Proteomic identification of a novel protein regulated in CA1 and CA3 hippocampal regions during intermittent hypoxia

Klein, Jon B.; Gozal, David; Pierce, William M.; Thongboonkerd, Visith; Scherzer, Janice A.; Sachleben, Leroy R.; Guo, Song; Cai, Jian; Gozal, Evelyne

doi:10.1016/s1569-9048(03)00074-0

Cited by 32 publications

(23 citation statements)

References 42 publications

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“…Brains were dissected on ice to retrieve the hippocampal formation bilaterally, and the CA1 and CA3 regions of the hippocampus were then dissected as described previously (7,43). Tissues corresponding to the CA1 region harvested from either normoxic or hypoxic animals were homogenized on ice with a tissue tearer in 300 l of sample buffer (58 mM dithiothreitol, 65 mM CHAPS, 7 M urea, 1.9 M thiourea, 1.75% pH 3-10 carrier ampholytes).…”

Section: Methodsmentioning

confidence: 99%

Akt-mediated Valosin-containing Protein 97 Phosphorylation Regulates Its Association with Ubiquitinated Proteins

Klein

Barati

et al. 2005

Journal of Biological Chemistry

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Hypoxia is a common environmental stress that influences signaling pathways and cell function. Previous studies from our laboratory have identified significant differences in cellular responses to sustained or intermittent hypoxia with the latter proving more cytotoxic. We hypothesized that differences in susceptibility of neurons to intermittent (IH) and sustained hypoxia (SH) are mediated by altered Akt signaling. SH, but not IH, induced a significant increase in Akt activation in rat CA1 hippocampal region extracts compared with room air controls. Most mammalian neurons are sensitive to hypoxia to a varying degree. However, the mechanisms underlying this heterogeneous sensitivity are not well understood. Heterotopic differences in hypoxic sensitivity are now well established in the brain during conditions of oxygen deprivation such as hypoxic ischemia and the oxidant stress that follows reperfusion or malonate-induced mitochondrial metabolic disruption (1-4). Previous studies from our laboratory have identified significant differences in the hypoxic susceptibility of the CA1 and CA3 hippocampal regions, with CA1 vulnerability being particularly prominent in response to intermittent mild hypoxic episodes (5-7). These findings suggest that the CA3 region is relatively resistant to hypoxia when compared with the CA1 region, and CA1 regional vulnerability to hypoxia varies depending on the mode, severity, and duration of the hypoxic insult (8 -10). The mechanisms underlying the greater susceptibility of the CA1 region to intermittent hypoxia (IH) 1 than to sustained hypoxia (SH) are unknown. We have recently shown in PC-12 cells that different signaling pathways are involved in sustained and intermittent hypoxia-induced cell death. Identifying these pathways may contribute to our understanding of differential brain susceptibility to sustained and intermittent hypoxia (11). In the present study, we found that in rat CA1, SH but not IH induces significant increase in Akt activation compared with room air (RA) control, suggesting that differences in Akt signaling maybe implicated in the differences in hypoxia sensitivity in IH versus SH. Proteomic analysis of Akt immunoprecipitates from RA, SH, and IH CA1 samples was performed to identify differences in the assembly of Akt-binding partners among different conditions. These studies demonstrated that SH but not IH increased expression and association of endoplasmic reticulum (ER)-proteasome interacting protein, velosin-containing protein 97 (VCP) with Akt, compared with RA-exposed animals. Additionally, VCP was identified as an Akt substrate in vitro and in vivo thereby potentially implicating the VCP-Akt complex in the regulation of protein quality control during SH.VCP is a member of the AAA (ATPases associated with various cellular activities) family and is divided into N (residues 1-187), D1 (residues 209 -460), D2 (residues 481-761), and C (residues 742-806) domains, and two linkers, N-D1 linker (residues 188 -208) and D1-D2 linker (residues 461-

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Section: Methodsmentioning

confidence: 99%

Akt-mediated Valosin-containing Protein 97 Phosphorylation Regulates Its Association with Ubiquitinated Proteins

Klein

Barati

et al. 2005

Journal of Biological Chemistry

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“…Using this method the authors compared post mortem hippocampal protein expression between controls, schizophrenia, and Alzheimer's disease patients reporting a series of differentially expressed proteins [8,9]. No other information on human hippocampus is available but in rat brain a series of proteins were identified in hippocampal subareas CA1 and CA3 [10,11] and 48 proteins were identified in mouse hippocampus [2].…”

Section: Introductionmentioning

confidence: 99%

Proteomic profiling of human hippocampus

Yang¹,

Czech

Lubec³

2004

Electrophoresis

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The hippocampus is an important brain region playing a central role in short-term memory formation. Information on protein expression in human hippocampus is limited and we therefore decided to generate a protein map of this pivotal brain structure thus providing protein building blocks and an analytical tool. Two-dimensional electrophoresis with in-gel digestion of protein spots followed by mass spectrometrical identification coupled to the Mascot search program was applied. Matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF) or in special cases MALDI-TOF/TOF was used in order to warrant unambiguous identification. 266 protein spots representing 165 different proteins and 101 probable isoforms or post-translational modifications were identified. The identified proteins were categorized into several protein groups as metabolic, cytoskeleton, antioxidant, chaperone, signaling, transcriptional, proteolytic, synaptosomal and neuronal, hypothetical and miscellaneous structures. This human hippocampus protein map provides an analytical tool for hydrophilic and even relatively small-molecular-weight proteins and contributes to the knowledge on protein building blocks thus forming the basis for protein-chemical work in this brain area in health and disease.

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“…Other hypothetical proteins with reduced levels in the fetal DS brain are KIAA1185, 55.2, 58.8 kDa, actin-related protein 3b (ARP3b), and putative GTP-binding protein PTD004, suggesting for abnormalities in cytoskeleton, signaling and folding machineries [169]. In an animal model of hypoxia, reduced levels for the hypothetical protein AK006737 were detected in CA1 and CA3 hippocampal regions [170].…”

Section: Proteins With Other Functionsmentioning

confidence: 99%

“…Liberatori et al [179] examined the expression pattern of VDAC isoforms in guinea pig brain synaptosomes, under both normoxic and hypoxic conditions. Hypoxia ischemia was also studied on rats in correlation with proteomics [68,127,170]. Proteins with deranged levels found in the various models are listed in Suppl.…”

Section: Animal Modelsmentioning

confidence: 99%

Proteomics‐driven progress in neurodegeneration research

Fountoulakis

Κοσσίδα

2006

Electrophoresis

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Proteomics-driven progress in neurodegeneration researchProteomics technologies have been widely used in the investigation of neurodegenerative and psychiatric disorders, and in particular in the detection of differences between healthy individuals and patients suffering from such diseases. Thus, brain and cerebrospinal fluid (CSF) samples from patients with Alzheimer's disease, Down syndrome, Pick's disease, Parkinson's disease, schizophrenia, and other disorders as well as brain and CSF from animals serving as models of neurological disorders have been analyzed by proteomics. 2-DE followed by MALDI-TOF-MS has been mainly applied as this proteomics approach provides the possibility of convenient quantification of protein levels and detection of post-translational modifications. About 330 unique proteins with deranged levels and modifications have been detected by proteomics approaches to be related to neurodegeneration and psychiatric disorders. They are mainly involved in metabolism pathways, cytoskeleton formation, signal transduction, guidance, detoxification, transport, and conformational changes. In this article, we provide a summary of the major contributions of proteomics technologies in the study of neurodegenerative and psychiatric diseases, in particular, in the detection of changes in protein levels and modifications related to these disorders. IntroductionNeurodegeneration is a major hallmark of a series of genetic and acquired disorders of the central nervous system (CNS), like Alzheimer's disease (AD), Down syndrome (DS), Pick's disease, Creutzfeldt-Jakob disease (CJD), and Parkinson's disease, which result in severe cognitive and motor deficits. A common characteristic of these diseases is that they are multifactorial.Their molecular basis is unknown and individual biomarkers are not reliable. Genomics and proteomics approaches have been applied to obtain a better understanding of neurodegeneration and psychiatric disorders. Genomics has been used to map and identify genes that are mutated in a wide variety of such disorders, like presenilins in AD. Next to genomics, which only provides a partial view of the biological problem, proteomics is widely used in clinical diagnosis as well and many changes in protein levels associated with specific diseases have been identified [1,2].Proteomics is a technology-driven science that studies the proteins of the various biological systems, their structures, interactions, post-translational modifications, and in particular the changes in their levels and modifications, which are the result of specific diseases or are induced by external factors.

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Proteomic identification of a novel protein regulated in CA1 and CA3 hippocampal regions during intermittent hypoxia

Cited by 32 publications

References 42 publications

Akt-mediated Valosin-containing Protein 97 Phosphorylation Regulates Its Association with Ubiquitinated Proteins

Akt-mediated Valosin-containing Protein 97 Phosphorylation Regulates Its Association with Ubiquitinated Proteins

Proteomic profiling of human hippocampus

Proteomics‐driven progress in neurodegeneration research

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