Use of BODIPY (493/503) to Visualize Intramuscular Lipid Dropletsin Skeletal Muscle

Spangenburg, Espen E.; Pratt, Stephen J.; Wohlers, Lindsay M.; Lovering, Richard M.

doi:10.1155/2011/598358

Cited by 107 publications

(78 citation statements)

References 16 publications

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“…Sections were stained with hematoxylin and eosin (H&E) for tissue evaluation. The nonpolar dye BODIPY (D3922; Molecular Probes, Life Technologies, Carlsbad, CA) was used to identify lipid depositions in skeletal muscle (60). To quantify the percentage of BODIPYpositive fibers, staining was visualized by fluorescence microscopy of the cryosections.…”

Section: Methodsmentioning

confidence: 99%

Early metabolic changes measured by ¹H MRS in healthy and dystrophic muscle after injury

Pratt

Spangenburg

et al. 2012

Journal of Applied Physiology

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Xu S, Pratt SJ, Spangenburg EE, Lovering RM. Early metabolic changes measured by 1 H MRS in healthy and dystrophic muscle after injury. J Appl Physiol 113: 808 -816, 2012. First published June 28, 2012 doi:10.1152/japplphysiol.00530.2012.-Skeletal muscle injury is often assessed by clinical findings (history, pain, tenderness, strength loss), by imaging, or by invasive techniques. The purpose of this work was to determine if in vivo proton magnetic resonance spectroscopy ( 1 H MRS) could reveal metabolic changes in murine skeletal muscle after contraction-induced injury. We compared findings in the tibialis anterior muscle from both healthy wild-type (WT) muscles (C57BL/10 mice) and dystrophic (mdx mice) muscles (an animal model for human Duchenne muscular dystrophy) before and after contraction-induced injury. A mild in vivo eccentric injury protocol was used due to the high susceptibility of mdx muscles to injury. As expected, mdx mice sustained a greater loss of force (81%) after injury compared with WT (42%). In the uninjured muscles, choline (Cho) levels were 47% lower in the mdx muscles compared with WT muscles. In mdx mice, taurine levels decreased 17%, and Cho levels increased 25% in injured muscles compared with uninjured mdx muscles. Intramyocellular lipids and total muscle lipid levels increased significantly after injury but only in WT. The increase in lipid was confirmed using a permeable lipophilic fluorescence dye. In summary, loss of torque after injury was associated with alterations in muscle metabolite levels that may contribute to the overall injury response in mdx mice. These results show that it is possible to obtain meaningful in vivo 1 H MRS regarding skeletal muscle injury.in vivo magnetic resonance spectroscopy; eccentric injury; mdx; muscle damage INTENSE PHYSICAL EXERCISE can cause skeletal muscle damage and inflammation, resulting in pain and a loss of maximal force production. Skeletal muscle injury resulting from high-force lengthening contractions, such as intense physical exercise, is clinically termed a "muscle strain" and is one of the most common ailments seen by physicians. A diagnosis of acute muscle strains is still typically made based on physical exam and patient history. Several direct and indirect indicators of skeletal muscle damage have been observed, including disruption of contractile tissue, cellular accumulation of calcium, delayed onset muscle soreness, a prolonged reduction in muscular strength and range of motion, and an increase in the appearance of muscle proteins in the blood, such as creatine (Cr) kinase (CK), lactate dehydrogenase, and myoglobin (2, 6, 19). Duchenne muscular dystrophy (DMD) is characterized clinically by severe, progressive, and irreversible loss of muscular function. The disease is caused by the lack of dystrophin, a large membrane-associated protein expressed in skeletal muscle and localized to the inner face of the sarcolemma (63). There are significant parallels between the damage incurred after lengthening ("eccentric") contractions ...

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Section: Methodsmentioning

confidence: 99%

Early metabolic changes measured by ¹H MRS in healthy and dystrophic muscle after injury

Pratt

Spangenburg

et al. 2012

Journal of Applied Physiology

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“…Myotubes were then stained with BODIPY (Molecular Probes, Carlsbad CA) as previously described ( 35 ). Myotubes were imaged with a Zeiss Axiovision 4 (Zeiss, Oberkochen, Germany) as previously described ( 34,36 ).…”

Section: Human Myotube Palmitate/oleate Incubation and Bodipy Imagingmentioning

confidence: 99%

BRCA1 is a novel regulator of metabolic function in skeletal muscle

Jackson

Gidlund

Norrbom

et al. 2014

Journal of Lipid Research

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This article is available online at http://www.jlr.org breast cancer and/or tumorigenesis in reproductive tissues ( 3 ). The BRCA1 gene produces either a full-length breast cancer type 1 susceptibility protein (BRCA1) or through alternatively splicing two documented variants, BRCA1 ⌬ 11 or BRCA1 ⌬ 11b, both of which lack a nuclear localization signal ( 4 ). Recently, BRCA1 was identifi ed as a regulator of lipid metabolism in human breast cancer cells (MCF7) as a result of direct interaction with the phosphorylated form of acetylCoA carboxylase (ACC-p) at the BRCA1 C-terminal (BRCT) domains ( 5, 6 ). The interaction encourages the maintenance of the phosphorylated state of ACC thereby altering lipid metabolism in the cancer cell line ( 5, 6 ).ACC has two isoforms, ACC1 or ACC2, with ACC2 containing an extra 146 amino acids in the NH 2 -terminal region. ACC activity is negatively regulated by phosphorylation of residue Ser 79 on ACC1 and Ser 221 on ACC2 ( 7,8 ). In the active form (i.e., dephosphorylated), ACC catalyzes the carboxylation of acetyl-CoA into malonyl-CoA (MaCoA). Changes in cellular MaCoA content alter intracellular lipid dynamics in two specifi c manners ( 9, 10 ). MaCoA directly contributes to de novo synthesis of palmitate via FAS and MaCoA also allosterically inhibits carnitine palmitoyltransferase-1 (CPT-1), a mitochondrial long chain fatty acid transporter ( 11 ). Thus, in mammary tissue the ability of BRCA1 to affect ACC activity alters cellular lipid concentrations by indirectly regulating rates of fatty acid synthesis and/or the fl ux of fatty acids into the mitochondria.Abstract Breast cancer type 1 (BRCA1) susceptibility protein is expressed across multiple tissues including skeletal muscle. The overall objective of this investigation was to defi ne a functional role for BRCA1 in skeletal muscle using a translational approach. For the fi rst time in both mice and humans, we identifi ed the presence of multiple isoforms of BRCA1 in skeletal muscle. In response to an acute bout of exercise, we found increases in the interaction between the native forms of BRCA1 and the phosphorylated form of acetyl-CoA carboxylase. Decreasing BRCA1 content using a shRNA approach in cultured primary human myotubes resulted in decreased oxygen consumption by the mitochondria and increased reactive oxygen species production. The decreased BRCA1 content also resulted in increased storage of intracellular lipid and reduced insulin signaling. These results indicate that BRCA1 plays a critical role in the regulation of metabolic function in skeletal muscle. Collectively, these data reveal BRCA1 as a novel target to consider in our understanding of metabolic function and risk for development of metabolic-based diseases. -Jackson, K. C., E-K. Gidlund, J. Abbreviations: ACC, acetyl-CoA carboxylase; ACC-p, phosphorylated form of acetyl-CoA carboxylase; AICAR, 5-amino-1-β-D-ribofuranosylimidazole-4-carboxamide; AMPK, AMP-activated protein kinase; BRCA1, Breast Cancer 1, early onset; BRCA1 MG KO, mammary gland tissue fr...

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“…Lipid droplets (LD) were visualized and quantified using BODIPY (493/503) (Invitrogen, Carlsbad, CA), as previously described by our group (52,63).…”

Section: Animalsmentioning

confidence: 99%

Ectopic lipid deposition and the metabolic profile of skeletal muscle in ovariectomized mice

Jackson

Wohlers

Lovering

et al. 2013

American Journal of Physiology-Regulatory, Integrative and Comp

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Disruptions of ovarian function in women are associated with increased risk of metabolic disease due to dysregulation of peripheral glucose homeostasis in skeletal muscle. Our previous evidence suggests that alterations in skeletal muscle lipid metabolism coupled with altered mitochondrial function may also develop. The objective of this study was to use an integrative metabolic approach to identify potential areas of dysfunction that develop in skeletal muscle from ovariectomized (OVX) female mice compared with age-matched ovary-intact adult female mice (sham). The OVX mice exhibited significant increases in body weight, visceral, and inguinal fat mass compared with sham mice. OVX mice also had significant increases in skeletal muscle intramyocellular lipids (IMCL) compared with the sham animals, which corresponded to significant increases in the protein content of the fatty acid transporters CD36/FAT and FABPpm. A targeted metabolic profiling approach identified significantly lower levels of specific acyl carnitine species and various amino acids in skeletal muscle from OVX mice compared with the sham animals, suggesting a potential dysfunction in lipid and amino acid metabolism, respectively. Basal and maximal mitochondrial oxygen consumption rates were significantly impaired in skeletal muscle fibers from OVX mice compared with sham animals. Collectively, these data indicate that loss of ovarian function results in increased IMCL storage that is coupled with alterations in mitochondrial function and changes in the skeletal muscle metabolic profile.

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Use of BODIPY (493/503) to Visualize Intramuscular Lipid Dropletsin Skeletal Muscle

Cited by 107 publications

References 16 publications

Early metabolic changes measured by ¹H MRS in healthy and dystrophic muscle after injury

Early metabolic changes measured by ¹H MRS in healthy and dystrophic muscle after injury

BRCA1 is a novel regulator of metabolic function in skeletal muscle

Ectopic lipid deposition and the metabolic profile of skeletal muscle in ovariectomized mice

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Use of BODIPY (493/503) to Visualize Intramuscular Lipid Dropletsin Skeletal Muscle

Cited by 107 publications

References 16 publications

Early metabolic changes measured by 1H MRS in healthy and dystrophic muscle after injury

Early metabolic changes measured by 1H MRS in healthy and dystrophic muscle after injury

BRCA1 is a novel regulator of metabolic function in skeletal muscle

Ectopic lipid deposition and the metabolic profile of skeletal muscle in ovariectomized mice

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Product

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Early metabolic changes measured by ¹H MRS in healthy and dystrophic muscle after injury

Early metabolic changes measured by ¹H MRS in healthy and dystrophic muscle after injury