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Comparative Protein Composition of the Brains of PACAP-Deficient Mice Using Mass Spectrometry-Based Proteomic Analysis

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Abstract

Pituitary adenylate cyclase-activating polypeptide (PACAP) is a widespread neuropeptide acting as a neurotransmitter, neuromodulator, or neurotrophic factor. The diverse biological actions provide the background for the variety of deficits observed in mice lacking endogenous PACAP. PACAP-deficient mice display several abnormalities, such as sudden infant death syndrome (SIDS)-like phenotype, decreased cell protection, and increased risk of Parkinson’s disease. However, the molecular and proteomic background is still unclear. Therefore, our aim was to investigate the differences in peptide and protein composition in the brains of PACAP-deficient and wild-type mice using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and mass spectrometric (MS)-based proteomic analysis. Brains from PACAP-deficient mice were removed, and different brain areas (cortex, hippocampus, diencephalon, mesencephalon, brainstem, and cerebellum) were separated. Brain pieces were weighed, homogenized, and further processed for electrophoretic analysis. Our results revealed several differences in diencephalon and mesencephalon. The protein bands of interest were cut from the gel, samples were digested with trypsin, and the tryptic peptides were measured by matrix-assisted laser desorption ionization time of flight (MALDI TOF) MS. Results were analyzed by MASCOT Search Engine. Among the altered proteins, several are involved in metabolic processes, energy homeostasis, and structural integrity. ATP-synthase and tubulin beta-2A were expressed more strongly in PACAP-knockout mice. In contrast, the expression of more peptides/proteins markedly decreased in knockout mice, like pyruvate kinase, fructose biphosphate aldolase-A, glutathione S-transferase, peptidyl propyl cis-trans isomerase-A, gamma enolase, and aspartate amino transferase. The altered expression of these enzymes might partially account for the decreased antioxidant and detoxifying capacity of PACAP-deficient mice accompanying the increased vulnerability of these animals. Our results provide novel insight into the altered biochemical processes in mice lacking endogenous PACAP.

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Acknowledgments

This research was supported by the European Union and the State of Hungary, co-financed by the European Social Fund in the framework of TÁMOP-4.2.4.A/2-11/1-2012-0001 ‘National Excellence Programs’ [A2-ACSJD-13-0302 (NKPR-2013-31609), A2-SZGYA-FOK-13-0003], OTKAK104984, PD109099, GVOP-3.2.1-2004-04-0172/3.0, TIOP 1.3.1-10/1-2010-0008, TIOP 1.3.1-07/1, TÁMOP-4.2.2.A-11/1/KONV-2012-0053, TÁMOP-4.2.2.A-11/1/KONV-2012-0024, NAP, PTE-MTA Lendület Program, and Arimura Foundation.

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Authors and Affiliations

  1. Department of Analytical Biochemistry Institute of Biochemistry and Medical Chemistry, University of Pecs, Pecs, Hungary

    G. Maasz, D. Petrovics, J. Schmidt, P. Avar, E. Jambor & L. Mark

  2. MTA-PTE Human Reproduction Research Group, Pecs, Hungary

    G. Maasz & L. Mark

  3. Imaging Center for Life and Material Sciences, University of Pecs, Pecs, Hungary

    G. Maasz, J. Schmidt & L. Mark

  4. Janos Szentagothai Research Center, University of Pecs, Pecs, Hungary

    G. Maasz, J. Schmidt & L. Mark

  5. Department of Anatomy PTE MTA Lendulet PACAP Research, University of Pecs, Pecs, Hungary

    Z. Pirger, D. Reglodi, P. Kiss, A. Rivnyak, A. Tamas & B. Gaszner

  6. Chemical Ecology and Neurobiology, Department of Experimental Zoology, Balaton Limnological Institute, Centre for Ecological Research, Hungarian Academy of Sciences, Tihany, Hungary

    Z. Pirger

  7. Laboratory of Molecular Neuropharmacology, Graduate School of Pharmaceutical Sciences, Osaka University, Osaka, Japan

    H. Hashimoto

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  1. G. Maasz
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  2. Z. Pirger
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  3. D. Reglodi
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  4. D. Petrovics
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  5. J. Schmidt
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  6. P. Kiss
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  7. A. Rivnyak
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  8. H. Hashimoto
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  9. P. Avar
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  10. E. Jambor
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  11. A. Tamas
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  12. B. Gaszner
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  13. L. Mark
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Corresponding author

Correspondence to D. Reglodi.

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Maasz, G., Pirger, Z., Reglodi, D. et al. Comparative Protein Composition of the Brains of PACAP-Deficient Mice Using Mass Spectrometry-Based Proteomic Analysis. J Mol Neurosci 54, 310–319 (2014). https://doi.org/10.1007/s12031-014-0264-0

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  • DOI: https://doi.org/10.1007/s12031-014-0264-0

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