Effects of NMDA receptor modulators on a blood–brain barrier in vitro model
| Autor: | Christian Pifl, Michael Freidl, Michael L. Berger, Franz Gabor, Johannes Winkler, Winfried Neuhaus, Phillip Szkokan, Christian R. Noe, Michael Wirth |
|---|---|
| Rok vydání: | 2011 |
| Předmět: |
Blotting
Western Glutamic Acid Context (language use) In Vitro Techniques Pharmacology Biology Blood–brain barrier Occludin Receptors N-Methyl-D-Aspartate Cell Line chemistry.chemical_compound medicine Humans Receptor Neurotransmitter Molecular Biology Reverse Transcriptase Polymerase Chain Reaction musculoskeletal neural and ocular physiology General Neuroscience Glutamate receptor Endothelial Cells Membrane Proteins Phosphoproteins Dizocilpine medicine.anatomical_structure 2-Amino-5-phosphonovalerate nervous system chemistry Blood-Brain Barrier Zonula Occludens-1 Protein NMDA receptor Calcium Neurology (clinical) Dizocilpine Maleate Excitatory Amino Acid Antagonists Neuroscience Developmental Biology medicine.drug |
| Zdroj: | Brain Research. 1394:49-61 |
| ISSN: | 0006-8993 |
| DOI: | 10.1016/j.brainres.2011.04.003 |
| Popis: | Changes of the functionality of the blood-brain barrier (BBB) have been reported in the context of several brain related diseases such as multiple sclerosis, epilepsy, Alzheimer's disease and stroke. Several publications indicated the presence and functionality of the NMDA receptor (NMDAR) at the brain endothelium and a possible involvement of the NMDAR in the above-mentioned diseases. Recently, it was shown that the application of the NMDAR antagonist MK801 can block several adverse effects at the BBB in vitro, but also that MK801 can significantly change the proteome of brain endothelial cells without simultaneous stimulation of NMDAR by glutamate. Based on these reports we investigated if NMDAR antagonists MK801 and D-APV can affect the intracellular calcium level (Ca²⁺i) of an in vitro BBB model based on human cell line ECV304 on their own and compared these results to effects mediated by NMDAR agonists glutamate and NMDA. Treatment of ECV304 cells for 30 min with glutamate resulted in no significant change of Ca²⁺i. On the contrary, application of NMDA and NMDAR antagonists D-APV and MK801 led to a significant and concentration dependent decrease of Ca²⁺i. Further studies revealed that glutamate was able to decrease the transendothelial electrical resistance (TEER) of the BBB in vitro model, whereas NMDA and D-APV were able to increase TEER. Analysis of the protein expression levels of tight junctional molecules ZO-1 and occludin showed a complex regulation after application of NMDAR modulators. In summary, it was shown that NMDAR antagonists can alter BBB key properties in vitro on their own. Moreover, although qPCR results confirmed the presence of NMDA receptor subunits NR1, NR2A, NR2B and NR2C, membrane binding studies failed to prove the typical plasma membrane localization and functionality in human BBB cell line ECV304. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|
Full Text from ScienceDirect