Name: visualization of mitochondrial respiratory function using cytochrome c oxidase / succinate dehydrogenase (cox/sdh) double-labeling histochemistry
Uploaded: 2011-11-23T12:00:00
Description: Watch this Scientific Journal Video about Visualization of Mitochondrial Respiratory Function using Cytochrome C Oxidase / Succinate Dehydrogenase COX/SDH Double-labeling Histochemistry at JoVE.com

Работа выполнена при поддержке Национального института старения (AG04418), Национальный институт по злоупотреблению наркотиками, Национальный Институт Здоровья-Каролинского института Высшей Программа партнерства Каролинского института, шведского исследовательского совета, шведский Питание мозга, и шведский фонд мозга. Большое спасибо Маттиас Карлен и доктор Джузеппе Coppotelli для творческой поддержки с рисунком 1 и 2 соответственно; Карин Pernold для оказания технической помощи, а также д-ра. Барри Дж. Хоффер, Ларс Олсон, и Нильс-Горан Ларссон на протяжении большей полезные советы и обсуждения.

1. Ткань подготовки к cryosectioning <ol><li> Жертва животного или шейки дислокации или обезглавливание, в соответствии с имеющимися этических разрешения. </li><li> Быстро собирать тканей интерес (например,. Головной мозг), и быстро заморозить на сухом льду (ткани может потребовать замораживани?...

<ol>
	<li>Larsson, N. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Somatic+mitochondrial+DNA+mutations+in+mammalian+aging.">Somatic mitochondrial DNA mutations in mammalian aging.</a> Annu. Rev. Biochem. 79, 683-706 (2010).</li><li>Cottrell, 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=Role+of+mitochondrial+DNA+mutations+in+disease+and+aging.">Role of mitochondrial DNA mutations in disease and aging.</a> Ann. NY Acad. Sci. 908, 199-207 (2000).</li><li>Harman, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+biologic+clock:+the+mitochondria.">The biologic clock: the mitochondria.</a> J. Am. Geriatr. Soc. 20, 145-147 (1972).</li><li>Wallace, D. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mitochondrial+genetics+-+a+paradigm+for+aging+and+degenerative+diseases.">Mitochondrial genetics - a paradigm for aging and degenerative diseases.</a> Science. 256, 628-632 (1992).</li><li>Trifunovic, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Premature+ageing+in+mice+expressing+defective+mitochondrial+DNA+polymerase.">Premature ageing in mice expressing defective mitochondrial DNA polymerase.</a> Nature. 429, 417-423 (2004).</li><li>Ameur, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ultra-deep+sequencing+of+mouse+mitochondrial+DNA:+mutational+patterns+and+their+origins.">Ultra-deep sequencing of mouse mitochondrial DNA: mutational patterns and their origins.</a> PLoS Genet. 7, e1002028-e1002028 (2011).</li><li>Safdar, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Endurance+exercise+rescues+progeroid+aging+and+induces+systemic+mitochondrial+rejuvenation+in+mtDNA+mutator+mice.">Endurance exercise rescues progeroid aging and induces systemic mitochondrial rejuvenation in mtDNA mutator mice.</a> Proc. Natl. Acad. Sci. U. S. A. 108, 4135-4140 (2011).</li><li>DiMauro, S., Bonilla, E., Zeviani, M., Nakagawa, M., DeVivo, D. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mitochondrial+myopathies.">Mitochondrial myopathies.</a> Ann. Neurol. 17, 521-538 (1985).</li><li>Old, S. L., Johnson, M. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Methods+of+microphotometric+assay+of+succinate+dehydrogenase+and+cytochrome+c+oxidase+activities+for+use+on+human+skeletal+muscle.">Methods of microphotometric assay of succinate dehydrogenase and cytochrome c oxidase activities for use on human skeletal muscle.</a> Histochem. J. 21, 545-555 (1989).</li><li>Chaturvedi, R. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Impaired+PGC-1alpha+function+in+muscle+in+Huntington's+disease.">Impaired PGC-1alpha function in muscle in Huntington's disease.</a> Hum. Mol. Genet. 18, 3048-3065 (2009).</li><li>Edgar, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Random+point+mutations+with+major+effects+on+protein-coding+genes+are+the+driving+force+behind+premature+aging+in+mtDNA+mutator+mice.">Random point mutations with major effects on protein-coding genes are the driving force behind premature aging in mtDNA mutator mice.</a> Cell. Metab. 10, 131-138 (2009).</li><li>Ross, J. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=High+brain+lactate+is+a+hallmark+of+aging+and+caused+by+a+shift+in+the+lactate+dehydrogenase+A/B+ratio.">High brain lactate is a hallmark of aging and caused by a shift in the lactate dehydrogenase A/B ratio.</a> Proc. Natl. Acad. Sci. U. S. A. 107, 20087-20092 (2010).</li><li>Crugnola, V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mitochondrial+respiratory+chain+dysfunction+in+muscle+from+patients+with+amyotrophic+lateral+sclerosis.">Mitochondrial respiratory chain dysfunction in muscle from patients with amyotrophic lateral sclerosis.</a> Arch. Neurol. 67, 849-854 (2010).</li><li>Nonaka, I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Muscle+pathology+in+cytochrome+c+oxidase+deficiency.">Muscle pathology in cytochrome c oxidase deficiency.</a> Acta. Neuropathol. 77, 152-160 (1988).</li><li>DiMauro, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mitochondrial+encephalomyopathies.">Mitochondrial encephalomyopathies.</a> Neurol. Clin. 8, 483-506 (1990).</li><li>Bonilla, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=New+morphological+approaches+to+the+study+of+mitochondrial+encephalomyopathies.">New morphological approaches to the study of mitochondrial encephalomyopathies.</a> Brain. Pathol. 2, 113-119 (1992).</li><li>Brierley, E. J., Johnson, M. A., Lightowlers, R. N., James, O. F., Turnbull, 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=Role+of+mitochondrial+DNA+mutations+in+human+aging:+implications+for+the+central+nervous+system+and+muscle.">Role of mitochondrial DNA mutations in human aging: implications for the central nervous system and muscle.</a> Ann. Neurol. 43, 217-223 (1998).</li><li>Borthwick, G. M., Johnson, M. A., Ince, P. G., Shaw, P. J., Turnbull, 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=Mitochondrial+enzyme+activity+in+amyotrophic+lateral+sclerosis:+implications+for+the+role+of+mitochondria+in+neuronal+cell+death.">Mitochondrial enzyme activity in amyotrophic lateral sclerosis: implications for the role of mitochondria in neuronal cell death.</a> Ann. Neurol. 46, 787-790 (1999).</li><li>Gellerich, F. N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mitochondrial+respiratory+rates+and+activities+of+respiratory+chain+complexes+correlate+linearly+with+heteroplasmy+of+deleted+mtDNA+without+threshold+and+independently+of+deletion+size.">Mitochondrial respiratory rates and activities of respiratory chain complexes correlate linearly with heteroplasmy of deleted mtDNA without threshold and independently of deletion size.</a> Biochim. Biophys. Acta. 1556, 41-52 (2002).</li><li>Larsson, N. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mitochondrial+transcription+factor+A+is+necessary+for+mtDNA+maintenance+and+embryogenesis+in+mice.">Mitochondrial transcription factor A is necessary for mtDNA maintenance and embryogenesis in mice.</a> Nat. Genet. 18, 231-236 (1998).</li><li>Ekstrand, M. I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Progressive+parkinsonism+in+mice+with+respiratory-chain-deficient+dopamine+neurons.">Progressive parkinsonism in mice with respiratory-chain-deficient dopamine neurons.</a> Proc. Natl. Acad. Sci. U. S. A. 104, 1325-1330 (2007).</li><li>Seligman, A. M., Karnovsky, M. J., Wasserkrug, H. L., Hanker, 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=Nondroplet+ultrastructural+demonstration+of+cytochrome+oxidase+activity+with+a+polymerizing+osmiophilic+reagent,+diaminobenzidine+(DAB).">Nondroplet ultrastructural demonstration of cytochrome oxidase activity with a polymerizing osmiophilic reagent, diaminobenzidine (DAB).</a> J. Cell. Biol. 38, 1-14 (1968).</li><li>Dubowitz, V., Brooke, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Muscle+Biopsy:+A+Modern+Approach.">Muscle Biopsy: A Modern Approach.</a> , (1973).</li><li>Cottrell, 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=Cytochrome+c+oxidase+deficient+cells+accumulate+in+the+hippocampus+and+choroid+plexus+with+age.">Cytochrome c oxidase deficient cells accumulate in the hippocampus and choroid plexus with age.</a> Neurobiol. Aging. 22, 265-272 (2001).</li><li>Blanco, C. E., Sieck, G. C., Edgerton, V. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Quantitative+histochemical+determination+of+succinic+dehydrogenase+activity+in+skeletal+muscle+fibres.">Quantitative histochemical determination of succinic dehydrogenase activity in skeletal muscle fibres.</a> Histochem. J. 20, 230-243 (1988).</li><li>Moraes, C. T., Ricci, E., Bonilla, E., DiMauro, S., Schon, E. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+mitochondrial+tRNA(Leu(UUR))+mutation+in+mitochondrial+encephalomyopathy,+lactic+acidosis,+and+strokelike+episodes+(MELAS):+genetic,+biochemical,+and+morphological+correlations+in+skeletal+muscle.">The mitochondrial tRNA(Leu(UUR)) mutation in mitochondrial encephalomyopathy, lactic acidosis, and strokelike episodes (MELAS): genetic, biochemical, and morphological correlations in skeletal muscle.</a> Am. J. Hum. Genet. 50, 934-949 (1992).</li><li>Petruzzella, V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Extremely+high+levels+of+mutant+mtDNAs+co-localize+with+cytochrome+c+oxidase-negative+ragged-red+fibers+in+patients+harboring+a+point+mutation+at+nt+3243.">Extremely high levels of mutant mtDNAs co-localize with cytochrome c oxidase-negative ragged-red fibers in patients harboring a point mutation at nt 3243.</a> Hum. Mol. Genet. 3, 449-454 (1994).</li><li>Tulinius, M. H., Holme, E., Kristiansson, B., Larsson, N. G., Oldfors, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mitochondrial+encephalomyopathies+in+childhood.+I.+Biochemical+and+morphologic+investigations.">Mitochondrial encephalomyopathies in childhood. I. Biochemical and morphologic investigations.</a> J. Pediatr. 119, 242-250 (1991).</li><li>Haas, R. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+in-depth+evaluation+of+suspected+mitochondrial+disease.">The in-depth evaluation of suspected mitochondrial disease.</a> Mol. Genet. Metab. 94, 16-37 (2008).</li></ol>

Комбинированный ЦОГ / SDH гистохимические метод позволяет визуализации клеток с митохондриальной дисфункцией. Этот метод, с ранних исследований, начиная с 1968 года, остается популярным, и многие считая его «золотым стандартом» для выявления митохондриальных заболеваний у пациентов, 1...

<table border="1"><tr><td> Название реагента </td><td> Компания </td><td> Номер по каталогу </td><td> Комментарии (необязательно) </td></tr><tr><td> Сухой лед </td><td> AGA AB газа </td><td> блочном виде </td><td></td></tr><tr><td> Изопентан (2-метилбутан) </td><td> Sigma-Aldrich </td><td> 277258 CAS: 78-78-4 </td><td></td></tr><tr><td> Cyrostat вложение решение </td><td> Сакура Finetek </td><td> Ткань Tek 4583 </td><td></td></tr><tr><td> Криостат </td><td> Microm </td><td> Microm Модель HM 500M </td><td></td></tr><tr><td> Слайды </td><td> Thermo Scientific </td><td> Супер Фрост Плюс Менцель Глезер J1800AMWZ </td><td></td></tr><tr><td> Обложка очки Боросиликатного стекла </td><td> VWR International </td><td> 16004-098 </td><td> 24 х 50 мм </td></tr><tr><td> Фильтровальная бумага </td><td> Munktell фильтра AB </td><td> Качество: 1350 СтатьяКоличество: 242 001 </td><td> 430 х 430 мм </td></tr><tr><td> 3,3 &#39;-диаминобензидина tetrahydrochloride (DAB) </td><td> Sigma-Aldrich </td><td> Sigma Система жидкостного субстрата, D7304 </td><td></td></tr><tr><td> Цитохром с (тип III, от лошадиного сердца) </td><td> Sigma-Aldrich </td><td> C2506 CAS: 9007-43-6 </td><td></td></tr><tr><td> Говядина каталазы (из печени) </td><td> Sigma-Aldrich </td><td> C9322 CAS: 9001-05-2 </td><td></td></tr><tr><td> Нитросинего тетразолия (NBT) </td><td> Sigma-Aldrich </td><td> N6876 CAS: 298-83-9 </td><td></td></tr><tr><td> Натрия сукцинат </td><td> Sigma-Aldrich </td><td> S2378 CAS: 6106-21-4 </td><td></td></tr><tr><td> Феназинметасульфат (ПМС) </td><td> Sigma-Aldrich </td><td> P9625 CAS: 299-11-6 </td><td> ПМС светочувствительным. Щит от света месте. </td></tr><tr><td> Азид натрия </td><td> Sigma-Aldrich </td><td> S8032 CAS: 26628-22-8 </td><td></td></tr><tг&gt; <td> Ксилол </td><td> VWR International </td><td> ЭМ-XX0060-4 </td><td></td></tr><tr><td> Entellan </td><td> VWR International </td><td> 100503-870 </td><td></td></tr><tr><td> Малонат (Малоновая кислоты) </td><td> Sigma-Aldrich </td><td> M1296 CAS: 141-82-2 </td><td></td></tr></table>

visualization of mitochondrial respiratory function using cytochrome c oxidase / succinate dehydrogenase (cox/sdh) double-labeling histochemistry

Mitochondrial DNA (mtDNA) defects are an important cause of disease and may underlie aging and aging-related alterations 1,2. The mitochondrial theory of aging suggests a role for mtDNA mutations, which can alter bioenergetics homeostasis and cellular function, in the aging process 3. A wealth of evidence has been compiled in support of this theory 1,4, an example being the mtDNA mutator mouse 5; however, the precise role of mtDNA damage in aging is not entirely understood 6,7.
Observing the activity of respiratory enzymes is a straightforward approach for investigating mitochondrial dysfunction. Complex IV, or cytochrome c oxidase (COX), is essential for mitochondrial function. The catalytic subunits of COX are encoded by mtDNA and are essential for assembly of the complex (Figure 1). Thus, proper synthesis and function are largely based on mtDNA integrity 2. Although other respiratory complexes could be investigated, Complexes IV and II are the most amenable to histochemical examination 8,9. Complex II, or succinate dehydrogenase (SDH), is entirely encoded by nuclear DNA (Figure 1), and its activity is typically not affected by impaired mtDNA, although an increase might indicate mitochondrial biogenesis 10-12. The impaired mtDNA observed in mitochondrial diseases, aging, and age-related diseases often leads to the presence of cells with low or absent COX activity 2,12-14. Although COX and SDH activities can be investigated individually, the sequential double-labeling method 15,16 has proved to be advantageous in locating cells with mitochondrial dysfunction 12,17-21.
Many of the optimal constitutions of the assay have been determined, such as substrate concentration, electron acceptors/donors, intermediate electron carriers, influence of pH, and reaction time 9,22,23. 3,3'-diaminobenzidine (DAB) is an effective and reliable electron donor 22. In cells with functioning COX, the brown indamine polymer product will localize in mitochondrial cristae and saturate cells 22. Those cells with dysfunctional COX will therefore not be saturated by the DAB product, allowing for the visualization of SDH activity by reduction of nitroblue tetrazolium (NBT), an electron acceptor, to a blue formazan end product 9,24. Cytochrome c and sodium succinate substrates are added to normalize endogenous levels between control and diseased/mutant tissues 9. Catalase is added as a precaution to avoid possible contaminating reactions from peroxidase activity 9,22. Phenazine methosulfate (PMS), an intermediate electron carrier, is used in conjunction with sodium azide, a respiratory chain inhibitor, to increase the formation of the final reaction products 9,25. Despite this information, some critical details affecting the result of this seemly straightforward assay, in addition to specificity controls and advances in the technique, have not yet been presented.

Watch this Scientific Journal Video about Visualization of Mitochondrial Respiratory Function using Cytochrome C Oxidase / Succinate Dehydrogenase COX/SDH Double-labeling Histochemistry at JoVE.com

Visualization of Mitochondrial Respiratory Function using Cytochrome C Oxidase / Succinate Dehydrogenase COX/SDH Double-labeling Histochemistry

Цитохром с оксидазы / натрия дегидрогеназы (ЦОГ / SDH) двойной маркировки метод позволяет для прямой визуализации митохондриальных дыхательных ферментов недостатки в свежезамороженных срезах тканей. Это просто гистохимические техники и полезен в расследовании митохондриальных болезней, старения и связанных со старением заболеваний.

Визуализация митохондриальной дыхательной функции использованием цитохрома C Оксидазы / сукцинатдегидрогеназы (ЦОГ / SDH) Дважды маркировки гистохимии

Mitochondrial DNA (mtDNA) defects are an important cause of disease and may underlie aging and aging-related alterations 1,2. The mitochondrial theory of ...

Research

JoVE Journal

Biology

Visualization of Mitochondrial Respiratory Function using Cytochrome C Oxidase / Succinate Dehydrogenase (COX/SDH) Double-labeling Histochemistry

Generation of Stable Transgenic C. elegans Using Microinjection

Генерация Стабильный Трансгенные Элеганс C. Использование Микроинъекция

Transgenic Caenorhabditis elegans can be readily created via microinjection of a DNA plasmid solution into the gonad 1.  The plasmid DNA rearranges to ...

Activation of Apoptosis by Cytoplasmic Microinjection of Cytochrome c

Activation of Apoptosis by Cytoplasmic Microinjection of Cytochrome c

Активация апоптоза Цитоплазматическая Микроинъекция цитохрома С</em

Apoptosis, or programmed cell death, is a conserved and highly regulated pathway by which cells die1.  Apoptosis can be triggered when cells encounter a ...

The ex vivo Isolated Skeletal Microvessel Preparation for Investigation of Vascular Reactivity

The ex vivo Isolated Skeletal Microvessel Preparation for Investigation of Vascular Reactivity

Бывший естественных условиях Изолированный скелета Подготовка микрососудов для исследования сосудистой реактивности

The isolated  microvessel preparation is an ex vivo preparation that allows for examination of the different contributions of  factors that control vessel ...

Assessment of Mitochondrial Functions and Cell Viability in Renal Cells Overexpressing Protein Kinase C Isozymes

Оценка митохондриальной функции и жизнеспособности клеток в клетках почек гиперэкспрессией протеинкиназы С Изоферменты

The protein kinase C (PKC) family of isozymes is involved in numerous physiological and pathological processes. Our recent data demonstrate that PKC ...

Analysis of Gene Function and Visualization of Cilia-Generated Fluid Flow in Kupffer's Vesicle

Анализ функций генов и визуализации реснички, созданного потока жидкости в Vesicle Купфера

Internal organs such as the  heart, brain, and gut develop left-right (LR) asymmetries that are critical for  their normal functions1. Motile cilia are ...

Visualization of Craniofacial Development in the sox10: kaede Transgenic Zebrafish Line Using Time-lapse Confocal Microscopy

Визуализация черепно-лицевого развития в Sox10: Kaede Трансгенные данио рерио Линия Использование Покадровый конфокальной микроскопии

Vertebrate palatogenesis is a highly choreographed and complex developmental process, which involves migration of cranial neural crest (CNC) cells, ...

Large-scale Gene Knockdown in C. elegans Using dsRNA Feeding Libraries to Generate Robust Loss-of-function Phenotypes

Large-scale Gene Knockdown in C. elegans Using dsRNA Feeding Libraries to Generate Robust Loss-of-function Phenotypes

Крупномасштабная Нокдаун гена в<em&gt; С. Элеганс</em&gt; Использование дцРНК кормления библиотек для генерации Прочные с потерей функции фенотипов

RNA interference by feeding worms bacteria expressing dsRNAs has been a useful tool to assess gene function in C. elegans. While this strategy works well ...

Measuring Respiratory Function in Mice Using Unrestrained Whole-body Plethysmography

Измерение функции внешнего дыхания у мышей, используя безудержного плетизмографии всего тела

Respiratory dysfunction is one of the leading causes of morbidity and mortality in the world and the rates of mortality continue to rise. Quantitative ...

Experimental Approaches to Study Mitochondrial Localization and Function of a Nuclear Cell Cycle Kinase, Cdk1

Экспериментальные подходы к изучению Митохондриальная локализации и функции цикла ядерных клеток киназа, CDK1

Although mitochondria possess their own transcriptional machinery, merely 1% of mitochondrial proteins are synthesized inside the organelle. The ...

High-resolution Respirometry to Assess Mitochondrial Function in Permeabilized and Intact Cells

Высокое разрешение респирометрии для оценки митохондриальной функции в проницаемыми и неповрежденных клеток

A high-resolution oxygraph is a device for measuring cellular oxygen consumption in a closed-chamber system with very high resolution and sensitivity in ...