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Intermittent Hypercapnic Hypoxia Induced Protein Changes in the Piglet Hippocampus Identified by MALDI-TOF-MS

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Abstract

Intermittent hypercapnic hypoxia (IHH) induces protein changes in the brainstem, but its effects on the hippocampus have not yet been studied. Using a proteomics-based approach, we tested the hypothesis that IHH up-regulates apoptotic promoters and down-regulates apoptotic inhibitors in the developing hippocampus. Male piglets aged 13–14 days were assigned to control (n = 6) or IHH (n = 5) groups. Using two-dimensional polyacrylamide gel electrophoresis, matrix-assisted laser desorption/ionisation-time of flight-mass spectrometry (MALDI-TOF-MS), a total of 26 protein spots were differentially expressed in IHH compared to control group. Thirteen of these (6 up-regulated, 7 down-regulated) were identified including 14-3-3θ/τ (increased), glial fibrillary acidic protein (increased) and α-internexin (decreased). Further analysis with western blot validated these proteins and immunohistochemistry showed specific regional changes in the subiculum, stratum radiatum and CA1 of the hippocampus. Most proteins identified were involved in promoting cell survival under apoptotic conditions. These findings improve our understanding of the cellular processes that occur in the hippocampus during IHH exposure, and have important implications in clinical settings where IHH is experienced, for example, during prone sleeping or with obstructive sleep apnea in an infant.

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Abbreviations

BDNF:

Brain-derived neurotrophic factor

DG:

Dentate gyrus

GFAP:

Glial fibrillary acidic protein

GRP94:

Glucose regulated protein-94

IHC:

Immunohistochemistry

IHH:

Intermittent hypercapnic hypoxia

NMDA:

N-methyl-d-aspartic acid

PP2A:

Serine/threonine protein phosphatase 2A

SIDS:

Sudden infant death syndrome

SR:

Stratum radiatum

Sub:

Subiculum

V-ATPase:

Vacuolar ATP synthase subunit B

WB:

Western blot

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Acknowledgments

Research was funded by NH&MRC #302006. Prof. Waters is supported by an NH&MRC Practitioner Fellowship (#206507). We thank Jonathan Krygier, a summer undergraduate research student for his contribution in quantification of the GFAP immunohistochemistry. We acknowledge the facilities as well as scientific and technical assistance from staff in the NANO Major National Research Facility at the Electron Microscope Unit, The University of Sydney.

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Authors and Affiliations

  1. Discipline of Pediatrics and Child Health, University of Sydney, Sydney, NSW, 2006, Australia

    Samantha Tang & Karen A. Waters

  2. Department of Medicine, University of Sydney, Room 206, Blackburn Building, DO6, Sydney, NSW, 2006, Australia

    Rita Machaalani & Karen A. Waters

  3. Discipline of Pathology, University of Sydney, Sydney, NSW, 2006, Australia

    Mohammad A. Kashem & Izuru Matsumoto

  4. Bosch Institute, University of Sydney, Sydney, NSW, 2006, Australia

    Samantha Tang, Rita Machaalani & Karen A. Waters

  5. The Children’s Hospital, Westmead Sydney, NSW, 2145, Australia

    Rita Machaalani & Karen A. Waters

  6. Department of Psychiatry, Asahiyama Hospital, 4-3-33 Futagoyama, Cyu-o-ku, Sapporo, Hokkaido, 064-0946, Japan

    Izuru Matsumoto

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  1. Samantha Tang
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  2. Rita Machaalani
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  3. Mohammad A. Kashem
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Correspondence to Rita Machaalani.

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Tang, S., Machaalani, R., Kashem, M.A. et al. Intermittent Hypercapnic Hypoxia Induced Protein Changes in the Piglet Hippocampus Identified by MALDI-TOF-MS. Neurochem Res 34, 2215–2225 (2009). https://doi.org/10.1007/s11064-009-0021-x

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  • DOI: https://doi.org/10.1007/s11064-009-0021-x

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