activation+initiates+early+odor+preference+learning+in+rat+pups:+support+for+the+mitral+cell/cAMP+model+of+odor+preference+learning.">Beta1-adrenoceptor or alpha1-adrenoceptor activation initiates early odor preference learning in rat pups: support for the mitral cell/cAMP model of odor preference learning.</a> Learn Mem. 13, 8-13 (2006).</li><li>Shakhawat, A. M., Harley, C. W., Yuan, Q. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Olfactory+bulb+alpha2-adrenoceptor+activation+promotes+rat+pup+odor-preference+learning+via+a+cAMP-independent+mechanism.">Olfactory bulb alpha2-adrenoceptor activation promotes rat pup odor-preference learning via a cAMP-independent mechanism.</a> Learn Mem. 19, 499-502 (2012).</li><li>Isaacson, 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=Odor+representations+in+mammalian+cortical+circuits.">Odor representations in mammalian cortical circuits.</a> Curr Opin Neurobiol. 20, 328-331 (2010).</li><li>Lethbridge, R., Hou, Q., Harley, C. W., Yuan, Q. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Olfactory+bulb+glomerular+NMDA+receptors+mediate+olfactory+nerve+potentiation+and+odor+preference+learning+in+the+neonate+rat.">Olfactory bulb glomerular NMDA receptors mediate olfactory nerve potentiation and odor preference learning in the neonate rat.</a> PLoS One. 7, e35024 (2012).</li><li>Yuan, Q., Harley, C. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=What+a+nostril+knows:+olfactory+nerve-evoked+AMPA+responses+increase+while+NMDA+responses+decrease+at+24-h+post-training+for+lateralized+odor+preference+memory+in+neonate+rat.">What a nostril knows: olfactory nerve-evoked AMPA responses increase while NMDA responses decrease at 24-h post-training for lateralized odor preference memory in neonate rat.</a> Learn Mem. 19, 50-53 (2012).</li><li>Schwob, J. E., Price, J. 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+development+of+axonal+connections+in+the+central+olfactory+system+of+rats.">The development of axonal connections in the central olfactory system of rats.</a> J Comp Neurol. 223, 177-202 (1984).</li><li>Kucharski, D., Hall, W. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=New+routes+to+early+memories.">New routes to early memories.</a> Science. 238, 786-788 (1987).</li><li>Kucharski, D., Johanson, I. B., Hall, W. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Unilateral+olfactory+conditioning+in+6-day-old+rat+pups.">Unilateral olfactory conditioning in 6-day-old rat pups.</a> Behav Neural Biol. 46, 472-490 (1986).</li><li>Cummings, D. M., Henning, H. E., Brunjes, P. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Olfactory+bulb+recovery+after+early+sensory+deprivation.">Olfactory bulb recovery after early sensory deprivation.</a> J Neurosci. 17, 7433-7440 (1997).</li><li>Kucharski, D., Hall, W. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Developmental+change+in+the+access+to+olfactory+memories.">Developmental change in the access to olfactory memories.</a> Behav Neurosci. 102, 340-348 (1988).</li><li>Brunjes, P. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Unilateral+odor+deprivation:+time+course+of+changes+in+laminar+volume.">Unilateral odor deprivation: time course of changes in laminar volume.</a> Brain Res Bull. 14, 233-237 (1985).</li><li>Kass, M. D., Pottackal, J., Turkel, D. J., McGann, J. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Changes+in+the+neural+representation+of+odorants+after+olfactory+deprivation+in+the+adult+mouse+olfactory+bulb.">Changes in the neural representation of odorants after olfactory deprivation in the adult mouse olfactory bulb.</a> Chem Senses. 38, 77-89 (2013).</li><li>Kim, H. H., Puche, A. C., Margolis, F. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Odorant+deprivation+reversibly+modulates+transsynaptic+changes+in+the+NR2B-mediated+CREB+pathway+in+mouse+piriform+cortex.">Odorant deprivation reversibly modulates transsynaptic changes in the NR2B-mediated CREB pathway in mouse piriform cortex.</a> J Neurosci. 26, 9548-9559 (2006).</li><li>Korol, D. L., Brunjes, P. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Rapid+changes+in+2-deoxyglucose+uptake+and+amino+acid+incorporation+following+unilateral+odor+deprivation:+a+laminar+analysis.">Rapid changes in 2-deoxyglucose uptake and amino acid incorporation following unilateral odor deprivation: a laminar analysis.</a> Brain Res Dev Brain Res. 52, 75-84 (1990).</li><li>Leung, C. H., Wilson, D. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Trans-neuronal+regulation+of+cortical+apoptosis+in+the+adult+rat+olfactory+system.">Trans-neuronal regulation of cortical apoptosis in the adult rat olfactory system.</a> Brain Res. 984, 182-188 (2003).</li></ol>

The authors have nothing to disclose.

首先由霍尔及其同事建立了幼鼠的单侧臭味学习和记忆模型中的一个关键的时间窗口。在一系列的研究中33,34,36，他们发现气味偏好的内存可以通过气味+豆浆配对的幼鼠进行单侧一个鼻孔在帕金森病6。偏好的记忆是强大的，当同样的鼻孔是在训练和测试开放的，但没有观察到，当闭塞鼻孔是畅通和测试。然而，在帕金森病12，当来自前嗅皮层前连合的连接成为功能32，未受过训练的鼻孔仅可以支持在帕金森病取得的气味偏好的6病变前联合的恢复单侧记忆的表达，因此不再访问由未经训练的鼻孔33。这种单侧的学习模式已成功复制了一种气味+抚摸范式24,25,31。气味+抚摸训练单鼻孔OCClusion到PD 6导致单侧内存PD 7 月31日 。对帕金森病3-6重复的单鼻孔闭塞导致较长的单侧记忆，持续至少48小时24。 在单侧臭味学习协议导致动物个体"裂脑"。这种模式有学习行为和支柱的生物生理变化很大的优势。例如，行为比较输出使用的是个体内的模型充分降低了变异动物之间的设计。幼仔在这么早的年龄有很大的不同在他们的活动和反应。帧内动物控制除去在性能和响应的固有的个体差异以及从分子和生理变化的评估在生物学特性差异。此外，单侧学习这个强大的婴儿模式借给我们与个人记忆的表现个人的生理和评估第一次机会不同持续时间24,25,31的回忆ê基础。用与体外比较在同一动物中的两个半球的生理学实验组合此单侧学习模型，它最近已表明，早期气味学习诱导突触可塑性，如在两个OB 31突触增加AMPA受体应答，并APC的24,25。增强的突触传递以下一些早期的气味学习转化为嗅网的增强输出申述24。 未来的研究应探讨气味记忆通过这种单侧学习模型的分子基础。这包括相关性的研究着眼于蛋白质和基因活化下列学习，和因果关系的研究看的和丢失的功能的蛋白质和特别感兴趣的基因的功能获得的效果。另一个令人兴奋的和重要的可能性是能够吨Ø相关的生理和分子变化的行为记忆的力量。对于每个小狗，它可以首先导出为开放和封闭鼻孔偏好存储器量度。随后的体外实验对相应的培训和未经培训的皮质会提供相关的生理变化。这是可能的，但是，偏好期间的气味再次接触测试本身会改变突触强度，或其他脑区显著记忆的表达促进。在我们目前的研究，生理和行为都进行了单独的同伙。这消除了有关行为测试的担忧影响的利益非常的参数。 用鼻子插一个需要注意的是带插头的插入和删除相关的潜在的神经组织损伤。为此，应注意已采取在插入鼻孔插头和幼崽与插入过程中出血应被排除，以避免任何可能的较长期的EF出血如炎症相关fects。长时间的阻塞（小时，天，月）鼻孔或嗅上皮永久消融导致长期嗅觉剥夺，神经损伤，并在OB降低神经元活动和PC 37-41，即使其中的一些影响由于长时间鼻孔闭塞是完全可逆的39,40。组织完整性急性鼻孔闭塞（〜15分钟）已经由区pCREB的免疫组织化学染色证实，一个活性依赖性神经元标记物，这已经显示出下列5天鼻孔闭塞39可以减少在年轻成年小鼠的个人电脑。区pCREB水平的鼻孔闭塞的同侧转播和PC均显著少在薄荷气味曝光，鼻孔闭塞时，确认在幼鼠嗅觉吞吐量的成功偏侧。然而，24小时以下的除去鼻子塞，在气味偏好测试的时候，区pCREB水平可比ıÑ ​​两个半球，提示急性闭塞的效果是完全可逆的24。因此，缺少优先薄荷与前面闭塞鼻孔测试的是，由于缺乏记忆，但不是由于改动或缺乏与在测试过程中的组织损伤相关的气味的感知。此外，控制电生理记录的O / S -动物（与一个鼻孔气味曝光期间遮挡而不抚摸）显示在fEPSPs没有区别用钙成像激活锥体细胞或数-也确认存在于梨状皮质功能上没有变化由于这些短期可逆的鼻孔闭塞24,25。

嗅觉是初级感觉方式在啮齿类动物中，没有它们就不能成功地浏览或在其环境中生存。这是新生儿的幼崽，它可以既看不见也听不到在第一产后一周，用嗅觉，以找到他们的母亲喂养1尤其重要。因此，新生儿幼鼠能条件反射地偏爱气味与简单的实验操作。各种刺激已被用来作为非条件刺激（UCS）以诱导调节的反应，新颖的气味（条件刺激，CS）的新生儿，包括嵌套环境2,3，乳乳4-6，抚摸或触觉刺激7- 12，尾巴捏13，产妇的唾液13，轻度电击足底14-18，和颅内脑刺激19。本研究采用了一种行之有效的早期气味偏好的模式，其中的气味，在这种情况下薄荷，我š以产生偏爱薄荷24小时后10,11,20结合触觉刺激。这些气味的记忆是依赖于完整的嗅觉电路，主要包括嗅球（OB）21-23和前梨状皮质（APC）24,25。 早期的气味偏好的学习实验研究，加深和拓宽了我们的哺乳动物记忆的分子和生理基础的理解。这种哺乳动物模型具有学习记忆机制的几大优势。首先，UCS信号的神经来源已经确定。如上面提到的各种刺激刺激蓝斑的去甲肾上腺素释放26，从而激活多个肾上腺素能受体中的转播和APC，引起支持学习22,27,28细胞和生理效应。第二，存储器 - 支持机制发生在良好定义的层的神经结构。该新生大鼠嗅觉电路简洁为研究人员提供与发掘相关的突触可塑性的复杂过程的理想框架。嗅觉的感觉神经元（OSN）中的嗅觉上皮项目上二尖瓣/簇绒细胞中的转播和这些二尖瓣/簇绒细胞反过来项目同侧经由横向嗅束（LOT）梨状皮层（PC），其他结构29之间。无论是在OB 30,31的OSN突触和突触地段24,25参阅APC已被确定为关键位点为支持学习记忆的突触变化和。第三，在幼年大鼠，嗅觉输入可以很容易地单侧。每个APC有一次，通过这种白色物质完全形成，在产后第12天（PD12）32前联合访问两国的气味信息。帕金森病12日前，异味输入可隔离通过单鼻孔闭塞24,25,31,33,34 &lt;到ipisilateral转播和APC/ SUP&gt;。单鼻孔闭塞允许从开启鼻孔的气味记忆的形成，并且防止在同一存储器从现有的闭塞鼻孔到PD 12 33。气味存储器分离到同侧半球包括OB和APC。因此，每只大鼠的小狗可以自己控制学习和支撑生理学。 在本研究中，单侧早期气味偏好学习协议被引入。该方法可作为一种强大的工具用于研究通过提供一种帧内动物控制24,25,31，从而减少了所需的动物数量和一般的变异支撑气味学习神经机制。鼻孔闭塞是可逆的，所述润滑脂或鼻子塞可以应用和以最小的应力或者损坏的动物中移除。在这里，首先，早期气味偏好的培训和测试的详细过程进行了描述，重点是使用单鼻孔闭塞了号的单侧协议ê插头。然后结果被提出来证明在分离的气味输入并产生单侧气味存储器单鼻孔闭塞的有效性。最后，利用该单侧学习模型来研究生理变化的嗅觉系统，这两个生成学习和支撑记忆表达的潜力进行了讨论。

<table><tbody><tr><td>Polythylene 20 tubing</td><td>Intramedic</td><td>427406</td><td>Non-radiopaque, Non-toxic</td></tr><tr><td>3-0 Silk suture thread</td><td>Syneture</td><td>Sofsilk</td><td>Non-absorbent</td></tr><tr><td>Silicone grease</td><td>Warner Instrument</td><td>64-0378</td><td>Odorless</td></tr><tr><td>2% Xylocaine gel</td><td>AstraZeneca</td><td>Prod. No 061</td><td>Lidocaine hydrochloride jelly, purchased at local pharmacy</td></tr><tr><td>Paint brush</td><td>Dynasty</td><td>206R</td><td>Similar size/other brands work too</td></tr><tr><td>Peppermint extract</td><td>Sigma-Aldrich</td><td>W284807</td><td>Other brands should be okay too</td></tr><tr><td>Training box</td><td>Custom-made</td><td>N/A</td><td>Acrylic box (20 x 20 x 5 cm&lt;sup&gt;3&lt;/sup&gt;), see &lt;b&gt;Figure 2A&lt;/b&gt;. Parameters and material for the box are not critical and can be modified. Material used should be odorless and does not absorb odors</td></tr><tr><td>Testing chamber</td><td>Custom-made</td><td>N/A</td><td>Stainless steel (30 x 20 x 18 cm&lt;sup&gt;3&lt;/sup&gt;), see &lt;b&gt;Figure 2B&lt;/b&gt;. Parameters and material for the chamber are not critical and can be modified. For example, an acrylic chamber instead of a stainless steel one can be used</td></tr><tr><td>pCREB antibody</td><td>Cell Signaling</td><td>9198</td><td>Ser 133 (87G3) Rabbit mAb</td></tr><tr><td>Chloral hydrate</td><td>Sigma-Aldrich</td><td>C8383</td><td>N/A</td></tr><tr><td>Paraformaldehype</td><td>Sigma-Aldrich</td><td>P6148</td><td>N/A</td></tr><tr><td>Sucrose</td><td>Sigma-Aldrich</td><td>S9378</td><td>N/A</td></tr></tbody></table>

a lateralized odor learning model in neonatal rats for dissecting neural circuitry underpinning memory formation

Rat pups during a critical postnatal period (&#8804; 10 days) readily form a preference for an odor that is associated with stimuli mimicking maternal care. Such a preference memory can last from hours, to days, even life-long, depending on training parameters. Early odor preference learning provides us with a model in which the critical changes for a natural form of learning occur in the olfactory circuitry. An additional feature that makes it a powerful tool for the analysis of memory processes is that early odor preference learning can be lateralized via single naris occlusion within the critical period. This is due to the lack of mature anterior commissural connections of the olfactory hemispheres at this early age. This work outlines behavioral protocols for lateralized odor learning using nose plugs. Acute, reversible naris occlusion minimizes tissue and neuronal damages associated with long-term occlusion and more aggressive methods such as cauterization. The lateralized odor learning model permits within-animal comparison, therefore greatly reducing variance compared to between-animal designs. This method has been used successfully to probe the circuit changes in the olfactory system produced by training. Future directions include exploring molecular underpinnings of odor memory using this lateralized learning model; and correlating physiological change with memory strength and durations.

在这里，我们回顾一些先前建立的结果24，以显示在分离的气味输入和学 ​​习到一个半球的鼻孔闭塞的有效性，并且该方法的可逆性。 在早期的气味偏好训练单鼻孔闭塞导致单侧气味记忆24。该存储器被限制在幸免鼻孔（ 图3）。当幼崽进行了测试气味的喜好与同鼻孔闭塞的训练时，他们表现出偏爱空调异味（ 如薄荷）。当幼崽与封闭相反的鼻孔进行测试，他们没有表现出偏爱的空调异味。总之，这些结果表明，一种气味偏好存储器仅形成并通过幸免鼻孔将经历了气味+抚摸缔调节表达。 <img alt="图3" fo:content-WIDTH ="5英寸"SRC ="/文件/ ftp_upload / 51808 / 51808fig3highres.jpg"WIDTH ="500"/&gt; 图3:单鼻孔闭塞诱导单侧气味学习的行为协议中示出的上面板。气味+抚摸着（O / S +）或气味只有（O / S - ）动物单鼻孔期间封闭训练，首先进行的气味偏好测试使用相同的鼻孔阻塞，然后用相反的鼻孔闭塞。下图显示的时间在两个选择，气味测试花了薄荷香味的床上用品不同群体之间的百分比。 * P &lt;0.05。误差线，平均值±标准差。从方丹等人转载。J.神经科学 （2013年）的许可。 在嗅觉系统的单侧激活单侧气味的训练成果气味曝光（ 图4）24时。单鼻孔闭塞防止激活的OB和同侧半球杜里的电脑ng的气味曝光。这表现在监测CREB的磷酸化的转播，和PC。如示于图4中 ，使用免疫组织化学，磷酸化CREB（区pCREB）是显著更少的包藏的半球下面的薄荷气味的曝光相比，对侧半球幸免。尼氏染色（ 图4A）表明，在OB的僧帽细胞层比较的胞体，并在两个半球的PC的锥体细胞层。然而，区pCREB是显著少在半球同侧包藏鼻孔（ 图4B）这两种细胞层。 <img alt="图4" fo:content-width="5in" src="/files/ftp_upload/51808/51808fig4highres.jpg" width="500" /> 图4单鼻孔闭塞导致嗅觉系统的单侧激活气味曝光时。 A）尼氏嗅球的染色（OB）和前梨状皮质（APC），B）以下的单鼻孔闭塞的小狗薄荷暴露在闭塞和逃得半球区pCREB表达。箭头表示在APC的转播和锥体细胞层僧帽细胞层。比例尺，500μm以下。从方丹等转载。神经科学杂志 （2013年）的许可。 单一试验（15-20分钟）鼻孔闭塞的效果是短暂的，可逆的，并且不会产生可能导致改变的气味感觉和减少神经元激活对气味测试24中可见较长期的神经元损伤。由于收录区pCREB染色的转播和PC（ 图5），在OB的僧帽细胞区pCREB的表达，而锥体细胞在PC异味曝光相媲美的闭塞和逃得半球之间，24小时以下的去除鼻子塞 - 同样的时间点气味偏好测试进行以下的早期气味偏好的培训。 <img alt="图5" fo:content-width="5in" src="/files/ftp_upload/51808/51808fig5highres.jpg" width="500" /> 图5。评估神经元的反应如下可逆的鼻孔闭塞，OB的区pCREB染色和APC，24小时以下的一个小狗去除鼻子塞。箭头表示在APC的转播和锥体细胞层僧帽细胞层。 MCL，僧帽细胞层。 PCL，锥体细胞层。比例尺，500μm以下为低倍率和100微米的较高放大倍数。

Watch this Scientific Journal Video about A Lateralized Odor Learning Model in Neonatal Rats for Dissecting Neural Circuitry Underpinning Memory Formation at JoVE.com

A Lateralized Odor Learning Model in Neonatal Rats for Dissecting Neural Circuitry Underpinning Memory Formation

This protocol introduces lateralized early odor preference learning in rats using acute single naris occlusion. Lateralized learning permits the examination of behavioral outcomes and underpinning biological mechanisms within the same animals, reducing variance induced by between-animal designs. This protocol can be used to investigate molecular mechanisms underpinning early odor learning.

新生大鼠于A气味偏侧学习模型解剖神经回路托换记忆的形成

Rat pups during a critical postnatal period (&#8804; 10 days) readily form a preference for an odor that is associated with stimuli mimicking maternal ...

a-lateralized-odor-learning-model-neonatal-rats-for-dissecting-neural

Nanyang Technological University

Research

JoVE Journal

Neuroscience

8.9K 次观看.  Faculty of Medicine, Memorial University. This protocol introduces lateralized early odor preference learning in rats using acute single naris occlusion. Lateralized learning permits the examination of behavioral outcomes and underpinning biological mechanisms within the same animals, reducing variance induced by between-animal designs. This protocol can be used to investigate molecular mechanisms underpinning early odor learning.

视频: A Lateralized Odor Learning Model in Neonatal Rats for Dissecting Neural Circuitry Underpinning Memory Formation

The Double-H Maze: A Robust Behavioral Test for Learning and Memory in Rodents

Spatial cognition research in rodents typically employs the use of maze tasks, whose attributes vary from one maze to the next. These tasks vary by their behavioral flexibility and required memory duration, the number of goals and pathways, and also the overall task complexity. A confounding feature in many of these tasks is the lack of control over the strategy employed by the rodents to reach the goal, e.g., allocentric (declarative-like) or egocentric (procedural) based strategies. The double-H maze is a novel water-escape memory task that addresses this issue, by allowing the experimenter to direct the type of strategy learned during the training period. The double-H maze is a transparent device, which consists of a central alleyway with three arms protruding on both sides, along with an escape platform submerged at the extremity of one of these arms.
Rats can be trained using an allocentric strategy by alternating the start position in the maze in an unpredictable manner (see protocol 1; &#167;4.7), thus requiring them to learn the location of the platform based on the available allothetic cues. Alternatively, an egocentric learning strategy (protocol 2; &#167;4.8) can be employed by releasing the rats from the same position during each trial, until they learn the procedural pattern required to reach the goal. This task has been proven to allow for the formation of stable memory traces.
Memory can be probed following the training period in a misleading probe trial, in which the starting position for the rats alternates. Following an egocentric learning paradigm, rats typically resort to an allocentric-based strategy, but only when their initial view on the extra-maze cues differs markedly from their original position. This task is ideally suited to explore the effects of drugs/perturbations on allocentric/egocentric memory performance, as well as the interactions between these two memory systems.

The goal of this protocol is to investigate spatial cognition in rodents. The double-H water maze is a novel test, which is particularly useful to elucidate the different components of learning, consolidation and memory, as well as the interplay of memory systems.

双-H迷宫:一个强大的行为测试学习和记忆的鼠害

Spatial cognition research in rodents typically employs the use of maze tasks, whose attributes vary from one maze to the next. These tasks vary by their ...

科研

行为学

A Method for Remotely Silencing Neural Activity in Rodents During Discrete Phases of Learning

This protocol describes how to temporarily and remotely silence neuronal activity in discrete brain regions while animals are engaged in learning and memory tasks. The approach combines pharmacogenetics (Designer-Receptors-Exclusively-Activated-by-Designer-Drugs) with a behavioral paradigm (sensory preconditioning) that is designed to distinguish between different forms of learning. Specifically, viral-mediated delivery is used to express a genetically modified inhibitory G-protein coupled receptor (the Designer Receptor) into a discrete brain region in the rodent. Three weeks later, when designer receptor expression levels are high, a pharmacological agent (the Designer Drug) is administered systemically 30 min prior to a specific behavioral session. The drug has affinity for the designer receptor and thus results in inhibition of neurons that express the designer receptor, but is otherwise biologically inert. The brain region remains silenced for 2-5 hr (depending on the dose and route of administration). Upon completion of the behavioral paradigm, brain tissue is assessed for correct placement and receptor expression. This approach is particularly useful for determining the contribution of individual brain regions to specific components of behavior and can be used across any number of behavioral paradigms.

This protocol describes how to temporarily and remotely silence neuronal activity in discrete brain regions while rats are engaged in learning and memory tasks. The approach combines pharmacogenetics (Designer-Receptors-Exclusively-Activated-by-Designer-Drugs) with a behavioral paradigm (sensory preconditioning) that is designed to distinguish between different components of learning.

为远程沉默的神经活动在鼠害的方法在学习独立的阶段

This protocol describes how to temporarily and remotely silence neuronal activity in discrete brain regions while animals are engaged in learning and ...

Appetitive Associative Olfactory Learning in Drosophila Larvae

In the following we describe the methodological details of appetitive associative olfactory learning in Drosophila larvae. The setup, in combination with genetic interference, provides a handle to analyze the neuronal and molecular fundamentals of specifically associative learning in a simple larval brain.
Organisms can use past experience to adjust present behavior. Such acquisition of behavioral potential can be defined as learning, and the physical bases of these potentials as memory traces1-4. Neuroscientists try to understand how these processes are organized in terms of molecular and neuronal changes in the brain by using a variety of methods in model organisms ranging from insects to vertebrates5,6. For such endeavors it is helpful to use model systems that are simple and experimentally accessible. The Drosophila larva has turned out to satisfy these demands based on the availability of robust behavioral assays, the existence of a variety of transgenic techniques and the elementary organization of the nervous system comprising only about 10,000 neurons (albeit with some concessions: cognitive limitations, few behavioral options, and richness of experience questionable)7-10.
Drosophila larvae can form associations between odors and appetitive gustatory reinforcement like sugar11-14. In a standard assay, established in the lab of B. Gerber, animals receive a two-odor reciprocal training: A first group of larvae is exposed to an odor A together with a gustatory reinforcer (sugar reward) and is subsequently exposed to an odor B without reinforcement 9. Meanwhile a second group of larvae receives reciprocal training while experiencing odor A without reinforcement and subsequently being exposed to odor B with reinforcement (sugar reward). In the following both groups are tested for their preference between the two odors. Relatively higher preferences for the rewarded odor reflect associative learning - presented as a performance index (PI). The conclusion regarding the associative nature of the performance index is compelling, because apart from the contingency between odors and tastants, other parameters, such as odor and reward exposure, passage of time and handling do not differ between the two groups9.

Appetitive Associative Olfactory Learning in Drosophila Larvae

Drosophila larvae are able to associate odor stimuli with gustatory reward. Here we describe a simple behavioral paradigm that allows the analysis of appetitive associative olfactory learning.

欲求联想嗅觉学习<em&gt;果蝇</em&gt;幼虫

In the following we describe the methodological  details of appetitive associative olfactory learning in Drosophila larvae. The setup, in combination with ...

神经科学

A Low-Cost, Odor-Reward Association Task for Tests of Learning and Memory

Robust and simple behavioral paradigms of appetitive, associative memory are crucial for researchers interested in cellular and molecular mechanisms of memory. In this paper, an effective and low-cost mouse behavioral protocol is described for examining the effects of physiological manipulation (such as the infusion of pharmacological agents) on the learning rate and duration of odor-reward memory. Representative results are provided from a study examining the differential role of tyrosine kinase receptor activity in short-term (STM) and long-term memory (LTM). Male mice were conditioned to associate a reward (sugar pellet) with one of the two odors, and their memory for the association was tested 2 or 48 h later. Immediately prior to the training, a tyrosine kinase (Trk) receptor inhibitor or vehicle infusions were delivered into the olfactory bulb (OB). Although there was no effect of the infusion on the learning rate, blockade of the Trk receptors in the OB selectively impaired LTM (48 h), and not short-term memory (STM; 2 h). The LTM impairment was attributed to the diminished odor selectivity as measured by the length of the digging time. The culmination of the results of this experiment showed that Trk receptor activation in the OB is the key in olfactory memory consolidation.

An odor-reward, associative learning task was used to investigate the differential effects of physiological manipulation on long-term and short-term memory.

Robust and simple behavioral paradigms of appetitive, associative memory are crucial for researchers interested in cellular and molecular mechanisms of ...

The Olfactory System as a Model to Study Axonal Growth Patterns and Morphology In Vivo

The olfactory system has the unusual capacity to generate new neurons throughout the lifetime of an organism. Olfactory stem cells in the basal portion of the olfactory epithelium continuously give rise to new sensory neurons that extend their axons into the olfactory bulb, where they face the challenge to integrate into existing circuitry. Because of this particular feature, the olfactory system represents a unique opportunity to monitor axonal wiring and guidance, and to investigate synapse formation. Here we describe a procedure for in vivo labeling of sensory neurons and subsequent visualization of axons in the olfactory system of larvae of the amphibian Xenopus laevis. To stain sensory neurons in the olfactory organ we adopt the electroporation technique. In vivo electroporation is an established technique for delivering fluorophore-coupled dextrans or other macromolecules into living cells. Stained sensory neurons and their axonal processes can then be monitored in the living animal either using confocal laser-scanning or multiphoton microscopy. By reducing the number of labeled cells to few or single cells per animal, single axons can be tracked into the olfactory bulb and their morphological changes can be monitored over weeks by conducting series of in vivo time lapse imaging experiments. While the described protocol exemplifies the labeling and monitoring of olfactory sensory neurons, it can also be adopted to other cell types within the olfactory and other systems.

The Olfactory System as a Model to Study Axonal Growth Patterns and Morphology In Vivo

We describe a protocol for in vivo labeling of olfactory sensory neurons by electroporation and subsequent confocal laser-scanning or multiphoton microscopy to visualize neuronal morphology and its development over time.

嗅觉系统作为模型来研究轴突生长的模式和形态<em&gt;在体内</em

The olfactory system has the unusual capacity to generate new neurons throughout the lifetime of an organism. Olfactory stem cells in the basal portion of ...

An Explant System for Time-Lapse Imaging Studies of Olfactory Circuit Assembly in Drosophila

~Neurons are precisely interconnected to form circuits essential for the proper function of the brain. The Drosophila olfactory system provides an excellent model to investigate this process since 50 types of olfactory receptor neurons (ORNs) from the antennae and maxillary palps project their axons to 50 identifiable glomeruli in the antennal lobe and form synaptic connections with dendrites from 50 types of second-order projection neurons (PNs). Previous studies mainly focused on identifying important molecules that regulate the precise targeting in the olfactory circuit using fixed tissues. Here, an antennae-brain explant system that recapitulates key developmental milestones of olfactory circuit assembly in culture is described. Through dissecting the external cuticle and cleaning opaque fat bodies covering the developing pupal brain, high quality images of single neurons from live brains can be collected using two-photon microscopy. This allows time-lapse imaging of single ORN axon targeting from live tissue. This approach will help reveal important cell biological contexts and functions of previously identified important genes and identify mechanisms underpinning the dynamic process of circuit assembly.

An Explant System for Time-Lapse Imaging Studies of Olfactory Circuit Assembly in Drosophila

This protocol describes the dissection procedure, culture condition, and live imaging of an antennae-brain explant system for the study of the olfactory circuit assembly.

一种用于果蝇嗅觉回路组装延时成像研究的外植体系统

~Neurons are precisely interconnected to form circuits essential for the proper function of the brain. The Drosophila olfactory system provides an ...

发育生物学

<meta charset="utf8"></head><body>为啮齿动物嗅觉实验设计灵活、经济高效的嗅觉计

Constructing an Olfactometer for Rodent Olfactory Behavior Studies

The use of olfactometers to study rodent behavior and brain activity during olfactory tasks is crucial for understanding brain circuits. These sophisticated devices allow researchers to precisely control and deliver odor stimuli, enabling the investigation of complex olfactory processes in rodents. Although commercially available olfactometers are convenient, they present challenges when technical issues arise, often requiring costly assistance and potentially disrupting research timelines. This article details the construction of a custom olfactometer specifically designed for mouse olfactory behavior experiments, providing a comprehensive list of parts and step-by-step instructions. The olfactometer is controlled through MATLAB, offering a user-friendly interface for researchers. Importantly, the open-source code allows users to modify and adapt the system, tailoring behavioral tasks to meet specific experimental needs. Building a customized olfactometer empowers users with the knowledge and capability to perform custom experimental design and troubleshooting independently, saving both time and resources. This approach not only enhances research flexibility but also fosters a deeper understanding of the equipment's functionality, ultimately leading to more robust and reliable olfactory studies in rodents.

<meta charset="utf8"></head><body>构建嗅觉计用于啮齿动物嗅觉行为研究 近红外光谱 心理咨询中的超扫描研究

This protocol describes the construction of an olfactometer for go/no-go olfactory behavior experiments. Step-by-step instructions, along with images, are provided to ensure the successful construction of the olfactometer. Information for troubleshooting issues encountered during the process is also included.

构建用于啮齿动物嗅觉行为研究的嗅觉计

The use of olfactometers to study rodent behavior and brain activity during olfactory tasks is crucial for understanding brain circuits. These ...

An Objective and Reproducible Test of Olfactory Learning and Discrimination in Mice

Olfaction is the predominant sensory modality in mice and influences many important behaviors, including foraging, predator detection, mating, and parenting. Importantly, mice can be trained to associate novel odors with specific behavioral responses to provide insight into olfactory circuit function. This protocol details the procedure for training mice on a Go/No-Go operant learning task. In this approach, mice are trained on hundreds of automated trials daily for 2&#8211;4 weeks and can then be tested on novel Go/No-Go odor pairs to assess olfactory discrimination, or be used for studies on how odor learning alters the structure or function of the olfactory circuit. Additionally, the mouse olfactory bulb (OB) features ongoing integration of adult-born neurons. Interestingly, olfactory learning increases both the survival and synaptic connections of these adult-born neurons. Therefore, this protocol can be combined with other biochemical, electrophysiological, and imaging techniques to study learning and activity-dependent factors that mediate neuronal survival and plasticity.

Here, we train mice on an associative learning task to test odor discrimination. This protocol also allows for studies on learning-induced structural changes in the brain.

新生大鼠于A气味偏侧学习模型解剖神经回路托换记忆的形成

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双-H迷宫:一个强大的行为测试学习和记忆的鼠害

为远程沉默的神经活动在鼠害的方法在学习独立的阶段

欲求联想嗅觉学习

A Low-Cost, Odor-Reward Association Task for Tests of Learning and Memory

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一种用于果蝇嗅觉回路组装延时成像研究的外植体系统

构建用于啮齿动物嗅觉行为研究的嗅觉计

构建用于啮齿动物嗅觉行为研究的嗅觉计

构建用于啮齿动物嗅觉行为研究的嗅觉计

小鼠嗅觉学习和鉴别的客观和可重复性试验