National Research, Development and Innovation Office (NKFIH) of Hungary(FK134751)
Thematic Excellence Program (Semmelweis University)(2020-4.1.1.-TKP2020) Funder: Ministry
for Innovation and Technology
(ÚNKP-23-4-I-SE-5)
(ÚNKP-23-II-SE-43)
(ÚNKP-23-5-SE-9)
(BO/00277/23/5)
(RRF-2.3.1-21-2022-00003)
(FK-146163)
Subjects:
Pharmacology and pharmacy
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.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).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.
