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The PPARδ ligand GW501516 reduces growth but not apoptosis in mouse inner medullary collecting duct cells.

Clark J, Nasrallah R, Hébert RL - PPAR Res (2009)

Bottom Line: The collecting duct (CD) expresses considerable amounts of PPARδ.High doses of GW501516 decreased both DNA and protein synthesis in these cells by 80%, but had no overall effect on cell viability.Although anisomycin treatment resulted in an increase of caspase-3 levels of about 2.59-fold of control, GW501516 did not affect anisomycin-induced changes in active caspase-3 levels.

View Article: PubMed Central - PubMed

Affiliation: Department of Cellular and Molecular Medicine, Kidney Research Centre, Faculty of Medicine, University of Ottawa, Ottawa, ON, Canada K1H 8M5.

ABSTRACT
The collecting duct (CD) expresses considerable amounts of PPARδ. While its role is unknown in the CD, in other renal cells it has been shown to regulate both growth and apoptosis. We thus hypothesized that PPARδ reduces apoptotic responses and stimulates cell growth in the mouse CD, and examined the effect of GW501516, a synthetic PPARδ ligand, on these responses in mouse IMCD-K2 cells. High doses of GW501516 decreased both DNA and protein synthesis in these cells by 80%, but had no overall effect on cell viability. Although anisomycin treatment resulted in an increase of caspase-3 levels of about 2.59-fold of control, GW501516 did not affect anisomycin-induced changes in active caspase-3 levels. These results show that a PPARδ ligand inhibits growth but does not affect anisomycin-apoptosis in a mouse IMCD cell line. This could have therapeutic implications for renal diseases associated with increased CD growth responses.

No MeSH data available.


Related in: MedlinePlus

GW501516 pretreatment has no effect onanisomycin-induced apoptosis. Serum-starved cells were stimulated with (a)–(c) serum-freemedia (control), (d)–(f) 10−6 M GW501516 (GW-6), (g)–(i) 10−8 M GW501516 (GW-8), (j)–(l) 250 ng/mLanisomycin (A250) for 24 hours, (m)–(o) 10−6 M, or (p)–(r) 10−8 M GW501516 for 3 hours pretreatment followed by 10−6 M or 10−8 M GW501516 for 24 hours with A250 (GW-6/A250 and GW-8/A250). The cells wereincubated with terminal transferase and biotin-16-dUTP for one hour, followedby two 45-minute incubations with streptavidin-Cy2 and anti-DAPI. The cellswere then mounted on slides, visualized with a fluorescent microscope, andcaptured with a camera. Apoptotic cells are shown in green, while the blue represents DAPI-positivecells.
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fig11: GW501516 pretreatment has no effect onanisomycin-induced apoptosis. Serum-starved cells were stimulated with (a)–(c) serum-freemedia (control), (d)–(f) 10−6 M GW501516 (GW-6), (g)–(i) 10−8 M GW501516 (GW-8), (j)–(l) 250 ng/mLanisomycin (A250) for 24 hours, (m)–(o) 10−6 M, or (p)–(r) 10−8 M GW501516 for 3 hours pretreatment followed by 10−6 M or 10−8 M GW501516 for 24 hours with A250 (GW-6/A250 and GW-8/A250). The cells wereincubated with terminal transferase and biotin-16-dUTP for one hour, followedby two 45-minute incubations with streptavidin-Cy2 and anti-DAPI. The cellswere then mounted on slides, visualized with a fluorescent microscope, andcaptured with a camera. Apoptotic cells are shown in green, while the blue represents DAPI-positivecells.

Mentions: In addition to looking at the effect ofGW501516 on caspase activity, TUNEL assays were performed to further evaluatethe apoptotic response. As shown in Figure 11, the proportion of TUNEL-positive cells increased while the totalnumber of cells decreases with the addition of 250 ng/mL anisomycin whencompared to control or the PPARδ agonist (at both concentrations). Pre/cotreatment of 10−6 M or 10−8 M GW501516 withanisomycin was comparable to anisomycin alone.


The PPARδ ligand GW501516 reduces growth but not apoptosis in mouse inner medullary collecting duct cells.

Clark J, Nasrallah R, Hébert RL - PPAR Res (2009)

GW501516 pretreatment has no effect onanisomycin-induced apoptosis. Serum-starved cells were stimulated with (a)–(c) serum-freemedia (control), (d)–(f) 10−6 M GW501516 (GW-6), (g)–(i) 10−8 M GW501516 (GW-8), (j)–(l) 250 ng/mLanisomycin (A250) for 24 hours, (m)–(o) 10−6 M, or (p)–(r) 10−8 M GW501516 for 3 hours pretreatment followed by 10−6 M or 10−8 M GW501516 for 24 hours with A250 (GW-6/A250 and GW-8/A250). The cells wereincubated with terminal transferase and biotin-16-dUTP for one hour, followedby two 45-minute incubations with streptavidin-Cy2 and anti-DAPI. The cellswere then mounted on slides, visualized with a fluorescent microscope, andcaptured with a camera. Apoptotic cells are shown in green, while the blue represents DAPI-positivecells.
© Copyright Policy - open-access
Related In: Results  -  Collection

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fig11: GW501516 pretreatment has no effect onanisomycin-induced apoptosis. Serum-starved cells were stimulated with (a)–(c) serum-freemedia (control), (d)–(f) 10−6 M GW501516 (GW-6), (g)–(i) 10−8 M GW501516 (GW-8), (j)–(l) 250 ng/mLanisomycin (A250) for 24 hours, (m)–(o) 10−6 M, or (p)–(r) 10−8 M GW501516 for 3 hours pretreatment followed by 10−6 M or 10−8 M GW501516 for 24 hours with A250 (GW-6/A250 and GW-8/A250). The cells wereincubated with terminal transferase and biotin-16-dUTP for one hour, followedby two 45-minute incubations with streptavidin-Cy2 and anti-DAPI. The cellswere then mounted on slides, visualized with a fluorescent microscope, andcaptured with a camera. Apoptotic cells are shown in green, while the blue represents DAPI-positivecells.
Mentions: In addition to looking at the effect ofGW501516 on caspase activity, TUNEL assays were performed to further evaluatethe apoptotic response. As shown in Figure 11, the proportion of TUNEL-positive cells increased while the totalnumber of cells decreases with the addition of 250 ng/mL anisomycin whencompared to control or the PPARδ agonist (at both concentrations). Pre/cotreatment of 10−6 M or 10−8 M GW501516 withanisomycin was comparable to anisomycin alone.

Bottom Line: The collecting duct (CD) expresses considerable amounts of PPARδ.High doses of GW501516 decreased both DNA and protein synthesis in these cells by 80%, but had no overall effect on cell viability.Although anisomycin treatment resulted in an increase of caspase-3 levels of about 2.59-fold of control, GW501516 did not affect anisomycin-induced changes in active caspase-3 levels.

View Article: PubMed Central - PubMed

Affiliation: Department of Cellular and Molecular Medicine, Kidney Research Centre, Faculty of Medicine, University of Ottawa, Ottawa, ON, Canada K1H 8M5.

ABSTRACT
The collecting duct (CD) expresses considerable amounts of PPARδ. While its role is unknown in the CD, in other renal cells it has been shown to regulate both growth and apoptosis. We thus hypothesized that PPARδ reduces apoptotic responses and stimulates cell growth in the mouse CD, and examined the effect of GW501516, a synthetic PPARδ ligand, on these responses in mouse IMCD-K2 cells. High doses of GW501516 decreased both DNA and protein synthesis in these cells by 80%, but had no overall effect on cell viability. Although anisomycin treatment resulted in an increase of caspase-3 levels of about 2.59-fold of control, GW501516 did not affect anisomycin-induced changes in active caspase-3 levels. These results show that a PPARδ ligand inhibits growth but does not affect anisomycin-apoptosis in a mouse IMCD cell line. This could have therapeutic implications for renal diseases associated with increased CD growth responses.

No MeSH data available.


Related in: MedlinePlus