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. 2021 Sep 30;11:19520. doi: 10.1038/s41598-021-98852-6

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© The Author(s) 2021

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

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Collagen cross-linking inhibitor BAPN affects collagen I, II and IV accumulation, appearance and distribution. Double immunofluorescence staining revealed the relationship between the new formed blood vessels and collagen I, II, IV distribution in control (left panel: a, c, e, e’, e’’) vs. BAPN-treated animals (right panel b, d, f, f’, f’’) at 7dpa. ECs are labeled green and collagen I, II, IV purple. In the control group, the capillaries have classical appearance and collagen I (a) and collagen II (c) are surrounding the blood vessels as well defined, almost continuous layer. Additionally, collagen I and II stripes are strongly present in the dermal region (arrows). BAPN-treated fish represent dramatically enlarged blood vessels with balloon-like appearance (asterisks). The collagen I (b) and collagen II (d), when present, appear sparse, as small solitary fragments randomly distributed and not associated with the vasculature and dermal region. In the control group collagen IV demonstrated classical co-localization with the EC as a major component of the basement membrane (e, e’, e’’). In the BAPN-treated animals, collagen IV appears disintegrated and diffuse, without co-localization with the ECs (f, f’, f’’). e’’ and f’’ display merged image of ECs and collagen IV corresponding to e, e’ and f, f’ respectively. Images are acquired by confocal microscope.