TY  - JOUR
AU  - Lourbopoulos, Athanasios
AU  - Müller, Stephan A
AU  - Jocher, Georg
AU  - Wick, Manfred
AU  - Plesnila, Nikolaus
AU  - Lichtenthaler, Stefan F
TI  - An Improved Method for Sampling and Quantitative Protein Analytics of Cerebrospinal Fluid of Individual Mice.
JO  - Molecular & cellular proteomics
VL  - 24
IS  - 5
SN  - 1535-9476
CY  - Bethesda, Md.
PB  - The American Society for Biochemistry and Molecular Biology
M1  - DZNE-2025-00672
SP  - 100958
PY  - 2025
AB  - The mouse is the species most commonly used in preclinical research, but protein analytics of murine cerebrospinal fluid (CSF) remains challenging because of low sample volumes (often <10 μl) and frequent contaminations with blood. We developed an improved CSF sampling method that allows routine collection of larger volumes (20-30 μl) of pure CSF from individual mice, enabling multiple protein analytical assays from a single sample. Based on cell counts and hemoglobin ELISAs, we provide an easy quality control workflow for obtaining cell- and blood-free murine CSF. Through mass spectrometry-based proteomics using an absolutely quantified external standard, we estimated concentrations for hundreds of mouse CSF proteins. While repeated CSF sampling from the same mouse was possible, it induced CSF proteome changes. Applying the improved method, we found that the mouse CSF proteome remains largely stable over time in wild-type mice, but that amyloid pathology in the 5xFAD mouse model of Alzheimer's disease massively changes the CSF proteome. Neurofilament light chain and TREM2, markers of neurodegeneration and activated microglia, respectively, were strongly upregulated and validated using immunoassays. In conclusion, our refined murine CSF collection method overcomes previous limitations, allowing multiple quantitative protein analyses for applications in biomedicine.
KW  - Animals
KW  - Mice
KW  - Proteomics: methods
KW  - Alzheimer Disease: cerebrospinal fluid
KW  - Cerebrospinal Fluid Proteins
KW  - Proteome: metabolism
KW  - Mice, Inbred C57BL
KW  - Disease Models, Animal
KW  - Biomarkers: cerebrospinal fluid
KW  - Mice, Transgenic
KW  - Neurofilament Proteins: cerebrospinal fluid
KW  - CSF collection (Other)
KW  - aging (Other)
KW  - cerebrospinal fluid (Other)
KW  - method (Other)
KW  - mouse (Other)
KW  - neurodegeneration (Other)
KW  - quantitative proteomics (Other)
KW  - trauma (Other)
KW  - Cerebrospinal Fluid Proteins (NLM Chemicals)
KW  - Proteome (NLM Chemicals)
KW  - Biomarkers (NLM Chemicals)
KW  - Neurofilament Proteins (NLM Chemicals)
KW  - neurofilament protein L (NLM Chemicals)
LB  - PUB:(DE-HGF)16
C6  - pmid:40157722
C2  - pmc:PMC12090247
DO  - DOI:10.1016/j.mcpro.2025.100958
UR  - https://pub.dzne.de/record/279042
ER  -