Mitotracker Green Is a <i>P</i>-Glycoprotein Substrate

Marques‐Santos, Luis Fernando; Oliveira, José Gustavo Pugliese de; Maia, Raquel Ciuvalschi; Rumjanek, Vivian M.

doi:10.1023/b:bire.0000007693.33521.18

Cited by 37 publications

(29 citation statements)

References 25 publications

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“…Mitochondrial mass was also measured in cells stably overexpressing IHG-1 by flow cytometric analysis of the fluorescence mitochondrial marker MitoTracker Red. 30,31 A significant increase in fluorescence (38%) was seen in IHG-1 overexpressing cells compared with controls, denoting an increase in mitochondrial mass ( Figure 4A IHG-1 was associated with a 1.4-fold increase in mtDNA (Figure 4B). We further confirmed increased mitochondrial mass in IHG-1-overexpressing cells by fluorescence imaging after incubation with MitoTracker Red ( Figure 4C).…”

Section: Loss Of Endogenous Ihg-1 Expression Results In Decreased Mitmentioning

confidence: 98%

IHG-1 Promotes Mitochondrial Biogenesis by Stabilizing PGC-1α

Hickey¹,

Corcoran²,

Docherty³

et al. 2011

Journal of the American Society of Nephrology

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Increased expression of Induced-by-High-Glucose 1 (IHG-1) associates with tubulointerstitial fibrosis in diabetic nephropathy. IHG-1 amplifies TGF-␤1 signaling, but the functions of this highly-conserved protein are not well understood. IHG-1 contains a putative mitochondrial-localization domain, and here we report that IHG-1 is specifically localized to mitochondria. IHG-1 overexpression increased mitochondrial mass and stabilized peroxisome proliferator-activated receptor ␥ coactivator-1␣ (PGC-1␣). Conversely, inhibition of IHG-1 expression decreased mitochondrial mass, downregulated mitochondrial proteins, and PGC-1␣-regulated transcription factors, including nuclear respiratory factor 1 and mitochondrial transcription factor A (TFAM), and reduced activity of the TFAM promoter. In the unilateral ureteral obstruction model, we observed higher PGC-1␣ protein expression and IHG-1 levels with fibrosis. In a gene-expression database, we noted that renal biopsies of human diabetic nephropathy demonstrated higher expression of genes encoding key mitochondrial proteins, including cytochrome c and manganese superoxide dismutase, compared with control biopsies. In summary, these data suggest that IHG-1 increases mitochondrial biogenesis by promoting PGC-1␣-dependent processes, potentially contributing to the pathogenesis of renal fibrosis.

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Section: Loss Of Endogenous Ihg-1 Expression Results In Decreased Mitmentioning

confidence: 98%

IHG-1 Promotes Mitochondrial Biogenesis by Stabilizing PGC-1α

Hickey¹,

Corcoran²,

Docherty³

et al. 2011

Journal of the American Society of Nephrology

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“…The utilization of ABC transporter protein fluorescent substrates has been extensively used to study the activity of these proteins in a wide range of cell types [25][26][27][28][29]. Using an efflux assay based on intracellular C-AM accumulation, we observed that embryonic cells of E. lucunter accumulated less C-AM than gametes (Figure 1).…”

Section: Discussionmentioning

confidence: 97%

Characterization of functional activity of ABCB1 and ABCC1 proteins in eggs and embryonic cells of the sea urchin Echinometra lucunter

et al. 2010

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ABC transporter (ATP-binding-cassette transporter) proteins have been strongly associated with the phenomenon of multidrug resistance in cancer cells. Furthermore, their physiological expression has been studied in many organisms, including bacteria, fungi, plants and vertebrate or invertebrate animals. Their widespread expression through the evolution demonstrates their relevance to the survival of living things. In the present study, we characterized the functional activity of ABCB1 and ABCC1 proteins in gametes and embryonic cells of the sea urchin Echinometra lucunter. The ABC transporter proteins' functional activity was up-regulated post-fertilization. Eggs and spermatozoa of E. lucunter accumulated more C-AM (calcein acetoxymethyl ester), a fluorescent substrate of ABCB1 and ABCC1 proteins, than embryonic cells. Verapamil, reversin 205 and indomethacin were able to increase C-AM influx in eggs and embryos. However, verapamil and reversin 205 were more efficient than indomethacin, suggesting a predominance of ABCB1 protein over ABCC1 protein activity. Multidrug resistance modulating agents, at the concentration range that inhibited ABC transporter proteins, did not block the embryonic development until blastula stage. However, inhibition of ABCB1-mediated efflux by reversin 205 circumvented resistance of embryos to the antimitotic vinca alkaloid vinblastine. Embryonic development was more efficiently blocked when colchicine was previously added to eggs than to embryos 5 min after fertilization. This set of results suggests that these proteins act as a fundamental biochemical barrier conferring a protective physiological role against toxic xenobiotics in E. lucunter embryos.

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“…This mitochondrial selective dye is a substrate of P-glycoprotein (Marques-Santos et al, 2003) and labels a subset of proteins found in mitochondrial enriched fractions (Presley et al, 2003). This dye was previously used for FCM analysis of mitochondria (Teranishi et al, 1999;Wakabayashi et al, 2000).…”

Section: Mitochondrial Stainingmentioning

confidence: 99%