Expression of cGMP-dependent protein kinase type I in mature white adipocytes

Leiss, Veronika; Illison, Julia; Domes, Katrin; Hofmann, Franz; Łukowski, Robert

doi:10.1016/j.bbrc.2014.08.071

Cited by 10 publications

(6 citation statements)

References 44 publications

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“…The uncoupling protein 1 (UCP1) expression pattern was compared between iWAT cryosections obtained from HFD-fed adipoqBK L1/L2 and adipoqBK +/L2 mice using a commercial antibody (dilution 1:400; Santa Cruz Biotechnology, Dallas, TX). BK immunodetection was performed in iWAT and eWAT sections derived from 10-week-old global and adipocyte-specific BK mutants and control litters using a primary antibody specific for the BK channel a-subunit (dilution 1:1,000) and hormone-sensitive lipase (dilution 1:400), according to a previously published protocol (27). In brief, antigen-antibody complexes were detected by a fluorescence-labeled secondary antibody (antirabbit AlexaFluor568; dilution 1:500) using the AxioCam MR system (Carl Zeiss) or by appropriate secondary antibodies conjugated to horseradish peroxidase (dilution 1:500).…”

Section: Adipose Tissue Histologymentioning

confidence: 99%

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Obesogenic and Diabetogenic Effects of High-Calorie Nutrition Require Adipocyte BK Channels

et al. 2016

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Elevated adipose tissue expression of the Ca- and voltage-activated K (BK) channel was identified in morbidly obese men carrying a BK gene variant, supporting the hypothesis that K channels affect the metabolic responses of fat cells to nutrients. To establish the role of endogenous BKs in fat cell maturation, storage of excess dietary fat, and body weight (BW) gain, we studied a gene-targeted mouse model with global ablation of the BK channel (BK) and adipocyte-specific BK-deficient (adipoqBK) mice. Global BK deficiency afforded protection from BW gain and excessive fat accumulation induced by a high-fat diet (HFD). Expansion of white adipose tissue-derived epididymal BK preadipocytes and their differentiation to lipid-filled mature adipocytes in vitro, however, were improved. Moreover, BW gain and total fat masses of usually superobese ob/ob mice were significantly attenuated in the absence of BK, together supporting a central or peripheral role for BKs in the regulatory system that controls adipose tissue and weight. Accordingly, HFD-fed adipoqBK mutant mice presented with a reduced total BW and overall body fat mass, smaller adipocytes, and reduced leptin levels. Protection from pathological weight gain in the absence of adipocyte BKs was beneficial for glucose handling and related to an increase in body core temperature as a result of higher levels of uncoupling protein 1 and a low abundance of the proinflammatory interleukin-6, a common risk factor for diabetes and metabolic abnormalities. This suggests that adipocyte BK activity is at least partially responsible for excessive BW gain under high-calorie conditions, suggesting that BK channels are promising drug targets for pharmacotherapy of metabolic disorders and obesity.

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Section: Adipose Tissue Histologymentioning

confidence: 99%

“…iWAT was dissected and processed as described above to generate total protein lysates for subsequent immunoblot analyses (27). Total protein was obtained from UD and DF 3T3-L1 fibroblasts.…”

Section: Immunoblot Analysis Of Iwat-and 3t3-l1-derived Protein Lysatesmentioning

confidence: 99%

Obesogenic and Diabetogenic Effects of High-Calorie Nutrition Require Adipocyte BK Channels

et al. 2016

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“…Western blot analysis was performed according to a previously established protocol (33)(34)(35). As a positive control, a PDE6H-hexahistidine fusion protein was loaded together with the protein samples derived from Pde6h Ϫ/Ϫ and WT retina.…”

Section: Methodsmentioning

confidence: 99%

Targeted Ablation of the Pde6h Gene in Mice Reveals Cross-species Differences in Cone and Rod Phototransduction Protein Isoform Inventory

Brennenstuhl¹,

Tanimoto

Burkard³

et al. 2015

Journal of Biological Chemistry

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Background: Phosphodiesterase-6 (PDE6) is a multisubunit enzyme essential for visual signal processing. Rare mutations in the human PDE6H gene result in incomplete color blindness. Results: Pde6h-deficient mice exhibit no signs of photoreceptor dysfunction. Conclusion: PDE6 configurations differ between species and are more interchangeable than previously thought. Significance: Presence of related isoforms in the retina may allow adjustments of the phototransduction components thereby preventing the occurrence of pathological conditions.

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“…In contrast, adipocytes expressed mostly PKG-Iβ with the PKG-Iα/PKG-Iβ ratio of 0.02. The expression of PKG-I in preadipocytes and adipocytes has been reported using Western blot assays [ 40 , 42 , 45 ]. However, differential expression of PKG-I isoforms in preadipocytes and adipocytes was reported in this study for the first time.…”

Section: Resultsmentioning

confidence: 99%

Capillary Isoelectric Focusing Immunoassay for Fat Cell Differentiation Proteomics

et al. 2015

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Profiling cellular proteome is critical to understanding signal integration during cell fate determination. In this study, the capability of capillary isoelectric focusing (cIEF) immunoassays to detect post-translational modifications (PTM) of protein isoforms is demonstrated. cIEF immunoassays exhibit protein detection sensitivity at up to 5 orders of magnitude higher than traditional methods. This detection ultra-sensitivity permits proteomic profiling of several nanograms of tissue samples. cIEF immunoassays are employed to simultaneously profile three protein kinases during fat cell differentiation: cGMP-dependent protein kinase type I (PKG-I) of the nitric oxide (NO) signaling pathway, protein kinase B (Akt) of the insulin signaling pathway, and extracellular signal-regulated kinase (ERK) of the mitogen-activated protein kinase (MAPK) signaling pathway. Interestingly, a switch in the expression level of PKG- isoforms is observed during fat cell differentiation. While both PKG-Iα and PKG-Iβ isoforms are present in preadipocytes, only PKG-Iβ isoform is expressed in adipocytes. On the other hand, the phosphorylation level increases for Akt while decreases for ERK1 and ERK2 following the maturation of preadipocytes into adipocytes. Taken together, cIEF immunoassay provides a highly sensitive means to study fat cell differentiation proteomics. cIEF immunoassay should be a powerful proteomics tool to study complex protein signal integration in biological systems.

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Expression of cGMP-dependent protein kinase type I in mature white adipocytes

Cited by 10 publications

References 44 publications

Obesogenic and Diabetogenic Effects of High-Calorie Nutrition Require Adipocyte BK Channels

Obesogenic and Diabetogenic Effects of High-Calorie Nutrition Require Adipocyte BK Channels

Targeted Ablation of the Pde6h Gene in Mice Reveals Cross-species Differences in Cone and Rod Phototransduction Protein Isoform Inventory

Capillary Isoelectric Focusing Immunoassay for Fat Cell Differentiation Proteomics

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