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

G. Maasz, Z. Pirger, D. Reglodi, D. Petrovics, J. Schmidt, P. Kiss, A. Rivnyak, H. Hashimoto, P. Avar, E. Jambor, A. Tamás, B. Gaszner, L. Márk

Research output: Contribution to journalArticle

9 Citations (Scopus)

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.

Original languageEnglish
JournalJournal of Molecular Neuroscience
DOIs
Publication statusAccepted/In press - Mar 19 2014

Fingerprint

Pituitary Adenylate Cyclase-Activating Polypeptide
Proteomics
Mass Spectrometry
Brain
Proteins
Diencephalon
Fructose-Bisphosphate Aldolase
Mesencephalon
Knockout Mice
Peptides
Neurotransmitter Agents
cis-trans-Isomerases
Biochemical Phenomena
Search Engine
Pyruvate Kinase
Sudden Infant Death
Cytoprotection
Phosphopyruvate Hydratase
Nerve Growth Factors
Tubulin

Keywords

  • Knockout
  • MALDI
  • PACAP
  • Proteomics

ASJC Scopus subject areas

  • Cellular and Molecular Neuroscience

Cite this

Comparative Protein Composition of the Brains of PACAP-Deficient Mice Using Mass Spectrometry-Based Proteomic Analysis. / Maasz, G.; Pirger, Z.; Reglodi, D.; Petrovics, D.; Schmidt, J.; Kiss, P.; Rivnyak, A.; Hashimoto, H.; Avar, P.; Jambor, E.; Tamás, A.; Gaszner, B.; Márk, L.

In: Journal of Molecular Neuroscience, 19.03.2014.

Research output: Contribution to journalArticle

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AU - Schmidt, J.

AU - Kiss, P.

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AU - Hashimoto, H.

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