Distinct proteomic profiles of amphetamine self-administration transitional states

Freeman, Willard M.; Brebner, Karen; Amara, Susan G.; Reed, Matthew S.; Pohl, Jan; Phillips, Anthony G.

doi:10.1038/sj.tpj.6500309

Cited by 41 publications

(44 citation statements)

References 49 publications

(65 reference statements)

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“…2D Gels-Quantitative 2D DIGE was performed as detailed previously (22). All 2D DIGE methods and results are provided in minimum information about a proteomics experiment-gel electrophoresis (MIAPE-GE)-compliant form.…”

Section: Methodsmentioning

confidence: 99%

The Retinal Proteome in Experimental Diabetic Retinopathy

Fort

Freeman

Losiewicz

et al. 2009

Molecular & Cellular Proteomics

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Diabetic retinopathy is the leading cause of blindness in working age persons. Targeted studies have uncovered several components of the pathophysiology of the disease without unveiling the basic mechanisms. This study describes the use of complementary proteomic and genomic discovery methods that revealed that the proteins of the crystallin superfamily are increased dramatically in early diabetic retinopathy. Orthogonal methods confirmed that the amplitude of the up-regulation is greater than other changes described so far in diabetic retinopathy. A detailed time course study during diabetes showed differential up-regulation of the different isoforms of the crystallins superfamily. ␣-and ␤-crystallins were regulated primarily at the translation level, whereas ␥-crystallins were also regulated transcriptionally. We also demonstrated cell-specific patterns of expression of the different crystallins in normal and diabetic rat retinas. Diabetic retinopathy is the leading cause of blindness in working age persons, and despite numerous studies, the pathophysiological mechanisms, especially during the early stages of diabetes, remain to be elucidated. Diabetic retinopathy develops, to some degree, in nearly all patients with diabetes and is the most common cause of new cases of blindness among adults. The predominant causes of vision loss are clinically significant macular edema and proliferative diabetic retinopathy, but vision impairment can be prevented or minimized if the retinopathy is identified in its early stages. Diabetic retinopathy includes microvascular and neuronal, glial, and microglial cell defects early in the course of the disease before clinically visible vascular lesions. Photoreceptor and ganglion cell death occurs as early as 2-4 weeks after the onset of diabetes (1-3).Because of the difficulty of studying the mechanisms of the early stages of diabetes in humans, rodent models of type 1 and type 2 diabetes have been developed. Those models have been used to study various specific aspects of early stages of diabetic retinopathy including blood retinal barrier leakage (4), growth factor signaling such as the insulin/insulinlike growth factor receptor and vascular endothelial cell receptor pathways (5, 6), and oxidative stress mechanisms such as reactive oxygen species and advanced glycation end product production (7,8). These targeted studies provide valuable information on different aspects of the pathology and are useful in new therapeutic development, but more detailed discovery research is also needed to understand the full range of metabolic dysregulation that leads to diabetes complications. Proteome profiling using two-dimensional (2D) 1 DIGE and/or isobaric tag for relative and absolute quantitation (iTRAQ) are methods that can characterize new pathophysiological components and potential therapeutic targets.Several studies have used 2D DIGE to gain a better understanding of diabetic complications in the heart and retina (9 -12). These global approaches revealed biochemical changes in the ret...

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Section: Methodsmentioning

confidence: 99%

The Retinal Proteome in Experimental Diabetic Retinopathy

Fort

Freeman

Losiewicz

et al. 2009

Molecular & Cellular Proteomics

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“…Tandem MALDI-ToF (MALDI-ToF/ToF) mass spectrometry was performed as described previously (10). Tandem mass spectrometry was performed on the 4800 Proteomics Analyzer (Applied Biosystems) and data were submitted to a MASCOT search engine for protein identification using GPS Explorer 3.6 software (Applied Biosystems).…”

Section: Methodsmentioning

confidence: 99%

“…2-DIGE and gel analysis was performed as previously detailed using samples (10,11). Depleted plasma was purified from lipids and nucleic acids by precipitation (2D-Cleanup, GE Healthcare) and quantified using the 2D-Quant protein assay (GE Healthcare).…”

Section: Methodsmentioning

confidence: 99%

Plasma Biomarkers in Pediatric Patients Undergoing Cardiopulmonary Bypass

et al. 2008

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ABSTRACT:It is critical to identify at-risk patients and minimize the deleterious effects of cardiopulmonary bypass (CPB) procedures in pediatric populations. The present study screened the plasma proteome of pediatric patients undergoing CPB procedures to identify potential clinical biomarkers related to tissue damage, inflammation, or other pathologies. Blood samples were collected at five different time points from 10 children undergoing a CPB procedure. Plasma was isolated and analyzed using two-dimensional differential in-gel electrophoresis and matrix-assisted laser desorption ionization time of flight mass spectrometry. Levels of differentially regulated proteins identified by two-dimensional differential in-gel electrophoresis, and related proteins were then measured in all time points and patients. As well, associated small molecules and ions were measured. The present study identified 13 proteins and protein isoforms altered in expression, including hemopexin, ceruloplasmin, inter-alpha inhibitor H4, and alpha-2-macroglobulin. Immunoblot analysis revealed significant decreases in each of these proteins during the CPB procedure. Significant changes in the levels of copper, iron, Hb, epinephrine, norepinephrine, and serotonin were observed. The potential markers of pathology (inflammation, oxidative stress) identified during this preliminary study may illuminate opportunities for preventative measures and/or treatments during and following CPB procedures in pediatric patients. (Pediatr Res 63: 638-644, 2008)A lthough cardiopulmonary bypass (CPB) methods are well optimized and mortality rates from pediatric CPB continue to decrease (1,2), surgery-related pathologies are still a major concern. In fact, changes have been observed in levels of cytokines (3), and CD antigens (4) in the blood following CPB procedures. Changes in such molecules may reflect tissue damage, inflammation, and other pathologies that could affect both the long-and short-term patient outcome. These markers of pathology are risk factors for in-hospital or post-discharge morbidity and mortality in pediatric CPB patients (5,6). A more complete catalog of the circulating changes accompanying pediatric CPB surgeries may enable better prevention and treatment of complications.Although survival with good acute outcomes after surgery is increasingly likely with pediatric CPB procedures, neurodevelopmental complications such as lower intelligence, motor deficits, and impaired language skills remain a concern (7). In addition, a higher incidence of behavioral and attention problems have been reported in these patients (8,9). Although the cause of such deficits has not been shown to result from the CPB procedure itself, it is important to identify the potential long-term effects of pediatric cardiac surgery on development.Previous studies by our laboratory have used proteomic techniques-specifically two-dimensional differential in-gel electrophoresis (2-DIGE)-to examine the proteomic profile of various tissues and body fluids (10 -12). We have p...

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“…The proteins were then subjected to trypsin digestion and mass spectrometric analysis (MALDI-TOF-MS/MS analysis) (33) at the microchemical and proteomics facility at Emory University as described previously (34,35). GPS Explorer 2.0 software (Applied Biosystems) and a MASCOT (͗www.matrixscience.com/͘) search engine were used for identification of peptide fragments.…”

Section: Identification Of Cationic Polypeptides Of Vaginal Fluidmentioning

confidence: 99%

Cationic Polypeptides Are Required for Anti-HIV-1 Activity of Human Vaginal Fluid

Venkataraman

Cole

Svoboda

et al. 2005

The Journal of Immunology

142

167

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Mucosal surfaces of the vagina are the portals for heterosexual transmission of HIV-1 and therefore play a fundamental role in the pathogenesis of primary infection. In the search for direct biological evidence for the role of human vaginal fluid in innate host defense, we characterized the anti-HIV-1 function of cationic polypeptides within minimally manipulated vaginal fluid. In the current study we revealed that vaginal fluid confers intrinsic anti-HIV-1 properties against both X4 and R5 strains of HIV-1 and could protect against HIV-1 infection and reduce proviral genome integration in organotypic cultures of human cervicovaginal tissue. The majority of this activity was contained in the cationic polypeptide fraction, and the depletion of cationic polypeptides using a selective cation exchange resin ablated most of the intrinsic activity against HIV-1. By adding the cationic polypeptide fraction to depleted vaginal fluid, we were able to restore activity against HIV-1. Using a proteomic approach, we identified 18 cationic polypeptides within vaginal fluid, nearly all of which are either known antimicrobials or have other purported roles in host defense. Interestingly, physiologic concentrations of 13 of the cationic polypeptides were not active alone against HIV-1, yet in concert they partially restored the anti-HIV-1 activity of cation-depleted vaginal fluid. These results suggest that synergism between cationic polypeptides is complex, and full anti-HIV-1 activity probably involves the aggregate of the cationic peptides and proteins in vaginal fluid.

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Distinct proteomic profiles of amphetamine self-administration transitional states

Cited by 41 publications

References 49 publications

The Retinal Proteome in Experimental Diabetic Retinopathy

The Retinal Proteome in Experimental Diabetic Retinopathy

Plasma Biomarkers in Pediatric Patients Undergoing Cardiopulmonary Bypass

Cationic Polypeptides Are Required for Anti-HIV-1 Activity of Human Vaginal Fluid

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