Capillary pericytes mediate coronary no-reflow after myocardial ischaemia
- University College London, United Kingdom
Figures
Figure 1
Cardiac pericyte morphology is appropriate for regulating capillary diameter.
(A) Coronary capillaries in rat left ventricle (dashed arrows in left panel show longitudinal capillaries, solid arrows show connector capillaries) labelled for basement membrane (FITC-isolectin B4) …
Figure 2
Ischaemia and reperfusion lead to no-reflow mediated by capillary block.
(A, B) Low power view of sham-operated heart (A) and a heart after LAD coronary artery occlusion and reperfusion (B), with perfusion volume assessed as intensity of FITC-albumin (green). In (B) …
Figure 3 with 1 supplement
No-reflow reflects blockage by pericyte constriction.
(A) Image of perfused and non-perfused capillaries in post-ischaemic left ventricle. Isolectin B4 labelling (white) defines positions of all vessels, while FITC-albumin labelling (green) shows …
Figure 3—figure supplement 1
Calculation of probability distribution for distance from a randomly placed block to a chosen pericyte soma.
(A) Diagram showing the calculation of the probabilities on the right of the panel. (B) Cumulative probability distribution for the position of a randomly placed block on this capillary. Figure 3H …
Tables
Key resources table
| Reagent type (species) or resource | Designation | Source or reference | Identifiers | Additional information |
|---|---|---|---|---|
| Rattus norvegicus (Sprague Dawley, male) | Rat | UCL Biological Services | ||
| genetic reagent (Mus musculus/spretus, male and female) | NG2-DsRed mice | doi: 10.1242/dev.004895 | JAX 008241 | |
| antibody | anti-NG2 (rabbit polyclonal) | Merck Millipore | AB5320 | (1:200) |
| antibody | anti-NG2 (mouse monoclonal) | AbCam | ab50009 | (1:200) |
| antibody | anti-PDGF receptor beta (rabbit polyclonal) | Santa Cruz Biotechnology | sc-432 | (1:200) |
| antibody | anti-tyrosine hydroxylase (sheep polyclonal) | Merck Millipore | AB1542 | (1:500) |
| antibody | anti-alpha smooth muscle actin (rabbit polyclonal) | AbCam | ab5694 | (1:100) |
| antibody | anti-beta actin (mouse monoclonal) | Abbiotec | 251815 | (1:100) |
| antibody | anti-gamma actin (mouse monoclonal) | AbCam | ab123034 | (1:100) |
| antibody | anti-glycophorin A (mouse monoclonal) | AbCam | ab9520 | (1:2000) |
| antibody | anti-neutrophil elastase (goat monoclonal) | Santa Cruz | sc9521 | (1:50) |
| antibody | anti-ICAM1 (mouse monoclonal) | AbCam | Ab171123 | (1:100) |
| antibody | Alexa Fluor 405 goat anti-rabbit (polyclonal) | Life Technologies | A31556 | (1:500) |
| antibody | Alexa Fluor 555 donkey anti-rabbit (polyclonal) | Life Technologies | A31572 | (1:500) |
| antibody | Alexa Fluor 555 donkey anti-mouse (polyclonal) | Life Technologies | A31570 | (1:500) |
| chemical compound | isolectin B4 - Alexa Fluor 647 | Molecular Probes | I32450 | (1:200) |
| chemical compound | isolectin B4 - FITC | Sigma-Aldrich | L2895 | (1:200) |
| chemical compound | adenosine | Sigma-Aldrich | A9251 | |
| chemical compound | gelatin | Sigma-Aldrich | G2625 | 5% in PBS |
| chemical compound | FITC-albumin | Sigma-Aldrich | A9771 | 1:200 in 5% gelatin |
| chemical compound | DAPI | Molecular Probes | D1306 | |
| chemical compound | terazosin | Sigma | T4680 | |
| software | Spike2 | www.ced.co.uk | in vivo data acquisition | |
| software | ImageJ | https://imagej.nih.gov/ij/ | image analysis | |
| software | OriginPro | www.originlab.com/Origin | statistical analysis |
Additional files
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Transparent reporting form
- https://doi.org/10.7554/eLife.29280.007
