Hemodynamic monitoring was performed using aseptic technique in rats anesthetized with Inactin (Sigma-Aldrich) 0.1 ml/100 g (125 mg/ml) unless otherwise specified. Briefly, a tracheostomy was created, the right internal jugular vein cannulated with PE50 tubing (Smiths Medical), and 0.9% NaCl instilled at 0.05 ml/100 g/min throughout surgery and then 0.03 ml/100 g/min during stabilization/monitoring. Rats received 5% inulin-FITC (Sigma-Aldrich) in 0.9% NaCl for GFR measurement. Body temperature was maintained at 37°C using a heat mat. Mean arterial pressure (MAP) and heart rate (HR) were measured via a right femoral artery PE50 catheter, portal pressure (PP) using a 24G cannula inserted into the portal vein, and RBF using a Doppler transit time probe (MA2PSB, ADInstruments) placed around the left renal artery proximal to the bifurcation, all connected to a Powerlab 4/35 system and analyzed using LabChart 7 Pro software (ADInstruments). Rats were stabilized for 20 min prior to data recording. The urinary bladder was catheterized with PE90 tubing. At the end of the experiment, blood was collected and tissues fixed in 10% buffered formalin or snap frozen.
Labchart 7 pro software
Manufactured by ADInstruments
Sourced in Australia, United Kingdom, United States
LabChart 7 Pro is a powerful data acquisition and analysis software for life science research. It provides a flexible and intuitive interface to record, visualize, and analyze physiological data from various instruments.
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Lab products found in correlation
Dam50 differential amplifier, by World Precision Instruments (5 mentions)
Powerlab 4 30, by ADInstruments (4 mentions)
Spike histogram software, by ADInstruments (3 mentions)
Mlt844, by ADInstruments (2 mentions)
Inactin, by Merck Group (1 mentions)
Pe50 tubing, by Smith & Nephew (1 mentions)
Fitc inulin, by Merck Group (1 mentions)
Powerlab 4 35 system, by ADInstruments (1 mentions)
Powerlab 8 30, by ADInstruments (1 mentions)
Pvr 1030, by Millar (1 mentions)
Mpvs 300, by Millar (1 mentions)
Ventri cath 507, by Millar (1 mentions)
Swan ganz catheter, by Edwards Lifesciences (1 mentions)
Powerlab acquisition system, by ADInstruments (1 mentions)
Sigmaplot 12, by Merck Group (1 mentions)
Labchart 7 pro, by ADInstruments (1 mentions)
Powerlab 4 30 data acquisition system, by ADInstruments (1 mentions)
Stereomicroscope, by Olympus (1 mentions)
Powerlabs16 35, by ADInstruments (1 mentions)
Matlab, by MathWorks (1 mentions)
24 protocols using labchart 7 pro software
1
Invasive Hemodynamic Monitoring in Rats
2
Extracellular Recording from Hoverfly Neurons
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We recorded from 13 male Eristalis tenax (Linnaeus 1758) hoverflies, 0.5–10 months old, reared and housed as described earlier (Nicholas et al., 2018 (link)). At the start of the experiment, the animal was immobilized ventral side up with a beeswax and resin mixture, and a small hole cut at the anterior end of the thorax. A sharp polyimide-insulated tungsten electrode (2 MΩ, Microprobes, Gaithersburg, MD, USA) was inserted into the cervical connective, with mechanical support given by a small wire hook. The animal was grounded via a silver wire inserted into the ventral cavity, which also served as the recording reference.
We recorded from type 2 optic flow-sensitive descending neurons, which were identified by their receptive field and physiological response properties (Nicholas et al., 2020 (link)). Extracellular signals were amplified at 1000× gain and filtered through a 10–3000 Hz bandwidth filter on a DAM50 differential amplifier (World Precision Instruments), with 50 Hz noise removed with a HumBug (Quest Scientific, North Vancouver, BC, Canada). The data were digitized via a Powerlab 4/30 (ADInstruments, Sydney, NSW, Australia) and acquired at 40 kHz with LabChart 7 Pro software (ADInstruments).
We recorded from type 2 optic flow-sensitive descending neurons, which were identified by their receptive field and physiological response properties (Nicholas et al., 2020 (link)). Extracellular signals were amplified at 1000× gain and filtered through a 10–3000 Hz bandwidth filter on a DAM50 differential amplifier (World Precision Instruments), with 50 Hz noise removed with a HumBug (Quest Scientific, North Vancouver, BC, Canada). The data were digitized via a Powerlab 4/30 (ADInstruments, Sydney, NSW, Australia) and acquired at 40 kHz with LabChart 7 Pro software (ADInstruments).
3
Ex Vivo Cardiac Function Assessment
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To evaluate the changes in heart function in the hearts independent of neurohormonal or loading conditions, we also characterised heart function ex vivo. After an intraperitoneal injection of pentobarbital, hearts were excised and perfused using a modified Krebs-Henseleit with added glucose (5mM) and palmitate (0.5 mM) bound to BSA (3%) [25 (link)]. Cardiac temperature was maintained at 37°C and data were obtained and analysed using LabChart 7Pro software (ADInstruments, Bella Vista, Australia).
One group of hearts was perfused in the working heart mode with an 8 mmHg preload and 50 mmHg afterload [25 (link)]. Left ventricular pressure changes were assessed using a 1.0-Fr conductance catheter (Millar Instruments, Houston, TX), inserted into the ventricle through the apex [26 (link)]. In another group, the hearts were perfused in the Langendorff mode [27 (link)], where left ventricular pressure was assessed using an intraventricular fluid-filled balloon. The volume of the balloon was adjusted so that the end-diastolic pressure was between 5–10 mmHg. To prevent build-up of fluid in the ventricle, a cannula (25 G) was inserted through the apex and into the lumen to allow drainage of fluid [27 (link)].
One group of hearts was perfused in the working heart mode with an 8 mmHg preload and 50 mmHg afterload [25 (link)]. Left ventricular pressure changes were assessed using a 1.0-Fr conductance catheter (Millar Instruments, Houston, TX), inserted into the ventricle through the apex [26 (link)]. In another group, the hearts were perfused in the Langendorff mode [27 (link)], where left ventricular pressure was assessed using an intraventricular fluid-filled balloon. The volume of the balloon was adjusted so that the end-diastolic pressure was between 5–10 mmHg. To prevent build-up of fluid in the ventricle, a cannula (25 G) was inserted through the apex and into the lumen to allow drainage of fluid [27 (link)].
4
In vivo Cardiac Pressure-Volume Measurements
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Immediately after echocardiography, animals were maintained under 1–2% isoflurane while performing right and LV catheterization via either the right internal carotid artery or the right external jugular vein. Pressure-volume (PV) data were collected by Millar ultra-miniature catheters (PVR-1045; mouse LV, PVR-1030; mouse RV: Millar Instruments, Inc., Houston, TX, USA) coupled to a Millar MPVS 300. Data were acquired using Powerlab 8/30 (AD Instruments, Oxfordshire, UK) and recorded using LabChart 7 Pro software (AD Instruments). PVAN v2.3 (Millar Instruments, Inc.) was used to analyze PV data as previously described.36 (link)
5
Pressure-Volume Analysis of Swine Heart
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Eight-weeks after AMI, upon anesthesia induction and mechanical ventilation as previously described, closed-chest LV pressure-volume (PV) recordings were performed with a 5-Fr pigtail multi-segment pressure-volume conductance catheter (Ventri-Cath™ 507; Millar Instruments, Inc., Houston, TX) inserted through the femoral artery and guided to the LV apex with a 7-Fr 90 cm sheath (Cordis®, Miami Lakes, FL). Evaluation of load independent parameters was performed by transient occlusion of the inferior vena cava with a 25 mm-diameter balloon catheter (NuCLEUS™, NuMED™, Hopkinton, NY) with ventilation suspended at end-expiration. Cardiac output was measured continuously by thermodilution using a Swan-Ganz catheter (Edwards LifeSciences™, CL) inserted through the right internal jugular vein (8-Fr). Data was continuously acquired and recorded with the PowerLab 16/35 16-channel acquisition system, and analyzed offline using the LabChart 7 Pro software (ADInstruments, Oxford, UK). Volume signal was calibrated for parallel conductance by 10 ml 10% saline injection and for slope factor α by simultaneous measurement of cardiac output. Volume data was calibrated for swine body surface area according to corrections for mini-pig strains (18 (link)). End-systolic and end-diastolic PV relationships were obtained by linear fitting and exponential fitting with a pressure asymptote, respectively.
6
Video-EEG Analysis of Epileptic Spikes
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For video‐EEG, electrodes were implanted between the skull and the surface of the left frontal cortex (reference) and both parietal cortices as described 46 . All EEG recordings were done at least 3 weeks after electrode implantation on freely behaving mice. For Fig 10 , digital EEG activity and video were recorded with Harmonie software (version 5.0b, Stellate Systems; Natus). Epileptic spikes were detected automatically with threshold Amp8. Artifactual spikes associated with movements of the recording wire were excluded from analysis. Spike frequency at rest was measured for 6 h during the light cycle and 6 h during the dark cycle and expressed as number of spikes per hour. Mice were then injected with PTZ (30 mg/kg) during the light cycle. The PTZ stock solution (5 mg/ml in phosphate‐buffered saline) was prepared from powder on the same day.
For Fig11 F, digital EEG activity and video were recorded with a PowerLab data acquisition system 16/35 and analyzed with LabChart 7 Pro software (AD Instruments) 85 . Spike frequency at rest was measured for 4 h during the light cycle and 3.2–4 h during the dark cycle and expressed as number of spikes per h.
For Fig
7
Wireless Cortical EEG Monitoring in Mice
8
Cardiac Function Assessment in Mice
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Mice were anesthetized using isoflurane inhalation (0.8–1.0 volume % in oxygen) and surface ECGs were recorded from subcutaneous 23-gauge needle electrodes attached to each limb using the Powerlab acquisition system (ADInstruments Ltd, Oxford, United Kingdom). ECG traces were signal averaged and analyzed for heart rate (RR-interval), P-wave, PR-, QRS- and QT-interval duration using the LabChart7Pro software (ADInstruments Ltd, Oxford, United Kingdom) RR interval was defined as the interval in ms between two consecutive R waves. PR-, P-wave, QRS- and QT-intervals were determined as indicated in Figure 4 A. QT-intervals were corrected for heart rate using the formula: QTc = QT/(RR/100)1/2 (RR in ms) [35 (link)].
9
In vivo Rat Electrocardiogram Recording
10
Magnetic Field Mapping for Coil Optimization
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Mapping of the magnetic field intensities was done to evaluate the influence of magnetic field shape over the coil relative to the rat central nervous system structures, and the effect of a magnetic shielding with a centered open window. Single magnetic pulses were recorded using a solenoid (Magprobe, Magventure, Farum, Denmark). The solenoid is a 2.5 cm diameter copper coil converting a magnetic field into a measurable electrical current (in Volts). The solenoid was placed as close as possible to the CB60 coil (570 different points of recording, 30×19 matrix) in all three dimensional directions of the magnetic field (x, y and z axes). The induced electrical current for each direction (x, y and z) was recorded by using the Powerlab device and the LabChart 7 Pro software (AD Instrument). The whole magnetic field recorded with the solenoid was calculated using the following formula: √(x2+y2+z2). Each dimension of the electrical field induced in the solenoid was reconstructed in 3D with a 2D Z-axis projection (Sigmaplot 12.5 software).
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