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Hanrieder, Jörg
Publications (10 of 11) Show all publications
Hanrieder, J., Ljungdahl, A. & Andersson, M. (2012). MALDI Imaging Mass Spectrometry of Neuropeptides in Parkinson's Disease. Journal of Visualized Experiments, 14(60), Article ID e3445.
Open this publication in new window or tab >>MALDI Imaging Mass Spectrometry of Neuropeptides in Parkinson's Disease
2012 (English)In: Journal of Visualized Experiments, ISSN 1940-087X, E-ISSN 1940-087X, Vol. 14, no 60, article id e3445Article in journal (Refereed) Published
Abstract [en]

MALDI imaging mass spectrometry (IMS) is a powerful approach that facilitates the spatial analysis of molecular species in biological tissue samples(2) (Fig.1). A 12 μm thin tissue section is covered with a MALDI matrix, which facilitates desorption and ionization of intact peptides and proteins that can be detected with a mass analyzer, typically using a MALDI TOF/TOF mass spectrometer. Generally hundreds of peaks can be assessed in a single rat brain tissue section. In contrast to commonly used imaging techniques, this approach does not require prior knowledge of the molecules of interest and allows for unsupervised and comprehensive analysis of multiple molecular species while maintaining high molecular specificity and sensitivity(2). Here we describe a MALDI IMS based approach for elucidating region-specific distribution profiles of neuropeptides in the rat brain of an animal model Parkinson's disease (PD). PD is a common neurodegenerative disease with a prevalence of 1% for people over 65 of age(3,4). The most common symptomatic treatment is based on dopamine replacement using L-DOPA(5). However this is accompanied by severe side effects including involuntary abnormal movements, termed L-DOPA-induced dyskinesias (LID)(1,3,6). One of the most prominent molecular change in LID is an upregulation of the opioid precursor prodynorphin mRNA(7). The dynorphin peptides modulate neurotransmission in brain areas that are essentially involved in movement control(7,8). However, to date the exact opioid peptides that originate from processing of the neuropeptide precursor have not been characterized. Therefore, we utilized MALDI IMS in an animal model of experimental Parkinson's disease and L-DOPA induced dyskinesia. MALDI imaging mass spectrometry proved to be particularly advantageous with respect to neuropeptide characterization, since commonly used antibody based approaches targets known peptide sequences and previously observed post-translational modifications. By contrast MALDI IMS can unravel novel peptide processing products and thus reveal new molecular mechanisms of neuropeptide modulation of neuronal transmission. While the absolute amount of neuropeptides cannot be determined by MALDI IMS, the relative abundance of peptide ions can be delineated from the mass spectra, giving insights about changing levels in health and disease. In the examples presented here, the peak intensities of dynorphin B, alpha-neoendorphin and substance P were found to be significantly increased in the dorsolateral, but not the dorsomedial, striatum of animals with severe dyskinesia involving facial, trunk and orolingual muscles (Fig. 5). Furthermore, MALDI IMS revealed a correlation between dyskinesia severity and levels of des-tyrosine alpha-neoendorphin, representing a previously unknown mechanism of functional inactivation of dynorphins in the striatum as the removal of N-terminal tyrosine reduces the dynorphin's opioid-receptor binding capacity(9). This is the first study on neuropeptide characterization in LID using MALDI IMS and the results highlight the potential of the technique for application in all fields of biomedical research.

Keywords
Medicine, Issue 60, Parkinson's disease, L-DOPA induced dyskinesia, striatum, opioid peptides, MALDI Imaging MS
National Category
Basic Medicine
Research subject
Neuroscience
Identifiers
urn:nbn:se:uu:diva-169644 (URN)10.3791/3445 (DOI)000209222700012 ()
Funder
Swedish Research Council, 522-2006-6416, 521-2007-5407Åke Wiberg FoundationThe Royal Swedish Academy of Sciences
Available from: 2012-03-05 Created: 2012-03-05 Last updated: 2018-01-12Bibliographically approved
Brittebo, E., Karlsson, O., Andersson, M., Berg, A.-L., Roman, E., Lindquist, N. G. & Hanrieder, J. (2012). Neurotoxin-induced fibril formation and protein changes in rodents. Toxicology Letters, 211(Suppl.), S193-193
Open this publication in new window or tab >>Neurotoxin-induced fibril formation and protein changes in rodents
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2012 (English)In: Toxicology Letters, ISSN 0378-4274, E-ISSN 1879-3169, Vol. 211, no Suppl., p. S193-193Article in journal, Meeting abstract (Other academic) Published
National Category
Neurosciences
Identifiers
urn:nbn:se:uu:diva-287605 (URN)10.1016/j.toxlet.2012.03.693 (DOI)
Available from: 2016-04-25 Created: 2016-04-25 Last updated: 2018-01-10Bibliographically approved
Brishammar, S., Hanrieder, J. & Bergquist, J. (2012). TMV-particle borne enhancer of a tobacco RNA-replicase. World Journal of Science and Technology, 2(7), 4-7
Open this publication in new window or tab >>TMV-particle borne enhancer of a tobacco RNA-replicase
2012 (English)In: World Journal of Science and Technology, ISSN 2231-2587, Vol. 2, no 7, p. 4-7Article in journal (Refereed) Published
Abstract [en]

According to separation studies it has been evident that a tobacco RNA-replicase after TMV-infection consists of two parts. The larger part is host-directed and will be combined with a virus borne small protein which considerably enhances the RNA-replicase activity and is therefore named replicase enhancer, Ree. This compound was found at HPLC-separations of TMV-coat proteins, and was detected using polymerase assay with a radioactive nucleotide involved. Molecular weight has been determined by mass spectrometry: with FT ICR MS to get the size – 6 023.3 - and with MALDI TOF MS to obtain a sequence of the 54 amino acids involved. Presumably Ree is fixed to the TMV-RNA at infection. The enzyme seems to produce minus-strands of the virus RNA.

Keywords
Enhancer, mass spectrometry, replicase, TMV, tobacco
National Category
Chemical Sciences
Identifiers
urn:nbn:se:uu:diva-184707 (URN)
Available from: 2012-11-15 Created: 2012-11-13 Last updated: 2012-11-15Bibliographically approved
Ljungdahl, A., Hanrieder, J., Fälth, M., Bergquist, J. & Andersson, M. (2011). Imaging Mass Spectrometry Reveals Elevated Nigral Levels of Dynorphin Neuropeptides in L-DOPA-Induced Dyskinesia in Rat Model of Parkinson's Disease. PLoS ONE, 6(9), e25653
Open this publication in new window or tab >>Imaging Mass Spectrometry Reveals Elevated Nigral Levels of Dynorphin Neuropeptides in L-DOPA-Induced Dyskinesia in Rat Model of Parkinson's Disease
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2011 (English)In: PLoS ONE, ISSN 1932-6203, E-ISSN 1932-6203, Vol. 6, no 9, p. e25653-Article in journal (Refereed) Published
Abstract [en]

L-DOPA-induced dyskinesia is a troublesome complication of L-DOPA pharmacotherapy of Parkinson's disease and has been associated with disturbed brain opioid transmission. However, so far the results of clinical and preclinical studies on the effects of opioids agonists and antagonists have been contradictory at best. Prodynorphin mRNA levels correlate well with the severity of dyskinesia in animal models of Parkinson's disease; however the identities of the actual neuroactive opioid effectors in their target basal ganglia output structures have not yet been determined. For the first time MALDI-TOF imaging mass spectrometry (IMS) was used for unbiased assessment and topographical elucidation of prodynorphin-derived peptides in the substantia nigra of a unilateral rat model of Parkinson's disease and L-DOPA induced dyskinesia. Nigral levels of dynorphin B and alpha-neoendorphin strongly correlated with the severity of dyskinesia. Even if dynorphin peptide levels were elevated in both the medial and lateral part of the substantia nigra, MALDI IMS analysis revealed that the most prominent changes were localized to the lateral part of the substantia nigra. MALDI IMS is advantageous compared with traditional molecular methods, such as radioimmunoassay, in that neither the molecular identity analyzed, nor the specific localization needs to be predetermined. Indeed, MALDI IMS revealed that the bioconverted metabolite leu-enkephalin-arg also correlated positively with severity of dyskinesia. Multiplexing DynB and leu-enkephalin-arg ion images revealed small (0.25 by 0.5 mm) nigral subregions with complementing ion intensities, indicating localized peptide release followed by bioconversion. The nigral dynorphins associated with L-DOPA-induced dyskinesia were not those with high affinity to kappa opioid receptors, but consisted of shorter peptides, mainly dynorphin B and alpha-neoendorphin that are known to bind and activate mu and delta opioid receptors. This suggests that mu and/or delta subtype-selective opioid receptor antagonists may be clinically relevant for reducing L-DOPA-induced dyskinesia in Parkinson's disease.

National Category
Neurosciences Chemical Sciences
Identifiers
urn:nbn:se:uu:diva-159673 (URN)10.1371/journal.pone.0025653 (DOI)000295941300046 ()
Funder
Swedish Research Council, 522-2006-6414 (MA), 521- 2007-5407 (MA), 342-2004-3944(JB) and 621-2008-3562(JB)
Available from: 2011-10-06 Created: 2011-10-06 Last updated: 2018-01-12Bibliographically approved
Hanrieder, J., Wicher, G., Bergquist, J., Andersson, M. & Fex-Svenningsen, A. (2011). MALDI mass spectrometry based molecular phenotyping of CNS glial cells for prediction in mammalian brain tissue. Analytical and Bioanalytical Chemistry, 401(1), 135-147
Open this publication in new window or tab >>MALDI mass spectrometry based molecular phenotyping of CNS glial cells for prediction in mammalian brain tissue
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2011 (English)In: Analytical and Bioanalytical Chemistry, ISSN 1618-2642, E-ISSN 1618-2650, Vol. 401, no 1, p. 135-147Article in journal (Refereed) Published
Abstract [en]

The development of powerful analytical techniques for specific molecular characterization of neural cell types is of central relevance in neuroscience research for elucidating cellular functions in the central nervous system (CNS). This study examines the use of differential protein expression profiling of mammalian neural cells using direct analysis by means of matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). MALDI-MS analysis is rapid, sensitive, robust, and specific for large biomolecules in complex matrices. Here, we describe a newly developed and straightforward methodology for direct characterization of rodent CNS glial cells using MALDI-MS-based intact cell mass spectrometry (ICMS). This molecular phenotyping approach enables monitoring of cell growth stages, (stem) cell differentiation, as well as probing cellular responses towards different stimulations. Glial cells were separated into pure astroglial, microglial, and oligodendroglial cell cultures. The intact cell suspensions were then analyzed directly by MALDI-TOF-MS, resulting in characteristic mass spectra profiles that discriminated glial cell types using principal component analysis. Complementary proteomic experiments revealed the identity of these signature proteins that were predominantly expressed in the different glial cell types, including histone H4 for oligodendrocytes and S100-A10 for astrocytes. MALDI imaging MS was performed, and signature masses were employed as molecular tracers for prediction of oligodendroglial and astroglial localization in brain tissue. The different cell type specific protein distributions in tissue were validated using immunohistochemistry. ICMS of intact neuroglia is a simple and straightforward approach for characterization and discrimination of different cell types with molecular specificity.

Keywords
Intact cell mass spectrometry (ICMS), MALDITOF- MS, Imaging mass spectrometry (IMS), Glial cells
National Category
Analytical Chemistry
Identifiers
urn:nbn:se:uu:diva-156047 (URN)10.1007/s00216-011-5043-y (DOI)000292156900012 ()21553124 (PubMedID)
Available from: 2011-07-08 Created: 2011-07-08 Last updated: 2017-12-11Bibliographically approved
Wetterhall, M., Zuberovic, A., Hanrieder, J. & Bergquist, J. (2010). Assessment of the partitioning capacity of high abundant proteins in human cerebrospinal fluid using affinity and immunoaffinity subtraction spin columns.. Journal of chromatography. B, 878(19), 1519-1530
Open this publication in new window or tab >>Assessment of the partitioning capacity of high abundant proteins in human cerebrospinal fluid using affinity and immunoaffinity subtraction spin columns.
2010 (English)In: Journal of chromatography. B, ISSN 1570-0232, E-ISSN 1873-376X, Vol. 878, no 19, p. 1519-1530Article in journal (Refereed) Published
Abstract [en]

The performance of three different affinity and immunoaffinity subtraction spin columns was investigated for the removal of the most abundant proteins in human cerebrospinal fluid (CSF). A pool of human CSF was processed with the spin columns and both the bound and flow through fractions were compared with each other and with intact CSF using 1D gel electrophoresis and nanoLC-MALDI-TOF/TOF-MS analysis. MASCOT MS/MS ionscores were compared before and after processing with the columns. The non-specific co-removal of proteins bound to the high abundant proteins, so called "sponge effect" was also examined for each spin column. The reproducibility of one of the spin columns, ProteomeLab IgY-12 proteome partitioning spin column, was further investigated by isobaric tags for relative and absolute quantification (iTRAQ) labeling and MS/MS analysis. Overall, 173 unique proteins were identified on a 95% MudPIT confidence scoring level. For all three spin columns, the number of proteins identified and their MASCOT scores were increased up to 10 times. The largest degree of non-specific protein removal was observed for a purely affinity based albumin removal column, where 28 other proteins also were present. The ProteomeLab IgY-12 proteome partitioning spin column showed very high reproducibility when combined with iTRAQ labeling and MS/MS analysis. The combined relative standard deviation (R.S.D.) for the high abundant protein removal, iTRAQ labeling and nanoLC-MALDI-TOF/TOF-MS analysis was less than 17.5%.

Keywords
Cerebrospinal fluid (CSF), mass spectrometry (MS), proteomics, immunoaffinity subtraction, protein fractionation, iTRAQ quantification
National Category
Analytical Chemistry
Research subject
Chemistry with specialization in Analytical Chemistry
Identifiers
urn:nbn:se:uu:diva-125425 (URN)10.1016/j.jchromb.2010.04.003 (DOI)000278649400013 ()20444656 (PubMedID)
Available from: 2010-05-19 Created: 2010-05-19 Last updated: 2017-12-12Bibliographically approved
Ekegren, T., Hanrieder, J. & Bergquist, J. (2008). Clinical perspectives of high-resolution mass spectrometry-based proteomics in neuroscience: Exemplified in amyotrophic lateral sclerosis biomarker discovery research. Journal of Mass Spectrometry, 43(5), 559-571
Open this publication in new window or tab >>Clinical perspectives of high-resolution mass spectrometry-based proteomics in neuroscience: Exemplified in amyotrophic lateral sclerosis biomarker discovery research
2008 (English)In: Journal of Mass Spectrometry, ISSN 1076-5174, E-ISSN 1096-9888, Vol. 43, no 5, p. 559-571Article in journal (Refereed) Published
Abstract [en]

Biomarker discovery is a central application in today's proteomic research. There is an urgent need for valid biomarkers to improve diagnostic tools and treatment in many disorders, such as the rapidly progressing neurodegenerative disorder amyotrophic lateral sclerosis (ALS) that has a fatal outcome in about 3 years and yet no curative treatment. Screening for clinically relevant biomarkers puts high demands on high-throughput, rapid and precise proteomic techniques. There is a large variety in the methods of choice involving mainly gel-based approaches as well as chromatographic techniques for multi-dimensional protein and peptide separations followed by mass spectrometry (MS) analysis. This special feature article will discuss some important aspects of MS-based clinical proteomics and biomarker discovery in the field of neuro degenerative diseases and ALS research respectively, with the aim to provide a prospective view on current and future research aspects in the field. Furthermore, examples for application of high-resolution MS-based proteomic strategies for ALS biomarker discovery will be demonstrated with two studies previously reported by our group. These studies include among others, utilization of capillary liquid chromatography-Fourier transform ion cyclotron resonance mass spectrometry (LC-FTICR-MS) for advanced protein pattern classification in cerebrospinal fluid (CSF) samples of ALS patients as well as highly sensitive protein identification in minimal amounts of postmortem spinal cord tissue and laser micro-dissected motor neurons using FT-ICR-MS in conjunction with nanoflow LC coupled to matrix-assisted laser desorption ionization time-of-flight tandem mass spectrometry (LC-MALDI-TOF-TOF-MS).

Keywords
Biomarker, Clinical proteomics, Amyotrophic lateral sclerosis, Neurodegeneration, FTICR-MS, MALDI-TOF-TOF MS, Cerebrospinal fluid, Nervous tissue
National Category
Chemical Sciences
Identifiers
urn:nbn:se:uu:diva-16225 (URN)10.1002/jms.1409 (DOI)000256451700001 ()
Available from: 2008-05-13 Created: 2008-05-13 Last updated: 2017-12-08Bibliographically approved
Hanrieder, J. (2008). Mass Spectrometry Based Protein Biomarker Discovery in Clinical Specimens. (Licentiate dissertation). Uppsala universitet
Open this publication in new window or tab >>Mass Spectrometry Based Protein Biomarker Discovery in Clinical Specimens
2008 (English)Licentiate thesis, comprehensive summary (Other academic)
Place, publisher, year, edition, pages
Uppsala universitet, 2008
National Category
Chemical Sciences
Identifiers
urn:nbn:se:uu:diva-153754 (URN)
Available from: 2011-06-23 Created: 2011-05-18 Last updated: 2011-06-23Bibliographically approved
Hanrieder, J., Nyakas, A., Naessén, T. & Bergquist, J. (2008). Proteomic Analysis of Human Follicular Fluid Using an Alternative Bottom-Up Approach. Journal of Proteome Research, 7(1), 443-449
Open this publication in new window or tab >>Proteomic Analysis of Human Follicular Fluid Using an Alternative Bottom-Up Approach
2008 (English)In: Journal of Proteome Research, ISSN 1535-3893, E-ISSN 1535-3907, Vol. 7, no 1, p. 443-449Article in journal (Refereed) Published
Abstract [en]

Human follicular fluid (hFF) is the in vivo environment of oocytes during follicular maturation in the ovaries. It contains a huge variety of compounds such as, e.g., proteins that might play an important role in follicular development and oocyte growth. Previous proteomic studies on follicular fluid have isolated and already identified a certain number of proteins. Nevertheless, only a small part of proteins present in follicular fluid have been covered so far and a large number have still not been identified. Therefore, the need for new, more resolving, and sensitive approaches in proteome research is evident. We utilized a proteomic setup based on in solution isoelectric focusing (IEF) and reversed-phase nanoliquid chromatography coupled to matrix-assisted laser desorption/ionization time-of-flight tandem mass spectrometry (nano-LC MALDI TOF/TOF MS) for in depth protein analysis of human follicular fluid samples of patients undergoing controlled ovarian hyper stimulation (COH) for in vitro fertilization therapy (IVF). This approach led to the significant identification of 69 proteins, where 32 have not been reported before to be found in human follicular fluid with proteomic methods. Among these findings, at least two relevant compounds essentially involved in hormone secretion regulation during the folliculogenetic process were identified: sex hormone binding globulin (SHBG) and inhibin A (INHA). To confirm these results, both proteins were further validated by immunoassays.

Keywords
human follicular fluid (hFF), proteomics, in solution isoelectric focusing (IEF), matrix-assisted laser desorption/ionization time-of-flight tandem mass spectrometry (MALDI TOF/TOF MS), sex hormone binding globulin (SHBG), inhibin A (INHA)
National Category
Medical and Health Sciences
Identifiers
urn:nbn:se:uu:diva-13650 (URN)10.1021/pr070277z (DOI)000252154200050 ()18047273 (PubMedID)
Available from: 2008-01-24 Created: 2008-01-24 Last updated: 2017-12-11Bibliographically approved
Fedulova, N., Hanrieder, J., Bergquist, J. & Emrén, L. O. (2007). Expression and purification of catalytically active human PHD3 in Escherichia coli. Protein Expression and Purification, 54(1), 1-10
Open this publication in new window or tab >>Expression and purification of catalytically active human PHD3 in Escherichia coli
2007 (English)In: Protein Expression and Purification, ISSN 1046-5928, E-ISSN 1096-0279, Vol. 54, no 1, p. 1-10Article in journal (Refereed) Published
Abstract [en]

Transcription factor HIF-1 is a key regulator in cellular adaptation to hypoxia. HIF prolyl hydroxylases (PHDs) control HIF-1 accumulation by hydroxylation dependent on molecular oxygen. Due to this regulation, PHDs have been pointed out as potential drug targets. We have purified catalytically active human PHD3 after heterologous expression in Escherichia coli. Histidine-tagged enzyme was isolated as monomer by immobilized Ni-affinity chromatography followed by gel filtration. Overexpression of bacterial chaperonins GroEL/ES at 30 °C substantially increased the yield of soluble PHD3. High concentrations of salt and reducing agent during purification prevented protein aggregation. The enzyme activity with peptide derived from HIF-1α was inhibited by Zn2+, desferrioxamine and imidazole. The hydroxylation activity was verified by mass spectrometry, and Pro567 in HIF-1α was discovered as a new site of hydroxylation.

Keywords
Chaperonine, dioxygenase, EGLN, hydroxyproline, ischemia, proline
National Category
Biochemistry and Molecular Biology
Research subject
Biochemistry
Identifiers
urn:nbn:se:uu:diva-10913 (URN)10.1016/j.pep.2007.02.018 (DOI)000246711200001 ()17434750 (PubMedID)
Available from: 2007-05-03 Created: 2007-05-03 Last updated: 2017-12-11Bibliographically approved
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