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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.

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AGS-499 increases telomerase activity in the FB of adult mice in a dose-dependent mannerMice were injected s.c. with AGS-499 (3, 6 and 12 mg/kg of body weight) or its vehicle DMSO. Twelve hours later whole cell protein extract was prepared from the FB. Telomerase activity was determined by TRAP assay. Two micrograms of whole cell protein extract was added to the TRAP specific reaction mixture. Negative control (NG) contained CHAPS buffer instead of protein extract. IS, internal standard.Quantification of the TRAP results by densitometric analysis using the EZquant software (mean ± s.e.m.; n = 10) and Student's t-test, p = *10.019, *20.011, *30.030.Telomerase activity by the real-time PCR-based TRAP assay kit. Whole cell protein extract was prepared from FB of mice treated with AGS-499 (3, 6 and 12 mg/kg of body weight). One microgram of protein was added to the reaction according to the manufacturer's instructions and telomerase activity products were calculated as moles DNA/µg proteins (mean ± s.e.m.; n = 6). Student's t-test p = *0.08. Symbols: UT, untreated mice; V, vehicle; NG, negative control.
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fig02: AGS-499 increases telomerase activity in the FB of adult mice in a dose-dependent mannerMice were injected s.c. with AGS-499 (3, 6 and 12 mg/kg of body weight) or its vehicle DMSO. Twelve hours later whole cell protein extract was prepared from the FB. Telomerase activity was determined by TRAP assay. Two micrograms of whole cell protein extract was added to the TRAP specific reaction mixture. Negative control (NG) contained CHAPS buffer instead of protein extract. IS, internal standard.Quantification of the TRAP results by densitometric analysis using the EZquant software (mean ± s.e.m.; n = 10) and Student's t-test, p = *10.019, *20.011, *30.030.Telomerase activity by the real-time PCR-based TRAP assay kit. Whole cell protein extract was prepared from FB of mice treated with AGS-499 (3, 6 and 12 mg/kg of body weight). One microgram of protein was added to the reaction according to the manufacturer's instructions and telomerase activity products were calculated as moles DNA/µg proteins (mean ± s.e.m.; n = 6). Student's t-test p = *0.08. Symbols: UT, untreated mice; V, vehicle; NG, negative control.

Mentions: Telomerase activity in the aforementioned protein extracts was assayed by TRAP. As can be seen in Fig 2A (n = 5 independent experiments), telomerase activity in the FB of untreated or vehicle-treated mice was very low, while significant telomerase activity was detected 12 h post AGS-499 treatment. Quantification of telomerase activity from the TRAP assay data of five independent experiments revealed an increase of 3- (p < 0.05), 3.3- (p < 0.01) and 2.2- (p < 0.05) fold in telomerase activity in mice treated with 3, 6 and 12 mg/kg, respectively (Fig 2B). To confirm the increase in telomerase activity in the mouse FB following AGS treatment, a real time PCR-based TRAP assay was used. The results revealed that treatment of mice with AGS-499 increased telomerase activity in a dose-dependent manner. An increase of 2.4-, 3-, and 2-fold was observed when 3, 6 and 12 mg/kg of AGS 499 were injected, respectively (Fig 2C). Among the examined AGS doses, 6 mg/kg exhibited the most potent effect and therefore was used henceforth.


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 FB of adult mice in a dose-dependent mannerMice were injected s.c. with AGS-499 (3, 6 and 12 mg/kg of body weight) or its vehicle DMSO. Twelve hours later whole cell protein extract was prepared from the FB. Telomerase activity was determined by TRAP assay. Two micrograms of whole cell protein extract was added to the TRAP specific reaction mixture. Negative control (NG) contained CHAPS buffer instead of protein extract. IS, internal standard.Quantification of the TRAP results by densitometric analysis using the EZquant software (mean ± s.e.m.; n = 10) and Student's t-test, p = *10.019, *20.011, *30.030.Telomerase activity by the real-time PCR-based TRAP assay kit. Whole cell protein extract was prepared from FB of mice treated with AGS-499 (3, 6 and 12 mg/kg of body weight). One microgram of protein was added to the reaction according to the manufacturer's instructions and telomerase activity products were calculated as moles DNA/µg proteins (mean ± s.e.m.; n = 6). Student's t-test p = *0.08. Symbols: UT, untreated mice; V, vehicle; NG, negative control.
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fig02: AGS-499 increases telomerase activity in the FB of adult mice in a dose-dependent mannerMice were injected s.c. with AGS-499 (3, 6 and 12 mg/kg of body weight) or its vehicle DMSO. Twelve hours later whole cell protein extract was prepared from the FB. Telomerase activity was determined by TRAP assay. Two micrograms of whole cell protein extract was added to the TRAP specific reaction mixture. Negative control (NG) contained CHAPS buffer instead of protein extract. IS, internal standard.Quantification of the TRAP results by densitometric analysis using the EZquant software (mean ± s.e.m.; n = 10) and Student's t-test, p = *10.019, *20.011, *30.030.Telomerase activity by the real-time PCR-based TRAP assay kit. Whole cell protein extract was prepared from FB of mice treated with AGS-499 (3, 6 and 12 mg/kg of body weight). One microgram of protein was added to the reaction according to the manufacturer's instructions and telomerase activity products were calculated as moles DNA/µg proteins (mean ± s.e.m.; n = 6). Student's t-test p = *0.08. Symbols: UT, untreated mice; V, vehicle; NG, negative control.
Mentions: Telomerase activity in the aforementioned protein extracts was assayed by TRAP. As can be seen in Fig 2A (n = 5 independent experiments), telomerase activity in the FB of untreated or vehicle-treated mice was very low, while significant telomerase activity was detected 12 h post AGS-499 treatment. Quantification of telomerase activity from the TRAP assay data of five independent experiments revealed an increase of 3- (p < 0.05), 3.3- (p < 0.01) and 2.2- (p < 0.05) fold in telomerase activity in mice treated with 3, 6 and 12 mg/kg, respectively (Fig 2B). To confirm the increase in telomerase activity in the mouse FB following AGS treatment, a real time PCR-based TRAP assay was used. The results revealed that treatment of mice with AGS-499 increased telomerase activity in a dose-dependent manner. An increase of 2.4-, 3-, and 2-fold was observed when 3, 6 and 12 mg/kg of AGS 499 were injected, respectively (Fig 2C). Among the examined AGS doses, 6 mg/kg exhibited the most potent effect and therefore was used henceforth.

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