Is β-Galactosidase Staining a Marker of Senescence in Vitro and in Vivo?

Severino, Joseph; Allen, Robert G.; Balin, Samuel J.; Balin, Arthur K.; Cristofalo, Vincent J.

doi:10.1006/excr.2000.4875

Cited by 341 publications

(231 citation statements)

References 36 publications

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“…In regenerative tissues and in vitro cultures, senescent cells may exhibit SA β‐gal activity, which is a measure of lysosomal galactosidase activity at pH 6.0 and indicative of altered/expanded lysosomal compartments (Severino, Allen, Balin, Balin, & Cristofalo, 2000). The examination of the gene that codes for the hydrolase enzyme, galactosidase beta (β) 1 ( Glb1 ), revealed that tau NFT mice expressed higher Glb1 gene expression than controls (Supporting Information Figure S3).…”

Section: Resultsmentioning

confidence: 99%

Tau protein aggregation is associated with cellular senescence in the brain

et al. 2018

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Tau protein accumulation is the most common pathology among degenerative brain diseases, including Alzheimer's disease (AD), progressive supranuclear palsy (PSP), traumatic brain injury (TBI), and over twenty others. Tau‐containing neurofibrillary tangle (NFT) accumulation is the closest correlate with cognitive decline and cell loss (Arriagada, Growdon, Hedley‐Whyte, & Hyman, 1992), yet mechanisms mediating tau toxicity are poorly understood. NFT formation does not induce apoptosis (de Calignon, Spires‐Jones, Pitstick, Carlson, & Hyman, 2009), which suggests that secondary mechanisms are driving toxicity. Transcriptomic analyses of NFT‐containing neurons microdissected from postmortem AD brain revealed an expression profile consistent with cellular senescence. This complex stress response induces aberrant cell cycle activity, adaptations to maintain survival, cellular remodeling, and metabolic dysfunction. Using four AD transgenic mouse models, we found that NFTs, but not Aβ plaques, display a senescence‐like phenotype. Cdkn2a transcript level, a hallmark measure of senescence, directly correlated with brain atrophy and NFT burden in mice. This relationship extended to postmortem brain tissue from humans with PSP to indicate a phenomenon common to tau toxicity. Tau transgenic mice with late‐stage pathology were treated with senolytics to remove senescent cells. Despite the advanced age and disease progression, MRI brain imaging and histopathological analyses indicated a reduction in total NFT density, neuron loss, and ventricular enlargement. Collectively, these findings indicate a strong association between the presence of NFTs and cellular senescence in the brain, which contributes to neurodegeneration. Given the prevalence of tau protein deposition among neurodegenerative diseases, these findings have broad implications for understanding, and potentially treating, dozens of brain diseases.

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Section: Resultsmentioning

confidence: 99%

Tau protein aggregation is associated with cellular senescence in the brain

et al. 2018

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“…The blue‐stained cells were counted in 10 fields under the microscope with 200× magnification and expressed as a percentage of positive cells. To avoid staining due to cell confluence rather than to proliferative senescence (Severino et al ., 2000), the assay was performed in subconfluent cultures displaying comparable cell density. Fat tissue β‐gal staining was performed immediately after mice were euthanized; tissues were cut into small pieces and stained as described by manufacturer's instructions.…”

Section: Methodsmentioning

confidence: 99%

Rapamycin inhibits the secretory phenotype of senescent cells by a Nrf2-independent mechanism

et al. 2017

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SummarySenescent cells contribute to age‐related pathology and loss of function, and their selective removal improves physiological function and extends longevity. Rapamycin, an inhibitor of mTOR, inhibits cell senescence in vitro and increases longevity in several species. Nrf2 levels have been shown to decrease with aging and silencing Nrf2 gene induces premature senescence. Therefore, we explored whether Nrf2 is involved in the mechanism by which rapamycin delays cell senescence. In wild‐type (WT) mouse fibroblasts, rapamycin increased the levels of Nrf2, and this correlates with the activation of autophagy and a reduction in the induction of cell senescence, as measured by SA‐β‐galactosidase (β‐gal) staining, senescence‐associated secretory phenotype (SASP), and p16 and p21 molecular markers. In Nrf2KO fibroblasts, however, rapamycin still decreased β‐gal staining and the SASP, but rapamycin did not activate the autophagy pathway or decrease p16 and p21 levels. These observations were further confirmed in vivo using Nrf2KO mice, where rapamycin treatment led to a decrease in β‐gal staining and pro‐inflammatory cytokines in serum and fat tissue; however, p16 levels were not significantly decreased in fat tissue. Consistent with literature demonstrating that the Stat3 pathway is linked to the production of SASP, we found that rapamycin decreased activation of the Stat3 pathway in cells or tissue samples from both WT and Nrf2KO mice. Our data thus suggest that cell senescence is a complex process that involves at least two arms, and rapamycin uses Nrf2 to regulate cell cycle arrest, but not the production of SASP.

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“…Senescent cells are characterized by telomere shortening and decreased telomerase activity. In addition, senescence-associated-beta-galactosidase (SA-ß-Gal) activity is used to identify the occurrence of senescence [16][17][18]. Recently, a few studies have started to report the association of senescence and intervertebral disc degeneration [19][20][21][22][23][24].…”

Section: Introductionmentioning

confidence: 99%

Accelerated premature stress-induced senescence of young annulus fibrosus cells of rats by high glucose-induced oxidative stress

Park

et al. 2014

International Orthopaedics (SICOT)

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Purposes Diabetes mellitus (DM) is thought to be an important aetiological factor in intervertebral disc degeneration. A glucose-mediated increase of oxidative stress is a major causative factor in development of diseases associated with DM. The aim of this study was to investigate the effect of high glucose on mitochondrial damage, oxidative stress and senescence of young annulus fibrosus (AF) cells. Methods AF cells were isolated from four-week-old young rats, cultured, and placed in either 10 % FBS (normal control) or 10 % FBS plus two different high glucose concentrations (0.1 M and 0.2 M) (experimental conditions) for one and three days. We identified and quantified the mitochondrial damage and reactive oxygen species (ROS) (oxidative stress). We also identified and quantified the occurrence of senescence and telomerase activity. Finally, the expressions of proteins were determined related to replicative senescence (p53-p21-pRB) and stress-induced senescence (p16-pRB). Results Two high glucoses enhanced the mitochondrial damage in young rat AF cells, which resulted in an excessive generation of ROS in a dose-and time-dependent manner for one and three days compared to normal control. Two high glucose concentrations increased the occurrence of senescence of young AF cells in a dose-and time-dependent manner. Telomerase activity declined in a dose-and timedependent manner. Both high glucose treatments increased the expressions of p16 and pRB proteins in young rat AF cells for one and three days. However, compared to normal control, the expressions of p53 and p21 proteins were decreased in young rat AF cells treated with both high glucoses for one and three days. Conclusions The present study demonstrated that high glucose-induced oxidative stress accelerates premature stress-induced senescence in young rat AF cells in a doseand time-dependent manner rather than replicative senescence. These results suggest that prevention of excessive generation of oxidative stress by strict blood glucose control could be important to prevent or to delay premature intervertebral disc degeneration in young patients with DM.

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Is β-Galactosidase Staining a Marker of Senescence in Vitro and in Vivo?

Cited by 341 publications

References 36 publications

Tau protein aggregation is associated with cellular senescence in the brain

Tau protein aggregation is associated with cellular senescence in the brain

Rapamycin inhibits the secretory phenotype of senescent cells by a Nrf2-independent mechanism

Accelerated premature stress-induced senescence of young annulus fibrosus cells of rats by high glucose-induced oxidative stress

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