Mitochondria Research

Science Note

[May 9, 2023]                                                                                                                                                                                                                            Previous Science Note
Mitochondrial unfolded protein response in neural stem cell aging

This article focusing on the mitochondrial protein folding stress is particularly significant in activated neural stem cells (NSCs) and neural progenitor cells (NPCs) within the neurogenic niche. This stress increases with age, leading to dysregulated cell cycles and mitochondrial activity in activated NSCs and NPCs in the dentate gyrus. 

The mitochondrial unfolded protein response regulates hippocampal neural stem cell aging    Chin-Ling Wang, et. al., Cell Metabolism (2023)

Point of Interest
- Elevated mitochondrial protein folding stress in aging neural stem cells
- SIRT7 safeguards neural stem cells by reducing mitochondrial protein folding stress
- SIRT7 overexpression enhances neurogenesis and cognitive function in aged mice
Related Techniques
           Glycolysis/Oxidative phosphorylation assay Glycolysis/OXPHOS Assay Kit
           Oxygen consumption rate assay Extracellular OCR Plate Assay Kit
           Mitochondrial membrane potential detection JC-1 MitoMP Detection Kit / MT-1 MitoMP Detection Kit
           Mitochondrial superoxide detection mtSOX Deep red - Mitochondrial superoxide Detection
           Mitophagy Detection Mitophagy Detection Kit and Mtphagy Dye
           Cellular senescence detection (Live cell imaging or FCM) Cellular Senescence Detection Kit
           Cellular senescence detection (Plate reader) Cellular Senescence Plate Assay Kit
           Lysosomal function assay Lysosomal pH and mass detection Kit
Related Applications

Metabolic shift to glycolysis in senescenct cells

 

NAD(+) levels decline during the aging process, causing defects in nuclear and mitochondrial functions and resulting in many age-associated pathologies*. Here, we try to redemonstrate this phenomenon in the doxorubicin (DOX)-induced cellular senescence model with a comprehensive analysis of our products.                       

*S. Imai, et al., Trends Cell Biol, 2014, 24, 464-471

Products in Use
   ① DNA Damage Detection Kit - γH2AX

   ② Cellular Senescence Detection Kit - SPiDER-βGal

   ③ NAD/NADH Assay Kit-WST

   ④ JC-1 MitoMP Detection Kit

   ⑤ Glycolysis/OXPHOS Assay Kit、Lactate Assay Kit-WST

2. Simultaneously detection of Lysosomal and Mitochondrial Dysfunction

We tried the simultaneous detection of lysosomal and mitochondrial dysfunction in Hela cells treated with CCCP or Antimycin (AN). CCCP and AN are well-known inducers of mitochondrial ROS regarding loss of mitochondrial membrane potential. Recent research showed the result that CCCP induces not only mitochondrial ROS but also lysosomal neutralization. To detect mitochondrial ROS, HeLa cells were labeled by mtSOX Deep Red - Mitochondrial Superoxide Detection, and the lysosomal mass and pH were detected separately with LysoPrime Green and pHLys Red. Co-staining with mtSOX and Lysosomal dyes demonstrated that CCCP causes lysosomal neutralization and mitochondrial ROS induction at the same time.

Products in Use
   - LysoPrime Green
   - pHLys Red
   - mtSOX Deep Red - Mitochondrial Superoxide Detection

 

Related Product
   - Lysosomal Acidic pH Detection Kit

 

 

 

Selection guide for mitochondria-related reagents

Mitochondria research is very multi-faceted, because the multi-functional organelle is not only involved in energy production in a cell, but other additional cellular functions. The active cycle of mitochondrial fusion and division induces morphological changes, which is called mitochondrial dynamics. Abnormalities in morphological control of mitochondria are associated with neurodegenerative diseases, metabolic disorders, aging, and so on. Therefore, the demand for long-term observation of mitochondrial dynamics has recently been increasing.

Selection Guide of Reagents

The following table lists reagents for mitochondrial research designed to stain and detect mitochondria (MitoBright LT, MitoTracker, etc.), mitochondrial membrane potential (JC-1, TMRM, TMRE, etc.), reactive oxygen species AKA ‘ROS’ (mtSOX, MitoSOX, etc.), mitophagy, and lipid peroxides.

Mitophagy

Mitophagy
Reagent Mtphagy Dye Keima-Red
Principle Mtphagy Dye (included in Mitophagy Detection Kit) is a pH-sensitive fluorescent probe that accumulates in mitochondria and emits red fluorescence due to acidic conditions in a lysosome. A pH-sentitive ratiometric fluorescent protein. The excitation spectrum changes accoring to pH. This protein shows high fluorescence ratio (Ex. 550 nm/440 nm) values in a lysozome.
Fixed cell staining – –
Live-cell staining Yes Yes
Fixation after live-cell staining – –
Staining time > 30 min –
Ex / Em 530 / 700 440, 550 / 620
Product code MD01, MT02 –

Lipophilic peroxide / Singlet oxygen / Superoxide

  Lipophilic peroxide Singlet oxygen Superoxide Superoxide
Reagent MitoPeDPP Si-DMA mtSOX Deep Red MitoSOX
Principle A cell-permeant fluorescent probe that accumulates in mitochondria and specifically reacts with lipophilic peroxides in mitochondria to emit fluorescence. A cell-permeant fluorescent probe that accumulates in mitochondria and specifically reacts with singlet oxigen generated in mitochondria to emit red fluorescence. A cell-permeant fluorescent probe that accumulates in mitochondria and reacts with superoxide generated in mitochondria to emit fluorescence. A cell-permeant fluorescent probe that accumulates in mitochondria and reacts with superoxide generated in mitochondria to emit red fluorescence.
Fixed cell staining – – – –
Live-cell staining Yes Yes Yes Yes
Fixation after live-cell staining – – – –
Staining time > 15 min > 45 min > 10 min > 10 min
Ex / Em 452 / 470 644 / 670 540 / 670 510 / 590
Product code M466 MT05 MT14 –

Membrane potential

Membrane potential
Reagent JC-1 MT-1 TMRM, TMRE
Principle A cell-permeant ratiometric fluorescent dye that accumulates in intact mitochondria due to the membrane potential. The excitation spectrum changes according to the mitochondria membrane potential. Cell-permeant fluorescent dyes that accumulate in intact mitochondria due to the membrane potential. MT-1 is extremely photostable and more sensitive than JC-1 and can provide equivalent detection sensitivity to TMRE. Cell-permeant fluorescent dyes that accumulate in intact mitochondria due to the membrane potential. Diffusion of the probes occurs in a damaged mitochondria that has the decreased membrane potential.
Fixed cell staining – – –
Live-cell staining Yes Yes Yes
Fixation after live-cell staining – Yes –
Staining time 30-60 min 30 min 30-60 min
Ex / Em Monomer: 514 / 529
J-aggregation: 585/590		 530-560 / 570-640 550 / 575
Product code MT09 MT13 –

Mitochondria staining

Mitochondria staining
Reagent MitoBright LT series MitoBright IM Red MitoTracker series Rhodamine 123
Principle Cell-permeant fluorescent dyes that accumulate in intact mitochondria due to the membrane potential. Cell-permeant fluorescent dyes that accumulate in intact mitochondria due to the membrane potential and covalently binds to proteins and other biomolecules. Cell-permeant fluorescent dyes that accumulate in intact mitochondria due to the membrane potential. Cell-permeant fluorescent dye that accumulates in intact mitochondria due to the membrane potential.
Fixed cell staining – – – –
Live-cell staining Yes Yes Yes Yes
Fixation after live-cell staining Yes Yes – –
Staining time 30 min 30 min 15-45 min > 15 min
Ex / Em 493 / 508, 547 / 563, 643 / 663 548 / 566 490 / 516 ~
644 / 665		 507 / 529
Product code MT10, MT11,MT12 MT15 – R233

Metal Ion Detection

   Iron ion (Fe2+) Calcium ion (Ca2+)
Reagent Mito-FerroGreen Rhod2-AM
Principle A cell-permeant fluorescent probe that accumulates in mitochondria and specifically reacts with ferrous ion in mitochondria to emit green fluorescence. A cell-permeant fluorescent probe that accumulates in mitochondria and specifically reacts with calcium ion in mitochondria to emit red fluorescence.
Fixed cell staining – –
Live-cell staining Yes Yes
Fixation after live-cell staining – –
Staining time 30 min 30-60 min
Ex / Em 505 / 535 553 / 576
Product code M489 R002 
Application Products
Mitophagy Detection Mitophagy Detection Kit
Mitochondrial Phospholipid Peroxidase Detection MitoPeDPP
Mitochondrial Ferrous Ion Detection Mito-FerroGreen
Mitochondrial Superoxide mtSOX Deep Red
Mitochondrial Singlet Oxygen Detection Si-DMA for Mitochondrial Singlet Oxygen Imaging
Mitochondrial Membrane Potential JC-1 MitoMP Detection Kit
MT-1 MitoMP Detection Kit
Mitochondria Staining MitoBright LT Green
MitoBright LT Red
MitoBright LT Deep Red
MitoBright IM Red for Immunostaining
-Cellstain- MitoRed
-Cellstain- Rh123
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