Apoptosis and Neurodegeneration

Towards the Development of Disease Specific Assays
Neurodegenerative diseases are becoming increasingly prevalent, especially in the Western societies, with larger percentage of members living to an older age. They have to be seen not only as a health problem, but since they are care-intensive, they also carry a significant economic burden.

Apoptotic pathways are induced in many of these diseases and are key culprits in disease progression.

Figure: Schematic representation of apoptotic pathways. Apoptosis triggered by internal (intrinsic) or external (extrinsic) stress signals that is activated by binding of ligands (e.g. FasL, APO-2L, TRAIL, TNF) to cell surface receptors (e.g. Fas, DR4, DR5, TNF-R1). The intrinsic apoptosis pathway might be triggered by p53 upon DNA damage following exposure to cellular stress. In the intrinsic pathway, death signal reaches mitochondria, leading to release of cytochrome c, which can binds to Apaf1. The cytochrome c/Apaf1 make a complex with pro-caspase-9 (in the presence of dATP), activates caspase-9, which promotes caspase-3 activation, eventually leading to cell death. The extrinsic pathway is initiated through the stimulation of the members of tumor necrosis factor receptor (TNF-R) family (transmembrane death receptors) by their respective ligands. These receptors activate pro-caspases-8, -10 by recruiting the endogenous adaptor protein FADD. Procaspase-8, -10 cleave themselves to form activated caspase-8 or -10. Ultimately, effector enzymes such as caspase-3, -6, -7 are activated in this cascade to mediate apoptosis. Likewise, there can be cross-talk between the intrinsic and extrinsic pathways. For example caspase-8 may cleave Bid to form tBid that is a strong activator of the intrinsic/mitochondrial apoptotic pathway. The intrinsic pathway is usually activated by the recruitment of BAX and BAK to outer mitochondrial membrane, causing cytochrome c release formation of apoptosome and subsequent activation of caspase-9. Activated caspase-9 proteolytically activates caspases-3, -6, and -7. Moreover, some of the effector caspases also can activate caspase-8, forming a positive amplification loop. doi:10.1016/j.pneurobio.2013.10.004.

Working with the Apoptosis Experts at Immunochemistry Technologies and Human Astroglial-Neuron Biosensors Experts at ArunA Biomedical, we plan on  developing disease specific assays. Here's a map of the general process.

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We welcome feedback and input on your interests. You can email rose@neuromics.com. We will continue to post updates on exciting new developments.

New Near Infrared Apoptosis Detection!

Detecting apoptosis in cells, tissue and living animals

In vitro and in vivo apoptosis detection assays are widely used in basic research and drug discovery. Neuromics has a wide range of kits providing capablilties to meet you unique requirements. We are pleased to offer even more options with our new Far Infrared Kits:
660 Polycaspase in vitro Apoptosis Detection Kit 25-50 Tests $199
660 Caspase-1  in vitro Apoptosis Detection Kit 25-50 Tests $199
NIR FLIVO™ 690 in vivo Apoptosis Tracer Kit 20 Tests-$399
NIR FLIVO™ 747 in vivo Apoptosis Tracer Kit 20 Tests-$399

Seeing is Believing

Images: To demonstrate the capabilities of the NIR-FLIVO® 747 apoptosis tracer, adult wild-type Balb/c mice were either inoculated with HSV-1 virus, which is known to induce apoptosis in the brain, or given a sham treatment. Seven days after viral inoculation, the mice were injected intravenously with either the NIR-FLIVO® 747apoptosis tracer (cat. KF17368), the NIR-FLIVO® 747 free dye (cat. KF17370, DyLight®747), or no reagent. Seven hours after reagent injection, the animals were imaged with a Carestream In-Vivo FX Pro imaging system. Strong caspase activity was located in the brain of the animal treated with HSV-1 and injected with NIR-FLIVO® 747 tracer (cat. KF17367). Minimal signal was detected in the liver region of the HSV-1-treated animal injected with the free dye control (HSV-Infected; NIR747FreeDye+) and of the uninfected mouse injected with NIR-FLIVO®747 tracer; the liver is the route of clearance for FLIVO® tracers. All reagent-injected mice show fluorescence signal in the tail where reagent is likely to pool after IV injection.

Images: Brain abscesses were induced in mice following the intracerebral inoculation of live S. aureus. Animals received intravenous injections of the pan-caspase tracer NIR-FLIVO™ 690 as an apoptosis probe or DyLight®690 (carboxylic acid form) as a control at 17 h post-infection, whereupon signals were acquired 1 h later from brain tissues immediately ex vivo using an IVIS Spectrum (Caliper Life Sciences). The same instrument settings were used to acquire both images. Strong caspase activity, detected with NIR-FLIVO™ 690 (arrows, image on right), was associated with brain abscesses, whereas minimal signal was detected in animals injected with the carboxylic acid control (image on left).

Check them all out
Apoptosis Research Reagents-Detection kits, antibodies and proteins