Limits...
Novel telomerase-increasing compound in mouse brain delays the onset of amyotrophic lateral sclerosis.

Eitan E, Tichon A, Gazit A, Gitler D, Slavin S, Priel E - EMBO Mol Med (2012)

Bottom Line: Hence, we and others hypothesized that increasing telomerase expression by pharmaceutical compounds may protect brain cells from death caused by damaging agents.The survival of telomerase-expressing cells (i.e. motor neurons), but not telomerase-deficient cells, exposed to oxidative stress was increased by AGS-499 treatment, suggesting that the AGS-499 effects are telomerase-mediated.Therefore, a controlled and transient increase in telomerase expression and activity in the brain by AGS-499 may exert neuroprotective effects.

View Article: PubMed Central - PubMed

Affiliation: The Shraga Segal Department of Immunology and Microbiology, Ben-Gurion University of the Negev, Beer-Sheva, Israel.

Show MeSH

Related in: MedlinePlus

AGS-499 increases telomerase activity in the forebrain, BS and SC in a time-dependent mannerMice were injected s.c. with AGS-499 (6 mg/kg) or its vehicle DMSO. Cytoplasmic (A) and nuclear (B) protein extracts from the FB, and whole cell protein extract from the BS or SC (C) at various intervals after compound injection were prepared.A-C. Telomerase activity by TRAP assay. Protein extracts (2 µg) were added to the TRAP-specific reaction mixture. Negative control (NG) contained CHAPS buffer instead of the protein extract. IS, internal standard.D. Quantification of the TRAP results by densitometric analysis using the EZquant software (mean ± s.e.m.; n = 10), Student's t-test p = *10.092, *20.007, *30.028, *40.005, *50.0002, *60.004, *70.045.E. Quantification of telomerase activity by the real-time PCR-based TRAP assay kit. Cytoplasmic and nuclear protein extracts (1 µg) derived from the FB of AGS-treated or untreated mice for various intervals, were added to the reaction according to the manufacturer's instructions and telomerase activity products were calculated as moles DNA/µg protein (mean ± s.e.m.; n = 6), Student's t-test p = *10.043, *20.026. Symbols: UT, untreated mice; V, vehicle.
© Copyright Policy
Related In: Results  -  Collection


getmorefigures.php?uid=PMC3376858&req=5

fig04: AGS-499 increases telomerase activity in the forebrain, BS and SC in a time-dependent mannerMice were injected s.c. with AGS-499 (6 mg/kg) or its vehicle DMSO. Cytoplasmic (A) and nuclear (B) protein extracts from the FB, and whole cell protein extract from the BS or SC (C) at various intervals after compound injection were prepared.A-C. Telomerase activity by TRAP assay. Protein extracts (2 µg) were added to the TRAP-specific reaction mixture. Negative control (NG) contained CHAPS buffer instead of the protein extract. IS, internal standard.D. Quantification of the TRAP results by densitometric analysis using the EZquant software (mean ± s.e.m.; n = 10), Student's t-test p = *10.092, *20.007, *30.028, *40.005, *50.0002, *60.004, *70.045.E. Quantification of telomerase activity by the real-time PCR-based TRAP assay kit. Cytoplasmic and nuclear protein extracts (1 µg) derived from the FB of AGS-treated or untreated mice for various intervals, were added to the reaction according to the manufacturer's instructions and telomerase activity products were calculated as moles DNA/µg protein (mean ± s.e.m.; n = 6), Student's t-test p = *10.043, *20.026. Symbols: UT, untreated mice; V, vehicle.

Mentions: Telomerase activity was determined by TRAP assay and by real time PCR TRAP using the same assay conditions as above. Telomerase activity in the FB, the BS and the SC increased with time following AGS treatment and exhibited a similar time-dependent activation to that found with TERT protein (Fig 4). As can be seen in Fig 4A–D for TRAP assay and Fig. .4E for real time PCR-based TRAP assay, telomerase activity in the cytoplasmic and nuclear extracts derived from FB and in the whole cell protein extract derived from the BS and SC exhibited a gradual increase with time, with a peak at 12 h (nucleus 2.54-fold, p <0.01; cytoplasm 2.63-fold, p <0.01), a decrease at 24 h (nucleus 2.44-fold, p <0.01; cytoplasm 1.54-fold, p <0.01) and reverting to the basal level 48 h after AGS treatment.


Novel telomerase-increasing compound in mouse brain delays the onset of amyotrophic lateral sclerosis.

Eitan E, Tichon A, Gazit A, Gitler D, Slavin S, Priel E - EMBO Mol Med (2012)

AGS-499 increases telomerase activity in the forebrain, BS and SC in a time-dependent mannerMice were injected s.c. with AGS-499 (6 mg/kg) or its vehicle DMSO. Cytoplasmic (A) and nuclear (B) protein extracts from the FB, and whole cell protein extract from the BS or SC (C) at various intervals after compound injection were prepared.A-C. Telomerase activity by TRAP assay. Protein extracts (2 µg) were added to the TRAP-specific reaction mixture. Negative control (NG) contained CHAPS buffer instead of the protein extract. IS, internal standard.D. Quantification of the TRAP results by densitometric analysis using the EZquant software (mean ± s.e.m.; n = 10), Student's t-test p = *10.092, *20.007, *30.028, *40.005, *50.0002, *60.004, *70.045.E. Quantification of telomerase activity by the real-time PCR-based TRAP assay kit. Cytoplasmic and nuclear protein extracts (1 µg) derived from the FB of AGS-treated or untreated mice for various intervals, were added to the reaction according to the manufacturer's instructions and telomerase activity products were calculated as moles DNA/µg protein (mean ± s.e.m.; n = 6), Student's t-test p = *10.043, *20.026. Symbols: UT, untreated mice; V, vehicle.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3376858&req=5

fig04: AGS-499 increases telomerase activity in the forebrain, BS and SC in a time-dependent mannerMice were injected s.c. with AGS-499 (6 mg/kg) or its vehicle DMSO. Cytoplasmic (A) and nuclear (B) protein extracts from the FB, and whole cell protein extract from the BS or SC (C) at various intervals after compound injection were prepared.A-C. Telomerase activity by TRAP assay. Protein extracts (2 µg) were added to the TRAP-specific reaction mixture. Negative control (NG) contained CHAPS buffer instead of the protein extract. IS, internal standard.D. Quantification of the TRAP results by densitometric analysis using the EZquant software (mean ± s.e.m.; n = 10), Student's t-test p = *10.092, *20.007, *30.028, *40.005, *50.0002, *60.004, *70.045.E. Quantification of telomerase activity by the real-time PCR-based TRAP assay kit. Cytoplasmic and nuclear protein extracts (1 µg) derived from the FB of AGS-treated or untreated mice for various intervals, were added to the reaction according to the manufacturer's instructions and telomerase activity products were calculated as moles DNA/µg protein (mean ± s.e.m.; n = 6), Student's t-test p = *10.043, *20.026. Symbols: UT, untreated mice; V, vehicle.
Mentions: Telomerase activity was determined by TRAP assay and by real time PCR TRAP using the same assay conditions as above. Telomerase activity in the FB, the BS and the SC increased with time following AGS treatment and exhibited a similar time-dependent activation to that found with TERT protein (Fig 4). As can be seen in Fig 4A–D for TRAP assay and Fig. .4E for real time PCR-based TRAP assay, telomerase activity in the cytoplasmic and nuclear extracts derived from FB and in the whole cell protein extract derived from the BS and SC exhibited a gradual increase with time, with a peak at 12 h (nucleus 2.54-fold, p <0.01; cytoplasm 2.63-fold, p <0.01), a decrease at 24 h (nucleus 2.44-fold, p <0.01; cytoplasm 1.54-fold, p <0.01) and reverting to the basal level 48 h after AGS treatment.

Bottom Line: Hence, we and others hypothesized that increasing telomerase expression by pharmaceutical compounds may protect brain cells from death caused by damaging agents.The survival of telomerase-expressing cells (i.e. motor neurons), but not telomerase-deficient cells, exposed to oxidative stress was increased by AGS-499 treatment, suggesting that the AGS-499 effects are telomerase-mediated.Therefore, a controlled and transient increase in telomerase expression and activity in the brain by AGS-499 may exert neuroprotective effects.

View Article: PubMed Central - PubMed

Affiliation: The Shraga Segal Department of Immunology and Microbiology, Ben-Gurion University of the Negev, Beer-Sheva, Israel.

Show MeSH
Related in: MedlinePlus