In regards to neurodegenerative disorders, it has been reported that the use of fluorescent markers of apoptosis in animal models of neurodegeneration could serve as method of in vivo disease progression monitoring.
In addition, Rodhe et al., found an association between cleaved caspase 3 and caspase 8 expression and the time of stroke onset in postmortem samples.
In the same line, previous findings showed that levels of plasma caspase-3, an apoptosis-related protein, after stroke correlated with infarct expansion and neurological outcome. It is suggested that the detection of apoptotic processes in patients with these types of neurological diseases could have potential clinical applications, including prediction of clinical outcome, effective monitoring of evolution of the disease and the selection of treatments that interfere with the apoptosis pathway. It is now becoming evident that an increasing number of pathological situations can be related to aberrant apoptosis, such as cerebrovascular and neurodegenerative diseases. However, necroptotic death, like cellular necrosis, culminates in cell lysis and release of the cytoplasmic content into the surrounding tissue, provoking immediate inflammation. Interestingly, an increasing number of studies have described a genetically programmed and regulated form of necrosis, termed necroptosis. Likewise, the process of necrosis, traditionally considered as an unprogrammed death resulting from an overwhelming insult, is also morphologically distinct from apoptosis in many of its characteristics such as loss of membrane integrity and rapid cell and organelle swelling. This dying process involves the engulfment of cytoplasmic material and intracellular organelles within double-membrane vesicles, called autophagosomes and occurs without unequivocal morphological manifestations of apoptosis and formation of apoptotic bodies. Recent studies have reported that besides apoptosis, programmed cell death also includes autophagic cell death. These small membranous vesicles, which have been released into their microenviroment and blood circulation, are removed by phagocytosis thus avoiding the inflammatory response, in the same manner as described for apoptotic cells. In the late phase of apoptosis, and as a result of cell fragmentation, apoptotic bodies are generated. During apoptosis, decrease in cell volume, cytoplasmic condensation, mitochondrial membrane permeabilization, DNA fragmentation, and chromatin condensation followed by nuclear fragmentation and cytoplasmic membrane blebbing takes place, thereby leading to the total desintegration of the cell. Three major mechanisms of cell death have been described: apoptosis (type I), cell death associated with autophagy (type II) and necrosis (type III). This easy, minimally time consuming and effective procedure for isolating and quantifying plasma apoptotic bodies could help physicians to implement the use of such vesicles as a non-invasive tool to monitor apoptosis in patients with cerebrovascular and neurodegenerative diseases for prognostic purposes and for monitoring disease activity. Electron microscopy, dynamic light scattering and proteomic characterization in combination with flow cytometry studies revealed that our isolation method achieves notable recovery rates of highly-purified intact apoptotic bodies. Authors, here, describe a reproducible centrifugation-based method combined with flow cytometry analysis to isolate and quantify plasma apoptotic bodies of patients with ischemic stroke, multiple sclerosis, Parkinson’s disease and also in healthy controls. Since there is no scientific literature establishing the most appropriate method for collecting and enumerating apoptotic bodies from human blood samples. We therefore propose to use circulating apoptotic bodies as biomarkers for measuring apoptotic death in patients with ischemic stroke and neurodegenerative diseases. Since apoptotic bodies are membrane vesicles that are released from fragmented apoptotic cells, it follows that the presence of these vesicles in the bloodstream is likely due to the apoptotic death of cells in tissues. The clinical application of the tissue sampling and imaging approaches to analyze apoptosis in neurological diseases is, however, limited. Consequently, in the last few years, there has been an ever-growing interest in the in vivo study of apoptosis. Improper regulation of apoptosis has been postulated as one of the main factors that contributes to the etiology and/or progression of several prevalent diseases, including ischemic stroke and neurodegenerative pathologies.