Cardiac electrographic and morphological changes following status epilepticus: Effect of clonidine  Morgayn I. Read, Anastasia A. Andreianova, Joanne C. Harrison, Chelsea S. Goulton, Ivan A. Sammut, D. Steven Kerr  Seizure - European Journal of Epilepsy  Volume 23, Issue 1, Pages 55-61 (January 2014) DOI: 10.1016/j.seizure.2013.09.012 Copyright © 2013 British Epilepsy Association Terms and Conditions

Fig. 1 Behavioural changes and electrographic spiking during KA induced seizure activity. (A) Seizure-related behavioural changes (cumulative seizure scores, wet dog shakes and Level 4 seizures) following KA (10mg/kg). (B) High amplitude ECoG spiking following KA. Each bar represents the number of spikes over a 15min recording period. Values represent the mean±SEM. #P<0.05 compared to baseline, *P<0.05 compared to saline-KA. Seizure - European Journal of Epilepsy 2014 23, 55-61DOI: (10.1016/j.seizure.2013.09.012) Copyright © 2013 British Epilepsy Association Terms and Conditions

Fig. 2 Effect of KA (dashed line) on ECG in rats. Values represent mean±SEM recorded over a 1min recording period every 5min. (A) Heart rate (beats per minute, b.p.m.). (B) Corrected QT interval in seconds (QTc=QT/(RR/100)1/2). (D) T wave elevation. *P<0.05 compared to saline-KA baseline, #P<0.05 compared to clonidine-KA baseline, +P<0.05 compared against respective time point in saline-KA group. Seizure - European Journal of Epilepsy 2014 23, 55-61DOI: (10.1016/j.seizure.2013.09.012) Copyright © 2013 British Epilepsy Association Terms and Conditions

Fig. 3 Time course of ECG, ECoG and behavioural changes during seizure induction. (A)–(E) Representative ECG traces during the 120min recording period in saline-KA animals. (A) Baseline (406b.p.m.). (B) Bradycardia (291b.p.m.). (C) Skipped beats. (D) Irregular heartbeat. (E) Tachycardia (592b.p.m.). (F) Representative traces showing combined heart rate (b.p.m.; black line trace), ECoG activity (mV; grey spectral trace) and behavioural score responses in individual saline-control, saline-KA and clonidine-KA rats following saline vehicle or KA (dashed line) administration. Seizure - European Journal of Epilepsy 2014 23, 55-61DOI: (10.1016/j.seizure.2013.09.012) Copyright © 2013 British Epilepsy Association Terms and Conditions

Fig. 4 Representative micrographs of left ventricular subendocardium collected at 48h following (A) saline administration to control animals showing normal representative myocardium; (B)–(F) Effects of KA administration showing: (B) Coagulative myocytolysis illustrated by hypercontraction band necrosis associated with fibre derangement (arrowheads), reversible degenerative cardiomyopathy, evidenced by myocyte vacuolisation (asterisks indicate intracellular vacuoles displacing nuclei) and oedema; (C) Fragmentation of necrotic myocytes with eosinophilic granular material and mononuclear inflammatory cell infiltrate (arrowheads), interstitial oedema, and loss of basement membrane; (D) Inflammatory necrosis encapsulated by collagen deposition (blue fibres) indicative of early restorative fibrosis. (E) Clonidine treatment prior to KA administration preserved normal cardiac morphology (MSB stain). Scale bar represents 20μm. (F) The percentage of tissue positively stained for fibrosis, recorded in 10 sections taken 5mm from the apex. *P<0.05 compared to control-saline, #P<0.05 compared to saline-KA. Seizure - European Journal of Epilepsy 2014 23, 55-61DOI: (10.1016/j.seizure.2013.09.012) Copyright © 2013 British Epilepsy Association Terms and Conditions

Figure 5 Representative micrograph (A) of a ventricular section at 48h post seizure in a saline-KA animal stained with MSB showing distribution of collagen deposition (blue staining, both infiltrative and perivascular (B; Scale bar=200μm) and inflammatory cell infiltrate (C; Scale bar=70μm) within the endo- and subendo-cardiac layers. Seizure - European Journal of Epilepsy 2014 23, 55-61DOI: (10.1016/j.seizure.2013.09.012) Copyright © 2013 British Epilepsy Association Terms and Conditions