6533b7d4fe1ef96bd1263555
RESEARCH PRODUCT
Simultaneous lipidomic and transcriptomic profiling in mouse brain punches of acute epileptic seizure model compared to controls
Raissa LernerD. R. Naomi VosLaura BindilaRon M. A. HeerenShane R. EllisBeat LutzJulia Maria Postsubject
0301 basic medicineBiochemistryTranscriptomechemistry.chemical_compoundEpilepsyMice0302 clinical medicineEndocrinologyTEMPORAL-LOBE EPILEPSYResearch Articlesmass spectrometrymessenger ribonucleic acidKainic AcidBrainNEUROLOGICAL DISORDERSQUANTITATIVE-ANALYSISEndocannabinoid systemLipidsCell biologyReal-time polymerase chain reactionmedicine.anatomical_structureAcute DiseaseArachidonic acidEpileptic seizuremedicine.symptomACID-INDUCED SEIZURESQD415-436BiologyMEMBRANE PHOSPHOLIPIDSENDOCANNABINOID SYSTEM03 medical and health sciencesCYTOPLASMIC PHOSPHOLIPASE A(2)SeizuresmedicineAnimalsendocannabinoidsphospholipidsGene Expression ProfilingRNACell BiologyMASS-SPECTROMETRYmedicine.diseaseDisease Models Animal030104 developmental biologychemistrynervous systemepilepsyLYSOPHOSPHATIDIC ACID030217 neurology & neurosurgeryTERT-BUTYL ETHERBasolateral amygdaladescription
In this study, we report the development of a dual extraction protocol for RNA and lipids, including phospholipids, endocannabinoids, and arachidonic acid, at high spatial resolution, e.g., brain punches obtained from whole frozen brains corresponding to four brain subregions: dorsal hippocampus, ventral hippocampus, basolateral amygdala, and hypothalamus. This extraction method combined with LC/multiple reaction monitoring for lipid quantification and quantitative PCR for RNA investigation allows lipidomic and transcriptomic profiling from submilligram amounts of tissue, thus benefiting the time and animal costs for analysis and the data reliability due to prevention of biological variability between animal batches and/or tissue heterogeneity, as compared with profiling in distinct animal batches. Moreover, the method allows a higher extraction efficiency and integrity preservation for RNA, while allowing concurrently quantitative analysis of low and high abundant lipids. The method was applied for brain punches obtained 1 h after kainic acid-induced epileptic seizures in mice (n = 10) compared with controls (n = 10), and enabled the provision of valuable new insights into the subregional lipid and RNA changes with epilepsy, highlighting its potential as a new viable tool in quantitative neurobiology.
| year | journal | country | edition | language |
|---|---|---|---|---|
| 2018-02-01 |