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Advanced echocardiography in adult zebrafish reveals delayed recovery of heart function after myocardial cryoinjury.

Hein SJ, Lehmann LH, Kossack M, Juergensen L, Fuchs D, Katus HA, Hassel D - PLoS ONE (2015)

Bottom Line: We show that functional recovery of cryoinjured hearts occurs in three distinct phases.Importantly, the regeneration process after cryoinjury extends far beyond the proposed 45 days described for ventricular resection with reconstitution of myocardial performance up to 180 days post-injury (dpi).The imaging modalities evaluated here allow sensitive cardiac phenotyping and contribute to further establish adult zebrafish as valuable cardiac disease model beyond the larval developmental stage.

View Article: PubMed Central - PubMed

Affiliation: Department of Medicine III, Cardiology, Heidelberg University Hospital, 69120 Heidelberg, Germany and DZHK (German Center for Cardiovascular Research), Partner Site Heidelberg/Mannheim, Heidelberg, Germany.

ABSTRACT
Translucent zebrafish larvae represent an established model to analyze genetics of cardiac development and human cardiac disease. More recently adult zebrafish are utilized to evaluate mechanisms of cardiac regeneration and by benefiting from recent genome editing technologies, including TALEN and CRISPR, adult zebrafish are emerging as a valuable in vivo model to evaluate novel disease genes and specifically validate disease causing mutations and their underlying pathomechanisms. However, methods to sensitively and non-invasively assess cardiac morphology and performance in adult zebrafish are still limited. We here present a standardized examination protocol to broadly assess cardiac performance in adult zebrafish by advancing conventional echocardiography with modern speckle-tracking analyses. This allows accurate detection of changes in cardiac performance and further enables highly sensitive assessment of regional myocardial motion and deformation in high spatio-temporal resolution. Combining conventional echocardiography measurements with radial and longitudinal velocity, displacement, strain, strain rate and myocardial wall delay rates after myocardial cryoinjury permitted to non-invasively determine injury dimensions and to longitudinally follow functional recovery during cardiac regeneration. We show that functional recovery of cryoinjured hearts occurs in three distinct phases. Importantly, the regeneration process after cryoinjury extends far beyond the proposed 45 days described for ventricular resection with reconstitution of myocardial performance up to 180 days post-injury (dpi). The imaging modalities evaluated here allow sensitive cardiac phenotyping and contribute to further establish adult zebrafish as valuable cardiac disease model beyond the larval developmental stage.

No MeSH data available.


Related in: MedlinePlus

Time-to-peak and maximal opposing wall delay analysis reveals considerable asynchronicity of injured to non-injured ventricular wall.Segmental strain traces recorded from Sham. (A-C) and cryoinjured, regenerating hearts at 14dpi (D-F) are shown. (A) Representative segmental strain traces of non-injured hearts exhibit only minor time-to-peak delay between opposing ventricular wall segments (OWD) as illustrated by the pink line. (For color-coding explanation see Fig 2B.) (B-C) Individual strain curves of anterior (B) and posterior wall segments (C). (D) Representative segmental strain curves derived from a regenerating heart at 14dpi demonstrating considerable OWD intervals. (E-F) Individual strain curves of injured anterior (E) and non-injured posterior wall segments (F). For each image displayed, the black line indicates the average curve. (G) 3D-reconstruction of regional strain analysis derived from a control (left) and injured (right) heart at 14dpi with the frontal view rotated 90° counterclockwise. Every peak represents consecutive systoles. The anterior wall (AW) is located in the front, while the posterior wall (PW) is situated in the back. In the control presentation (left) the AW and PW peaks are completely overlapping, presenting absolute synchronicity. However, at 14dpi (right) the residual AW activity is visible as a small peak offset to the right of the PW peak in the back. This offset represents the OWD. (H) Quantitative analysis of mean OWDs at depicted time points. Small numbers indicate number of animals measured. Values are expressed as mean ± SEM; *, p<0.05, unpaired student’s t-test and ANOVA with post hoc comparisons by Bonferroni’s multiple comparison test.
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pone.0122665.g006: Time-to-peak and maximal opposing wall delay analysis reveals considerable asynchronicity of injured to non-injured ventricular wall.Segmental strain traces recorded from Sham. (A-C) and cryoinjured, regenerating hearts at 14dpi (D-F) are shown. (A) Representative segmental strain traces of non-injured hearts exhibit only minor time-to-peak delay between opposing ventricular wall segments (OWD) as illustrated by the pink line. (For color-coding explanation see Fig 2B.) (B-C) Individual strain curves of anterior (B) and posterior wall segments (C). (D) Representative segmental strain curves derived from a regenerating heart at 14dpi demonstrating considerable OWD intervals. (E-F) Individual strain curves of injured anterior (E) and non-injured posterior wall segments (F). For each image displayed, the black line indicates the average curve. (G) 3D-reconstruction of regional strain analysis derived from a control (left) and injured (right) heart at 14dpi with the frontal view rotated 90° counterclockwise. Every peak represents consecutive systoles. The anterior wall (AW) is located in the front, while the posterior wall (PW) is situated in the back. In the control presentation (left) the AW and PW peaks are completely overlapping, presenting absolute synchronicity. However, at 14dpi (right) the residual AW activity is visible as a small peak offset to the right of the PW peak in the back. This offset represents the OWD. (H) Quantitative analysis of mean OWDs at depicted time points. Small numbers indicate number of animals measured. Values are expressed as mean ± SEM; *, p<0.05, unpaired student’s t-test and ANOVA with post hoc comparisons by Bonferroni’s multiple comparison test.

Mentions: We next assessed synchronicity of systolic anterior to posterior wall motion by investigating the time-to-peak contraction differences between fastest and slowest myocardial segment of opposite ventricular walls (opposing wall delay). As a reference point to distinguish systolic and diastolic phases during contraction, we used M-Mode recordings derived from LAX B-Mode-sequences. While in sham operated controls only minor delays between anterior and posterior wall contractions can be detected (Fig 6A-6C), the opposing wall delay after cryoinjury to the AW was significantly increased until 60dpi (Fig 6A-6H). This difference becomes even more pronounced when analyzing individual speckles in higher regional resolution and in 3D display (Fig 6G). At 14dpi asynchronism between anterior and posterior wall peaked and remained significant until 60dpi with complete recovery at least at 120dpi (Fig 6H). Detailed parameters are summarized in S3 Table.


Advanced echocardiography in adult zebrafish reveals delayed recovery of heart function after myocardial cryoinjury.

Hein SJ, Lehmann LH, Kossack M, Juergensen L, Fuchs D, Katus HA, Hassel D - PLoS ONE (2015)

Time-to-peak and maximal opposing wall delay analysis reveals considerable asynchronicity of injured to non-injured ventricular wall.Segmental strain traces recorded from Sham. (A-C) and cryoinjured, regenerating hearts at 14dpi (D-F) are shown. (A) Representative segmental strain traces of non-injured hearts exhibit only minor time-to-peak delay between opposing ventricular wall segments (OWD) as illustrated by the pink line. (For color-coding explanation see Fig 2B.) (B-C) Individual strain curves of anterior (B) and posterior wall segments (C). (D) Representative segmental strain curves derived from a regenerating heart at 14dpi demonstrating considerable OWD intervals. (E-F) Individual strain curves of injured anterior (E) and non-injured posterior wall segments (F). For each image displayed, the black line indicates the average curve. (G) 3D-reconstruction of regional strain analysis derived from a control (left) and injured (right) heart at 14dpi with the frontal view rotated 90° counterclockwise. Every peak represents consecutive systoles. The anterior wall (AW) is located in the front, while the posterior wall (PW) is situated in the back. In the control presentation (left) the AW and PW peaks are completely overlapping, presenting absolute synchronicity. However, at 14dpi (right) the residual AW activity is visible as a small peak offset to the right of the PW peak in the back. This offset represents the OWD. (H) Quantitative analysis of mean OWDs at depicted time points. Small numbers indicate number of animals measured. Values are expressed as mean ± SEM; *, p<0.05, unpaired student’s t-test and ANOVA with post hoc comparisons by Bonferroni’s multiple comparison test.
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Related In: Results  -  Collection

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pone.0122665.g006: Time-to-peak and maximal opposing wall delay analysis reveals considerable asynchronicity of injured to non-injured ventricular wall.Segmental strain traces recorded from Sham. (A-C) and cryoinjured, regenerating hearts at 14dpi (D-F) are shown. (A) Representative segmental strain traces of non-injured hearts exhibit only minor time-to-peak delay between opposing ventricular wall segments (OWD) as illustrated by the pink line. (For color-coding explanation see Fig 2B.) (B-C) Individual strain curves of anterior (B) and posterior wall segments (C). (D) Representative segmental strain curves derived from a regenerating heart at 14dpi demonstrating considerable OWD intervals. (E-F) Individual strain curves of injured anterior (E) and non-injured posterior wall segments (F). For each image displayed, the black line indicates the average curve. (G) 3D-reconstruction of regional strain analysis derived from a control (left) and injured (right) heart at 14dpi with the frontal view rotated 90° counterclockwise. Every peak represents consecutive systoles. The anterior wall (AW) is located in the front, while the posterior wall (PW) is situated in the back. In the control presentation (left) the AW and PW peaks are completely overlapping, presenting absolute synchronicity. However, at 14dpi (right) the residual AW activity is visible as a small peak offset to the right of the PW peak in the back. This offset represents the OWD. (H) Quantitative analysis of mean OWDs at depicted time points. Small numbers indicate number of animals measured. Values are expressed as mean ± SEM; *, p<0.05, unpaired student’s t-test and ANOVA with post hoc comparisons by Bonferroni’s multiple comparison test.
Mentions: We next assessed synchronicity of systolic anterior to posterior wall motion by investigating the time-to-peak contraction differences between fastest and slowest myocardial segment of opposite ventricular walls (opposing wall delay). As a reference point to distinguish systolic and diastolic phases during contraction, we used M-Mode recordings derived from LAX B-Mode-sequences. While in sham operated controls only minor delays between anterior and posterior wall contractions can be detected (Fig 6A-6C), the opposing wall delay after cryoinjury to the AW was significantly increased until 60dpi (Fig 6A-6H). This difference becomes even more pronounced when analyzing individual speckles in higher regional resolution and in 3D display (Fig 6G). At 14dpi asynchronism between anterior and posterior wall peaked and remained significant until 60dpi with complete recovery at least at 120dpi (Fig 6H). Detailed parameters are summarized in S3 Table.

Bottom Line: We show that functional recovery of cryoinjured hearts occurs in three distinct phases.Importantly, the regeneration process after cryoinjury extends far beyond the proposed 45 days described for ventricular resection with reconstitution of myocardial performance up to 180 days post-injury (dpi).The imaging modalities evaluated here allow sensitive cardiac phenotyping and contribute to further establish adult zebrafish as valuable cardiac disease model beyond the larval developmental stage.

View Article: PubMed Central - PubMed

Affiliation: Department of Medicine III, Cardiology, Heidelberg University Hospital, 69120 Heidelberg, Germany and DZHK (German Center for Cardiovascular Research), Partner Site Heidelberg/Mannheim, Heidelberg, Germany.

ABSTRACT
Translucent zebrafish larvae represent an established model to analyze genetics of cardiac development and human cardiac disease. More recently adult zebrafish are utilized to evaluate mechanisms of cardiac regeneration and by benefiting from recent genome editing technologies, including TALEN and CRISPR, adult zebrafish are emerging as a valuable in vivo model to evaluate novel disease genes and specifically validate disease causing mutations and their underlying pathomechanisms. However, methods to sensitively and non-invasively assess cardiac morphology and performance in adult zebrafish are still limited. We here present a standardized examination protocol to broadly assess cardiac performance in adult zebrafish by advancing conventional echocardiography with modern speckle-tracking analyses. This allows accurate detection of changes in cardiac performance and further enables highly sensitive assessment of regional myocardial motion and deformation in high spatio-temporal resolution. Combining conventional echocardiography measurements with radial and longitudinal velocity, displacement, strain, strain rate and myocardial wall delay rates after myocardial cryoinjury permitted to non-invasively determine injury dimensions and to longitudinally follow functional recovery during cardiac regeneration. We show that functional recovery of cryoinjured hearts occurs in three distinct phases. Importantly, the regeneration process after cryoinjury extends far beyond the proposed 45 days described for ventricular resection with reconstitution of myocardial performance up to 180 days post-injury (dpi). The imaging modalities evaluated here allow sensitive cardiac phenotyping and contribute to further establish adult zebrafish as valuable cardiac disease model beyond the larval developmental stage.

No MeSH data available.


Related in: MedlinePlus