Panel of synaptic protein ELISAs for evaluating neurological phenotype

Paul E. Gottschall, Joanne M. Ajmo, Autumn K. Eakin, Matthew D. Howell, Hina Mehta, Lauren A. Bailey

Research output: Contribution to journalArticle

19 Scopus citations

Abstract

The purpose of this study was to develop ELISAs for key neural proteins, three synaptic and one glial, that exist in different intracellular compartments, which would be used as a measure of synaptic phenotype. These assays would be valuable to neurologically phenotype transgenic mouse models of human disease and also human disease itself using minimal amounts of post-mortem tissue. We showed that supernatant from crude brain tissue homogenates extracted in RIPA buffer containing 0.1% SDS bind to synaptophysin, synaptosome-associated protein of 25 kDa (SNAP-25), post-synaptic density-95 (PSD-95), and glial fibrillary acidic protein (GFAP) antibody pairs with high affinity and selectivity. Overall, RIPA + 0.1% SDS were more efficient than RIPA + 2% SDS or a buffer containing only 1% Triton-X-100. Diluting the brain extracts resulted in dose-dependent binding to the antibody pairs for each neural protein, with EC50s that varied from 8.6 μg protein for PSD-95 to 0.23 μg for GFAP. The assays were used to measure synaptic marker protein levels at various times during mouse development and GFAP in a model of disease accompanied by neuroinflammation. Comparison of ELISAs with Western blots by measuring marker levels in brain extract from developing mice showed a greater relative difference in values derived from ELISA. These ELISAs should be valuable to phenotype the synapse in neurological disease and their rodent models.

Original languageEnglish (US)
Pages (from-to)885-893
Number of pages9
JournalExperimental Brain Research
Volume201
Issue number4
DOIs
StatePublished - Apr 1 2010

    Fingerprint

Keywords

  • Enzyme-linked immunoassay
  • Glial fibrillary acidic protein (GFAP)
  • Post-synaptic density-95 (PSD-95)
  • Synapse
  • Synaptophysin
  • Synaptosome-associated protein of 25 kDa (SNAP-25)

ASJC Scopus subject areas

  • Neuroscience(all)

Cite this

Gottschall, P. E., Ajmo, J. M., Eakin, A. K., Howell, M. D., Mehta, H., & Bailey, L. A. (2010). Panel of synaptic protein ELISAs for evaluating neurological phenotype. Experimental Brain Research, 201(4), 885-893. https://doi.org/10.1007/s00221-010-2182-x