Table 1 a Summary of the Most Relevant Studies on CSD Experimental Models Regarding The

Table 1 A summary of the most relevant studies on CSD experimental models regarding the effects of currently used drugs and drugs under investigation for migraine prophylaxis

Drug * Animals Model Administration Outcome Effects Comments Authors [Ref] and dosage (s) Measurements DL-propranolol Isoflurane- KCl-induced CSD Chronic IP Number of CSDs Significant dose-dependent No effect of D- Ayata et al., 2006 [154] anaesthetised injection reduction propranolol rats 20mg/kg/day CSD propagation Significant dose-dependent reduction

Electrically- Threshold for Significant rise stimulated CSD CSD Single doses administered 1 hour before electrical threshold determination were ineffective

Amytriptyline Isoflurane- KCl-induced CSD Chronic IP Number of CSDs Significant dose-dependent Ayata et al., 2006 [154] anaesthetised injection reduction rats 10 or 20mg/kg/d Single doses CSD propagation Significant dose-dependent administered 1 hour reduction before electrical threshold Electrically- Threshold for Significant rise determination were stimulated CSD CSD ineffective

Flunarizine Alfentanil- KCl-induced CSD Acute IP. injection Depression of Reversal Reid et al., 1987 [166] anaesthetised 40 mg/Kg EEG activity rats negative shift in Block DC potential

threshold for the Enhancement elicitation of SD Flunarizine Uretane- KCl-induced CSD Acute IV injection Cortical Inhibition No effect 5-HT2 and Shimazawa et al., 1995 [169] anaesthetised 1 mg/kg hypoperfusion histamine H1 rats antagonist dimetotiazine

C fos expression Significant attenuation in ipsilateral Similar effect of the 1 frontoparietal Ca2+ channel blockers, Cortex induced by lomerizine (KB-2796) CSD

Methysergide Isoflurane- KCl-induced CSD Chronic IP Number of CSDs Significant dose-dependent Ayata et al., 2006 [154] anaesthetised injection reduction rats 0.1 mg and 1mg/kg/ CSD propagation Significant dose-dependent day reduction

Electrically- Threshold for Significant rise stimulated CSD CSD Valproate Alpha- Mechanically- Acute IP injection Speed of CSD Significant reduction No effects of DHE, Kaube & Goadsby, 1994 [153] chloralose- induced CSD 60 mg/Kg propagation acethylsalicylic acid, anaesthetised (needle prick) metoprolol, cats (females) Cortical blood No change clonazepam, lignocaine flow increase during the hyperemic phase Valproate Isoflurane- KCl-induced CSD Chronic IP Number of CSDs Significant dose-dependent Ayata et al., 2006 [154] anaesthetised injection reduction rats 25, 50, 100, or 200mg/kg/ CSD propagation Significant dose-dependent day reduction

Electrically- Threshold for Significant rise stimulated CSD CSD Valproate Chloral KCl-induced CSD Daily IP CSD frequency at Reduced at the anterior Bogdanov et al., 2011 [163] hydrate- injections for 1 a distal (parieto- recoding site only anaesthetised month occipital) and a rats proximal (frontal) 200 mg/kg/die electrode

CSD propagation Suppressed at the anterior between the two recoding site only by 32% electrode sites and slowed propagation velocity

Number of Fos- Decreased immunoreactive nuclei in frontal cortex.

2 Topiramate Alpha- Mechanically- Acute Cortical Inhibited Inhibition was shown Akerman & Goadsby, 2005 [154] chloralose- induced CSD administration by depolarization in all rats and 8 out of anaesthetised (needle prick) needle plunge into 13 cats rats and cats the cortex CSD propagation Inhibited

30 mg/kg regional cerebral hypoperfusion Inhibited Topiramate KCl-induced CSD Chronic IP Number of CSDs Significant dose-dependently Ayata et al., 2006 [154] injection reduced Long-term suppression 40, 60, or 80 CSD propagation mg/kg/day Significantly dose- Electrically- dependently reduced stimulated CSD Threshold for CSD Significantly raised

Topiramate Rats§ KCl-induced CSD In vitro CSD area Reduced No effect of Tozzi et al., 2012 [72] application 10 min carbamazepine after the first Intrinsic optical Reduced episode of CSD signals (IOS) Similar effects of 100 μM intensity NM DAR antagonist L -A PV) and Na+Channel blocker TTX Topiramate Isoflurane- KCl-induced CSD Once-daily peroral Frequency of Significantly dose- Unekawa et al., 2012 [155] anaesthetised treatment CSDs dependently reduced male rats 50, 100, 200 or Long-term suppression 600 mg/kg CSD propagation Significantly dose- dependently reduced

Interval between Lengthened CSD episodes Gabapentin Isoflurane- KCl-induced CSD Single IV injection Frequency of Dose-dependently reduced the Hoffmann et al., 2010 [157] anaesthetised (100 or 200mg/kg) CSDs frequency of CSDs by up to rats 30% within 1 hrs

Electrically- Speed of CSD Not changed stimulated CSD propagation 3 Threshold for CSD Significantly raised Lamotrigine Chloral- KCl-induced CSD Daily I.P. Frequency of Reduced at the proximal and No effect of riboflavin Bogdanov et al., 2011 [163] hydrate- injections for 1 CSDs at a distal distal electrodes anaesthetised month (parieto-occipital) rats 15 mg/kg/die and a proximal (frontal) electrode

CSD propagation Suppressed by 37% and 60% between the two at proximal and distal electrode sites electrodes.

Number of Fos- Decreased immunoreactive nuclei in frontal cortex. Magnesium sulphate Fentanyl citrate KCl-induced CSD Acute IV injection Frequency of Significantly reduced Similar effect of MK- van der Hel et al., 1998 [176] and fluanisone- Given. at 90 min CSDs 801 anaesthetised after onset of rats CSDs 10/mg/kg Novel potential therapeutic options Tonabersat Halotane- KCl-induced CSD Acute IP injection Cortical Significantly reduced No effect of Read et al., 2001 [151] anaesthetised 10 mg/ kg extracellular field sumatriptan rats potential depolarisations

Cortical and brain Brain stem cGMP was stem cGMP by abolished tonabersat Tonabersat Halotane KCl-induced CSD Acute IP and IV Apparent diffusion Significantly reduced ADC Changes of ADC Bradley et al., 2001 [183] anaesthetised injection coefficient (ADC) express reduced of cats, 10 mg/ kg measured in vivo magnitude and number subsequently by diffusion of CSD events initiated maintained weighted images and duration of CSD with alpha- activity . chloralose # These effects are partially shared by sumatriptan CGRP antagonists Isoflurane- KCl-induced CSD Incubation with CSD area Significantly reduced Exogenous CGRP Tozzi et al., 2012 [72] anaesthetised solutions of the partially reverted the rats antagonists of Intrinsic optical Significantly reduced inhibition neocortical slices signals (IOS) evoked by MK-8825 4 (2 hrs before CSD intensity induction)

CGRP 8-37 10 μ M

MK-8825 10 μ M

BIBN4096BS 0.1 μ M Other drugs of interest for migraine Memantine Isoflurane- KCl-induced CSD Acute CSD event dose-dependently decreased Similar results were Peeters et al., 2007 [51] anesthesized administration number obtained for two [pan-NMDA-R rats antagonists with blocker] 1, 3, and 10 mg/kg CSD amplitude Significantly reduced selectivity for NMDA- R containing the NR2B subunit (CP-101,606 and Ro 25-6981) Amiloride Propofol or Mechanically- Acute IV injection CSD event Significantly inhibited The selective ASIC1a Holland et al., 2012 [185] isoflurane induced CSD number blocker psalmotoxin [Acid-Sensing Ion anesthesized (needle prick) 10mg/kg inhibited CSD Channel 1 (ASIC 1) rats (males) blocker] Neurogenic Significantly reduced CSD was blocked only Electrical vasodilation of in 1 of 8 ASIC 1 stimulation of the middle meningeal knockout (ASIC1-/-) thinned cranium artery mice with deletion of the ACCN2gene Neuronal Significantly inhibited response of A- fiber input to dural electrical stimulation Legend * The order of the drugs presented in the Table has been adopted from current guidelines: beta-blockers, tricyclic antidepressants, Ca 2+ channel blockers, antiepileptics, magnesium and other potentially useful anti-migraine molecules § This was an in vitro study and therefore the effects from the anesthesia (isoflurane) were most likely dissipated at the time of cortical slice testing # Alpha-chloralose was used to maintain anesthesia due to the known effects of alotane on CSD events Abbreviations : ADC: Apparent diffusion coefficient; cGMP: cyclic guanosine monophosphate; ASIC 1: Acid-Sensing Ion Channel 1; DHE: Dihydroergotamine; IP: intraperitoneal; IOS: Intrinsic optical signals; IV: intravenous; NMDA-R : N-methyl-D-Aspartate Receptor