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| 1. |
Wruck, C.J., Götz, M.E., Herdegen, T., Varoga, D., Brandenburg, L-O. and Pufe, T.
(2008)
Kavalactones protect neural cells against amyloid β peptide-induced neurotoxicity via ERK1/2-dependent Nrf2-activation
Molecular Pharmacology Fast Forward
March 11, 2008
,
epub ahead of print
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Notes:
The accumulation of the toxic form of the Amyloid-β peptide is known to induce oxidative damage in the brain. Although treatment with antioxidants has not proven effective at controlling AD symptoms, inducing the natural systems in the brain that protect from oxidative damage may provide a possible therapeutic approach. A host of antioxidant and detoxifying enzymes are upregulated by binding of the Nrf2 transcription factor to the ARE (antioxidant response element) regulatory sequence. The authors used a Dual Luciferase® Reporter Assay to assess modulation of gene activity through ARE by kavalactones. Kavalactones are compounds found in the roots and rhizomes of Kava (Piper methysticum), a plant cultivated an used in some Pacific societies for medicinal and social uses. The ARE1 region from the rat NAD(P)H:quinone oxidoreductase-1 gene was placed upstream of a pGL3 firefly luciferase reporter construct and cotransfected along with a pRL-TK Renilla control construct into PC12 or C6 cells. The data show induction of luciferase activity by kavalactones. Further investigation shows that the kavalactones promote Nrf2 stabilization possibly through the ERK1/2 pathway.
(0003859) |
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Products: Dual-Luciferase® Reporter Assay System | pGL3-Basic Vector | pGL3-Control Vector | pGL3-Enhancer Vector | pGL3-Promoter Vector | pRL-TK Vector |
| 2. |
He, W., Shi, Q., Hu, X. and Yan, R.
(2007)
The membrane topology of RTN3 and its effect on binding of RTN3 to BACE1
J. Biol. Chem.
282
,
29144-29151
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Notes:
The authors of this study determined the membrane topology of reticulon 3 (RTN3), an integral membrane protein that is expressed at high levels in neruons and has been show to negatively regulate the activity of BACE1 (Beta site APP-Cleaving Enzyme). Disruption of RTN3 is associated with incidence of dystrophic neurites in AD brain. RTN3 was translated using the TNT® Quick Coupled Transcription/Translation System in the presence of Canine Microsomal Membranes and labeled using the Transcend™ Non-Radioactive Translation Detection System.
(0003860) |
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Products: Canine Pancreatic Microsomal Membranes | TNT® SP6 Quick Coupled Transcription/Translation System | TNT® SP6 Quick Coupled Transcription/Translation System, Trial Size | TNT® T7 Quick Coupled Transcription/Translation System | TNT® T7 Quick Coupled Transcription/Translation System, Trial Size | Transcend™ Chemiluminescent Non-Radioactive Translation Detection System | Transcend™ Colorimetric Non-Radioactive Translation Detection System |
| 3. |
Tanaka, M., Chock, P.B., and Stadtman, E.R.
(2007)
Oxidized messenger RNA induces translation errors.
Proc. Natl. Acad. Sci. U S A
104
,
66-71
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Notes:
Oxidative damage has been associated with a range of age-related neurological conditions. In this study, the effect of mRNA oxidation was investigated. A direct correlation was observed between the extent of oxidation and the frequency of translation errors. The authors excised the firefly luciferase (luc2) gene from the pGL4.14 Vector, attached a FLAG tag to the 5´ terminus and a Myc tag to the 3´ terminus, and subcloned the gene into a pGEM-4Z Vector that had been modified to append a poly(A) sequence. The construct was transfected into HEK293 cells, which were then cultured in the presence of an oxidizing agent. The occurrence of truncated protein fragments and short peptides increased in the presence of the oxidizing agent in a concentration-dependent manner. The effects of oxidation of mRNA were also investigated in in vitro translation experiments using mRNA treated with an iron-ascorbate mixture and hydrogen peroxide. Translation in vitro was performed using rabbit reticulocyte lysate supplemented with protease inhibitors. The translation products were detected using anti-FLAG and anti-c-Myc antibodies.
(0003630) |
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Products: pGEM®-4Z Vector | pGL4.14[luc2/Hygro] Vector | Rabbit Reticulocyte Lysate System, Nuclease Treated |
| 4. |
Zhang, Y-w., Wang, R., Liu, Q., Zhang, H., Liao, F-F. and Xu, H.
(2007)
Presenilin/γ-secretase-dependent processing of β-amyloid precursor protein regulates EGF receptor expression
Proc. Natl. Acad. Sci. U S A
104
,
10613-10618
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Notes:
The authors of this study investigated the downstream effects of the release of the intracellular domain of the Amyloid-β precursor protein (AICD) on cellular activities. They amplified the 5′ region of the mouse EGFR gene and cloned it into a pGL3 vector. This construct was cotransfected into embryonic fibroblasts derived from APP/APLP2 DKO mice along with a vector expressing AICD, AICD and the multidomain protein Fe65, Fe65 alone or NotchΔE, along with a Renilla control vector to normalize data for transfection efficiency. The data indicate that AICD negatively regulates transcription of the EGF Receptor gene.
(0003861) |
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Products: pGL3-Promoter Vector | pRL-SV40 Vector |
| 5. |
Renaud, S., Loukinov, D., Abdullaev, Z., Guilleret, I., Bosman, F.T., Lobanenkov, V. and Benhattar, J.
(2007)
Dual role of DNA methylation inside and outside of CTCF-binding regions in the transcriptional regulation of the telomerase hTERT gene
Nucleic Acids Research
35
,
1245-1256
.
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Notes:
Telomeres shorten by 50–100 bases with each cell division, making the telomere a "mitotic counter" that can limit cellular lifespan. Telomerase is a two-component protein consisting of a reverse transcriptase (hTERT) bound to its own RNA template that can act to maintain telomere length in dividing cells. Telomerase is highly active in dividing cells such as germ cells, stem cells and many cancers. This paper investigated the role of methylation of the hTERT promoter and the transcription factor CTCF in regulation of telomerase activity. LacZ reporter plasmids driven by the hTERT minimal promoter were transiently transfected into HeLa cells, and reporter assays were performed on lysate generated using Passive Lysis Buffer. The hTERT minimal promoter did not show activity if all of the CpG sites were methylated. The promoter and first exon of hTERT were amplified using PCR Master Mix from sodium bisulfite-treated genomic DNA isolated from telomerase-positive cell lines and tissues. The resulting fragments were cloned using the pGEM®-T Vector System II. For the methylation cassette assay, methylated and unmethylated fragments were cloned into a methylated or unmethylated vector using the LigaFast™ Rapid DNA Ligation System. The authors conclude that methylation plays a dual role in regulating hTERT expression. CTCF will bind to the first exon of hTERT when the hTERT CpG island is not methylated, resulting in downregulation of hTERT expression. Although CTCF cannot bind the hTERT promoter when the DNA is completely methylated, the methylation itself completely represses transcription. In situations where there is partial methylation of the promoter, such as in tumor cells, CTCF cannot bind to the promoter, but the partial methylation is not enough to repress transcription, and hTERT is expressed.
(0003641) |
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Products: LigaFast™ Rapid DNA Ligation System | MspI | Passive Lysis 5X Buffer | PCR Master Mix | pGEM®-T Vector System II |
| 6. |
Witkowski, J.M., Soroczynska-Cybula, M., Bryl, E., Smolenska, Z., and Jozwik, A.
(2007)
Klotho—a Common Link in Physiological and Rheumatoid Arthritis-Related Aging of Human CD4 Lymphocytes
J. Immunology
178
,
771–777
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Notes:
Klotho knockout mice exhibit a phenotype of precocious aging, organ failure, osteoporosis. Humans with specific Klotho alleles are at increased risk for osteoporosis, atherosclerosis and decreased lifespan. The authors of this study looked at the expression of Klotho in CD4+ lymphocytes in patients suffering from rheumatoid arthritis and age-matched healthy individuals. cDNA was prepared from total RNA isolated from purified CD4+ lymphocytes using the ImProm-II™ Reverse Transcription System. Klotho expression, protein level and activity was decreased in the lymphocytes from the RA patients.
(0003654) |
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Products: ImProm-II™ Reverse Transcriptase | ImProm-II™ Reverse Transcription System |
| 7. |
Liu, Y., Kern, J.T., Walker, J.R., Johnson, J.A., Schultz, P.G., and Luesch, H.
(2007)
A genomic screen for activators of the antioxidant response element.
Proc. Natl. Acad. Sci. U S A
104
,
5205-5210
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Notes:
These authors screened a library of 15,000 expression cDNAs in neuroblastoma IMR-32 cells searching for genes that activate the antioxidant response element, ARE. ARE is a cis-acting enhancer element found in the 5´ flanking region of many genes that are involved in protection from oxidative stress. The library was screened using a luciferase reporter construct under the control of an ARE-containing promoter. Luminescence, indicating the presence of cDNA-activating ARE, was measured using the Bright-Glo™ Luciferase Assay System. cDNA clones showing reproducible activation were selected for further analysis. The authors tested the effect of over expression of these ARE activators on the ability to resist oxidative stress. IMR-32 cells expressing the various cDNAs were exposed to hydrogen peroxide or rotenone, and the effect on cell viability was measured using the CellTiter-Glo® Assay. Cells overexpressing the ARE-activators were more resistant to oxidative stress than controls.
(0003629) |
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Products: Bright-Glo™ Luciferase Assay System | CellTiter-Glo® Luminescent Cell Viability Assay |
| 8. |
Vessal, M., Mishra, S., Moulik, S. and Murphy, L.J.
(2006)
Prohibitin attenuates insulin-stimulated glucose and fatty acid oxidation in adipose tissue by inhibition of pyruvate carboxylase
FEBS Journal
273
,
568-576
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Notes:
Prohibitin (PHB-1) is a multifunctional protein that is located in the mitochondria and plasma membrane and is secreted by adipocytes. Previously PHB-1 was shown to function as a chaperone protein for newly made subunits of mitochondrial respiratory enzymes. This paper describes a role for exogenous PHB-1 in the modulation of insulin-stimulated glucose and fatty acid oxidation. To identify PHB-1 binding partners, His-tagged PHB-1 was incubated with adipocytes. Membranes were solubilized and proteins separated by electrophoresis. Silver-stained bands were excised and destained before treatment with sequencing grade Trypsin. The tryptic peptides were analyzed by HPLC and Mass Spectrometry.
(0003634) |
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Products: Sequencing Grade Modified Trypsin | Sequencing Grade Modified Trypsin, Frozen |
| 9. |
Mourikis, P., Hurlbut, G.D. and Artavanis-Tsakonas, S.
(2006)
Enigma, a mitochondrial protein affecting lifespan and oxidative stress response in Drosophila
Proc. Natl. Acad. Sci. USA
103
,
1307–1312
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Notes:
The authors of this study describe mutations in a gene, Enigma (Egm), that increase lifespan and protect against oxidative stress. They mapped the gene and showed that it encodes a mitochontrial protein that is homologous to enzymes involved in the β-oxidation of fatty acids. To determine what other genes might be affected by loss of function of Egm, the authors performed an RNAi experiment using Drosophila Kc-167 cells. Double-stranded RNA was produced using the RiboMax™ Large Scale RNA Production System, and a portion of the Egm cDNA was amplified from that RNA. Some of the genes affected by the Egm RNA include a homolog of a mammalian enzyme known to be involved in fatty acid β-oxidation and enzymes involved in detoxification including glutathione-S-transferase and a cytochrome P450.
(0003668) |
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Products: RiboMAX™ Large Scale RNA Production System—SP6 | RiboMAX™ Large Scale RNA Production System—T7 |
| 10. |
Luo, L., Chen, H., Trush, M.A., Show, M.D., Anway, M.D. and Zirkin, B.R.
(2006)
Aging and the Brown Norway Rat Leydig Cel Antioxidant Defense System
Journal of Andrology
27
,
240-247
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Notes:
Rat cDNA fragments of the genes encoding copper-zinc superoxide dismutase, manganese superoxide dismutase and glutathione peroxidase were amplified and cloned into the pGEM®-T Easy Vector. The clones were used to probe Northern blots to examine the effect of age on the expression of these antioxidant defense genes.
(0003632) |
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Products: pGEM®-T Easy Vector System I | pGEM®-T Easy Vector System II |
| 11. |
Payne, A.M., Zheng, Z., Messi, M.L., Milligan, C.E., González, E. and Delbono, O.
(2006)
Motor neurone targeting of IGF-1 prevents specific force decline in ageing mouse muscle
J. Physiol.
570
,
283-294
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Notes:
Overexpression of IGF-1 can delay or prevent aging problems in motor neurons and skeletal muscle. The authors of this paper were able to target IGF-1 to motor neurons using a fusion protein containing tetanus toxin fragment C (TTC). Motor neurons will bind, take up and transport the TTC fragment with no toxicity to the neurons. Full-length human IGF-1 cDNA was generated by PCR and inserted into the pGEM®-T Easy Vector. TTC amplified from Clostridium tetani CN655 genomic DNA was inserted into the vector. The new IFG-1-TTC insert was used for PCR to eventually produce the fusion protein for the studies.
(0003635) |
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Products: pGEM®-T Easy Vector System I | pGEM®-T Easy Vector System II |
| 12. |
Zirah, S., Kozin, S.A., Mazur, A.K., Blond, A., Cheminant, M., Ségalas-Milazzo, I., Debey, P., and Rebuffat, S.
(2006)
Structural changes of region 1-16 of the Alzheimer disease amyloid beta-peptide upon zinc binding and in vitro aging.
J. Biol. Chem.
281
,
2151–61
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Notes:
Aspartyl modifications of the amyloid peptide Aβ are related to protein conformation changes associated with amyloid deposits found in Alzheimer disease patients. The authors investigated region 1–16 of Aβ, the minimal zinc-binding domain, to determine if in vitro aging of Aβ-(1–16) results in aspartyl modifications. The level of isoaspartic acid was quantitated using the ISOQUANT® Isoaspartate Detection Kit. Reactions were performed with 10µl of a 7.5µM peptide solution at 30°C for 30 minutes, and reaction products were quantitated by reverse phase HPLC analysis.
(0003665) |
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Products: ISOQUANT® Isoaspartate Detection Kit |
| 13. |
Tumer, N., Scarpace, P.J., Dogan, M.D., Broxson, C.S., Matheny, M., Yurek, D.M., Peden, C.S., Burger, C., Muzyczka, N. and Mandel, R.J.
(2006)
Hypothalamic rAAV-mediated GDNF gene delivery ameliorates age-related obesity.
Neurobiol. Aging
27
,
459–70
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Notes:
To examine the effect of GDNF expression on body weight, recombinant adeno-associated virus (rAAV) expressing GDNF was injected intrahypothalamically into young and senescent rats. Hypothalamuses were harvested, lysed and used undiluted, and at dilutions of 1:50 and 1:500 in the GDNF Emax® ImmunoAssay System. The amount of GDNF was expressed as a concentration of GDNF protein per milligram of hypothalamic tissue sample.
(0003342) |
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Products: GDNF Emax® ImmunoAssay System |
| 14. |
Shukla, V., Coumoul, X., Cao, L., Wang, R.-H., Xiao, C., Xu, X., Andò, S., Yakar, S., LeRoith, D. and Deng, C.
(2006)
Absence of the full-length breast cancer-associated gene-1 leads to increased expression of insulin-like growth factor signaling axis members.
Cancer Res.
66
,
7151-7157
.
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Notes:
The authors of this study investigated the influence of BRCA1 on insulin-like growth factor 1 (IGF-1) signaling. In mice lacking full-length BRCA, they observed increased IGF-1 expression as well as changes in expression of other proteins within the IGF-1 signaling pathway including Irs-I. They observed increases in IGF-I and Irs-I expression in mammary tumors from these same mice. To understand better the relationship between BRCA1 and IGF, they transfected a mouse mammary tumor cell line with a small hairpin RNA directed against BRCA1 and showed that Irs-1 mRNA and promoter activity increases. Similar results were observed in human UBR60 cells. Irs-1 promoter activity was assessed using the Dual-Luciferase® Reporter Assay System.
(0003604) |
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Products: Dual-Luciferase® Reporter Assay System | Dual-Luciferase® Reporter Assay System 10-Pack |
| 15. |
Kunieda, T., Minamino,T., Katsuno, T., Tateno,K., Nishi, J-i., Miyauchi, H., Orimo, M., Okada, S. and Komuro, I.
(2006)
Cellular senescence impairs circadian expressionof clock genes in vitro and in vivo
Circulation Research
98
,
532–539
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Notes:
The authors of this study investigated the effect of cellular senescence on expression of the genes that regulate circadian rhythms, specifically Per2 and BmalI. The Dual-Luciferase® Assay was used to determine activity of a PER gene reporter in senescent and young primary cultured human aortic vascular smooth muscle cells. Senescent cells activated CREB, but much more weakly than dividing cells did.
(0003672) |
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Products: Dual-Luciferase® Reporter Assay System | Dual-Luciferase® Reporter Assay System 10-Pack |
| 16. |
Seehuus, S-C., Norberg, K., Gimsa, U., Krekling, T. and Amdam, G.V.
(2006)
Reproductive protein protects functionally sterile honey bee workers from oxidative stress
Proc. Natl. Acad. Sci. U S A
103
,
962-967
.
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Notes:
The authors of this paper demonstrate that the pathways controlling lifespan and senescence are linked to the pathways controlling fertility in the honey bee. Using RNAi experiments, the authors demonstrate that vitellogenin protects honey bee workers from oxidative stress. Double-stranded RNA was synthesized from the vitellogenin clone AP4a5 using the RiboMax® T7 System. In additional experiments the authors assessed apoptosis in brain tissue sections using the DeadEND™ Fluorometric TUNEL system.
(0003636) |
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Products: DeadEnd™ Fluorometric TUNEL System | RiboMAX™ Large Scale RNA Production System—T7 |
| 17. |
Hu, D., Serrano, F., Oury, T.D. and Klann, E.
(2006)
Aging-Dependent alterations in synaptic plasticity and memory in mice that overexpress extracellular superoxide dismutase
J. Neuroscience
26
,
3993–3941
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Notes:
Reactive oxygen species (ROS) are considered neurotoxic and contribute to age-related cognitive decline; however, ROS are also required components of the signal transduction pathways involved in synaptic plasticity and memory. The authors of this study investigated the effect of overexpression of extracellular superoxide dismutase (EC-SOD) in mice. They specifically looked at the activation of stress-related signaling pathways involving ERK, JNK and p38. The AntiACTIVE® MAPK pAb and the Anti-ERK 1/2 pAb were used to probe Western blots of protein isolated from aged wildtype and transgenic mice. The transgenic mice did not show the same age-related upregulation of these proteins as the wildtype mice.
(0003671) |
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Products: Anti-ACTIVE® MAPK Family Sampler | Anti-ERK 1/2 pAb, Rabbit |
| 18. |
Bakshi, P., Liao, Y-F., Gao, J., Ni, J., Stein, R., Yeh, L-A., Wolfe, M.S.
(2005)
A high-throughput screen to identifiy inhibitors of amyloid beta-protein precursor processing
J. Biomol. Screen.
10
,
1-12
.
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Notes:
A key component in the pathogenesis of Alzheimer’s disease is cerebral accumulation of amyloid-beta protein (Aβ). Aβ is produced by proteolysis of amyloid-β-protein precursor (APP) by ß- and gamma-secretases, thus these enzymes are considered important drug targets for Alzheimer’s disease. Existing assays for assessing inhibition of alpha-, beta- and gamma-secretases include HPLC or ELISA assays that are cumbersome, expensive and not well-suited to high-throughput screening. The authors developed a luciferase reporter system to identify new molecules that inhibit APP processing. They then successfully interfaced this sensitive, specific and quantitative assay with a high-throughput screen, useful for identifying both inhibitors and stimulators of APP processing.
(0003775) |
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Products: Bright-Glo™ Luciferase Assay System | CytoTox-ONE™ Homogeneous Membrane Integrity Assay | CytoTox-ONE™ Homogeneous Membrane Integrity Assay, HTP | Dual-Luciferase® Reporter Assay System | Dual-Luciferase® Reporter Assay System 10-Pack | Passive Lysis 5X Buffer | pRL-TK Vector |
| 19. |
Yamamoto, M., Clark, J.D., Pastor, J.V., Gurnani, P., Nandi, A., Kurosu, H., Miyoshi, M., Ogawa, Y., Castrillon, D.H., Rosenblatt, K.P. and Kuro-o, M.
(2005)
Regulation of Oxidative Stress by the Anti-aging Hormone Klotho
J. Biol. Chem.
280
,
38029–38034
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Notes:
Mice that overexpress Klotho exhibit an extended lifespan and delayed aging. In this study, the authors show that Klotho protein protects against oxidative stress and activates the FoxO transcription factors, inducing expression of manganese superoxide dismutase. Two luciferase constructs were made, one with luciferase under the control of the Fas ligand promoter and one under the control of the human SOD2 promoter. HeLa cells were transfected with one of the two luciferase constructs and the pRL-CMV Renilla control vector. Transfected cells were treated with or without Klotho protein, and cell lysates were analyzed using the Dual-Luciferase® Assay. Klotho protein stimulated the activity of the Fas ligand gene promoter and the SOD2 promoter.
(0003667) |
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Products: Dual-Luciferase® Reporter Assay System | Dual-Luciferase® Reporter Assay System 10-Pack | pRL-CMV Vector |
| 20. |
Al-Regaiey, K.A., Masternak, M.M., Bonkowski, M., Sun, L. and Bartke, A.
(2005)
Long-lived growth hormone receptor knockout mice: Interaction of reduced insulin-like growth factor I/insulin signaling and caloric restriction
Endocrinology
146
,
851–860
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Notes:
The authors of this study examined the relationship between pathways through which growth hormone (GH), insulin-like growth factor I (IGF-I), and caloric restriction regulate lifespan. Normal and long-lived GH-receptor knockout (GHRKO) mice were compared with transgenic mice overexpressing bovine GH (short lived). When all mice were subjected to caloric restriction, the Normal and GHRKO mice showed decreases in phosphorylation of hepatic Akt, increases in PPAR-gamma and increases in FOXO I transcription. The AntiACTIVE® p38 pAb was used to examine p38 activity in all three types of mice subjected to caloric restriction. Active p38 was increased in normal and GHRKO mice compared to the transgenic mice and normal mice fed ad lib. The authors show that the Akt/FOXO pathway plays a major role in regulating longevity and provides a link between IGF-1, caloric restriction and growth hormone regulation of lifespan.
(0003652) |
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Products: Anti-ACTIVE® p38 pAb, Rabbit, (pTGpY) |
| 21. |
Debacq-Chainiaux,F., Borlon, C., Pascal, T., Royer, V., Eliaers,F., Ninane, N., Carrard, G., Friguet, B., de Longueville, F., Boffe, S., Remacle, J. and Toussaint, O.
(2005)
Repeated exposure of human skin fibroblasts to UVB at subcytotoxic level triggers premature senescence through the TGF-β1 signaling pathway
J. Cell Science
118
,
743–758
.
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Notes:
The authors of this study developed a model for UVB-induced premature senescence of skin human diploid fibroblasts. Markers of senescence were expressed in the system, and the authors were able to detect a common mitochondrial DNA 4,977-bp deletion that is associated with oxidative damage. After a series of ten UVB stresses, total RNA was prepared from the cells using the RNAgents® Total RNA Isolation System and used for RT-PCR to detect differentially expressed genes. Forty-four stress or senescence-associated genes were identified that were differentially expressed between UVB irradiated and untreated cells including c-fos, c-jun, insulin-like growth factor binding protein 3, several HSPs, genes involved in protection from oxidative stress, and the type II receptor of TGF-β. The Anti-ACTIVE® Caspase-3 pAb was used to assess whether the UVB treatment or incubation with TGF-β1 led to apoptosis in this system.
(0003670) |
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Products: Anti-ACTIVE® Caspase-3 pAb | RNAgents® Total RNA Isolation System |
| 22. |
Baba, T., Shimizy, T., Suzuki, Y-i., Ogawara, M., Isono, K-i., Kosek, H., Kurosawa, H. and Shirasawa, T.
(2005)
Estrogen, insulin, and dietary signals cooperatively regulate longevity signals to enhance resistance to oxidative stress in mice
J. Biol. Chem.
280
,
16417-16426
.
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Notes:
The authors of this study created a mouse model of a C. elegans insulin receptor mutation. The Altered Sites® II in vitro Mutagenesis System was used to create a transgenic mouse containing a Pro1195Leu substitution in the insulin receptor. Mice homozygous for this mutation died as neonates. Heterozygous mice grew normally, but lifespan was increased by 33% in females and 18% in males. Manganese superoxide dismutase was upregulated in both male and female heterozygotes. Estrogen further increased longevity of the mutant mice as did caloric restriction.
(0003669) |
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Products: Altered Sites® II in vitro Mutagenesis System |
| 23. |
Sarkar, D., Lebedeva, I.V., Emdad, L., Kang, D-c., Baldwin, A.S. and Fisher, P.B.
(2004)
Human polynucleotide phosphorylase (hPNPase0ld-35): A potential link between aging and inflammation
Cancer Research
64
,
7473-7478
.
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Notes:
Human polynucleotide phosphorylase (hPNPaseold-35) was originally identified as a gene that is upregulated during cellular differentiation and senescence. The authors of this study show that overexpression of hPNPaseold-35 results in the increased production reactive oxygen species (ROS) and the activation of the NF-κB pathway, resulting in expression of the inflammatory cytokines IL-6 and IL-8. HeLa cells were transfected with either empty pGL3-Basic or 3kB-Luc (pGL3-Basic containing three tandem NF-κB binding sites). Transfected cells were infected with an empty adenoviral vector or one containing hPNPaseold-35. Luciferase assays were conducted using the Luciferase Assay System, and signal was normalized to a pSV-βgal control. A 10- to 12-fold increase in luciferase activity was observed in the cells infected with the adenovirus containing hPNPaseold-35 compared to the control cells. This activation was inhibited by compounds that reduced ROS. The authors suggest that hPNPaseold-35 plays an important role in producing age-related inflammation.
(0003643) |
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Products: Luciferase Assay System | pGL3-Control Vector | pSV-β-Galactosidase Control Vector |
| 24. |
Jia, K., Chen, D. and Riddle, D.L.
(2004)
The TOR pathway interacts with the insulin signaling pathway to regulate C. elegans larval development, metabolism and life span
Development
131
,
3897-3906
.
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Notes:
The authors of this study investigated the function of the daf-15 gene in C. elegans. The daf-15 gene encodes the C. elegans ortholog of raptor, a protein involved in the TOR signaling pathway. In C. elegans, daf-15 transcription is regulated by DAF-16, a FOXO transcription factor which is itself regulated by the insulin/IGF-1 signaling pathway. This work links regulation of the TOR pathway, which controls cell growth, to the insulin/IGF-1 pathway, known to affect lifespan, development and metabolism. Three candidate daf-15 genes were amplified by PCR and cloned into the pGEM®-T Vector. The Riboprobe® SP6/T7 transcription system was used to transcribe RNA from the candidate clones. Semiquantitative RT-PCR was performed to compare daf-15 mRNA levels in daf-2 and daf-16; daf-2 mutant animals. The mRNA was purified from total worm RNA using the PolyATtract® mRNA Isolation System.
(0003633) |
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Products: pGEM®-T Vector System I | pGEM®-T Vector System II | PolyATtract® mRNA Isolation System I (Refill for Z5200) | PolyATtract® mRNA Isolation System II with Magnetic Stand | PolyATtract® mRNA Isolation System III with Magnetic Stand | PolyATtract® mRNA Isolation System IV (Refill for Z5300) | Riboprobe® Combination System—SP6/T7 RNA Polymerase |
| 25. |
Jezierski, M.K., and Sohrabji, F.
(2001)
Neurotrophin expression in the reproductively senescent forebrain is refractory to estrogen stimulation.
Neurobiol. Aging
22
,
309-319
.
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Notes:
The regulation of brain-derived neurotrophic factor, nerve growth factor and other neurotrophin ligands and receptors by estrogen was characterized in young adult and reproductively senescent rat brains. BNDF, NT-4, and NGF protein levels were monitored using Promega's BDNF Emax® ImmunoAssay, NT-4 Emax® ImmunoAssay System, and NGF Emax® ImmunoAssay Systems, respectively .
(0002320) |
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Products: BDNF Emax® ImmunoAssay System | NGF Emax® ImmunoAssay System |