Name: the c-fos protein immunohistological detection: a useful tool as a marker of central pathways involved in specific physiological responses in vivo and ex vivo
Uploaded: 2016-04-25T14:00:00
Description: Watch this Scientific Journal Video about The c-FOS Protein Immunohistological Detection: A Useful Tool As a Marker of Central Pathways Involved in Specific Physiological Responses In Vivo and Ex Vivo

The University Paris 13 supported this work. ASPT was supported by a University Paris 13 fellowship and the "Association Fran&#231;aise pour le Syndrome d'Ondine". FJ was supported by a Laboratory of Excellence GR-Ex fellowship. The GR-Ex (ref ANR-11-LABX-0051) is funded by the program "Investissement d'avenir" of the French National Research agency (ref ANR-11-IDEX-0005-02).

注：C-FOSの検出は、いくつかのステップ（ 図1）を含む標準化された手順です。全ての実験は、ラットまたはマウスで行いました。実験プロトコルは、動物実験チャールズ・ダーウィン（CE5 / 2011/05）に倫理委員会によって承認された動物のケアのために9月22日の欧州共同体理事会、2010（63分の2010 / EU）に基づいて行われ、に従って行いました動物のケアのためのフランスの法律に...

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	<li>Curran, T., Teich, N. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Identification+of+a+39,000-dalton+protein+in+cells+transformed+by+the+FBJ+murine+osteosarcoma+virus.">Identification of a 39,000-dalton protein in cells transformed by the FBJ murine osteosarcoma virus.</a> Virology. 116, 221-235 (1982).</li><li>Curran, T., MacConnell, W. P., van Straaten, F., Verma, I. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Structure+of+the+FBJ+murine+osteosarcoma+virus+genome:+molecular+cloning+of+its+associated+helper+virus+and+the+cellular+homolog+of+the+v-fos+gene+from+mouse+and+human+cells.">Structure of the FBJ murine osteosarcoma virus genome: molecular cloning of its associated helper virus and the cellular homolog of the v-fos gene from mouse and human cells.</a> Mol Cell Biol. 3, 914-921 (1983).</li><li>Curran, T., Miller, A. D., Zokas, L., Verma, I. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Viral+and+cellular+fos+proteins:+a+comparative+analysis.">Viral and cellular fos proteins: a comparative analysis.</a> Cell. 36, 259-268 (1984).</li><li>Herdegen, T., Leah, J. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Inducible+and+constitutive+transcription+factors+in+the+mammalian+nervous+system:+control+of+gene+expression+by+Jun,+Fos+and+Krox,+and+CREB/ATF+proteins.">Inducible and constitutive transcription factors in the mammalian nervous system: control of gene expression by Jun, Fos and Krox, and CREB/ATF proteins.</a> Brain Res Brain Res Rev. 28, 370-490 (1998).</li><li>Dragunow, M., Faull, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+use+of+c-fos+as+a+metabolic+marker+in+neuronal+pathway+tracing.">The use of c-fos as a metabolic marker in neuronal pathway tracing.</a> J Neurosci Methods. 29, 261-265 (1989).</li><li>Bullitt, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Expression+of+c-fos-like+protein+as+a+marker+for+neuronal+activity+following+noxious+stimulation+in+the+rat.">Expression of c-fos-like protein as a marker for neuronal activity following noxious stimulation in the rat.</a> J Comp Neurol. 296, 517-530 (1990).</li><li>Greenberg, M. E., Ziff, E. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Stimulation+of+3T3+cells+induces+transcription+of+the+c-fos+proto-oncogene.">Stimulation of 3T3 cells induces transcription of the c-fos proto-oncogene.</a> Nature. 311, 433-438 (1984).</li><li>Greenberg, M. E., Greene, L. A., Ziff, E. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Nerve+growth+factor+and+epidermal+growth+factor+induce+rapid+transient+changes+in+proto-oncogene+transcription+in+PC12+cells.">Nerve growth factor and epidermal growth factor induce rapid transient changes in proto-oncogene transcription in PC12 cells.</a> J Biol Chem. 260, 14101-14110 (1985).</li><li>Muller, R., Bravo, R., Burckhardt, J., Curran, T. <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+c-fos+gene+and+protein+by+growth+factors+precedes+activation+of+c-myc.">Induction of c-fos gene and protein by growth factors precedes activation of c-myc.</a> Nature. 312, 716-720 (1984).</li><li>Teppema, L. J., Berkenbosch, A., Veening, J. G., Olievier, C. N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hypercapnia+induces+c-fos+expression+in+neurons+of+retrotrapezoid+nucleus+in+cats.">Hypercapnia induces c-fos expression in neurons of retrotrapezoid nucleus in cats.</a> Brain Res. 635, 353-356 (1994).</li><li>Teppema, L. 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=Expression+of+c-fos+in+the+rat+brainstem+after+exposure+to+hypoxia+and+to+normoxic+and+hyperoxic+hypercapnia.">Expression of c-fos in the rat brainstem after exposure to hypoxia and to normoxic and hyperoxic hypercapnia.</a> J Comp Neurol. 388, 169-190 (1997).</li><li>Larnicol, N., Wallois, F., Berquin, P., Gros, F., Rose, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=c-fos-like+immunoreactivity+in+the+cat's+neuraxis+following+moderate+hypoxia+or+hypercapnia.">c-fos-like immunoreactivity in the cat's neuraxis following moderate hypoxia or hypercapnia.</a> J Physiol Paris. 88, 81-88 (1994).</li><li>Bodineau, L., Larnicol, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Brainstem+and+hypothalamic+areas+activated+by+tissue+hypoxia:+Fos-like+immunoreactivity+induced+by+carbon+monoxide+inhalation+in+the+rat.">Brainstem and hypothalamic areas activated by tissue hypoxia: Fos-like immunoreactivity induced by carbon monoxide inhalation in the rat.</a> Neuroscience. 108, 643-653 (2001).</li><li>Erickson, J. T., Millhorn, D. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hypoxia+and+electrical+stimulation+of+the+carotid+sinus+nerve+induce+Fos-like+immunoreactivity+within+catecholaminergic+and+serotoninergic+neurons+of+the+rat+brainstem.">Hypoxia and electrical stimulation of the carotid sinus nerve induce Fos-like immunoreactivity within catecholaminergic and serotoninergic neurons of the rat brainstem.</a> J Comp Neurol. 348, 161-182 (1994).</li><li>Bodineau, L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Consequences+of+in+utero+caffeine+exposure+on+respiratory+output+in+normoxic+and+hypoxic+conditions+and+related+changes+of+Fos+expression:+a+study+on+brainstem-spinal+cord+preparations+isolated+from+newborn+rats.">Consequences of in utero caffeine exposure on respiratory output in normoxic and hypoxic conditions and related changes of Fos expression: a study on brainstem-spinal cord preparations isolated from newborn rats.</a> Pediatr Res. 53, 266-273 (2003).</li><li>Voituron, N., Frugiere, A., Gros, F., Macron, J. M., Bodineau, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Diencephalic+and+mesencephalic+influences+on+ponto-medullary+respiratory+control+in+normoxic+and+hypoxic+conditions:+an+in+vitro+study+on+central+nervous+system+preparations+from+newborn+rat.">Diencephalic and mesencephalic influences on ponto-medullary respiratory control in normoxic and hypoxic conditions: an in vitro study on central nervous system preparations from newborn rat.</a> Neuroscience. 132, 843-854 (2005).</li><li>Voituron, N., Frugiere, A., Champagnat, J., Bodineau, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hypoxia-sensing+properties+of+the+newborn+rat+ventral+medullary+surface+in+vitro.">Hypoxia-sensing properties of the newborn rat ventral medullary surface in vitro.</a> J Physiol. 577, 55-68 (2006).</li><li>Voituron, N., 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+kreisler+mutation+leads+to+the+loss+of+intrinsically+hypoxia-activated+spots+in+the+region+of+the+retrotrapezoid+nucleus/parafacial+respiratory+group.">The kreisler mutation leads to the loss of intrinsically hypoxia-activated spots in the region of the retrotrapezoid nucleus/parafacial respiratory group.</a> Neuroscience. 194, 95-111 (2011).</li><li>Suzue, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Respiratory+rhythm+generation+in+the+in+vitro+brain+stem-spinal+cord+preparation+of+the+neonatal+rat.">Respiratory rhythm generation in the in vitro brain stem-spinal cord preparation of the neonatal rat.</a> J Physiol. 354, 173-183 (1984).</li><li>Gage, G. J., Kipke, D. R., Shain, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Whole+animal+perfusion+fixation+for+rodents.">Whole animal perfusion fixation for rodents.</a> J Vis Exp. , e3564 (2012).</li><li>Rousseau, J. P., Caravagna, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Electrophysiology+on+isolated+brainstem-spinal+cord+preparations+from+newborn+rodents+allows+neural+respiratory+network+output+recording.">Electrophysiology on isolated brainstem-spinal cord preparations from newborn rodents allows neural respiratory network output recording.</a> J Vis Exp. , e53071 (2015).</li><li>Start, R. D., Layton, C. M., Cross, S. S., Smith, 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=Reassessment+of+the+rate+of+fixative+diffusion.">Reassessment of the rate of fixative diffusion.</a> J Clin Pathol. 45, 1120-1121 (1992).</li><li>Paxinos, G., Watson, C. <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 rat brain in stereotaxic coordinates. , (1998).</li><li>Paxinos, G., Franklin, K. B. <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 stereotaxic coordinates. , (2001).</li><li>Berquin, P., Bodineau, L., Gros, F., Larnicol, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Brainstem+and+hypothalamic+areas+involved+in+respiratory+chemoreflexes:+a+Fos+study+in+adult+rats.">Brainstem and hypothalamic areas involved in respiratory chemoreflexes: a Fos study in adult rats.</a> Brain Res. 857, 30-40 (2000).</li><li>Berquin, P., Cayetanot, F., Gros, F., Larnicol, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Postnatal+changes+in+Fos-like+immunoreactivity+evoked+by+hypoxia+in+the+rat+brainstem+and+hypothalamus.">Postnatal changes in Fos-like immunoreactivity evoked by hypoxia in the rat brainstem and hypothalamus.</a> Brain Res. 877, 149-159 (2000).</li><li>Bodineau, L., Cayetanot, F., Frugiere, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Fos+study+of+ponto-medullary+areas+involved+in+the+in+vitro+hypoxic+respiratory+depression.">Fos study of ponto-medullary areas involved in the in vitro hypoxic respiratory depression.</a> Neuroreport. 12, 3913-3916 (2001).</li><li>Takakura, A. C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Peripheral+chemoreceptor+inputs+to+retrotrapezoid+nucleus+(RTN)+CO2-sensitive+neurons+in+rats.">Peripheral chemoreceptor inputs to retrotrapezoid nucleus (RTN) CO2-sensitive neurons in rats.</a> J Physiol. 572, 503-523 (2006).</li><li>Mulkey, D. K., 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=Respiratory+control+by+ventral+surface+chemoreceptor+neurons+in+rats.">Respiratory control by ventral surface chemoreceptor neurons in rats.</a> Nat Neurosci. 7, 1360-1369 (2004).</li><li>Finley, J. C., Katz, D. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+central+organization+of+carotid+body+afferent+projections+to+the+brainstem+of+the+rat.">The central organization of carotid body afferent projections to the brainstem of the rat.</a> Brain Res. 572, 108-116 (1992).</li><li>Bodineau, L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Data+supporting+a+new+physiological+role+for+brain+apelin+in+the+regulation+of+hypothalamic+oxytocin+neurons+in+lactating+rats.">Data supporting a new physiological role for brain apelin in the regulation of hypothalamic oxytocin neurons in lactating rats.</a> Endocrinology. 152, 3492-3503 (2011).</li><li>Okada, Y., Chen, Z., Jiang, W., Kuwana, S., Eldridge, 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=Anatomical+arrangement+of+hypercapnia-activated+cells+in+the+superficial+ventral+medulla+of+rats.">Anatomical arrangement of hypercapnia-activated cells in the superficial ventral medulla of rats.</a> J Appl Physiol (1985). 93, 427-439 (2002).</li><li>Saadani-Makki, F., Frugiere, A., Gros, F., Gaytan, S., Bodineau, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Involvement+of+adenosinergic+A1+systems+in+the+occurrence+of+respiratory+perturbations+encountered+in+newborns+following+an+in+utero+caffeine+exposure.+a+study+on+brainstem-spinal+cord+preparations+isolated+from+newborn+rats.">Involvement of adenosinergic A1 systems in the occurrence of respiratory perturbations encountered in newborns following an in utero caffeine exposure. a study on brainstem-spinal cord preparations isolated from newborn rats.</a> Neuroscience. 127, 505-518 (2004).</li><li>Morgan, J. I., Cohen, D. R., Hempstead, J. L., Curran, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mapping+patterns+of+c-fos+expression+in+the+central+nervous+system+after+seizure.">Mapping patterns of c-fos expression in the central nervous system after seizure.</a> Science. 237, 192-197 (1987).</li><li>Sagar, S. M., Sharp, F. R., Curran, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Expression+of+c-fos+protein+in+brain:+metabolic+mapping+at+the+cellular+level.">Expression of c-fos protein in brain: metabolic mapping at the cellular level.</a> Science. 240, 1328-1331 (1988).</li><li>Herdegen, T., Kovary, K., Leah, J., Bravo, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Specific+temporal+and+spatial+distribution+of+JUN,+FOS,+and+KROX-24+proteins+in+spinal+neurons+following+noxious+transsynaptic+stimulation.">Specific temporal and spatial distribution of JUN, FOS, and KROX-24 proteins in spinal neurons following noxious transsynaptic stimulation.</a> J Comp Neurol. 313, 178-191 (1991).</li><li>Marina, N., Morales, T., Diaz, N., Mena, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Suckling-induced+activation+of+neural+c-fos+expression+at+lower+thoracic+rat+spinal+cord+segments.">Suckling-induced activation of neural c-fos expression at lower thoracic rat spinal cord segments.</a> Brain Res. 954, 100-114 (2002).</li></ol>

C-FOSの前初期遺伝子であり、その生成物の検出は、C-FOSタンパク質は、古典的なインビボ 11,13,25,28 内の特定の呼吸応答に関与する神経細胞集団を同定するために使用され、 エクスビボで 16-18、 27,32,33。 議定書の中で重要なステップ 灌流工程​​の間には注意してください。 4％PF...

C-fosの遺伝子は、1980 1,2の初めに初めて同定し、その産物は、遺伝子活性化剤の特性3,4を有する核タンパク質として1984年に特徴づけられました。これは、ニューロンの刺激に関連した長期的なメカニズムに参加しています。実際に、ニューロンの活性の変化は、転写因子のc-FOSの産生を誘導する前初期遺伝子c-FOSの発現を誘導する二次メッセンジャーシグナル伝達カスケードにつながります。後者は、後期遺伝子の発現を開始し、したがって、刺激4多くの異なる種類の神経系の適応応答に関与しています。したがって、1980 5,6の端部から、C-FOSタンパク質の検出は、しばしば、ニューロン経路をマッピングするための一般的な4および中枢神経系（CNS）の活性に遺伝子転写の外因性因子の効果を研究するために使用されています異なる生理学的に関与アル条件。 基底のc-fosの発現は、マウス、ラット、ネコ、サル、およびヒトの4を含む様々な種において研究されています。これにより、その発現の動態は比較的よく知られています。転写活性化は、（5〜20分）7,8迅速であり、かつmRNA蓄積を刺激9の発症後30〜45分の間で最大に達し、12分の短い半減期で低下します。 C-FOSタンパク質合成は、mRNAの蓄積に続く、20〜90分後に刺激6で免疫組織化学によって検出することができました。 c-fosの発現の分析は、古典的低酸素症または高炭酸ガス10-14への換気応答に関与する中心的な呼吸ネットワークを識別するために、in vivo試験で使用されています。最近では、このツールはまた、低酸素症またはhに中央呼吸ネットワーク適応を探索するためにex vivoで脳幹の製剤で使用されましたypercapnia 15-18。確かに、これらの製剤は、古典的に中央呼吸ドライブ19に同化リズミカルな活動を生成します。従って、製剤のこのタイプは、完全に求心路遮断であるという利点を有し、従って、C-fosの発現についての結果は、周辺構造の介入なしに、中央の刺激の結果を反映します。 C-FOSの検出は、免疫組織化学的または免疫組織蛍光アプローチによって行うことができます。間接的な免疫は、c-FOSに対する一次抗体と一次抗体が産生された種に対して指向二次抗体の使用を必要とします。免疫組織化学法のために、二次抗体は、基質（ペルオキシダーゼH 2 O 2）に作用する酵素（例えばペルオキシダーゼ）にコンジュゲートされています。酵素反応の生成物は、色原体（3.3ジアミノベンジジンtetrahydrochloridによって開発されていますE）、それを染色し、光学顕微鏡下で観察することができます。反応は、ニッケル硫酸アンモニウムを用いて強化することができます。これらの方法は、異なる生理学的な課題中の活性物質の神経細胞の検出、したがって、同定および/または連続した生理的応答に関与末梢および中枢経路のマッピングを可能にします。

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			<td height="21" style="height:21px;width:216px;">Cell culture plate 12-Well</td>
			<td style="width:71px;">Costa</td>
			<td style="width:119px;">35/3</td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="106">
			<td height="106" style="height:107px;width:216px;">15 mm Netwell inserts with mesh polyester membrane</td>
			<td style="width:71px;">Corning</td>
			<td style="width:119px;">3477</td>
			<td style="width:167px;">The 15mm diameter well inserts have 74&#181;m polyester mesh bottoms attached to polystyrene inserts</td>
		</tr>
		<tr height="85">
			<td height="85" style="height:85px;width:216px;">primary antibody (rabbit polyclonal antibody against the c-Fos protein)</td>
			<td style="width:71px;">Santa Cruz Biotechnology</td>
			<td style="width:119px;">sc-52</td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:216px;">Vectastain Elite ABC KIT&#160;</td>
			<td rowspan="2" style="width:71px;">Vector laboratories</td>
			<td rowspan="2" style="width:119px;">PK-6101</td>
			<td rowspan="2" style="width:167px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">(Rabbit IgG-secondary antibody)</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">NaH2PO4*2H2O</td>
			<td style="width:71px;">Sigma</td>
			<td style="width:119px;">71505</td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="21">
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			<td height="21" style="height:21px;width:216px;">Xylene</td>
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</table>

the c-fos protein immunohistological detection: a useful tool as a marker of central pathways involved in specific physiological responses in vivo and ex vivo

多くの研究は、特定の生理的な規制に関与する脳領域を特定し、マップしようとしています。癌原遺伝子C-FOS、前初期遺伝子は、種々の刺激に応答してニューロンで発現されます。タンパク質生成物は、容易にそれらの活性の変化を表示するニューロンのグループをマップするC-FOSの検出の使用につながる、免疫組織化学的技術を用いて検出することができます。この記事では、低酸素症または高炭酸ガスに換気の適応に関与脳幹ニューロン集団の同定に焦点を当てました。 2つのアプローチが、動物の生体内で関与する神経細胞集団を同定することが記載され、ex vivoでの求心路遮断脳幹準備インチインビボ 、動物は。高炭酸ガスまたは低酸素ガス混合物にさらされたex vivoで 、求心路遮断準備が低酸素や高炭酸ガス人工脳脊髄液で灌流しました。両方の場合において、いずれかの制御インビボ動物またはE でX vivoでの製剤は、正常酸素とnormocapnic条件下で維持しました。これら二つのアプローチの比較は、インビボにおいて 、末梢および/ ​​または中枢ニューロンの活性化、すなわちの起源の決意を可能にし、 エクスビボ 、brainstemsを集め、固定し、切片にスライスしました。切片を調製した後、C-FOSタンパク質の免疫組織化学的検出は、低酸素または高炭酸ガス刺激によって活性化された細胞の脳幹のグループを識別するためになされました。標識された細胞は、脳幹の呼吸器の構造で計数しました。対照条件と比較して、低酸素症または高炭酸ガス血症は、c-FOSの数を増加させ、したがって、中央呼吸ドライブの適応に関与するニューロン経路の構成されているいくつかの特定の脳幹部位で細胞を標識しました。

c-FOSの検出は、 生体内 （ 図2A） における低酸素および高炭酸ガス血症として、またはこれらの条件のex vivo（ 図2B）を模倣する状況では、特定の条件下で活性化された細胞の識別グループを可能にする便利なツールです。 インビボ 、新生児、若いです、または成人の齧歯類は、ガス環境が継続的に正確に30〜180分<...

Watch this Scientific Journal Video about The c-FOS Protein Immunohistological Detection: A Useful Tool As a Marker of Central Pathways Involved in Specific Physiological Responses In Vivo and Ex Vivo

The c-FOS Protein Immunohistological Detection: A Useful Tool As a Marker of Central Pathways Involved in Specific Physiological Responses In Vivo and Ex Vivo

C-FOSタンパク質免疫組織学的検出：特定の生理反応に関与する中央経路のマーカーとして有用ツール<em&gt;インビボ</em&gt;と<em&gt; ex vivoで</em

Here, we present a protocol based on c-FOS protein immunohistological detection, a classical technique used for the identification of neuronal populations involved in specific physiological responses in vivo and ex vivo.

Many studies seek to identify and map the brain regions involved in specific physiological regulations. The proto-oncogene c-fos, an immediate early gene, ...

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Biology

The c-FOS Protein Immunohistological Detection: A Useful Tool As a Marker of Central Pathways Involved in Specific Physiological Responses In Vivo and Ex Vivo

Single Cell Electroporation in vivo within the Intact Developing Brain

無傷の開発脳内の生体内の単一細胞のエレクトロポレーション

Single-cell electroporation (SCE) is a specialized technique allowing the delivery of DNA or other macromolecules into individual cells within intact ...

Method for Culture of Early Chick Embryos ex vivo (New Culture)

Method for Culture of Early Chick Embryos ex vivo New Culture

初期のニワトリ胚をex vivo（新文化）の文化のための方法

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マウス網膜神経節細胞における遺伝子発現のための子宮内および生体外でのエレクトロポレーションで

The retina and its sole output neuron, the retinal ganglion cell (RGC), comprise an excellent model in which to examine biological questions such as cell ...

In-vivo Detection of Protein-protein Interactions on Micro-patterned Surfaces

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マイクロパターン化表面でのタンパク質間相互作用の in vivo検出

Unraveling the interaction network of molecules in-vivo is key to understanding the mechanisms that regulate cell function and metabolism. A multitude of ...

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Extensive research has provided ample evidences suggesting that protein folding in the cell is a co-translational process1-5. However, the exact pathway ...

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小説<em&gt;エクスビボ</emマウス胎児ハート&gt;文化法

Developmental studies in the mouse are hampered  by the inaccessibility of the embryo during gestation. Thus, protocols to  isolate and culture individual ...

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The folding and assembly of proteins is essential for protein function, the long-term health of the cell, and longevity of the organism. Historically, the ...

Chromatin Immunoprecipitation Assay for the Identification of Arabidopsis Protein-DNA Interactions In Vivo

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シロイヌナズナタンパク質-DNA相互作用の同定のための免疫沈降アッセイクロマチン<em&gt;インビボ</em

Intricate gene regulatory networks orchestrate biological processes and developmental transitions in plants. Selective transcriptional activation and ...

Measuring the Stiffness of Ex Vivo Mouse Aortas Using Atomic Force Microscopy

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の剛性を測定します<em&gt; ex vivoで</em原子間力顕微鏡を使用&gt;マウス大動脈

Arterial stiffening is a significant risk factor and biomarker for cardiovascular disease and a hallmark of aging. Atomic force microscopy (AFM) is a ...

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生体内でRb タンパク質 sumo 化ひと細胞の検出と解析

The post-translational modifications of proteins are critical for the proper regulation of intracellular signal transduction. Among these modifications, ...