We would like to thank Dr. Harry Orr at the University of Minnesota for his generous gift of transgenic mice. We would also like to thank the following Skidmore College alum for their work performing the preceding experiments: Monica Villegas, Porter Hall, Mitchell Spring, Nicholas Toker, Jenny Zhang, Chloe Larson and Cheyanne Slocum. Furthermore, we would like to thank Skidmore College for funding the development of these methods.

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저자는 공개 아무것도 없어.

설명 된 바와 같이 이러한 방법을 사용하는 경우, 이들은 검출 및 소뇌 신경 퇴행성 질환의 마우스 모델에서 산화 적 인산화 매개 미토콘드리아 기능 부전을 경감 할 수있다. 결합 된 생화학 적 및 행동 분석은 소뇌 신경 퇴행성 질환의 병리에 미토콘드리아 기여의 정도를 결정하기위한 다각적 인 방법입니다. 신진 대사를 자극하고 미토콘드리아 기능을 강화하기 위해 숙신산과 쥐를 치료함으로?...

미토콘드리아의 전자 전달계를 이용하여 산화 적 인산화 (OXPHOS)를 통해 생성 된 미토콘드리아 ATP의 대부분과, 아데노신 삼인산 (ATP), 세포 에너지의 필수 조효소의 주요 생산자이다. 뇌는 높은 신진 대사 요구와 신경 활동에 전력을 공급하기위한 산화 적 인산화에 대한 의존도를 부여, 미토콘드리아 기능 장애에 매우 민감하다. 그 결과, 미토콘드리아 장애는 노화 과정 트리거 (1) 및 다수의 신경 퇴행성 질환은 2, 3, 4의 병인에 연루되어있다. 따라서 미토콘드리아는 신경 퇴행에 대한 매력적인 치료 대상이되는 것을 다음과 같습니다. 이 프로토콜에서, 우리는 미토콘드리아의 연구를위한 모델 신경 퇴행성 질환과 같은 척수 소 뇌성 실조증 유형 1 (SCA1)의 사용을 채택리터 부전과 미토콘드리아 - 표적으로 한 치료의 개발. SCA1 다른 뇌 영역의 조롱박 소뇌의 뉴런과 뉴런의 진보적 인 변성을 트리거 ataxin-1 유전자 산물의 폴리 글루타민 (polyQ) 반복 확장 돌연변이에 의해 발생합니다. 조롱박 세포 특이 적 프로모터의 제어하에 polyQ 돌연변이 ataxin-1 유전자를 발현합니다 (SCA1 마우스로 지정) 여기서 사용 된 트랜스 제닉 마우스 선은 SCA1 5 조롱박 세포 성분의 표적 분석을 허용한다. SCA1 마우스는 점진적 조롱박 세포의 퇴화를 받아야하고 운동 실조증 보행 (6)을 개발한다. 미토콘드리아 복잡한 기능 장애 및 미토콘드리아 타겟 치료 효능은 분자 및 행동 분석의 배터리로 평가 될 수있다. 복잡한 미토콘드리아 장애는 소뇌 조직 안에서 변화된 산소 소비량을 검출 호흡 분석하여 생체 측정전자 전달계 기판 및 억제제 (7)의 존재. 호흡 분석은 이전에 투과성이 조직, 미토콘드리아 분리하고, 전체 조직 7, 8, 9로 사용되어왔다. 이들은 예컨대 투과 전자 현미경 또는 면역 형광 염색법과 같은 형태 학적 데이터 수집 방법과 달리 미토콘드리아 기능의 직접적인 평가를 허용한다. 전체 조직보다는 격리 미토콘드리아의 사용은 분리 과정 중에 발생할 수 7 건강한 미토콘드리아 바이어스 선택을 방지한다. 도시 된 바와 같이 프로토콜에 적응되면, 호흡 분석은 소뇌 신경 퇴행성 질환 상태에서 미토콘드리아 기능 장애를 검출하기위한 유용한 방법이다. 대사 비특이적 활성제 퇴행성 diseas의 형질 전환 마우스 모델에서 미토콘드리아 기능 부전을 추론하는데 사용될 수있다전자 및 새로운 치료법의 개발에 도움. 퀘르세틴 보효소 Q10과 크레아틴 모든 환자 및 신경 퇴행성 질환 10, 11, 12, 13, 14, 15, 16의 동물 모델에서 신경 퇴행성 질환의 병리를 개선하는 것으로 나타났다. 여기에서 우리는 신진 대사를 자극하고 신경 퇴행성 질환에서 미토콘드리아의 기능을 높일 수있는 새로운 신진 대사 활성화, 숙신산을 제시한다. 활성제가 혈액 뇌 장벽을 가로 지르는 것을 보장하기 위해, HPLC 처리 된 마우스는 17 신경 조직에 배달을 검출하도록 사용 하였다. 숙신산과 같은 대사 표적 수용성 화합물의 치료 효과를 평가하기 위해, 행동 패러다임과 immunopathological 연구 전지가 사용될 수있다. 뒤소뇌 신경 퇴행성 질환, 풋 프린트 활주로 분석, 빔 분석 및 가속 회전로드 분석에서 발견 된 모터 조정 적자에 전자는 행동 병리학 6, 18, 19의 구조를 검색하는 데 사용됩니다. 이러한 조치는 소뇌 조직 6, 20, 21의 정의 소엽 내에서 분자 (조롱박 세포 돌기의 아버 길이로 정의) 층의 두께 및 조롱박 세포 소마 수를 평가하여 소뇌 cytoarchitecture의 immunopathological 평가로 보충된다. 여기에서 우리는 대사 대상으로 수용성 화합물 검출 및 미토콘드리아 기능 장애의 치료를위한 여러 신경 병리학 및 행동 방법을 제시한다. 우리는 SCA1의 트란에 미토콘드리아 기능 장애를 분석하는 미토콘드리아 호흡의 생체 분석을 사용sgenic 마우스입니다. 더욱이, 우리는 질병 증상과 병리 더 SCA1 질병 진행에서 미토콘드리아 기능 부전을 연루 수용성 미토콘드리아 부스터 숙신산 향상되어 있음을 보여준다.

<table><tbody><tr><td>Adenosine diphosphate</td><td>Sigma Aldrich</td><td>A2754</td><td>ADP</td></tr><tr><td>Ascorbate</td><td>Sigma Aldrich</td><td>A7631</td><td></td></tr><tr><td>Bovine serum albumin</td><td>Sigma Aldrich</td><td>A2153</td><td>BSA</td></tr><tr><td>4&#039;,6-Diamidino-2-phenylindole</td><td>Sigma Aldrich</td><td>D9542</td><td>DAPI</td></tr><tr><td>Digitonin</td><td>Sigma Aldrich</td><td>D141</td><td></td></tr><tr><td>Dithiothreitol</td><td>Sigma Aldrich</td><td>D0632</td><td>DTT</td></tr><tr><td>Donkey serum</td><td>Sigma Aldrich</td><td>D9663</td><td></td></tr><tr><td>Glutamate</td><td>Sigma Aldrich</td><td>1446600</td><td></td></tr><tr><td>Malate</td><td>Sigma Aldrich</td><td>6994</td><td></td></tr><tr><td>Mannitol</td><td>Sigma Aldrich</td><td>M4125</td><td></td></tr><tr><td>Paraformaldehyde</td><td>Sigma Aldrich</td><td>P6148</td><td></td></tr><tr><td>Potassium-lactobionate</td><td>Bio-Sugars</td><td>69313-67-3</td><td></td></tr><tr><td>Rotenone</td><td>Sigma Aldrich</td><td>R8875</td><td></td></tr><tr><td>Saponin</td><td>Sigma Aldrich</td><td>47036</td><td></td></tr><tr><td>Succinic Acid</td><td>Sigma Aldrich</td><td>S3674</td><td></td></tr><tr><td>&lt;em&gt;N,N,N&amp;prime;,N&amp;prime;&lt;/em&gt;-Tetramethyl-&lt;em&gt;p&lt;/em&gt;-phenylenediamine</td><td>Sigma Aldrich</td><td>T7394</td><td>TMPD</td></tr><tr><td>Triton X-100</td><td>Sigma Aldrich</td><td>T9284</td><td></td></tr><tr><td>Urea</td><td>Sigma Aldrich</td><td>U0631</td><td></td></tr><tr><td>Vectashield mounting medium</td><td>Vector Labs</td><td>H-1000</td><td></td></tr><tr><td>&lt;strong&gt;Antibodies&lt;/strong&gt;</td><td></td><td></td><td></td></tr><tr><td>11NQ antibody (anti-ataxin-1 )</td><td></td><td></td><td>Servadio, &lt;i&gt;et al.&lt;/i&gt; 1995, PMID: 7647801</td></tr><tr><td>Alexa Fluor 488 anti-mouse secondary antibody</td><td>Life Technologies</td><td>A-11015</td><td></td></tr><tr><td>Alexa Fluor 594 anti-rabbit secondary antibody</td><td>Life Technologies</td><td>A-11012</td><td></td></tr><tr><td>Calbindin antibody (goat)</td><td>Santa Cruz</td><td>C-20</td><td></td></tr><tr><td>&lt;strong&gt;Animals&lt;/strong&gt;</td><td></td><td></td><td></td></tr><tr><td>Control transgenic mice</td><td>Harry Orr, Ph.D.</td><td>A02</td><td>Burright, &lt;i&gt;et al.&lt;/i&gt; 1997, PMID: 9217978</td></tr><tr><td>SCA1 mice</td><td>Harry Orr, Ph.D.</td><td>B05</td><td>Burright, &lt;i&gt;et al.&lt;/i&gt; 1997, PMID: 9217978</td></tr><tr><td>Wildtype mice</td><td>The Jackson Laboratory</td><td>001800</td><td></td></tr><tr><td>&lt;strong&gt;Equipment&lt;/strong&gt;</td><td></td><td></td><td></td></tr><tr><td>ESM-100L microtome</td><td>ERMA</td><td></td><td>Sledge microtome</td></tr><tr><td>Fluoview FV1200 Confocal Microscope</td><td>Olympus</td><td></td><td></td></tr><tr><td>Glycerol-gelatin slides</td><td>FD Neuro Technologies</td><td>PO101</td><td></td></tr><tr><td>Hamilton syringe</td><td>Sigma Aldrich</td><td>VCAT 80465</td><td></td></tr><tr><td>OXYT1 Oxytherm Electrode Control Unit</td><td>Hansatech Instruments</td><td></td><td></td></tr><tr><td>P.T.F.E. paper</td><td>Cole-Parmer</td><td>UX-08277-15</td><td></td></tr><tr><td>Rotallion Rotarod</td><td>PPP&amp;amp;G</td><td></td><td>contact corresponding author for information</td></tr><tr><td>Ultimate 3000 HPLC</td><td>Dionex</td><td></td><td></td></tr><tr><td>&lt;strong&gt;Software&lt;/strong&gt;</td><td></td><td></td><td></td></tr><tr><td>ImageJ</td><td>National Institute of Health</td><td></td><td>&lt;a href=&quot;http://imagej.nih.gov/ij/&quot;&gt;http://imagej.nih.gov/ij/&lt;/a&gt;</td></tr><tr><td>Cell counter plugin (for ImageJ)</td><td>National Institute of Health</td><td></td><td>&lt;a href=&quot;http://rsb.info.nih.gov/ij/plugins/cell-counter.html&quot;&gt;http://rsb.info.nih.gov/ij/plugins/cell-counter.html&lt;/a&gt;</td></tr><tr><td>3P&amp;amp;G Rota-Rod v3.3.3 (rotarod software)</td><td>PPP&amp;amp;G</td><td></td><td>contact corresponding author for information</td></tr><tr><td>Phidget21.dll (required for rotarod software)</td><td>DLL-Files.com</td><td></td><td>&lt;a href=&quot;https://www.dll-files.com/phidget21.dll.html&quot;&gt;https://www.dll-files.com/phidget21.dll.html&lt;/a&gt;</td></tr></tbody></table>

treating sca1 mice with water-soluble compounds to non-specifically boost mitochondrial function

미토콘드리아 기능 장애는 노화 과정에서 여러 유전 척수 소 뇌성 운동 실조증과 소뇌의 진보적 인 변성에 의해 표시 다른 운동 장애를 포함한 신경 퇴행성 질환에 중요한 역할을한다. 이 프로토콜의 목적은 척수 소 뇌성 실조증 유형 1 (SCA1)에서 미토콘드리아 기능 부전을 평가하고, 병의 진행을 느리게하는 수용성 화합물 숙신산 통해 호흡 대사 약물 표적의 효능을 평가하는 것이다. 이 접근은 다른 소뇌 질환에 적용 할 수있다 수용성 요법 숙주에 적응 될 수있다. 미토콘드리아 호흡의 생체 분석을 감지하고 미토콘드리아 기능에서 질병 관련 변경 사항을 정량화하는 데 사용됩니다. 유전 적 증거 (게시되지 않은 데이터)와 SCA1 마우스 모델에서 미토콘드리아 기능 장애의 단백체 증거, 우리는 수용성 대사 부스터의 치료의 효능을 평가홈 케이지 마시는 물에 직접 화합물을 용해하여 uccinic 산. 혈액 뇌 장벽을 통과하는 약물의 능력은 고성능 액체 크로마토 그래피 (HPLC)를 사용하여 추론 될 수있다. 이들 화합물의 효능은 다음 가속 로타로드 (rotarod), 밸런스 빔 테스트 및 공간 분석 등 다양한 행동 패러다임을 사용하여 테스트 할 수있다. 소뇌 Cytoarchitectural 무결성 조롱박 세포핵 및 조롱박 수지상 세포 및 소마 검출 면역 분석법을 이용하여 평가할 수있다. 이러한 방법은 미토콘드리아 장애와 소뇌 신경 퇴행성 질환의 수용성 화합물 치료의 효능을 결정하기위한 견고한 기술이다.

숙신산과 소뇌 미토콘드리아의 약물 학적 타겟팅을 통해 우리는 소뇌 신경 퇴행성 질환 SCA1의 마우스 모델에서 미토콘드리아 기능 장애를 방지 할 수 있습니다. 숙신산 탈수소 효소, 숙신산,의 표준 전자 공여체 치료 (그림 1A) 다음과 같은 치료 및 신경 병리학 적 평가의 두 번째 주 동안 행동 평가의 시작과 함께, 한 달 동안 SCA1 마우스의 홈 케이?...

Watch this Scientific Journal Video about Treating SCA1 Mice with Water-Soluble Compounds to Non-Specifically Boost Mitochondrial Function at JoVE.com

Treating SCA1 Mice with Water-Soluble Compounds to Non-Specifically Boost Mitochondrial Function

We present a biochemical and behavioral protocol to evaluate the efficacy of mitochondria-targeted water-soluble compounds for the treatment of Spinocerebellar ataxia type 1 (SCA1) and other cerebellar neurodegenerative diseases.

에 수용성 화합물로 SCA1 마우스 치료가 아닌 것은 - 특히 미토콘드리아 기능 향상

Mitochondrial dysfunction plays a significant role in the aging process and in neurodegenerative diseases including several hereditary spinocerebellar ...

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10.5K 조회수.  Skidmore College. We present a biochemical and behavioral protocol to evaluate the efficacy of mitochondria-targeted water-soluble compounds for the treatment of Spinocerebellar ataxia type 1 (SCA1) and other cerebellar neurodegenerative diseases. 

비디오: Treating SCA1 Mice with Water-Soluble Compounds to Non-Specifically Boost Mitochondrial Function

Experimental Protocol for Detecting Mitochondrial Function in Hepatocytes Exposed to Organochlorine Pesticides

This paper presents detailed methods on detecting hepatic mitochondrial function for a better understanding the cause of metabolic disorders caused by environmental organochlorine pesticides (OCPs) in hepatocytes. HepG2 cells were exposed to &#946;-hexachlorocyclohexane (&#946;-HCH) for 24 h at an equivalent dose of internal exposure in general population. Ultrastructure in hepatocytes was examined by transmission electron microscopy (TEM) to show the damage of mitochondria. Mitochondrial function was further evaluated by mitochondrial fluorescence intensity, adenosine 5'-triphosphate (ATP) levels, oxygen consumption rate (OCR) and mitochondrial membrane potential (MMP) in HepG2 cells incubated with &#946;-HCH. The mitochondria fluorescence intensity after stained by mitochondrial green fluorescent probe was observed with a fluorescence microscopy. The luciferin-luciferase reaction was used to determine ATP levels. The MMP was detected by the cationic dye JC-1 and analyzed under flow cytometry. OCR was measured with an extracellular flux analyzer. In summary, these protocols were used in detecting mitochondrial function in hepatocytes with to investigate mitochondria damages.

Understanding the influence of environmental organochlorine pesticides (OCPs) on mitochondrial function in hepatocytes is important in exploring the mechanism of OCPs causing metabolic disorders. This paper presents detailed methods on detecting hepatic mitochondrial function.

유기체 살충제에 노출된 간세포에서 미토콘드리아 기능을 검출하기 위한 실험 프로토콜

This paper presents detailed methods on detecting hepatic mitochondrial function for a better understanding the cause of metabolic disorders caused by ...

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환경과학

Application of Chronic Stimulation to Study Contractile Activity-induced Rat Skeletal Muscle Phenotypic Adaptations

Skeletal muscle is a highly adaptable tissue, as its biochemical and physiological properties are greatly altered in response to chronic exercise. To investigate the underlying mechanisms that bring about various muscle adaptations, a number of exercise protocols such as treadmill, wheel running, and swimming exercise have been used in the animal studies. However, these exercise models require a long period of time to achieve muscle adaptations, which may be also regulated by humoral or neurological factors, thus limiting their applications in studying the muscle-specific contraction-induced adaptations. Indirect low frequency stimulation (10 Hz) to induce chronic contractile activity (CCA) has been used as an alternative model for exercise training, as it can successfully lead to muscle mitochondrial adaptations within 7 days, independent of systemic factors. This paper details the surgical techniques required to apply the treatment of CCA to the skeletal muscle of rats, for widespread application in future studies.

This protocol describes the use of the chronic contractile activity model of exercise to observe stimulation-induced skeletal muscle adaptations in the rat hindlimb.

수축 성 활동 유도 된 쥐 골격 근육 Phenotypic 적응 연구 만성 자극에의 응용

Skeletal muscle is a highly adaptable tissue, as its biochemical and physiological properties are greatly altered in response to chronic exercise. To ...

발생학

Exploring Mitochondrial Energy Metabolism of Single 3D Microtissue Spheroids Using Extracellular Flux Analysis

Three-dimensional (3D) cellular aggregates, termed spheroids, have become the forefront of in vitro cell culture in recent years. In contrast to culturing cells as two-dimensional, single-cell monolayers (2D culture), spheroid cell culture promotes, regulates, and supports physiological cellular architecture and characteristics that exist in vivo, including the expression of extracellular matrix proteins, cell signaling, gene expression, protein production, differentiation, and proliferation. The importance of 3D culture has been recognized in many research fields, including oncology, diabetes, stem cell biology, and tissue engineering. Over the last decade, improved methods have been developed to produce spheroids and assess their metabolic function and fate.
Extracellular flux (XF) analyzers have been used to explore mitochondrial function in 3D microtissues such as spheroids using either an XF24 islet capture plate or an XFe96 spheroid microplate. However, distinct protocols and the optimization of probing mitochondrial energy metabolism in spheroids using XF technology have not been described in detail. This paper provides detailed protocols for probing mitochondrial energy metabolism in single 3D spheroids using spheroid microplates with the XFe96 XF analyzer. Using different cancer cell lines, XF technology is demonstrated to be capable of distinguishing between cellular respiration in 3D spheroids of not only different sizes but also different volumes, cell numbers,&#160;DNA content and type.
The optimal mitochondrial effector compound concentrations of oligomycin, BAM15, rotenone, and antimycin A are used to probe specific parameters of mitochondrial energy metabolism in 3D spheroids. This paper also discusses methods to normalize data obtained from spheroids and addresses many considerations that should be considered when exploring spheroid metabolism using XF technology. This protocol will help drive research in advanced in vitro spheroid models.

These protocols will help users probe mitochondrial energy metabolism in 3D cancer cell-line-derived spheroids using Seahorse extracellular flux analysis.

세포외 플럭스 분석을 이용한 단일 3D 미세조직 구상체의 미토콘드리아 에너지 대사 탐구

Three-dimensional (3D) cellular aggregates, termed spheroids, have become the forefront of in vitro cell culture in recent years. In contrast to culturing ...

암 연구학

Assessing Mitochondrial Function in Sciatic Nerve by High-Resolution Respirometry

Mitochondrial dysfunction in peripheral nerves accompanies several diseases associated with peripheral neuropathy, which can be triggered by multiple causes, including autoimmune diseases, diabetes, infections, inherited disorders, and tumors. Assessing mitochondrial function in mouse peripheral nerves can be challenging due to the small sample size, a limited number of mitochondria present in the tissue, and the presence of a myelin sheath. The technique described in this work minimizes these challenges by using a unique permeabilization protocol adapted from one used for muscle fibers, to assess sciatic nerve mitochondrial function instead of isolating the mitochondria from the tissue. By measuring fluorimetric reactive species production with Amplex Red/Peroxidase and comparing different mitochondrial substrates and inhibitors in saponin-permeabilized nerves, it was possible to detect mitochondrial respiratory states, reactive oxygen species (ROS), and the activity of mitochondrial complexes simultaneously. Therefore, the method presented here offers advantages compared to the assessment of mitochondrial function by other techniques.

High-resolution respirometry coupled to fluorescence sensors determines mitochondrial oxygen consumption and reactive oxygen species (ROS) generation. The present protocol describes a technique to assess mitochondrial respiratory rates and ROS production in the permeabilized sciatic nerve.

고해상도 호흡 측정법에 의한 좌골 신경의 미토콘드리아 기능 평가

Mitochondrial dysfunction in peripheral nerves accompanies several diseases associated with peripheral neuropathy, which can be triggered by multiple ...

생화학

DHODH 및 SDH 활성을 측정하기 위한 안정 동위원소 추적 및 LC-MS 분석

Oxygen-Independent Assays to Measure Mitochondrial Function in Mammals

The flow of electrons in the mitochondrial electron transport chain (ETC) supports multifaceted biosynthetic, bioenergetic, and signaling functions in mammalian cells. As oxygen (O2) is the most ubiquitous terminal electron acceptor for the mammalian ETC, the O2 consumption rate is frequently used as a proxy for mitochondrial function. However, emerging research demonstrates that this parameter is not always indicative of mitochondrial function, as fumarate can be employed as an alternative electron acceptor to sustain mitochondrial functions in hypoxia. This article compiles a series of protocols that allow researchers to measure mitochondrial function independently of the O2 consumption rate. These assays are particularly useful when studying mitochondrial function in hypoxic environments. Specifically, we describe methods to measure mitochondrial ATP production, de novo pyrimidine biosynthesis, NADH oxidation by complex I, and superoxide production. In combination with classical respirometry experiments, these orthogonal and economical assays will provide researchers with a more comprehensive assessment of mitochondrial function in their system of interest.

저자 스포트라이트: 포유류의 미토콘드리아 기능을 측정하기 위한 산소 독립적 분석  

Here, we present a compilation of assays to directly measure mitochondrial function in mammalian cells independently of their ability to consume molecular oxygen.

포유류의 미토콘드리아 기능을 측정하기 위한 산소 비의존적 분석

The flow of electrons in the mitochondrial electron transport chain (ETC) supports multifaceted biosynthetic, bioenergetic, and signaling functions in ...

Mitochondrial Transfer from Mouse Adipose-Derived Mesenchymal Stem Cells into Aged Mouse Oocytes

Due to the decline in the quantity and quality of oocytes related to age, the fertility of women over 35 years of age has declined sharply. The molecular mechanisms that maintain oocyte quality remain unclear, thus it is difficult to increase the birth rate of women over 35 years old at present. Oocytes contain more mitochondria than any type of cell in the body, and any mitochondrial dysfunction can lead to reduced oocyte quality. In the 1990s, oocyte cytoplasmic transfer resulted in great success in human reproduction but was accompanied by ethical controversies. Autologous mitochondrial transplantation is expected to be a useful technique to increase the quality of oocytes that have decreased due to age. In the present study, we used adipose-derived stem cells from aged mice as a mitochondria donor to increase the quality of oocytes of aged mice. Further development of autologous mitochondrial transfer technology will provide a new and effective treatment for infertility in aged women.

<meta charset="utf8"></head><body>마우스 지방 유래 중간엽 줄기 세포에서 노화된 마우스 난모세포로의 미토콘드리아 전달

Here, we describe the isolation of mitochondria from mouse adipose-derived mesenchymal stem cells, and then transfer the mitochondria into aged mouse oocytes to improve the quality of the oocytes.

마우스 지방 유래 중간엽 줄기 세포에서 노화된 마우스 난모세포로의 미토콘드리아 전달

Due to the decline in the quantity and quality of oocytes related to age, the fertility of women over 35 years of age has declined sharply. The molecular ...

Supercomplex 분석을 위한 작은 조직 샘플의 미토콘드리아 분획

Isolation of Mitochondria for Mitochondrial Supercomplex Analysis from Small Tissue and Cell Culture Samples

Over the last decades, the evidence accumulated about the existence of respiratory supercomplexes (SCs) has changed our understanding of the mitochondrial electron transport chain organization, giving rise to the proposal of the "plasticity model." This model postulates the coexistence of different proportions of SCs and complexes depending on the tissue or the cellular metabolic status. The dynamic nature of the assembly in SCs would allow cells to optimize the use of available fuels and the efficiency of electron transfer, minimizing reactive oxygen species generation and favoring the ability of cells to adapt to environmental changes. 
More recently, abnormalities in SC assembly have been reported in different diseases such as neurodegenerative disorders (Alzheimer's and Parkinson's disease), Barth Syndrome, Leigh syndrome, or cancer. The role of SC assembly alterations in disease progression still needs to be confirmed. Nevertheless, the availability of enough amounts of samples to determine the SC assembly status is often a challenge. This happens with biopsy or tissue samples that are small or have to be divided for multiple analyses, with cell cultures that have slow growth or come from microfluidic devices, with some primary cultures or rare cells, or when the effect of particular costly treatments has to be analyzed (with nanoparticles, very expensive compounds, etc.). In these cases, an efficient and easy-to-apply method is required. This paper presents a method adapted to obtain enriched mitochondrial fractions from small amounts of cells or tissues to analyze the structure and function of mitochondrial SCs by native electrophoresis followed by in-gel activity assays or western blot.

저자 스포트라이트: Unveiling Oxidative Phosphorylation System Dynamics and Mitochondrial Roles in Health and Disease

This protocol describes a technique for the analysis of respiratory supercomplexes when only small amounts of samples are available.

Isolation of Mitochondria for Mitochondrial Supercomplex Analysis from Small Tissue and Cell Culture Samples(소조직 및 세포 배양 샘플에서 미토콘드리아 초복합체 분석을 위한 미토콘드리아 분리) 

Over the last decades, the evidence accumulated about the existence of respiratory supercomplexes (SCs) has changed our understanding of the mitochondrial ...

에 수용성 화합물로 SCA1 마우스 치료가 아닌 것은 - 특히 미토콘드리아 기능 향상

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유기체 살충제에 노출된 간세포에서 미토콘드리아 기능을 검출하기 위한 실험 프로토콜

수축 성 활동 유도 된 쥐 골격 근육 Phenotypic 적응 연구 만성 자극에의 응용

세포외 플럭스 분석을 이용한 단일 3D 미세조직 구상체의 미토콘드리아 에너지 대사 탐구

고해상도 호흡 측정법에 의한 좌골 신경의 미토콘드리아 기능 평가

포유류의 미토콘드리아 기능을 측정하기 위한 산소 비의존적 분석

포유류의 미토콘드리아 기능을 측정하기 위한 산소 비의존적 분석

포유류의 미토콘드리아 기능을 측정하기 위한 산소 비의존적 분석

마우스 지방 유래 중간엽 줄기 세포에서 노화된 마우스 난모세포로의 미토콘드리아 전달

마우스 지방 유래 중간엽 줄기 세포에서 노화된 마우스 난모세포로의 미토콘드리아 전달

Isolation of Mitochondria for Mitochondrial Supercomplex Analysis from Small Tissue and Cell Culture Samples(소조직 및 세포 배양 샘플에서 미토콘드리아 초복합체 분석을 위한 미토콘드리아 분리)