Disrupting KATP channels diminishes the estrogen-mediated protection in female mutant mice during ischemia-reperfusion

Clinical Proteomics

Nian-Qing Shi¹  Wei Xu⁶  Hector Valdivia⁵  Grzegorz Sabat⁴  Dong Xu²  Jianjiong Gao³ 

[1] Department of Medicine, University of Wisconsin, Room 8418, WIMR II, 1111 Highland Ave., Madison, WI 53705, USA;Department of Computer Science and CS Bond Life Sciences Center, University of Missouri, Columbia, MO 65211, USA;Computational Biology Center and Center for Molecular Oncology, Memorial Sloan-Kettering Cancer Center, New York, NY 10065, USA;Biotechnology Center, Mass Spectrometry Facility, University of Wisconsin, Madison, WI 53706, USA;Department of Internal Medicine, University of Michigan, 2800 Plymouth Ave., 26-235 N, Ann Arbor, MI 48105, USA;McArdle Laboratory for Cancer Research, University of Wisconsin, 1400 University Ave., Madison, WI 53706, USA

关键词: Glycosylation;    Estrogen receptor;    Estrogen;    Gender difference;    Myocardial infarction;    Sulfonylurea receptor;    KATP channel;   

Others  :  802558
DOI  :  10.1186/1559-0275-11-19

 received in 2013-10-01, accepted in 2014-02-19,  发布年份 2014

【 摘 要 】

Background

Estrogen has been shown to mediate protection in female hearts against ischemia-reperfusion (I-R) stress. Composed by a Kir6.2 pore and an SUR2 regulatory subunit, cardiac ATP-sensitive potassium channels (KATP) remain quiescent under normal physiological conditions but they are activated by stress stimuli to confer protection to the heart. It remains unclear whether KATP is a regulatory target of estrogen in the female-specific I-R signaling pathway. In this study, we aimed at delineating the molecular mechanism underlying estrogen modulation on KATP channel activity during I-R.

Materials and methods

We employed KATP knockout mice in which SUR2 is disrupted (SUR2KO) to characterize their I-R response using an in vivo occlusion model. To test the protective effects of estrogen, female mice were ovariectomized and implanted with 17β-estradiol (E2) or placebo pellets (0.1 μg/g/day, 21-day release) before receiving an I-R treatment. Comparative proteomic analyses were performed to assess pathway-level alterations between KO-IR and WT-IR hearts.

Results and discussion

Echocardiographic results indicated that KO females were pre-disposed to cardiac dysfunction at baseline. The mutant mice were more susceptible to I-R stress by having bigger infarcts (46%) than WT controls (31%). The observation was confirmed using ovariectomized mice implanted with E2 or placebo. However, the estrogen-mediated protection was diminished in KO hearts. Expression studies showed that the SUR2 protein level, but not RNA level, was up-regulated in WT-IR mice relative to untreated controls possibly via PTMs. Our antibodies detected different glycosylated SUR2 receptor species after the PNGase F treatment, suggesting that SUR2 could be modified by N-glycosylation. We subsequently showed that E2 could further induce the formation of complex-glycosylated SUR2. Additional time-point experiments revealed that I-R hearts had increased levels of N-glycosylated SUR2; and DPM1, the first committed step enzyme in the N-glycosylation pathway. Comparative proteomic profiling identified 41 differentially altered protein hits between KO-IR and WT-IR mice encompassing those related to estrogen biosynthesis.

Conclusions

Our findings suggest that KATP is likely a downstream regulatory target of estrogen and it is indispensable in female I-R signaling. Increasing SUR2 expression by N-glycosylation mediated by estrogen may be effective to enhance KATP channel subunit expression in I-R.

【 授权许可】

   
2014 Gao et al.; licensee BioMed Central Ltd.

【 图 表 】

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