Aging in a dish: Age‐dependent changes of neuronal survival, protein oxidation, and creatine kinase BB expression in long‐term hippocampal cell culture

Abstract

Results from different experimental systems demonstrate that increased oxidative damage plays a role in normal aging and age-associated pathology. In the current study, long-term cultures of hippocampal neurons were examined as a model system. It was established that neuronal survival in long-term culture decreases according to the Gompertz law and that neuronal "aging in the dish" is associated with increased oxidative damage of cell proteins. The increase of protein carbonyl formation in aged neurons was demonstrated both by Western blot analysis for oxidized proteins and by in situ immunocytochemical method, which was developed to analyze protein oxidation in fixed cells. In aging neuronal cultures, a gradual increase in creatine kinase (CK) content but decreased activity of enzyme per immunoreactive protein was found, suggesting the accumulation of inactive CK molecules. The increase in CK content was not a result of generalized protein elevation, since analysis of beta-actin content showed a time-dependent loss, probably reflecting decreased number of cellular processes with aging. These findings, showing "aging in a dish," consistent with the notion that aging is associated with increased protein oxidation, provide a system for study of age-related neurodegenerative disorders associated with oxidative stress.

Is senescence-associated β-galactosidase a marker of neuronal senescence?

“…Some research on neuron senescence has been carried out in long-term cultures of primary neurons and collected data seem to suggest that such cultures can be used as a model of neuronal aging [6, 7]. Overall, neurons can be kept alive for as long as 60 days, during which time, they display signs of development, maturation and finally aging and death.…”

Polychlorinated Biphenyl (PCBs)-Induced Oxidative Stress Plays a Critical Role on Cerebellar Dopaminergic Receptor Expression: Ameliorative Role of Quercetin

“…Andres et al (2008) studied that creatine kinase/phosphocreatine system plays a key role in cellular energy buffering and energy transport particularly in cells with high and fluctuating energy requirements like neurons. The ATP binding domain of brain-specific creatine kinase contains essential arginine, histidine, lysine, and cysteine residues that may be targeted by ROS (Aksenova et al 1999). Koufen and Stark (2000) demonstrated that direct oxidation of sulphydryl groups in cysteines located in the active site of creatine kinase also may inactivate the enzyme.…”

Xy99-5038 Promotes Long-Term Survival of Cultured Retinal Neurons

“…Age-dependent increases in the density of Ca 2+ channels in long-term culture of hippocampal cells have been reported to be analogous to patterns seen in neurons of in vivo aged animals (Porter et al, 1997). Age-dependent changes of neuronal survival, protein oxidation and creatine kinase BB expression in long-term hippocampal cell culture have also been described as aging in dish (Aksenova et al, 1999). Using long-term mesencephalic culture, Troadec and coworkers demonstrated that noradrenaline promotes survival and function of dopaminergic neurons (Troadec et al, 2001).…”

“…A complex of genetic, programmed, and variable factors producing the age-related changes affect retinal neuronal survival and can result in retinal neuronal degeneration. Long-term neuronal culture has been used as an aging and neurological research tool for understanding the changes of aging, their impact on disease, and for the development of agents able to modify agerelated changes and to prevent neurodegeneration (Kuroda et al, 1995;Puttfarcken et al, 1996;Aksenova et al, 1999;Troadec et al, 2001). Age-dependent increases in the density of Ca 2+ channels in long-term culture of hippocampal cells have been reported to be analogous to patterns seen in neurons of in vivo aged animals (Porter et al, 1997).…”

“…During the last decade, significant progress was made in culturing of highly differentiated cell, such as neurons. It became possible to consider the application of long-term primary neuronal cultures to study age-related changes in vitro (Kuroda et al, 1995;Puttfarcken et al, 1996;Porter et al, 1997;Aksenova et al, 1999;Potter & DeMarse, 2001;Troadec et al, 2001). Longterm neuronal culture has been used as aging and neurological research tool to study the effects of long-term exposure to aluminum, a risk factor for Alzheimer's disease, on the cultured cortical neurons (Kuroda et al, 1995).…”