Cortical Spreading Depression Can Be Triggered by Sensory Stimulation in Primed Wild Type Mouse Brain: a Mechanistic Insight to Migraine Aura Generation

Abstract Background: Unlike the spontaneously appearing aura in migraineurs, experimentally, cortical spreading depression (CSD), the neurophysiological correlate of aura is induced by non-physiological stimuli. Consequently, neural mechanisms involved in spontaneous CSD generation, which may provide insight how migraine starts in an otherwise healthy brain, remains largely unclear. We hypothesized that CSD can be physiologically induced by sensory stimulation in primed mouse brain. Methods: Cortex was made susceptible to CSD with partial inhibition of Na+/K+-ATPase by epidural application of a low dose of Na+/K+-ATPase blocker ouabain that does not induce repetitive CSDs or by knocking-down α2 subunit of Na+/K+-ATPase, which is crucial for K+ and glutamate re-uptake by astrocytes, with shRNA. Stimulation-induced CSDs and extracellular K+ changes were monitored in vivo electrophysiologically or with a K+-sensitive fluoroprobe (IPG-4). Results: After priming with ouabain, photic stimulation increased the CSD incidence compared with non-stimulated animals (44.0 vs. 4.9%, p<0.001). Whisker stimulation was less effective (14.9 vs. 2.4%, p=0.02). Knocking-down Na+/K+-ATPase (50% decrease in mRNA) lowered the CSD threshold in all mice tested but triggered stimulus-induced CSDs in 14.3% and 16.7% of mice with photic and whisker stimulation, respectively. Confirming Na+/K+-ATPase hypofunction, extracellular K+ significantly rose during stimulation after subthreshold ouabain or shRNA treatment unlike controls. In line with higher CSD susceptibility, K+ rise was more prominent after ouabain. To gain insight to preventive mechanisms reducing the incidence of stimulus-induced CSDs, we applied an A1-receptor (DPCPX) or GABA-A (bicuculine) antagonist over the occipital cortex, because adenosine formed during stimulation or inhibitory interneuron activity can reduce CSD susceptibility. DPCPX induced CSDs or CSD-like small-DC shifts during photic stimulation, whereas bicuculine was not effective. Conclusions: Our findings indicate that normal brain is well protected against CSD generation. For CSD to be ignited under physiological conditions, priming and predisposing factors are required as seen in migraine patients. Intense sensory stimulation has the potential to trigger a CSD when co-existing conditions can bring extracellular K+ and glutamate concentrations over threshold via reduced uptake of K+ and glutamate (e.g. inefficient fueling of α2-Na+/K+-ATPase due to reduced glycogen breakdown) or facilitated glutamate release (e.g. reduced presynaptic adenosinergic inhibition).

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Src family kinases activity is required for transmitting purinergic P2X7 receptor signaling in cortical spreading depression and neuroinflammation

The Journal of Headache and Pain ◽

10.1186/s10194-021-01359-8 ◽

2021 ◽

Vol 22 (1) ◽

Author(s):

Lingdi Nie ◽

Dongqing Ma ◽

John P. Quinn ◽

Minyan Wang

Keyword(s):

Gene Expression ◽

Mouse Brain ◽

P2x7 Receptor ◽

Cortical Spreading Depression ◽

Spreading Depression ◽

Glutamate Release ◽

Brain Slices ◽

Src Family Kinases ◽

Receptor Signaling ◽

Purinergic P2x7 Receptor

Abstract Background Purinergic P2X7 receptor plays an important role in migraine pathophysiology. Yet precise molecular mechanism underlying P2X7R signaling in migraine remains unclear. This study explores the hypothesis that P2X7 receptor transmits signaling to Src family kinases (SFKs) during cortical spreading depression (CSD) and neuroinflammation after CSD. Methods CSD was recorded using electrophysiology in rats and intrinsic optical imaging in mouse brain slices. Cortical IL-1β and TNFα mRNA levels were detected using qPCR. Glutamate release from mouse brain slices was detected using glutamate assay. Results The data showed that deactivation of SFKs by systemic injection of PP2 reduced cortical susceptibility to CSD in rats and CSD-induced IL-1β and TNF-α gene expression in rat ipsilateral cortices. Consistently, in mouse brain slices, inhibition of SFKs activity by saracatinib and P2X7 receptor by A740003 similarly reduced cortical susceptibility to CSD. When the interaction of P2X7 receptor and SFKs was disrupted by TAT-P2X7, a marked reduction of cortical susceptibility to CSD, IL-1β gene expression and glutamate release after CSD induction were observed in mouse brain slices. The reduced cortical susceptibility to CSD by TAT-P2X7 was restored by NMDA, and disrupting the Fyn-NMDA interaction using TAT-Fyn (39-57) but not disrupting Src-NMDA receptor interaction using TAT-Src (40-49) reduced cortical susceptibility to CSD. Furthermore, activation of P2X7 receptor by BzATP restored the TAT-Fyn (39-57)-reduced cortical susceptibility to CSD. Conclusion This study reveals that SFKs activity transmits P2X7 receptor signaling to facilitate CSD propagation via glutamatergic pathway and promote neuroinflammation, which is of particular relevance to migraine.

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Src Family Kinases Activity is Required for Transmitting Purinergic P2X7 Receptor Signaling in Cortical Spreading Depression and Neuroinflammation

10.21203/rs.3.rs-907798/v1 ◽

2021 ◽

Author(s):

Lingdi Nie ◽

Dongqing Ma ◽

John P Quinn ◽

Minyan Wang

Keyword(s):

Gene Expression ◽

Mouse Brain ◽

P2x7 Receptor ◽

Cortical Spreading Depression ◽

Spreading Depression ◽

Glutamate Release ◽

Brain Slices ◽

Src Family Kinases ◽

Purinergic P2x7 Receptor ◽

Precise Molecular Mechanism

Abstract Background Purinergic P2X7 receptor plays a key role in migraine pathophysiology. Yet precise molecular mechanism underlying P2X7R signaling in migraine remains unclear. This study explores the hypothesis that P2X7 receptor transmits signaling to Src family kinases (SFKs) during cortical spreading depression (CSD) and CSD-induced neuroinflammation. Methods CSD was recorded using electrophysiology in rats, and intrinsic optical imaging in mouse brain slices. Cortical IL-1β and TNFα mRNA expression were detected using qPCR. Glutamate release in mouse brain slices was detected using glutamate assay. Results The data showed that systematic deactivation of SFKs by PP2 reduced cortical susceptibility to CSD in rats and CSD-induced IL-1b and TNF-a gene expression in rat ipsilateral cortices. Consistently, in mouse brain slices, inhibition of SFKs activity by saracatinib and P2X7 receptor by A740003 similarly reduced cortical susceptibility to CSD. When the interaction of P2X7 receptor-SFKs was disrupted by TAT-P2X7, a marked reduction of cortical susceptibility to CSD, CSD-induced IL-1b gene expression and glutamate release were observed in mouse brain slices. The reduced cortical susceptibility to CSD by TAT-P2X7 was restored by NMDA and disrupting Fyn-NMDA interaction using TAT-Fyn (39-57), but not disrupting Src-NMDA receptor using TAT-Src (40-49), reduced cortical susceptibility to CSD. Furthermore, activation of P2X7 receptor by BzATP restored the TAT-Fyn (39-57)-reduced cortical susceptibility to CSD. Conclusion This study reveals that SFKs activity mediates P2X7 receptor pore formation facilitating CSD propagation, CSD-induced neuroinflammation and glutamate release, of particular relevance to migraine.

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In Vivo Uptake of [3H]Nimodipine into Brain during Cortical Spreading Depression

Journal of Cerebral Blood Flow & Metabolism ◽

10.1097/00004647-199705000-00014 ◽

1997 ◽

Vol 17 (5) ◽

pp. 586-590 ◽

Cited By ~ 5

Author(s):

Sachiko Osuga ◽

Antoine M. Hakim ◽

Hitoshi Osuga ◽

Matthew J. Hogan

Keyword(s):

Rat Brain ◽

Cortical Spreading Depression ◽

Spreading Depression ◽

Control Group ◽

Similar Measurement ◽

Contralateral Hemisphere ◽

Radiotracer Uptake ◽

Uptake Studies ◽

In Vivo Uptake

We report autoradiographic measurements of the in vivo uptake of [3H]nimodipine during the nonischemic depolarization of cortical spreading depression (CSD) in rat brain. [3H]Nimodipine uptake in brain was determined regionally in rats undergoing CSD (n = 8) and was significantly increased in cortex (14 ± 7%) and hippocampus (10 ± 6%) on the stimulated side relative to the contralateral hemisphere when compared with the same measurements in a control group (n = 8). A similar measurement using the physiologically inert radiotracer [14C]iodoantipyrine to control for potential effects of CSD on radioligand distribution showed a minimal increase (2.4 ± 0.7%) of radiotracer uptake in cortex after CSD. This increase was significantly less than that observed in the [3H]nimodipine uptake studies. We hypothesize that increased in vivo [3H]nimodipine uptake in CSD identifies regions of depolarization and thus infers activation of the L-type voltage sensitive calcium channels.

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Diffusion-Weighted MRI Used to Detect in Vivo Modulation of Cortical Spreading Depression: Comparison of Sumatriptan and Tonabersat

Experimental Neurology ◽

10.1006/exnr.2001.7809 ◽

2001 ◽

Vol 172 (2) ◽

pp. 342-353 ◽

Cited By ~ 32

Author(s):

Daniel P. Bradley ◽

Martin I. Smith ◽

Chaiyapoj Netsiri ◽

Justin M. Smith ◽

Kurt H.J. Bockhorst ◽

...

Keyword(s):

Cortical Spreading Depression ◽

Spreading Depression ◽

Diffusion Weighted Mri ◽

Diffusion Weighted

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Development of a neural probe to measure potassium during a cortical spreading depression in-vivo

2013 Transducers & Eurosensors XXVII: The 17th International Conference on Solid-State Sensors, Actuators and Microsystems (TRANSDUCERS & EUROSENSORS XXVII) ◽

10.1109/transducers.2013.6627226 ◽

2013 ◽

Author(s):

E. J. van der Wouden ◽

E. A. Tolner ◽

M. Odijk ◽

W. Olthuis ◽

A. M. J. M. van den Maagdenburg ◽

...

Keyword(s):

Cortical Spreading Depression ◽

Spreading Depression ◽

Neural Probe

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Multiple Irradiation Affects Cellular and Extracellular Components of the Mouse Brain Tissue and Adhesion and Proliferation of Glioblastoma Cells in Experimental System In Vivo

International Journal of Molecular Sciences ◽

10.3390/ijms222413350 ◽

2021 ◽

Vol 22 (24) ◽

pp. 13350

Author(s):

Maxim O. Politko ◽

Alexandra Y. Tsidulko ◽

Oxana A. Pashkovskaya ◽

Konstantin E. Kuper ◽

Anastasia V. Suhovskih ◽

...

Keyword(s):

Brain Tissue ◽

Mouse Brain ◽

Glial Cells ◽

Ex Vivo ◽

Experimental System ◽

Normal Brain ◽

Biosynthetic Activity ◽

Normal Brain Tissue

Intensive adjuvant radiotherapy (RT) is a standard treatment for glioblastoma multiforme (GBM) patients; however, its effect on the normal brain tissue remains unclear. Here, we investigated the short-term effects of multiple irradiation on the cellular and extracellular glycosylated components of normal brain tissue and their functional significance. Triple irradiation (7 Gy*3 days) of C57Bl/6 mouse brain inhibited the viability, proliferation and biosynthetic activity of normal glial cells, resulting in a fast brain-zone-dependent deregulation of the expression of proteoglycans (PGs) (decorin, biglycan, versican, brevican and CD44). Complex time-point-specific (24–72 h) changes in decorin and brevican protein and chondroitin sulfate (CS) and heparan sulfate (HS) content suggested deterioration of the PGs glycosylation in irradiated brain tissue, while the transcriptional activity of HS-biosynthetic system remained unchanged. The primary glial cultures and organotypic slices from triple-irradiated brain tissue were more susceptible to GBM U87 cells’ adhesion and proliferation in co-culture systems in vitro and ex vivo. In summary, multiple irradiation affects glycosylated components of normal brain extracellular matrix (ECM) through inhibition of the functional activity of normal glial cells. The changed content and pattern of PGs and GAGs in irradiated brain tissues are accompanied by the increased adhesion and proliferation of GBM cells, suggesting a novel molecular mechanism of negative side-effects of anti-GBM radiotherapy.

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Effects of Antiepileptic Drugs on K+-Induced Glutamate Release and Spreading Depression in the Hippocampus Determined with an In Vivo Microdialysis Glutamate Biosensor (MGB)

Epilepsia ◽

10.1111/j.1528-1157.1998.tb01910.x ◽

1998 ◽

Vol 39 (S5) ◽

pp. 56-56

Author(s):

Motohiro Okada ◽

Yuko Kawata ◽

Kazuhiro Kiryu ◽

Kazuhisa Mizuno ◽

Kazumaru Wada ◽

...

Keyword(s):

Antiepileptic Drugs ◽

Spreading Depression ◽

Glutamate Release ◽

In Vivo Microdialysis

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Sarcoma Family Kinase-Dependent Pannexin-1 Activation after Cortical Spreading Depression Is Mediated by NR2A-Containing Receptors

International Journal of Molecular Sciences ◽

10.3390/ijms21041269 ◽

2020 ◽

Vol 21 (4) ◽

pp. 1269 ◽

Cited By ~ 1

Author(s):

Fan Bu ◽

Lingdi Nie ◽

John P Quinn ◽

Minyan Wang

Keyword(s):

Mouse Brain ◽

Protein Interactions ◽

Cortical Spreading Depression ◽

Spreading Depression ◽

Regulatory Mechanism ◽

Brain Slices ◽

Pannexin 1 ◽

Migraine Headaches ◽

Aspartate Receptor ◽

Channel Opening

Cortical spreading depression (CSD) is a propagating wave of depolarization followed by depression of cortical activity. CSD triggers neuroinflammation via the pannexin-1 (Panx1) channel opening, which may eventually cause migraine headaches. However, the regulatory mechanism of Panx1 is unknown. This study investigates whether sarcoma family kinases (SFK) are involved in transmitting CSD-induced Panx1 activation, which is mediated by the NR2A-containing N-methyl-D-aspartate receptor. CSD was induced by topical application of K+ to cerebral cortices of rats and mouse brain slices. SFK inhibitor, PP2, or NR2A–receptor antagonist, NVP–AAM077, was perfused into contralateral cerebral ventricles (i.c.v.) of rats prior to CSD induction. Co-immunoprecipitation and Western blot were used for detecting protein interactions, and histofluorescence for addressing Panx1 activation. The results demonstrated that PP2 attenuated CSD-induced Panx1 activation in rat ipsilateral cortices. Cortical susceptibility to CSD was reduced by PP2 in rats and by TAT-Panx308 that disrupts SFK–Panx1 interaction in mouse brain slices. Furthermore, CSD promoted activated SFK coupling with Panx1 in rat ipsilateral cortices. Moreover, inhibition of NR2A by NVP–AAM077 reduced elevation of ipsilateral SFK–Panx1 interaction, Panx1 activation induced by CSD and cortical susceptibility to CSD in rats. These data suggest NR2A-regulated, SFK-dependent Panx1 activity plays an important role in migraine aura pathogenesis.

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