4.5 Article

A Simple and Effective Sample Preparation Strategy for MALDI-MS Imaging of Neuropeptide Changes in the Crustacean Brain Due to Hypoxia and Hypercapnia Stress

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AMER CHEMICAL SOC
DOI: 10.1021/jasms.9b00107

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资金

  1. National Science Foundation [CHE-1710140]
  2. National Institutes of Health (NIH) [1R01DK071801, R56 MH110215]
  3. NIH [NIH-NCRR S10RR029531]
  4. University of Wisconsin Carbone Cancer Center Pancreas Cancer Task Force
  5. NIH General Medical Sciences F31 Fellowship [1F31GM119365]
  6. National Institute of General Medical Sciences of the NIH [T32GM008349]
  7. National Institutes of Health under Ruth L. Kirschstein National Research Service Award from the National Heart Lung and Blood Institute [T32 HL 007936]
  8. Madison Cardiovascular Research Center
  9. National Institutes of Health-General Medical Sciences F31 National Research Service Award [1F31GM126870]
  10. Wisconsin Alumni Research Foundation
  11. University of Wisconsin.Madison School of Pharmacy

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Matrix-assisted laser desorption/ionization (MALDI)-MS imaging has been utilized to image a variety of biomolecules, including neuropeptides. Washing a tissue section is an effective way to eliminate interfering background and improve detection of low concentration target analyte molecules; however, many previous methods have not been tested for neuropeptide analysis via MALDI-MS imaging. Using crustaceans as a neurological model organism, we developed a new, simple washing procedure and applied this method to characterize neuropeptide changes due to hypoxia stress. With a 10 s 50:50 EtOH:H2O wash, neuropeptide coverage was improved by 1.15-fold, while normalized signal intensities were increased by 5.28-fold. Specifically, hypoxia and hypercapnia stress conditions were investigated due to their environmental relevance to marine invertebrates. Many neuropeptides, including RFamides, pyrokinin, and cardioactive peptides, showed distinct up- and down-regulation for specific neuropeptide isoforms. Since crustacean neuropeptides are homologous to those found in humans, results from these studies can be applied to understand potential roles of neuropeptides involved in medical hypoxia and hypercapnia.

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