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Matrix-assisted laser desorption/ionization mass spectrometry imaging of cardiolipins in rat organ sections

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Abstract

Cardiolipin (CL) is a class of phospholipid tightly associated with the mitochondria functions and a prime target of oxidative stress. Peroxidation of CL dissociates its bound cytochrome C, a phenomenon that reflects oxidative stress sustained by the organ and a trigger for the intrinsic apoptotic pathway. However, CL distribution in normal organ tissues has yet to be documented. Fresh rat organs were snap-frozen, cut into cryosections that were subsequently desalted with ammonium acetate solution, and vacuum-dried. CL distribution in situ was determined using matrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI-MSI) technique on sections sublimed with 2,5-dihydroxybenzoic acid. CL images in rat cardiac ventricular section showed a homogeneous distribution of a single m/z 1447.9 ion species that was confirmed as the (18:2)4 CL by tandem mass spectrometry. The presence of low abundant (18:2)3(18:1) CL with the bulk (18:2)4 CL in quadriceps femoris rendered the muscle CL exhibiting a slightly deviated isotopic pattern from that of cardiac muscle. In rat liver, MALDI-MSI unveiled three CL-containing mass ranges, each with a unique in situ distribution pattern. Co-registration of the CL ion images with its stained liver section image further revealed the association of CLs in each mass range with the functional zones in the liver parenchyma and suggests the participation of in situ CLs with localized hepatic functions such as oxidation, conjugation, and detoxification. The advances in CL imaging offer an approach with molecular accuracy to reveal potentially dysregulated metabolic machineries in acute and chronic diseased states.

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Acknowledgments

This study was sponsored by National Scientific Council, Taiwan (grant No: NSC-99-2320-B-110-001-MY3), by Veterans General Hospital-Kaohsiung-National Sun Yat-Sen University Joint Research Consortium (grant No: VGHNSU101-002 and VGHNSU 102–004), and by "Aim for the Top University Plan" of National Sun Yat-Sen University, Taiwan. The authors acknowledge the generous support of Dr. Jentaie Shiea, Department of Chemistry, National Sun Yat-Sen University, Taiwan, for the use of MALDI-TOF/TOF instrument.

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Authors and Affiliations

  1. Department of Biological Sciences, National Sun Yat-Sen University, 70 Lien-Hai Rd., Kaohsiung, 80424, Taiwan

    Hay-Yan J. Wang, Hsuan-Wen Wu, Ping-Ju Tsai, Cheng Bin Liu & Zhi-Fu Zheng

  2. Department of Surgery, Yuan’s General Hospital, Kaohsiung, 80249, Taiwan

    Ping-Ju Tsai

  3. Department of Obstetrics and Gynecology, Veterans General Hospital-Kaohsiung, Kaohsiung, 81362, Taiwan

    Cheng Bin Liu

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  1. Hay-Yan J. Wang
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Correspondence to Hay-Yan J. Wang.

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Wang, HY.J., Wu, HW., Tsai, PJ. et al. Matrix-assisted laser desorption/ionization mass spectrometry imaging of cardiolipins in rat organ sections. Anal Bioanal Chem 406, 565–575 (2014). https://doi.org/10.1007/s00216-013-7492-y

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