Proteomic analysis of the porcine platelet proteome and alterations induced by thrombin activation

Esteso, Gloria; Mora, María I.; Garrido, Juan J.; Corrales, Fernando J.; Moreno, A.

doi:10.1016/j.jprot.2008.08.002

Cited by 14 publications

(8 citation statements)

References 32 publications

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“…Organs from slaughtered pigs represent an unlimited, reliable and inexpensive resource of viable cell material for all fields of applied research. Many papers have been published where tissue samples from slaughtered pigs have been successfully used for research and testing 58, 63, 93, 97, 136–141. This makes it a convenient, relevant and a cost‐effective model system.…”

Section: Advantages and Disadvantages Of The Porcine Modelmentioning

confidence: 99%

“…Mass‐based searching such as MASCOT often gives the incorrect identification number (ID) simply because the actual sequence of the protein is not available in the database, as the swine sequence database is still incomplete, and homologous sequences from other species are not sufficiently identical to achieve a high enough score to produce a match. At this juncture, available porcine protein sequence databases are few and, if available, are somewhat fragmented and sparse (Table 3, 66, 136–140, 144–155). Many different platforms are being used and data is not always submitted to a common repository.…”

Section: Advantages and Disadvantages Of The Porcine Modelmentioning

confidence: 99%

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Recent advances in the use of Sus scrofa (pig) as a model system for proteomic studies

2011

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Of the numerous animal models available for proteomic studies only a small number have been successfully used in understanding human biology. To date, rodents have been widely employed in proteomic and genomic studies but often these models do not truly mimic the relevant human conditions. On the other hand, the pig shows similarity in size, shape and physiology to human and has been used as a major mammalian model for many studies concerning xenotransplantation, cardiovascular diseases, blood dynamics, nutrition, general metabolic functions, digestive-related disorders, respiratory diseases, diabetes, kidney and bladder diseases, organ-specific toxicity, dermatology and neurological sequelae. With the substantially improved knowledge of the structure and function of the pig genome in the last two decades it has been found that this animal shares a high sequence and chromosomal structure homology with humans. Nevertheless, in comparison to other available model organisms, very little work has been devoted to pig proteomics until recently. Keeping this in mind, the present review will highlight some of the advantages and disadvantages of pig as a model system for proteomic studies.

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Section: Advantages and Disadvantages Of The Porcine Modelmentioning

confidence: 99%

Section: Advantages and Disadvantages Of The Porcine Modelmentioning

confidence: 99%

Recent advances in the use of Sus scrofa (pig) as a model system for proteomic studies

2011

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“…This system can generate the functional analysis and networks that are most significant to the data set. IPA compares proteins in the input group and displays a rank-ordered list of pathways and networks, whose activities are most likely affected [36]. Networks are displayed graphically as nodes (proteins) and edges (the biological relationship between the nodes).…”

Section: Systems Biology Analysismentioning

confidence: 99%

Quantitative proteomics by 2‐DE, ¹⁶O/¹⁸O labelling and linear ion trap mass spectrometry analysis of lymph nodes from piglets inoculated by porcine circovirus type 2

et al. 2011

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Porcine circovirus type 2 (PCV2) has been identified as the essential causal agent of postweaning multisystemic wasting syndrome. However, little is known regarding the mechanism(s) underlying the pathogenesis of PCV2-induced disease and the interaction of the virus with the host immune system. Here, we present a proteomics study on inguinal lymph nodes of piglets inoculated with PCV2, in order to better understand the pathogenesis of postweaning multisystemic wasting syndrome and the pathways might be affected after infection. We used two proteomics strategies, 2-DE and 1-DE followed by (16)O/(18)O peptide labelling and peptide identification and quantification by MS. More than 100 proteins were found to be differentially regulated and the results obtained by the two strategies were fairly concordant but also complementary, the (18)O labelling approach being a more robust alternative. Analysis of these proteins by systems biology tools revealed the implication of acute phase response and NrF2-mediated oxidative stress, suggesting a putative role for these pathways in the pig immune response. Besides, CD81 was found to be up-regulated, suggesting a possible role in the internalization of the virus. The use of proteomics technologies together with biology analysis systems opens up the way to gain more exhaustive and systematic knowledge of virus-pathogen interactions.

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“…Image Analysis: Silver stained gels were scanned (GS-800 calibrated imaging densitometer, Bio-Rad) and analyzed for differences in protein patterns between groups with the 8.0.1 PDQuest software (BioRad), using a single master that included all gels of each independent experiment 17 . Each spot was assigned a relative value that corresponded to the single spot volume compared to the volume of all spots in the gel, following background extraction.…”

Section: Proteomic Analysismentioning

confidence: 99%

“…As platelets are anucleated, activation is mainly controlled by translocation of proteins, altered interactions, and posttranslational modifications, such as phosphorylation and glycosylation. Whole platelet proteome in human 16 and swine platelets, 17 as well as several platelet subproteomes 15 of resting and stimulated platelets, have been analyzed. Studies examining protein changes, such as translocation in response to platelet stimulus, are highly helpful, as they allow the identification of key proteins in platelet function.…”

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confidence: 99%

Proteomic Signature of Thrombin-Activated Platelets After In Vivo Nitric Oxide–Donor Treatment

Peña¹,

Padró²,

Molins³

et al. 2011

ATVB

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Objective-Growing insight into the antiplatelet properties of new nitric oxide (NO) donors has expanded their potential use in cardiovascular diseases. As such, we reported that oral administration of a new exogenous NO donor (LA419) induced significant inhibition of platelet deposition on damaged vascular wall without provoking hypotension in an in vivo experimental model. Thrombin is one of the major triggers of platelet deposition and thrombosis on injured vessels; however, the effects of NO on thrombin-induced platelet activation are not fully known. Here, our aim was to investigate the inhibitory effects of exogenous NO administration on the major changes in platelet proteins induced by thrombin. Methods and Results-Platelets were obtained from a group of swine orally treated with LA419 (0.9 mg kg Ϫ1 ) or placebo for 8 days. Washed platelets were incubated with thrombin (0.4 NIH U/mL). Platelet proteins were then sequentially extracted based on differential solubility and studied by two-dimensional electrophoresis, mass spectrometry (matrixassisted laser desorption ionization/time of flight), Western blot, and confocal immunofluorescence. NO treatment abrogated thrombin effects on 24 proteins involved in actin assembly, signaling, and metabolic activity. NO treatment prevented thrombin-induced translocation of gelsolin, filamin, 14-3-3, phosphatidylinositol 3-kinase-␥ isoform, and growth factor receptor-bound protein 2 (Grb2). Conclusion-Our results show that exogenous NO donor treatment renders platelets less sensitive to thrombin activation and inhibits thrombosis by interfering with the platelet shape change machinery. Key Words: platelets Ⅲ thrombosis Ⅲ cell signaling Ⅲ nitric oxide donor Ⅲ proteomics P latelets are one of the main targets for drug development in the prevention and treatment of cardiovascular and cerebrovascular atherothrombosis. 1 Advances in the understanding of the mechanisms involved in platelet activation and aggregation have provided new avenues for drug development. 2,3 Inactive platelets brought by the circulation are recruited to the site of vascular injury by the combined effects of a number of molecules (thrombin, collagen, fibrinogen, von Willebrand factor, ADP, thromboxane A2, epinephrine, and others) that trigger platelet signaling events and promote adhesion and aggregation. 4,5 These signaling events arise from multimolecular complexes formed between integrin cytoplasmic tails, intracellular signaling molecules, and structural cytoskeleton proteins. 6 Nitric oxide (NO) released from platelets and endothelium under physiological conditions is involved in several vascular processes, including reduction of cytokine-induced expression of tissue factor, 7 monocyte adhesion to the endothelium, vasodilation, and inhibition of platelet adhesion and aggregation. 8 -11 NO regulates platelet activation mainly through cGMP-dependent mechanisms. 12 NO-induced inhibition of platelet function has long been known; however, the signaling pathways underlying the inhibitory action of ...

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Proteomic analysis of the porcine platelet proteome and alterations induced by thrombin activation

Cited by 14 publications

References 32 publications

Recent advances in the use of Sus scrofa (pig) as a model system for proteomic studies

Recent advances in the use of Sus scrofa (pig) as a model system for proteomic studies

Quantitative proteomics by 2‐DE, ¹⁶O/¹⁸O labelling and linear ion trap mass spectrometry analysis of lymph nodes from piglets inoculated by porcine circovirus type 2

Proteomic Signature of Thrombin-Activated Platelets After In Vivo Nitric Oxide–Donor Treatment

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Proteomic analysis of the porcine platelet proteome and alterations induced by thrombin activation

Cited by 14 publications

References 32 publications

Recent advances in the use of Sus scrofa (pig) as a model system for proteomic studies

Recent advances in the use of Sus scrofa (pig) as a model system for proteomic studies

Quantitative proteomics by 2‐DE, 16O/18O labelling and linear ion trap mass spectrometry analysis of lymph nodes from piglets inoculated by porcine circovirus type 2

Proteomic Signature of Thrombin-Activated Platelets After In Vivo Nitric Oxide–Donor Treatment

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Quantitative proteomics by 2‐DE, ¹⁶O/¹⁸O labelling and linear ion trap mass spectrometry analysis of lymph nodes from piglets inoculated by porcine circovirus type 2