In Vivo Analysis of Cell Proliferation and Apoptosis in the CNS

Lossi, Laura; Mioletti, Silvia; Aimar, Patrizia; Bruno, R.; Merighi, Adalberto

doi:10.1007/978-0-387-22460-2_14

Cited by 11 publications

(7 citation statements)

References 120 publications

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“…In addition, the number of cellular mechanisms that have been put into play for explaining the numerous purported actions of resveratrol and other vegetal PPs is so high that virtually all the major cell pathways appeared to be involved [82], at times with opposite effects [20]. In addition, PI is widely used to stain dead cells as it is extruded from live cells with an intact membrane irrespective of the mechanism and type of death [83] and PI-based experiments thus offered prompt and, very importantly, comparable readouts of the effect of each PP onto NOND, independently from its mechanism(s) of action.…”

Section: Preparation Of Pps-containing Media Incubation Of Cultures mentioning

confidence: 99%

Protective Effects of Some Grapevine Polyphenols against Naturally Occurring Neuronal Death

et al. 2020

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The interest in the biological properties of grapevine polyphenols (PPs) in neuroprotection is continuously growing in the hope of finding translational applications. However, there are several concerns about the specificity of action of these molecules that appear to act non-specifically on the permeability of cellular membranes. Naturally occurring neuronal death (NOND) during cerebellar maturation is a well characterized postnatal event that is very useful to investigate the death and rescue of neurons. We here aimed to establish a baseline comparative study of the potential to counteract NOND of certain grapevine PPs of interest for the oenology. To do so, we tested ex vivo the neuroprotective activity of peonidin- and malvidin-3-O-glucosides, resveratrol, polydatin, quercetin-3-O-glucoside, (+)-taxifolin, and (+)-catechin. The addition of these molecules (50 μM) to organotypic cultures of mouse cerebellum explanted at postnatal day 7, when NOND reaches a physiological peak, resulted in statistically significant (two-tailed Mann–Whitney test—p < 0.001) reductions of the density of dead cells (propidium iodide+ cells/mm2) except for malvidin-3-O-glucoside. The stilbenes were less effective in reducing cell death (to 51–60%) in comparison to flavanols, (+)-taxifolin and quercetin 3-O-glucoside (to 69–72%). Thus, molecules with a -OH group in ortho position (taxifolin, quercetin 3-O-glucoside, (+)-catechin, and peonidin 3-O-glucoside) have a higher capability to limit death of cerebellar neurons. As NOND is apoptotic, we speculate that PPs act by inhibiting executioner caspase 3.

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Section: Preparation Of Pps-containing Media Incubation Of Cultures mentioning

confidence: 99%

Protective Effects of Some Grapevine Polyphenols against Naturally Occurring Neuronal Death

et al. 2020

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“…Apoptosis is accompanied by a specific pattern of DNA damage that leads to the formation of low-molecular weight DNA oligomers. These can be visualized after DNA isolation and electrophoresis in tissue extracts, or directly in situ by the use of the TUNEL technique [49]. The other main molecular features of apoptosis are the release of cytochrome c from mitochondria, and the activation of caspases, including caspase-3, in the cytosol [50,51].…”

Section: Caspase-3 In Cerebellar Developmentmentioning

confidence: 99%

Caspase-3 Mediated Cell Death in the Normal Development of the Mammalian Cerebellum

Lossi

Merighi

2018

IJMS

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135

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Caspase-3, onto which there is a convergence of the intrinsic and extrinsic apoptotic pathways, is the main executioner of apoptosis. We here review the current literature on the intervention of the protease in the execution of naturally occurring neuronal death (NOND) during cerebellar development. We will consider data on the most common altricial species (rat, mouse and rabbit), as well as humans. Among the different types of neurons and glia in cerebellum, there is ample evidence for an intervention of caspase-3 in the regulation of NOND of the post-mitotic cerebellar granule cells (CGCs) and Purkinje neurons, as a consequence of failure to establish proper synaptic contacts with target (secondary cell death). It seems possible that the GABAergic interneurons also undergo a similar type of secondary cell death, but the intervention of caspase-3 in this case still remains to be clarified in full. Remarkably, CGCs also undergo primary cell death at the precursor/pre-migratory stage of differentiation, in this instance without the intervention of caspase-3. Glial cells, as well, undergo a process of regulated cell death, but it seems possible that expression of caspase-3, at least in the Bergmann glia, is related to differentiation rather than death.

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“…In particular, caspase 3 cleavage triggers apoptosis in CGCs as the endpoint of a cascade that involves other caspases in vitro (Allsopp et al 2000). Characterization of programmed cell death in vertebrate (mammalian) neurons mainly relies on in vitro approaches (Rao & White 1997, Selimi et al 1997, Sastry & Rao 2000, Denecker et al 2001, Shastry et al 2001, also as a consequence of a series of inherent technical difficulties in studying apoptosis in vivo (Lossi et al 2002a).…”

Section: Introductionmentioning

confidence: 99%

Molecular morphology of neuronal apoptosis: Analysis of caspase 3 activation during postnatal development of mouse cerebellar cortex

Lossi¹,

Tamagno²,

Merighi³

2004

Histochem J

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We have used the mammalian post-natal cerebellar cortex as a model to dissect out the molecular morphology of neuronal apoptosis in a well-defined population of central neurons: the cerebellar granule cells. By immunocytochemistry, in situ labeling of apoptotic cells, and analysis of cerebellar slices following particle-mediated gene transfer (biolistics), we have studied the relationship of cell death and cleavage of caspase 3, a key molecule in the execution of apoptosis, and monitored caspase 3 activation in living cells. Our results demonstrate the existence of caspase dependent and independent apoptotic pathways affecting the cerebellar granule cells at different stages of their life. Apoptosis of proliferating precursors and young pre-migratory cells occurs in the absence of caspase 3 cleavage, whereas cell death of post-mitotic post-migratory neurons is directly linked to caspase 3 activation. Data obtained from cerebellar cortex can be generalized to outline a more comprehensive picture of the cellular and molecular mechanisms of neuronal death not only in development, but also in a number of pathological conditions leading to neuronal loss.

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In Vivo Analysis of Cell Proliferation and Apoptosis in the CNS

Cited by 11 publications

References 120 publications

Protective Effects of Some Grapevine Polyphenols against Naturally Occurring Neuronal Death

Protective Effects of Some Grapevine Polyphenols against Naturally Occurring Neuronal Death

Caspase-3 Mediated Cell Death in the Normal Development of the Mammalian Cerebellum

Molecular morphology of neuronal apoptosis: Analysis of caspase 3 activation during postnatal development of mouse cerebellar cortex

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