This work is supported in part by American Heart Association Scientist Development Grant and NARSAD Young Investigator Award to Yingwei Mao.

<ol>
	<li>Schwenk, F., Baron, U., Rajewsky, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+Cre-transgenic+mouse+strain+for+ubiquitous+deletion+of+loxP-flanked+gene+segments+including+deletion+in+germ+cells.">A Cre-transgenic mouse strain for ubiquitous deletion of loxP-flanked gene segments including deletion in germ cells.</a> Nucleic Acids Research. 23 (24), 5080-5081 (1995).</li><li>Hirrlinger, 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=Split-cre+complementation+indicates+coincident+activity+of+different+genes+in+vivo.">Split-cre complementation indicates coincident activity of different genes in vivo.</a> PLoS One. 4 (1), 1-10 (2009).</li><li>Fenno, L. 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=Targeting+cells+with+single+vectors+using+multiple-feature+Boolean+logic.">Targeting cells with single vectors using multiple-feature Boolean logic.</a> Nature Protocols. 11 (7), 763-772 (2014).</li><li>Paxinos, G., Franklin, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> The mouse brain in sterotaxic coordinates. , (2001).</li><li>Gould, T. D., Dao, D. T., Kovacsics, 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=Mood+and+anxiety+related+phenotypes+in+mice:+the+open+field+test.">Mood and anxiety related phenotypes in mice: the open field test.</a> Humana Press. 42, 1-20 (2009).</li><li>Denenberg, V. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Open-field+behavior+in+the+rat:+W.0hat+does+it+mean.">Open-field behavior in the rat: W.0hat does it mean.</a> Annals of the New York Academy of Sciences. 159, 852-859 (1969).</li><li>Bogdanova, O. V., Kanekar, S., D&#8217;Anci, K. E., Renshaw, P. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Factors+influencing+behavior+in+the+forced+swim+test.">Factors influencing behavior in the forced swim test.</a> Physiology &amp; Behavior. 118, 227-239 (2013).</li><li>Porsolt, R. D., Anton, G., Blavet, N., Jalfre, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Behavioral+despair+in+rats:+A+new+model+sensitive+to+antidepressant+treatments.">Behavioral despair in rats: A new model sensitive to antidepressant treatments.</a> European Journal of Pharmacology. 47 (4), 379-391 (1978).</li><li>Hunsberger, J., Dunman, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Animal+models+for+depression-like+and+anxiety-like+behavior.">Animal models for depression-like and anxiety-like behavior.</a> Nature Protocol Exchange. , (2007).</li><li>Hommel, J. D., Sears, R. M., Georgescu, D., Simmons, D. L., DiLeone, R. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Local+gene+knockdown+in+the+brain+using+viral-mediated+RNA+interference.">Local gene knockdown in the brain using viral-mediated RNA interference.</a> Nature Medicine. 9 (12), 1539-1544 (2003).</li><li>Mao, 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=Disrupted+in+schizophrenia+1+regulates+neuronal+progenitor+proliferation+via+modulation+of+GSK3beta/beta-catenin+signaling.">Disrupted in schizophrenia 1 regulates neuronal progenitor proliferation via modulation of GSK3beta/beta-catenin signaling.</a> Cell. 136 (6), 1017-1031 (2009).</li><li>Clark, M. S., Sexton, T. J., McClain, M., Root, D., Kohen, R., Neumai, J. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Overexpression+of+5-HT1B+receptor+in+dorsal+raphe+nucleus+using+herpes+simplex+virus+gene+transfer,+increases+anxiety+behavior+after+inescapable+stress.">Overexpression of 5-HT1B receptor in dorsal raphe nucleus using herpes simplex virus gene transfer, increases anxiety behavior after inescapable stress.</a> Journal of Neuroscience. 2 (11), 4550-4562 (2002).</li><li>Tasan, R. O., 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=The+central+and+basolateral+amygdala+are+critical+sites+of+neuropeptide+Y/Y2+receptor-mediated+regulation+of+anxiety+and+depression.">The central and basolateral amygdala are critical sites of neuropeptide Y/Y2 receptor-mediated regulation of anxiety and depression.</a> Journal of Neuroscience. 30 (18), 6282-6290 (2010).</li><li>Salmon, P., Trono, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Production+and+titration+of+lentiviral+vectors.">Production and titration of lentiviral vectors.</a> Current Protoc Hum Gen. 12 (12.10), (2007).</li><li>Carter, 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=Anesthetizing+mice.">Anesthetizing mice.</a> Methods Mol Biol. 18, 135-136 (1993).</li><li>Alachkar, A., Jiang, D., Harrison, M., Zhou, Y., Chen, G., Mao, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=An+EJC+factor+RBM8a+regulates+anxiety+behaviors.">An EJC factor RBM8a regulates anxiety behaviors.</a> Current Molecular Medicine. 13 (6), 887-899 (2013).</li><li>Bernal, J., Baldwin, M., Gleason, T., Kuhlman, S., Moore, G., Talcott, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Guidelines+for+rodent+survival+surgery.">Guidelines for rodent survival surgery.</a> Journal of Investigative Surgery. 22, 445-451 (2009).</li><li>Wahlsten, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Getting+ready+for+testing.+Mouse+Behavioral+Testing:+How+to+use+mice+in+behavioral+neuroscience.">Getting ready for testing. Mouse Behavioral Testing: How to use mice in behavioral neuroscience.</a> Academic Press. , 133-141 (2011).</li><li>Kheirbek, M. 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=Differential+control+of+learning+and+anxiety+along+the+dorso-ventral+axis+of+the+dentate+gyrus.">Differential control of learning and anxiety along the dorso-ventral axis of the dentate gyrus.</a> Neuron. 6 (77), 955-968 (2013).</li><li>Snyder, J. S., Soumier, A., Brewer, M., Pickel, J., Cameron, H. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Adult+hippocampal+neurogenesis+buffers+stress+responses+and+depressive+behaviors.">Adult hippocampal neurogenesis buffers stress responses and depressive behaviors.</a> Nature. 476 (7361), 458-461 (2011).</li><li>Sik-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=Induction+of+neuronal+vascular+endothelial+growth+factor+expression+by+cAMP+in+the+dentate+gyrus+of+the+hippocampus+is+required+for+antidepressant-like+behaviors.">Induction of neuronal vascular endothelial growth factor expression by cAMP in the dentate gyrus of the hippocampus is required for antidepressant-like behaviors.</a> Journal of Neuroscience. 29 (26), 8493-8505 (2009).</li><li>Miller, D. G., Adam, M. A., Miller, A. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Gene+transfer+by+retrovirus+vectors+occurs+only+in+cells+that+are+actively+replicating+at+the+time+of+infection.">Gene transfer by retrovirus vectors occurs only in cells that are actively replicating at the time of infection.</a> Molecular and Cellular Biology. 10 (8), 4239-4242 (1990).</li><li>Rubinson, D. 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=A+lentivirus-based+system+to+functionally+silence+genes+in+primary+mammalian+cells,+stem+cells+and+transgenic+mice+by+RNA+interference.">A lentivirus-based system to functionally silence genes in primary mammalian cells, stem cells and transgenic mice by RNA interference.</a> Nature Genetics. 33, 401-406 (2003).</li></ol>

The authors have nothing to disclose.

حقن المجسم الناجحة تعتمد على ثلاثة عوامل هي: الحفاظ على الماوس على قيد الحياة، وتحديد نقطة الصفر الصحيحة للإحداثيات (رأس الإبرة على تقاطع bregma والخط interaural)، وتحديد عمق المناسب لنقطة الصفر (رأس الإبرة مجرد لمس السطح الخارجي للأنسجة المخ). الجدوى من الفئران هو المهم. الب...

وقد الماوس تستخدم على نطاق واسع في علم الأعصاب لأنه من السهل التلاعب وراثيا. تقنيات الجينات خروج المغلوب تسمح للباحثين للتحقيق في كيفية عمل كل عامل وراثي الأشكال السلوكيات الماوس. وعلاوة على ذلك، يوفر نظام لجنة المساواة العرقية loxp أداة قيمة لنسيجيا وخلية type- بالضربة القاضية جين معين في الفئران، والتي تمكن الباحثين لدراسة وظيفة الجينات في الأنسجة المختلفة. 1 ومع ذلك، من الناحية العملية، وأنماط التعبير من المروجين لجنة المساواة العرقية يصعب السيطرة ولقد لذلك العديد من السائقين حتى الآن لجنة المساواة العرقية التي أنشئت لم يتحقق التعبير منطقة محددة. 2،3 بدلا من ذلك، حقن المجسم هو الأسلوب الذي يستهدف مناطق محددة في الدماغ من الماوس الكبار. عن طريق حقن الفيروسات المعدلة وراثيا معربا عن كدنا] أو shRNA، لا يمكن أن يتحقق التشكيل منطقة محددة في التعبير الجيني. على الرغم من أن حجم الدماغ من كل فأر يختلف، موقع مناطق محددة في الدماغ يمكن تحديدها باستخدام coordi المجسمتعيين المتحكمة من المعالم على الجمجمة من مخ الفأر. المعالم الأكثر شيوعا هي bregma، لامدا، والخط interaural. باستخدام الإحداثيات التي تم الحصول عليها من أطلس الدماغ، 4 في المكان المحدد من كل منطقة في الدماغ يمكن تحديدها من قبل انتيرو-الخلفي (A / P)، وسطي الاطراف (M / L)، و (D / V) محاور الظهري البطني من bregma / interaural خط التقاطع. عادة الفيروسات حقنها في أدمغة الفئران الموسومة هي إما أحمر أو أخضر بروتين فلوري (RFP أو GFP)، بحيث الحقن يمكن التأكد عن طريق الفحص المجهري الفلورسنت. تقييم السلوكية للفئران ضرورية خصوصا للبحوث الأساسية من الاضطرابات النفسية. أعراض الاضطرابات النفسية لدى المرضى الذين عادة ما تشمل السلوكيات الشاذة. بعض هذه السلوكيات البشرية هي الحفظ تطويريا ويمكن تحاكي مباشرة، ولاحظ في الماوس. على سبيل المثال، والاكتئاب يمكن أن تكون على غرار في الماوس عن طريق قياس اليأس السلوكي. الأشخاص الذين يعانون من الاكتئاب كثيراأون تشعر وكأن شيئا لم يفعلوا سيساعد أي وقت مضى، من أعراض التي يمكن أن تؤدي في النهاية إلى الانتحار. في القوارض، وهذا يمكن أن تكون على غرار باستخدام اختبار السباحة القسري (FST)، والذي يقيس مقدار الوقت ماوس السباحة مقابل عائمة في بركة من الماء (ينظر إليها على التخلي). يتم التحقق من صحة هذا النموذج من قبل إنقاذ النمط الظاهري مع عقاقير مضادة للاكتئاب. 7،8،9 الفئران التي تلقت عقاقير مضادة للاكتئاب سوف تنفق أقل كثيرا من الوقت متحرك مقارنة مع الضوابط غير المعالجة. تم تصميم اختبار السلوكي آخر، الاختبار الميداني المفتوحة (OFT) لتقييم الحركة في الفئران، وبالإضافة إلى ذلك يمكن استخدامها لتحليل النمط الظاهري حريصة في الفئران. 5،6 يقوم هذا الاختبار على أساس أن الفئران يشعرون أكثر أمانا عندما تكون قريبة على الحائط في حقل مفتوح الرواية. سوف الفئران نوع البرية استكشاف في النهاية بيئة جديدة، كما أنها حيوانات غريبة. ومع ذلك، تنفق وقتا أقل في وسط الميدان يدل على القلق في الماوس، والماوس سوف لن تكون قادرة على التغلب على initiaل الخوف يأتي نتيجة للبيئة جديدة. القلق من الفأرة، وكمية من مقدار الوقت الذي يقضيه في وسط حقل مفتوح، يمكن مقارنة القلق السريرية في البشر، والتي هي موجودة في العديد من الاضطرابات النفسية. الجمع بين الحقن المجسم مع النماذج السلوكية هو وسيلة جديدة لتغيير التعبير عن جين معين في منطقة الدماغ المستهدفة. ومن ثم يمكن تحديد أثر هذان التعبير الجيني على السلوكيات الماوس. وعلى النقيض من خروج المغلوب الدماغ كله، وهذه الطريقة مفيدة بشكل خاص لأنها تستهدف مناطق محددة في الدماغ فقط. بالإضافة إلى ذلك، عادة ما يتم إجراء حقن المجسم في الكبار النوع البري الماوس، لذلك، وقد تم الحفاظ على الذاتية التعبير الجيني في جميع أنحاء مراحل النمو. هذه الطريقة سوف تجنب تأثير الخلط إذا كنت بحاجة الجينات من أجل البقاء خلال المرحلة الجنينية أو بعد الولادة للتنمية. أحد أوجه القصور الرئيسية هو أن فئران التجارب تحتاج إلى اجتيازهاح عملية جراحية الغازية، التي جماجم من الفئران لا بد من فتح. وعلاوة على ذلك، يتم تحديد درجة التشكيل الجيني من عيار وكفاءة الفيروس. يحتاج الفيروس ليتم حقنه في المنطقة الصحيحة باستخدام الإحداثيات المجسم، الأمر الذي يتطلب أدوات خاصة. التحقق من موقع الحقن الصحيح لا يمكن إلا أن تكتمل بعد الوفاة. تم عرض هذه الطريقة تستخدم سابقا لاختبار إشراك جين معين في مختلف الأمراض العصبية. على سبيل المثال، فيروسية بوساطة رني استهداف الجين ث (الذي يسمح الدوبامين أن يتم تصنيعها) تم حقنها في المكتنزة المادة السوداء، وأجريت تحليل السلوك الحركي. استخدمت 10 دراسة أخرى حقن المجسم لإسكات DISC1 الفيروسة البطيئة لتقييم سلوك الماوس في العلاقة إلى الفصام. ضربة قاضية من DISC1 أدى إلى زيادة الحركة ردا على الجدة (التشابه الأعراض الإيجابية في انفصام الشخصية)، وزيادة الجمود فيFST. 11 وبالمثل، وجدت دراسة إضافية 5-HT 1B overexpression أدى إلى زيادة السلوك الاستكشافي في التجارة المشروعة، بما يتفق مع النمط الظاهري المضادة للقلق باستخدام هذا الأسلوب. يمكن 12 حقن المجسم تقديم فيروس لجنة المساواة العرقية للحث على إعادة التركيب في لجنة المساواة العرقية loxp الفئران . وقد استخدم هذا الأسلوب لحذف انتقائي مستقبلات Y2 في اللوزة ونواة السرير من الانتهائي السطور. بناء على التحليل السلوكي، وقد وجدت هذه الفئران إلى النمط الظاهري المضادة للاكتئاب عندما تم حذف هذا الجين في اللوزة المركزية، ولكن لا النمط الظاهري عندما تم حذف هذا الجين في اللوزة basolateral أو نواة السرير من الانتهائي السطور. 13 وهكذا، هذه التقنية يوفر أداة فريدة من نوعها لدراسة تأثير الجينات على سلوكيات الحيوان.

<table border="0" cellpadding="0" cellspacing="0" width="608">
	<colgroup>
		<col />
		<col />
		<col />
		<col />
	</colgroup>
	<tbody>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Name of Material/ Equipment</td>
			<td style="width:107px;">Company</td>
			<td style="width:119px;">Catalog Number</td>
			<td style="width:167px;">Comments/Description</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Stereotactic Apparatus item 51725</td>
			<td style="width:107px;">Stoelting co.</td>
			<td align="right" style="width:119px;">51725</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Quintessential stereotaxic pump&#160;</td>
			<td style="width:107px;">Stoelting co.</td>
			<td align="right" style="width:119px;">53311</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Injection Styringe, 65 RN</td>
			<td style="width:107px;">Hamilton</td>
			<td style="width:119px;">7633-01</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">DMEM</td>
			<td style="width:107px;">Sigma-Aldrich</td>
			<td style="width:119px;">D5796</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">PEI&#160;</td>
			<td style="width:107px;">Polysciences Inc.</td>
			<td align="right" style="width:119px;">23966</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Scissors</td>
			<td style="width:107px;">Fine Surgical Tools</td>
			<td style="width:119px;">14084-08</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Blunt end forceps</td>
			<td style="width:107px;">Fine Surgical Tools</td>
			<td style="width:119px;">11002-12</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Needle holder&#160;</td>
			<td style="width:107px;">Fine Surgical Tools</td>
			<td style="width:119px;">12001-13</td>
			<td></td>
		</tr>
		<tr height="84">
			<td height="84" style="height:84px;width:216px;">Drill</td>
			<td style="width:107px;">Ram Products Inc</td>
			<td style="width:119px;">Microtorque control box, Tech2000 handpiece</td>
			<td>with pedal</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Glass bead sterilizer</td>
			<td style="width:107px;">Inotech&#160;</td>
			<td style="width:119px;">IS-400</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Absorbable sutures&#160;</td>
			<td style="width:107px;">Unify</td>
			<td style="width:119px;">M-K518r19</td>
			<td>Absorbable, reverse cutting</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Cotton swabs</td>
			<td style="width:107px;">VWR</td>
			<td style="width:119px;">89031-270</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Heating pad</td>
			<td style="width:107px;">Gaymar</td>
			<td style="width:119px;">T-pump TP-500 PN11184-000</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Artificial tears</td>
			<td style="width:107px;">Rugby</td>
			<td style="width:119px;">NDC 0536-6550-91</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Disposable syringe</td>
			<td style="width:107px;">BD Syringe</td>
			<td align="right" style="width:119px;">309623</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Cloth</td>
			<td style="width:107px;"></td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Gloves&#160;</td>
			<td style="width:107px;">Ansell</td>
			<td style="width:119px;">Senseitouch #7823</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Avertin or other anesthetic</td>
			<td style="width:107px;">see recipe citation</td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Ketoprofen or other analgesic</td>
			<td style="width:107px;">see veternarian&#160;</td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Tracking software</td>
			<td style="width:107px;">Noldus</td>
			<td style="width:119px;">Ethovision XT</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Tracking Camera</td>
			<td style="width:107px;">Noldus</td>
			<td style="width:119px;">Media Recorder</td>
			<td></td>
		</tr>
	</tbody>
</table>

applying stereotactic injection technique to study genetic effects on animal behaviors

حقن المجسم هو تقنية مفيدة لتقديم lentiviruses عيار عالية لمناطق الدماغ المستهدفة في الفئران. Lentiviruses يمكن إما بإفراط أو التعبير الجيني ضربة قاضية في منطقة مركزة نسبيا دون أضرار جسيمة للأنسجة الدماغ. بعد الشفاء، ويمكن اختبار الماوس حقنه في مختلف المهام السلوكية مثل اختبار الحقل المفتوح (OFT) والسباحة الاختبار القسري (FST). تم تصميم OFT لتقييم الحركة والنمط الظاهري حريصة في الفئران عن طريق قياس مقدار الوقت الذي يقضيه الماوس في وسط حقل مفتوح الرواية. سوف فأر أكثر قلقا يمضون وقتا أقل بكثير في وسط الميدان رواية مقارنة بالمجموعة الضابطة. وFST يقيم النمط الظاهري المضادة للاكتئاب عن طريق قياس مقدار الوقت الذي تنفق الفئران غير متحرك عند وضعها في دلو من الماء. وهناك الفأر مع النمط الظاهري المضادة للاكتئاب تنفق أقل بكثير متحرك الوقت مقارنة للسيطرة على الحيوانات. والهدف من هذا البروتوكول هو استخدام ster يسودحقن eotactic من الفيروسة البطيئة بالتزامن مع الاختبارات السلوكية لتقييم مدى عوامل وراثية تعدل سلوك الحيوان.

<html> حقن المجسم دقيق يعتمد بشكل كبير على تحديد الإحداثيات الصحيحة. يجب تعيين رأس الإبرة المستخدمة لحقن الفيروس مباشرة على تقاطع bregma والخط interaural (الشكل 1). فمن المفيد استخدام stereomicroscope للتأكد ما إذا كان أو لم يتم وضع الإبرة بشكل صحيح. عند النظر من خلال المجهر، وي?...

Watch this Scientific Journal Video about Applying Stereotactic Injection Technique to Study Genetic Effects on Animal Behaviors at JoVE.com

Applying Stereotactic Injection Technique to Study Genetic Effects on Animal Behaviors

حقن المجسم من lentiviruses معربا عن cDNAs أو shRNAs يمكن أن تعدل التعبير الجيني في مناطق محددة في الدماغ لدى الفئران. هنا، نقدم بروتوكول الجمع بين الحقن المجسم مع المهام السلوكية، مثل الاختبار الميداني المفتوحة (OFT) والسباحة الاختبار القسري (FST).

تطبيق المجسم حقن التقنية لدراسة التأثيرات الوراثية على سلوكيات الحيوان

Stereotactic injection is a useful technique to deliver high titer lentiviruses to targeted brain areas in mice. Lentiviruses can either overexpress or ...

applying-stereotactic-injection-technique-to-study-genetic-effects-on

Research

JoVE Journal

Behavior

14.0K Views.  The Pennsylvania State University.  حقن المجسم من lentiviruses معربا عن cDNAs أو shRNAs يمكن أن تعدل التعبير الجيني في مناطق محددة في الدماغ لدى الفئران. هنا، نقدم بروتوكول الجمع بين الحقن المجسم مع المهام السلوكية، مثل الاختبار الميداني المفتوحة (OFT) والسباحة الاختبار القسري (FST). 

Video: Applying Stereotactic Injection Technique to Study Genetic Effects on Animal Behaviors

Stereotactic Injection of MicroRNA-expressing Lentiviruses to the Mouse Hippocampus CA1 Region and Assessment of the Behavioral Outcome

MicroRNAs (miRNAs) are small regulatory single-stranded RNA molecules around 22 nucleotides long that may each target numerous mRNA transcripts and dim an entire gene expression pathway by inducing destruction and/or inhibiting translation of these targets. Several miRNAs play key roles in maintaining neuronal structure and function and in higher-level brain functions, and methods are sought for manipulating their levels for exploring these functions. Here, we present a direct in vivo method for examining the cognitive consequences of enforced miRNAs excess in mice by stereotactic injection of miRNA-encoding virus particles. Specifically, the current protocol involves injection into the hippocampal CA1 region, which contributes to mammalian memory consolidation, learning, and stress responses, and offers a convenient injection site. The coordinates are measured according to the mouse bregma and virus perfusion is digitally controlled and kept very slow. After injection, the surgery wound is sealed and the animals recover. Lentiviruses encoding silencers of the corresponding mRNA targets serve to implicate the specific miRNA/target interaction responsible for the observed effect, with na&iuml;ve mice, mice injected with saline and mice injected with "empty" lentivirus vectors as controls. One month post-injection, the animals are examined in the Morris Water Maze (MWM) for assessing their navigation learning and memory abilities. The MWM is a round tank filled with colored water with a small platform submerged 1 cm below the water surface. Steady visual cues around the tank allow for spatial navigation (sound and the earth's magnetic field may also assist the animals in navigating). Video camera monitoring enables measuring the route of swim and the time to find and amount the platform. The mouse is first taught that mounting the hidden platform offers an escape from the enforced swimming; it is then tested for using this escape and finally, the platform is removed and probe tests examine if the mouse remembers its previous location. Repeated tests over several consecutive days highlight improved performance of tested mice at shorter latencies to find and mount the platform, and as more direct routes to reach the platform or its location. Failure to show such improvement represents impaired learning and memory and/or anxiety, which may then be tested specifically (e.g. in the elevated plus maze). This approach enables validation of specific miRNAs and target transcripts in the studied cognitive and/or stress-related processes.

MicroRNAs have significant roles in brain structure and function. Here we describe a method to enforce hippocampal miRNA over-expression using stereotactic injection of an engineered miRNA-expressing lentivirus. This approach can serve as a relatively rapid way to assess the in vivo effects of over-expressed miRNAs in specific brain regions.

حقن المجسم من الرنا الميكروي، معربا عن Lentiviruses إلى CA1 الحصين ماوس المنطقة وتقييم حصيلة السلوكية

MicroRNAs (miRNAs) are small regulatory single-stranded RNA molecules around 22 nucleotides long that may each target numerous mRNA transcripts and dim an ...

A Within-subjects Experimental Protocol to Assess the Effects of Social Input on Infant EEG

Despite the importance of social interactions for infant brain development, little research has assessed functional neural activation while infants socially interact. Electroencephalography (EEG) power is an advantageous technique to assess infant functional neural activation. However, many studies record infant EEG only during one baseline condition. This protocol describes a paradigm that is designed to comprehensively assess infant EEG activity in both social and nonsocial contexts as well as tease apart how different types of social inputs differentially relate to infant EEG. The within-subjects paradigm includes four controlled conditions. In the nonsocial condition, infants view objects on computer screens. The joint attention condition involves an experimenter directing the infant's attention to pictures. The joint attention condition includes three types of social input: language, face-to-face interaction, and the presence of joint attention. Differences in infant EEG between the nonsocial and joint attention conditions could be due to any of these three types of input. Therefore, two additional conditions (one with language input while the experimenter is hidden behind a screen and one with face-to-face interaction) were included to assess the driving contextual factors in patterns of infant neural activation. Representative results demonstrate that infant EEG power varied by condition, both overall and differentially by brain region, supporting the functional nature of infant EEG power. This technique is advantageous in that it includes conditions that are clearly social or nonsocial and allows for examination of how specific types of social input relate to EEG power. This paradigm can be used to assess how individual differences in age, affect, socioeconomic status, and parent-infant interaction quality relate to the development of the social brain. Based on the demonstrated functional nature of infant EEG power, future studies should consider the role of EEG recording context and design conditions that are clearly social or nonsocial.

This novel protocol is designed to assess the neural bases of social interaction in infants. The paradigm is designed to tease apart how various social inputs such as language, joint attention, and face-to-face interaction relate to infant neural activation. Infant EEG power is recorded during both social and nonsocial conditions.

وضمن موضوعات-البروتوكول التجريبي لتقييم آثار إدخال الاجتماعي على EEG الرضع

Despite the importance of social interactions for infant brain development, little research has assessed functional neural activation while infants ...

Testing Animal Anxiety in Rats: Effects of Open Arm Ledges and Closed Arm Wall Transparency in Elevated Plus Maze Test

The elevated plus maze test is a behavioral test for assessing animal anxiety in rodents. Although this test is widely applied in the field of behavioral science, conflicting outcomes are often provided from different laboratories. To identify reasons for the different outcomes, we previously focused on arm features, which differ between laboratories, most notably the presence/absence of ledges at the sides of open arms and the transparency/opaqueness of closed arm walls. In a previous report, we used a custom designed container to compare rat behavior on different combinations of open and closed arm designs under otherwise identical experimental conditions, and showed that differences in arm features interfere with experimental outcomes. In brief, open arm ledges significantly increased anxiety-like behavior in rats, while transparent arms may also have decreased this behavior. Furthermore, we verified a higher detection sensitivity of the effect of an anxiolytic drug in a combination of no-ledged open arm + opaque walled closed arm compared with a combination of ledged open arm + transparent walled closed arm. In this report, we introduce our protocol for the elevated plus maze test, together with discussion of the key results from the previous report and our experimental experience. We believe this report will provide useful information for researchers who have employed or who plan to use the elevated plus maze in their studies.

Rats' anxiety-like behavior tested by the elevated plus maze varies highly depending on the testing environment. Here we show how apparatus settings, especially arm structure, affect behavior on the maze, and we explain a reliable protocol to detect effects of anxiolytic drugs. This report will contribute to anxiety research.

اختبار القلق الحيوان في الفئران: آثار الحواف ذراع مفتوحة ومغلقة الذراع الجدار الشفافية في مرتفعة بالإضافة إلى اختبار المتاهة

The elevated plus maze test is a behavioral test for assessing animal anxiety in rodents. Although this test is widely applied in the field of behavioral ...

Assessment of Social Transmission of Food Preferences Behaviors

Olfactory recognition deficits are suggested to be able to serve as clinical marker to differentiate Alzheimer&#39;s disease (AD) subjects from healthy aging groups. For example, olfactory dysfunction in AD can present as impairment in olfactory recognition, emerging during early stages of the disease and worsening while the disease progresses. The social transmission of food preferences (STFP) task is based on a rudimentary form of communication between rodents concerning distant foods dependent on the transmission of olfactory cues. Healthy wild-type mice would prefer to eat a novel, flavored food that was previously cued by a conspecific, and this food preference would be hampered in transgenic AD mice, such as the APP/PS1 model. Indeed, a strong preference for the cued food in C57Bl6/J mice of 3 months of age was found, and this was reduced in 3 months old transgenic APP/PS1 mice. In summary, STFP task could be a powerful measure to be integrated in present subclinical detection assays of AD.

This paper presents a protocol for the investigation of social transmission of food preference in mice. The advantages and possible applications for this procedure, for instance, in detecting early changes in AD mouse models, are highlighted. To conclude, interpretation of the results in light of critical details are discussed.

تقييم انتقال اجتماعية من السلوكيات التفضيلات الغذائية

Olfactory recognition deficits are suggested to be able to serve as clinical marker to differentiate Alzheimer&#39;s disease (AD) subjects from healthy ...

Tickling, a Technique for Inducing Positive Affect When Handling Rats

Handling small animals such as rats can lead to several adverse effects. These include the fear of humans, resistance to handling, increased injury risk for both the animals and the hands of their handlers, decreased animal welfare, and less valid research data. To minimize negative effects on experimental results and human-animal relationships, research animals are often habituated to being handled. However, the methods of habituation are highly variable and often of limited effectiveness. More potently, it is possible for humans to mimic aspects of the animals&#39; playful rough-and-tumble behavior during handling. When applied to laboratory rats in a systematic manner, this playful handling, referred to as tickling, consistently gives rise to positive behavioral responses. This article provides a detailed description of a standardized rat tickling technique. This method can contribute to future investigations into positive affective states in animals, make it easier to handle rats for common husbandry activities such as cage changing or medical/research procedures such as injection, and be implemented as a source of social enrichment. It is concluded that this method can be used to efficiently and practicably reduce rats&#39; fearfulness of humans and improve their welfare, as well as reliably model positive affective states.

This article demonstrates the standardized application of playful handling, a tickling technique designed to mimic rat rough-and-tumble play. This technique is effective at reducing fearful reactions to humans and generating positive affect when rats are handled for common husbandry activities and medical and research procedures such as injection.

دغدغة، تقنية لإحداث تأثير إيجابي عند التعامل مع الفئران

Handling small animals such as rats can lead to several adverse effects. These include the fear of humans, resistance to handling, increased injury risk ...

Using a Classroom-Based Deese Roediger McDermott Paradigm to Assess the Effects of Imagery on False Memories

Associated word list procedures can elicit false memories in predictable ways by inducing associative processing, thus making it harder to monitor the accuracy of memories. The purpose of the method presented here was to induce false memories using lists of either semantically or phonologically related words and to assess the effects of imagery instructions on the recall and recognition of those false memories. To do this, we used a modified version of the Deese Roediger McDermott (DRM) paradigm. We adapted word lists from previous DRM studies to suit imagery procedures and created an automated presentation to present the word lists in classroom settings. We then recruited undergraduate classes and instructed some of the classes to create mental images of the list words as they were being presented, while instructing the other classes to simply remember the words. The automated presentation presented word lists to participants, one word at a time, alternating between phonologically and semantically related lists. Participants used paper-pencil recall packets to immediately recall list items, complete a distractor activity, and take a subsequent final recognition test. Often, participants immediately recalled and later recognized words that were related to the list items but were not actually presented; these are known as critical lures and indicate a false memory. The protocol detailed here describes a four-step procedure - list presentation, immediate recall, distractor phase, and final recognition - that can assess the effects of list type and imagery instruction within the DRM paradigm on memory. The automated nature of the list presentation provides the ability to systematically vary variables of interest, and the paper and pencil method of data collection affords an easily accessible method for collecting data in classroom settings. The protocol also offers options to modify the procedure to a more traditional DRM paradigm without imagery and/or list type manipulations. The use of this protocol can provide results relevant to both classroom learning and cognitive science principles.

The method presented here induced false memories using lists of related words and also assessed the effects of imagery instructions on the recall and recognition of those false memories. This protocol details a modified version of the Deese Roediger McDermott (DRM) paradigm.

باستخدام نموذج مكديرموت روديغر ديس المستندة إلى الفصول الدراسية لتقييم آثار الصور الملتقطة في ذكريات كاذبة

Associated word list procedures can elicit false memories in predictable ways by inducing associative processing, thus making it harder to monitor the ...

Assessing the Coherence of Parents' Short Narratives Regarding their Child Using the Five-Minute Speech Sample Procedure

Valid and efficient measures for assessing the quality of parent-child relationships are needed to facilitate research and evidence-based practice with parents. This paper focuses on such a method, namely Five-Minute Speech Sample-Coherence (FMSS-Coherence). In this method, a parent is asked to speak for five uninterrupted minutes about her/his child and their relationship. The resulted narrative is coded for coherence, namely the extent to which the parent provides&#160;in the narrative&#160;a clear, consistent, multidimensional and well-supported portrayal of the child. FMSS-Coherence is based on attachment research that shows that the coherence of parents&#8217; narratives is indicative of the quality of the parent-child relationship and child adjustment. It overcomes the limitations of attachment narrative measures which are typically labor intensive. FMSS-Coherence is less nuanced than extant attachment narrative measures. Yet, studies of families from different cultural backgrounds, across different child ages and in the context of typically developing children as well as children with special needs suggest that coherence can be reliably evaluated using the FMSS procedure. Furthermore, parents&#8217; FMSS-Coherence is associated with parenting quality and children&#8217;s socio-emotional adjustment. Thus, it holds promise for researchers and practitioners who seek a relatively time- and cost-effective method for assessing the coherence of parents&#8217; narratives regarding their child.

Assessing the Coherence of Parents&#039; Short Narratives Regarding their Child Using the Five-Minute Speech Sample Procedure

This paper introduces a method for assessing the coherence of parents' short narratives regarding their child and their relationship using the five-minute speech sample procedure.

تقييم اتساق الروايات القصيرة للوالدين فيما يتعلق بأطفالهم باستخدام إجراء عينة الكلام لمدة خمس دقائق

Valid and efficient measures for assessing the quality of parent-child relationships are needed to facilitate research and evidence-based practice with ...

Applying an eMASS Customization Program as a Research Tool to Evaluate Consumer Benefits

As many scholars and practitioners study personalization and relationship marketing, it is important to provide personalization such as mass customization through marketing technology. The purpose of this study is to examine how to conduct consumer research using an online survey and analysis of data. This study examines consumers&#39;&#160;perceived benefits while customizing a product as well as emotional product attachment, attitudes toward a customization program, and loyalty intentions in the context of online retailing. In addition, this study investigates how consumer responses are different based on individual characteristics such as fashion innovativeness. An online survey company in South Korea recruited 290 female apparel shoppers who purchased apparel online. To enhance external validity, this study used an existing retail website with a well-established mass customization program. After completing the customization program, participants complete the online questionnaire. Structural equation modeling (SEM) and latent mean analyses (LMAs) are then performed for analyses. This study stresses the importance of testing measurement invariance for mean comparisons. Before the SEM and LMA, this study follows the hierarchy of invariance tests (configural invariance test, metric invariance test, and scalar invariance test), which are not considered by traditional approaches such as ANOVA. These statistical analyses provide applicability of the invariance test procedures and LMA to consumer behaviors. The conclusions of mean differences have integrity and validity because they are guided by a sophisticated statistical procedure to ensure measurement invariance.

Presented here is a protocol to examine consumer responses toward mass customization in the context of online retailing. The protocol details the online survey procedure and how to analyze data using structural equation modeling and group differences using latent mean analyses.

تطبيق برنامج تخصيص eMASS كاداه بحثيه لتقييم فوائد المستهلك

As many scholars and practitioners study personalization and relationship marketing, it is important to provide personalization such as mass customization ...

An Experimental Paradigm for Measuring the Effects of Ageing on Sentence Processing

Previous studies have found that older adults have greater difficulties in processing syntactically complex sentences than younger adults. However, the exact regions where the difficulties arise have not been fully identified. In this study, a maze task was implemented to investigate how older adults and younger adults differentially processed two types of sentences with different levels of syntactic complexity, namely subject relative clauses and object relative clauses. Participants were asked to choose between two alternatives at each segment of the sentences. The task required participants to engage in a strictly incremental mode of processing. The reading times for each segment were recorded, allowing the quantification of the difficulty of sentence reading. The task allowed us to identify the exact locations of the processing difficulty and thus facilitate a more accurate assessment of the age-related decline in sentence processing. The results indicate that the effect of ageing was found mainly at the head nouns, but not at other regions of the sentences, a finding which suggests that the maze task is an effective method to identify the exact location of the ageing effect on sentence processing. The implications of this experimental paradigm for investigating the effect of ageing on sentence processing are discussed.

Here, we present a protocol to examine the age-related decline in sentence processing using a maze task that allowed us to localize the processing difficulty at each word of the sentences during reading.

نموذج تجريبي لقياس اثار الشيخوخة علي تجهيز الجمل

Previous studies have found that older adults have greater difficulties in processing syntactically complex sentences than younger adults. However, the ...

The Use of Traditional Fear Tests to Evaluate Different Emotional Circuits in Cattle

Animal temperament is complex and has implications for productivity and economic profitability. Quantifying an animal&#39;s response to differing stimuli may facilitate breeding selections and identify animals that are better suited to specific management strategies. Multiple tests have been developed to evaluate cattle temperament (e.g., exit velocity, chute score, pen score, open field test, startle test, bovine zero maze), but each of these tests evaluates the animal&#39;s response to different stimuli (e.g., isolation, novel environment, startle, willingness to enter an enclosed area). Cattle temperament has been observed to be relatively stable over time. However, the evaluation of temperament has the potential to be influenced by current conditions, previous experiences, and observer bias. Many of these temperament tests have been improperly categorized as fear tests and have also been criticized for being subjective. This paper provides a framework for standardizing behavioral tests for cattle and suggests that these different evaluations assess different aspects of the animal&#39;s overall temperament.

Here, we present a protocol to conduct a variety of behavioral tests in cattle that have been designed to evaluate emotions. A battery of behavioral tests (open field test, startle test, bovine zero maze, exit velocity, pen score, and chute score) were conducted to evaluate different components of animal temperament.

استخدام اختبارات الخوف التقليدية لتقييم الدوائر العاطفية المختلفة في الماشية

Animal temperament is complex and has implications for productivity and economic profitability. Quantifying an animal&#39;s response to differing stimuli ...