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CardiLect: A combined cross-species lectin histochemistry protocol for the automated analysis of cardiac remodelling
Tamás G Gergely1,2,3,4, Tamás Kovács1,2,3,4, Andrea Kovács1,2,3,4
1Department of Pharmacology and Pharmacotherapy, Semmelweis University, Budapest, Hungary.
Insights
This study introduces a cost-effective, cross-species lectin histochemistry protocol for assessing cardiac remodelling. The new method accurately measures cardiomyocyte size and microvascular density, correlating well with heart failure indicators.
Area of Science:
- Cardiovascular Research
- Histology and Pathology
- Biomedical Engineering
Background:
- Cardiac remodelling is key in heart failure, often studied using cardiomyocyte cross-sectional area (CSA) and microvascular density (MVD).
- Current histological methods like immunohistochemistry require species-specific antibodies and optimized protocols.
- Lectin histochemistry offers a cost-effective, cross-species alternative, but direct comparisons are limited.
Purpose of the Study:
- To compare antibody-based and lectin-based methods for assessing cardiac CSA and MVD.
- To develop and validate a combined lectin histochemistry protocol ('CardiLect') for cardiac remodelling assessment.
- To provide an automated image analysis tool for histological data.
Main Methods:
- Compared anti-laminin/wheat germ agglutinin (WGA) for CSA and anti-CD31/isolectin B4 (ILB4) for MVD.
- Developed the 'CardiLect' protocol using WGA and ILB4, tested across species (zebrafish, rodents, large animals, humans).
- Created an ImageJ script ('CardiLect Analyser') for automated analysis and validated it against echocardiographic findings.
Main Results:
- The 'CardiLect' protocol demonstrated efficacy across multiple species for assessing cardiac remodelling.
- Automated analysis showed significant correlations: CSA with left ventricular (LV) mass and ejection fraction (EF); MVD with LV mass and EF.
- Histological parameters correlated significantly with echocardiographic indicators of cardiac function and mass.
Conclusions:
- The 'CardiLect' protocol and 'CardiLect Analyser' offer an easy-to-use, cost-effective, and cross-species solution for histological assessment of cardiac remodelling.
- This approach simplifies and enhances the study of cardiac remodelling in diverse preclinical and clinical models.
- The validated protocol and analysis tool facilitate accurate and efficient evaluation of cardiac structural changes.
Background:
Cardiac remodelling, a crucial aspect of heart failure, is commonly investigated in preclinical models by quantifying cardiomyocyte cross-sectional area (CSA) and microvascular density (MVD) via histological methods, such as immunohistochemistry. To achieve this, optimized protocols are needed, and the species specificity is dependent on the antibody used. Lectin histochemistry offers several advantages compared to antibody-based immunohistochemistry, including as cost-effectiveness and cross-species applicability. Direct comparisons between the two methods are lacking from the literature.
Methods And Results:
In this study, we compared antibody- and lectin-based methods for the histological assessment of cardiomyocyte CSA (with the use of anti-laminin and wheat germ agglutinin [WGA]) and microvascular density (utilizing anti-CD31 and isolectin B4 [ILB4]) using different embedding and antigen/carbohydrate retrieval techniques. Here, we describe a detailed, easy-to-use combined lectin histochemistry protocol (WGA and ILB4, 'CardiLect' protocol) for the histological assessment of cardiac remodelling. The lectin-based approach has been evaluated on a cross-species basis, and its efficacy has been demonstrated in zebrafish, rodents, large animals and human samples. We provide an ImageJ script ('CardiLect Analyser') for automated image analysis, validated in a preclinical heart failure model by correlating histological parameters with echocardiographic findings. CSA showed a significant positive correlation with left ventricular (LV) mass (P = 0.0098, rS = 0.7545) and significant negative correlation with markers of systolic function, such as ejection fraction (EF) (P = 0.0402, rS = -0.6364). Microvascular density showed significant negative correlation with LV mass (P = 0.0055, rS = -0.7622) and significant positive correlation with EF (P = 0.0106, rS = 0.7203).
Conclusions:
The described combined lectin histochemistry protocol with the provided ImageJ script is an easy-to-use, cost-effective, cross-species approach for the histological assessment of cardiac remodelling.
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