| Metadata |
| datasetIdentifier | PASS01349 |
| datasetType | SWATH |
| submitter | Meghan May <mmay3@une.edu> |
| submitter_organization | University of New England |
| lab_head_full_name | Karen Houseknecht |
| lab_head_email | khouseknecht@une.edu |
| lab_head_organization | University of New England |
| lab_head_country | United States |
| datasetTag | Antipsych_Drug_Exp |
| datasetTitle | Proteomic Changes in Heart and Liver during Atypical Antipsychotic Drug Treatment in a Mouse Model |
| publicReleaseDate | 2019-03-11 00:00:00 |
| finalizedDate | |
| summary | |
| contributors | Megan Beauchemin
Calvin Vary
Christine Lary
Kieran Wynne
Barbara Conley
Kathy Nevola
Deborah Barlow
Karen Houseknecht |
| publication | Unpublished |
| growth | 8 week old male C57BL/6J mice were treated once daily with vehicle or a low, clinically relevant doses of risperidone or olanzapine for 4 weeks. At the end of each treatment period, mice were sacrificed by CO2 asphyxiation (5 day treatment) or avertin (2.5% in PBS) injection (4 week treatment). |
| treatment | Mice were administered vehicle (VEH; 0.1% acetic acid, PO), risperidone (RIS); or olanzapine (OLAN) (Sigma; 1 mg/kg, PO) by oral gavage. This drug dose reflects plasma drug exposure consistent with what is observed clinically. |
| extraction | After necropsy, whole hearts and portions of liver from RIS-treated (N=5), OLAN-treated (N=5), and VEH-treated (N=4) mice were homogenized in HTNG lysis buffer (20 mM HEPES, 150mM NaCl, 1.5mM MgCl2, 10% glycerol, 1% Triton‐X 100 1mM EDTA, protease‐inhibitor cocktail (Calbiochem). Protein concentration from the tissue homogenate was determined using a bicinchonic acid assay (BCA, Pierce). |
| separation | Tryptic peptides were resolved on a Dionex Ultimate 3000 (RSLCnano) chromatography system (ThermoFisher Scientific). |
| digestion | 40 µg of protein from each sample was trypsin digested for MS analysis (ProteoExtract digestion kit, Calbiochem). |
| acquisition | The mass spectrometer was operated using data dependent acquisition (DDA) to create a library and sequential window acquisition of all theoretical spectra (SWATH) was implemented for relative quantitation. For both modes of operation (DDA and SWATH) the mass spectrometer used an ion spray voltage floating (ISVF) of 2400 V, curtain gas (CUR) 25, interface heater temperature (IHT) 150 °C, ion source gas 1 of 6, and a declustering potential (DP) 100 V
All data acquired in data dependent acquisition (DDA) mode used a high resolution MS scan from 400-1250 m/z to select the 15 most intense ions prior to MS/MS analysis using CID (Collision Induced Dissociation). |
| informatics | For identification of peptides, multiple fragment ion chromatograms were retrieved from the spectral library for each peptide of interest. These spectra were compared with the extracted fragment ion traces for the corresponding isolation window to identify the transitions that best identify the target peptide. SWATH analysis was performed using PeakView software, and MarkerView software was utilized for principal component analysis and T-test comparisons. The proteomic data was analyzed in the following pairwise comparisons: heart RIS to heart VEH control; heart OLAN to heart VEH control; liver RIS to liver VEH control; liver OLAN to liver VEH control. An adjusted p-value was calculated for each protein in each pairwise data set using the FDR method. Two significant protein lists were then made for each pairwise data set using the raw and adjusted p-values with a threshold of 0.05. |
| instruments | AB SCIEX TripleTOF 5600 |
| species | Mouse |
| massModifications | None |
