Vevo 3100 ultrasound

Mice (n=4) were anaesthetised with 3% isoflurane in O₂. Temperature was monitored and maintained at 37.0 ± 0.5°C throughout with a feedback heating system. Respiration was monitored and maintained between 30 and 80 breaths per minute. An electrocardiogram was also acquired. Using a Vevo 3100 ultrasound (Visualsonics, Amsterdam, Netherlands) and a mouse cardiology probe (MX550D), a respiration-gated 3D ultrasound image was acquired across the whole neck. The left carotid artery was located, and 10 Power Doppler blood flow clips of 30 s duration were acquired per mouse at respiration rates across the range of 30–80 breaths per minute, achieved by varying the concentration of isoflurane between 3 and 1%. Carotid blood flow values from the Power Doppler trace were extracted using an open-source plot digitiser [31 ], and values representing the systolic and diastolic flow velocities were used in pCASL simulations to determine labelling efficiency.

At the first and last week of SPRC treatment, the hair was removed from the chest of mice using depilatory cream. The mice were then anesthetized with 1.5% isoflurane and placed in a supine position on the test bench, with ultrasound gel applied onto the chest. Mouse two-dimensional echocardiography was performed using a Vevo3100 ultrasound device (VisualSonics Inc., Canada), as previously described (18 (link)). B-mode, M-mode, and Power Doppler Mode ultrasound images of the left ventricle were recorded. All measurements were averaged for five consecutive cardiac cycles.

The mice were anesthetized and imaged using a Vevo 3100 Ultrasound (Visual Sonics, Toronto, Ontario, Canada) after TAC or Sham surgery. The instrument instructions were followed to detect the end-diastolic diameter (LVID; d), left ventricular end-systolic diameter (LVID; s), left ventricular ejection fraction (EF%), and fractional shortening (FS%) of the mouse hearts.

The UK4b compound used was synthesized as previously discussed^{36 (link)}. All animal experiments described here were performed with the approval of the University of Kentucky Institutional Animal Care and Use Committee (IACUC #2019-3373) and in accordance with the Guide for the Care and Use of Laboratory Animals from National Institutes of Health/Office of Laboratory Animal Welfare (NIH/OLAW), and were in fact also consistent with the ARRIVE (Animal Research: Reporting of In Vivo Experiments) guidelines (https://arriveguidelines.org). Animals were housed in the University of Kentucky DLAR facilities inside clean cages with ad libitum access to normal chow and water. Buprenorphine, ketamine, and xylazine were purchased from Covetrus (Dublin, Ohio, USA). Angiotensin II (AngII) was purchased from VWR (Radnor, PA, USA). ALZET Osmotic Pumps (model 2004) were purchased from Braintree Scientific (Braintree, MA, USA).
In vivo ultrasound imaging was done using a Vevo 3100 Ultrasound (Fujifilm VisualSonics, Inc.) and data analyzed using VevoLAB software (Fujifilm Visualsonics, Inc.) whose use was provided by the University of Kentucky Cardiovascular Research Priority Area. PGE₂ was measured using the commercially available enzyme linked immunosorbent assay (ELISA) kit (catalog #514531) from Cayman Chemical (Ann Arbor, MI).