Action Potential Duration Restitution Portraits of Mammalian Ventricular Myocytes: Role of Calcium Current  Elena G. Tolkacheva, Justus M.B. Anumonwo,

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Action Potential Duration Restitution Portraits of Mammalian Ventricular Myocytes: Role of Calcium Current  Elena G. Tolkacheva, Justus M.B. Anumonwo, José Jalife  Biophysical Journal  Volume 91, Issue 7, Pages 2735-2745 (October 2006) DOI: 10.1529/biophysj.106.083865 Copyright © 2006 The Biophysical Society Terms and Conditions

Figure 1 Stimulation protocol (A) that was used to record restitution portraits of isolated rabbit (B) and guinea pig (C) myocytes with ruptured patch AP clamp technique. In A, only one segment of PDP for one value of BCL (B) is shown. In B and C, three different responses are present at each value of BCL: APD accommodation (black), dynamic RC (blue), and local S1-S2 RCs (purple). Note that different colors in stimulation protocol A match with recorded responses in B and C. In C, gray traces represent conventional S1-S2 RCs recorded for different values of S1 (S1=900ms and 500ms). Insets demonstrate rabbit and guinea pig AP profiles. Biophysical Journal 2006 91, 2735-2745DOI: (10.1529/biophysj.106.083865) Copyright © 2006 The Biophysical Society Terms and Conditions

Figure 2 (A) Dynamic RCs for rabbit and guinea pig myocytes obtained using ruptured patch AP clamp technique for different stimulation protocols: PDP I (solid squares) and PDP II (open squares) for rabbit and PDP III (open circles) for guinea pig myocytes. (B) Time constant, τ, of APD accommodation as a function of BCL for rabbit (solid squares) and guinea pig (open circles) isolated myocytes. Biophysical Journal 2006 91, 2735-2745DOI: (10.1529/biophysj.106.083865) Copyright © 2006 The Biophysical Society Terms and Conditions

Figure 3 Slopes Sdyn (solid symbols) and S12 (open symbols) measured from the restitution portraits of the (A) rabbit and (C) guinea pig myocytes as functions of BCL. In A, two different modifications of PDP were used: PDP I (squares) and PDP II (circles). The relationship between Sdyn and S12 for (B) rabbit and (D) guinea pig myocytes. Note that this relationship does not depend on pacing history: in B, data obtained using different PDPs could be fitted with a single curve. Biophysical Journal 2006 91, 2735-2745DOI: (10.1529/biophysj.106.083865) Copyright © 2006 The Biophysical Society Terms and Conditions

Figure 4 (A) Segment of the restitution portrait showing transition between 1:1 responses and alternans: dynamic RC (solid circles), local S1-S2 RC for the shortest BCL (open squares), stable alternans (stars). (Inset) Traces of APs for the stable alternans. (B) Values of different slopes measured at the onset of alternans: Sdyn and S12 were measured at the steady-state point, and S12max was measured at the point of small perturbation (A). Biophysical Journal 2006 91, 2735-2745DOI: (10.1529/biophysj.106.083865) Copyright © 2006 The Biophysical Society Terms and Conditions

Figure 5 (A) Dynamic RCs for rabbit myocytes obtained using perforated (open circles) and ruptured (solid circles) patch techniques. Examples of AP profiles (B) (BCL=1000ms) and dynamic RCs (C) for rabbit myocytes obtained using perforated patch technique under control (solid squares), RY/TG (open squares), and nisoldipine (open stars) conditions. Biophysical Journal 2006 91, 2735-2745DOI: (10.1529/biophysj.106.083865) Copyright © 2006 The Biophysical Society Terms and Conditions

Figure 6 (A) Slopes Sdyn, (B) slopes S12, and (D) APD measured from the restitution portraits of the rabbit myocytes (obtained using perforated patch technique) under control (solid squares), nisoldipine (open stars), and RY/TG (open squares) conditions. (C) Time course of APD accommodation under these conditions. Biophysical Journal 2006 91, 2735-2745DOI: (10.1529/biophysj.106.083865) Copyright © 2006 The Biophysical Society Terms and Conditions

Figure 7 (A) Representative AP profiles for BCL=800 and 200ms, and step pulse that were used to measure peak ICa. (B) Peak ICa measurements under these conditions. Biophysical Journal 2006 91, 2735-2745DOI: (10.1529/biophysj.106.083865) Copyright © 2006 The Biophysical Society Terms and Conditions

Figure 8 AP clamp peak ICa measurements. APDs from the restitution portrait recorded for BCL=350ms (A) and 200ms (B) using steps II–VI of PDP II. (C) Normalized peak ICa at BCL=350ms. (D) Normalized peak ICa at BCL=200ms. Biophysical Journal 2006 91, 2735-2745DOI: (10.1529/biophysj.106.083865) Copyright © 2006 The Biophysical Society Terms and Conditions

Figure 9 (A) Durations of APs and step pulse as functions of DIs for conventional voltage-clamp step protocol (open circles) and for APs voltage-clamp protocols (solid circles). Broken lines represent equal BCLs for 800 and 500ms. (B) Peak ICa recorded during step (open circles) and APs (solid circles) voltage-clamp protocols. Biophysical Journal 2006 91, 2735-2745DOI: (10.1529/biophysj.106.083865) Copyright © 2006 The Biophysical Society Terms and Conditions