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Distinguishing mouse strains by proteomic analysis of pelage hair.

ABSTRACT: AKR/J mice display a hair interior defect (hid) phenotype for which the molecular basis is unknown. To investigate the application of hair-shaft proteomics to the study of such diseases, pelage from AKR/J and two other mouse strains without this defect was analyzed by shotgun proteomics. The results permitted the identification of 111 proteins from tryptic digests of total hair from AKR/J-hid/hid mice, which were predominantly keratins (Krts) and Krt-associated proteins (Krtaps). From the non-solubilizable (crosslinked) fraction of the hair remaining after extensive detergent extraction, 58 proteins were identified. The majority were Krts and Krtaps, but junctional and other membrane proteins, cytoplasmic proteins, and histones were also identified. The results indicate the incorporation of a multitude of proteins into highly crosslinked material. Comparison of unique peptides generated among hair samples from AKR/J-hid/hid, FVB/NJ+/+, and LP/J+/+ mice indicated that these inbred strains could be distinguished by their proteomic patterns. Transmission electron microscopy after mild treatment in detergent and reducing agent permitted the visualization of projections of cortex cells, with characteristic filament patterns, into adjoining medulla cells. Hair shafts from AKR/J mice were deficient in these projections and also exhibited relatively low levels of trichohyalin, a possible contributor to or marker for the hid phenotype.

SUBMITTER: Rice RH

PROVIDER: S-EPMC2853740 | biostudies-literature | 2009 Sep

REPOSITORIES: biostudies-literature

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Distinguishing mouse strains by proteomic analysis of pelage hair.

Rice Robert H RH Rocke David M DM Tsai Hua-Sheng HS Silva Kathleen A KA Lee Young Jin YJ Sundberg John P JP

The Journal of investigative dermatology 20090319 9

AKR/J mice display a hair interior defect (hid) phenotype for which the molecular basis is unknown. To investigate the application of hair-shaft proteomics to the study of such diseases, pelage from AKR/J and two other mouse strains without this defect was analyzed by shotgun proteomics. The results permitted the identification of 111 proteins from tryptic digests of total hair from AKR/J-hid/hid mice, which were predominantly keratins (Krts) and Krt-associated proteins (Krtaps). From the non-so ...[more]

PMID: 19295613

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Project description:BACKGROUND:The differentiation between pancreatic head cancer (PHC) and distal cholangiocarcinoma (DCC) can be challenging because of their anatomical and histopathological similarity. This is an important problem, because the distinction has important implications for the treatment of these malignancies. However, there are no biomarkers for the differential diagnosis of PHC and DCC. The present study aimed to identify novel diagnostic immunohistochemical biomarkers to distinguish PHC from DCC. METHODS:Liquid chromatography tandem mass spectrometry (LC-MS/MS) was employed to detect candidate proteins. Ten PHC and 8 DCC specimens were analyzed by LC-MS/MS. Selected proteins were evaluated, using immunohistochemical analysis, to determine whether they would be appropriate biomarkers. Finally, we generated biomarker panels to improve diagnostic accuracy. We applied these panels to clinically difficult cases (cases in which different diagnoses were made before and after operation). RESULTS:Consequently, 1820 proteins were detected using LC-MS/MS. Fifteen differentially expressed proteins were selected as candidates based on semi-quantitative comparison. We first performed immunohistochemical staining on samples from the small cohort group (12 PHCs and 12 DCCs) using 15 candidates. KRT17, ANXA10, TMEM109, PTMS, and ATP1B1 showed favorable performances and were tested in the next large cohort group (72 PHCs and 74 DCCs). Based on immunohistochemical analysis, KRT17 performed best for the diagnosis of PHC as a single marker; additionally, PTMS exhibited good performance for the diagnosis of DCCs. Moreover, we indicated the KRT17+/ANXA10+/PTMS- staining pattern as a biomarker panel for the correct diagnosis of PHC and KRT17-/ANXA10-/PTMS+ for the diagnosis of DCC. After immunohistochemical staining for examining samples from the clinically difficult cases, these panels showed satisfactory diagnostic performance with 85.7% (6/7) accuracy. CONCLUSIONS:We conclude that 5 proteins and 2 biomarker panels are promising for distinguishing PHC from DCC, and patients with an equivocal diagnosis would benefit from the application of these biomarkers. Confirmatory studies are needed to generalize these findings to other populations.

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Distinguishing mouse strains by proteomic analysis of pelage hair.

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Distinguishing mouse strains by proteomic analysis of pelage hair.

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