Preparation of a cell-free extract from rat brain which can initiate protein synthesis in vitro

Cosgrove, James W.; Rapoport, Stanley I.

doi:10.1007/bf00966123

Cited by 16 publications

(15 citation statements)

References 64 publications

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“…1). The sucrose [6,7,10]. In this analysis, we observed that there was a profound loss of 60S-polyribosome complexes in MCI and AD subjects, relative to control individuals (Fig.…”

Section: Isolation and Characterization Of Ribosomesmentioning

confidence: 83%

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Decreased RNA, and Increased RNA Oxidation, in Ribosomes from Early Alzheimer’s Disease

et al. 2006

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All cells rely on efficient protein synthesis in order to maintain cellular homeostasis. Recent studies from our laboratory indicate that declines in protein synthesis and ribosome function occur in the earliest stage of Alzheimer's disease (AD). Additional studies indicate a potential role for ribosomal RNA oxidation as a potential mediator of decreased protein synthesis in AD. The ribosome is a complex of proteins and nucleic acids that mediates all protein synthesis. At present it is unclear if significant alterations in ribosomal RNA occurs within the ribosome complex during the progression of AD. In this study we examined the amount of ribosomal RNA in the different ribosomal fractions generated from control subjects, individuals with mild cognitive impairment (MCI), and individuals with AD. Studies were conducted in the inferior parietal lobule of each subject. Together, these data demonstrate that during the progression of AD there is a gross decline in the amount of ribosomal RNA within the ribosome complex. Additionally, these studies provide evidence for gross elevations in RNA oxidation within the ribosome complex of MCI and AD. Together, these data strongly suggest a role for RNA alterations within the ribosome as a mediator of decreased protein synthesis in both MCI and AD.

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“…1). The sucrose [6,7,10]. In this analysis, we observed that there was a profound loss of 60S-polyribosome complexes in MCI and AD subjects, relative to control individuals (Fig.…”

Section: Isolation and Characterization Of Ribosomesmentioning

confidence: 83%

“…The different ribosomal complexes were purified as described previously by our laboratory and others [4,6,7]. Briefly, frozen brain specimens from the inferior parietal lobule were placed in buffer A containing following (in mM): 320 sucrose, 50 HEPES, 140 potassium acetate, 2.5 dithiotheritol, pH 7.5.…”

Section: Purification and Analysis Of Ribosomesmentioning

confidence: 99%

Decreased RNA, and Increased RNA Oxidation, in Ribosomes from Early Alzheimer’s Disease

et al. 2006

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“…Polyribosomes were purified and analyzed for their ability to translate proteins as described previously (Cosgrove and Rapoport, 1986;Aloni et al, 1992). Briefly, frozen brain specimens from the indicated brain region were placed in buffer A [containing the following (in mM): 320 sucrose, 50 HEPES, 140 potassium acetate, 4 magnesium acetate, 2.5 dithiotheritol, pH 7.5], using a 2.5 vol of buffer A.…”

Section: Methodsmentioning

confidence: 99%

Ribosome Dysfunction Is an Early Event in Alzheimer's Disease

Qian¹,

Markesbery²,

Chen³

et al. 2005

J. Neurosci.

388

333

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Alzheimer's disease (AD) is a progressive and devastating disorder that is often preceded by mild cognitive impairment (MCI). In the present study, we report that in multiple cortical areas of MCI and AD subjects, there is a significant impairment in ribosome function that is not observed in the cerebellum of the same subjects. The impairment in ribosome function is associated with a decreased rate and capacity for protein synthesis, decreased ribosomal RNA and tRNA levels, and increased RNA oxidation. No alteration in the level of initiation factors was observed in the brain regions exhibiting impairments in protein synthesis. Together, these data indicate for the first time that impairments in protein synthesis may be one of the earliest neurochemical alterations in AD and directly demonstrate that the polyribosome complex is adversely affected early in the development of AD. These data have important implications for AD studies involving proteomics and studies analyzing proteolysis in AD, indicate that oxidative damage may contribute to decreased protein synthesis, and suggest a role for alterations in protein synthesis as a novel contributor to the onset and development of AD.

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“…The protein synthesis rate was measured using a cell-free system validated for the brain (Burda et al 1994 ;Cosgrove & Rapoport, 1986 ;. Tissues from frontal cortices were homogenized in 2.5 vol.…”

Section: Translation Assays In Vitromentioning

confidence: 99%

Effects of neonatal MK-801 treatment on p70S6K-S6/eIF4B signal pathways and protein translation in the frontal cortex of the developing rat brain

Kim

Park

Kim

et al. 2010

Int. J. Neuropsychopharm.

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Systemic injections of MK-801, a selective NMDAR antagonist, into neonatal rats induces long-term neurochemical and behavioural changes. It has been suggested that these changes form the neurodevelopmental basis for schizophrenia-like behaviour in rats. In this study, 7-d-old rats were treated with MK-801, and their frontal cortices were examined to investigate the effects on p70S6K-S6 signal pathway and on protein translation, which play crucial roles in the neurodevelopmental process. MK-801, in doses of 0.5 and 1.0 mg/kg, induced a decrease in phosphorylation of p70S6K and its substrates, S6 and eIF4B, in the first 8 h, and no change at 24 and 48 h. These effects were more prominent after two injections of MK-801 than one. Decreased S6 phosphorylation by MK-801 was evident in the prefrontal, cingulate, and insular cortex. In two representative upstream p70S6K-S6 pathways related to ERK1/2 and Akt, changes in ERK1/2-p90RSK phosphorylation were accompanied by changes of p70S6K-S6. Although two MK-801 injections induced a dose-dependent decrease in phosphorylation of Akt and mTOR at 4 and 8 h, a single injection did not produce a significant effect. Protein synthesis rate, measured by [3H]leucine incorporation in frontal cortical tissue, was reduced until 24 h after two MK-801 (1.0 mg/kg) injections. In summary, this study found that neonatal MK-801 treatment induced dysregulation in the p70S6K-S6/eIF4B pathway and protein translation in the frontal cortex of the developing rat brain, which may suggest an important role of protein translation machinery in the MK-801 neurodevelopmental animal model of schizophrenia.

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Preparation of a cell-free extract from rat brain which can initiate protein synthesis in vitro

Cited by 16 publications

References 64 publications

Decreased RNA, and Increased RNA Oxidation, in Ribosomes from Early Alzheimer’s Disease

Decreased RNA, and Increased RNA Oxidation, in Ribosomes from Early Alzheimer’s Disease

Ribosome Dysfunction Is an Early Event in Alzheimer's Disease

Effects of neonatal MK-801 treatment on p70S6K-S6/eIF4B signal pathways and protein translation in the frontal cortex of the developing rat brain

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