Mural cells protect the adult brain from hemorrhage but do not control the blood–brain barrier in developing zebrafish

Mammalian neurovascular endothelial cells (ECs) have unique functional and structural properties to regulate substance exchange between the blood and the brain, establishing the blood–brain barrier (BBB) (Zhao et al., 2015; Armulik et al., 2011). Loss of normal BBB function is associated with neuropathological states, including neurodegeneration, infection, and cancer (Zhao et al., 2015; Armulik et al., 2011). Mural cells are vascular support cells that include pericytes and vascular smooth muscle cells (vSMCs) and control vascular development, function and BBB permeability (Levéen et al., 1994; Lindahl et al., 1997; Armulik et al., 2010; Daneman et al., 2010; Eilken et al., 2017; Simonavicius et al., 2012; Benjamin et al., 1998). PDGFB/PDGFRβ signaling is required for the development of mural cells (Levéen et al., 1994; Lindahl et al., 1997; Hellström et al., 1999; Hirschi et al., 1998; Soriano, 1994) and their depletion in PDGFB- or PDGFRβ-deficient rodents causes altered angiogenesis, aneurysm, and vessel leakiness (Levéen et al., 1994; Lindahl et al., 1997; Armulik et al., 2010; Daneman et al., 2010; Ando et al., 2021b). Strikingly, rodent models of pericyte loss have an open BBB, with free transport of tracer dyes from the blood stream into the brain parenchyma but an otherwise intact vasculature (Armulik et al., 2010; Daneman et al., 2010). These and other observations have led to a current model whereby pericytes control BBB function by suppressing EC transcytosis, via a mechanism not yet fully understood. Such a mechanism holds significant translational promise to safely ‘open’ the BBB in therapeutic settings, by discovering ways to disrupt pericyte control of ECs (Armulik et al., 2010; Mäe et al., 2021).

Studies of the BBB using invertebrates like Drosophila and some basal vertebrate fishes (e.g. some Elasmobranchii and Chondrostei) have suggested that the makeup of the BBB can vary significantly across the phylogeny. These species display a primarily glial cell barrier, rather than an EC barrier (Paredes-González et al., 2025; Limmer et al., 2014). However, in the modern teleost Danio rerio (zebrafish) the BBB cellular composition is more similar to that seen in mammals (Jeong et al., 2008; Xie et al., 2010; Quiñonez-Silvero et al., 2020; Umans et al., 2017; O’Brown et al., 2019). As in mammals, the zebrafish BBB is controlled by tightly adherent vascular ECs that share many characteristics with the ECs of the mammalian brain. Zebrafish BBB endothelium expresses conserved molecular markers (e.g. glut1, cldn5, and mfsd2a) (Xie et al., 2010; Umans et al., 2017; O’Brown et al., 2019; Li et al., 2022; Vanhollebeke et al., 2015) and displays similar tight cell–cell junctions as observed at the mammalian BBB (Jeong et al., 2008; Umans et al., 2017; Li et al., 2022). The membrane transport protein Mfsd2a has been suggested to have a conserved role in control of BBB function in zebrafish and mice (O’Brown et al., 2019; Guemez-Gamboa et al., 2015), likely acting to reduce the transfer of substances between bloodstream and parenchyma by suppressing EC transcytosis (Ben Zvi et al., 2014; Andreone et al., 2017). Furthermore, during development the mechanisms that control the formation of BBB ECs appear to show a high level of conservation between zebrafish and mice. Recent studies have identified major roles for Wnt–β-catenin signaling, brain EC specific MMPs, Vegfs, and chemokine signaling in early BBB vascular EC development (Vanhollebeke et al., 2015; Eubelen et al., 2018; Parab et al., 2023; Parab et al., 2021).

After the initial development of zebrafish BBB vasculature, mural cell populations develop and mature. Transcriptionally, developing pericytes and vSMCs express many genes that are highly conserved with mammals, for example zebrafish pericytes express well known markers such as pdgfrb, kcne4, and abcc9 (Shih et al., 2021). As in mice, pdgfrb mutants fail to form brain pericytes, exhibit loss of vSMCs and display aneurysms as adults (Ando et al., 2021b; Ando et al., 2016). As well as conserved pdgfrb control of mural cell development, Notch signaling (Ulrich et al., 2016) also plays a role in mural cell development in zebrafish and in mice, with Notch 1 and 3 controlling the formation of pericytes in mice and Notch 2 and 3 paralogs important in zebrafish (Wang et al., 2014; Ando et al., 2019). Overall, studies have suggested a high level of conservation of the molecular control of mural cell development, yet how mural cells control BBB function has not been interrogated in detail in zebrafish.

BBB function is first established in zebrafish between 3 days post fertilization (dpf, no barrier) and 5 dpf (established barrier) (O’Brown et al., 2019). During this period, developing mural cells (pericytes and vSMCs) are establishing and expanding their interactions with brain ECs (Ando et al., 2016; Ando et al., 2019; Ando et al., 2021a). The first glial cell neurovascular interactions are also observed during this period (Gall et al., 2025). Throughout subsequent larval, juvenile and adult development vSMC, pericyte, and glial cell coverage of the vasculature increases (Ando et al., 2016; Gall et al., 2025; Chen et al., 2020). This is similar to mammals, albeit with some differences in the relative timing that lineages arise, and the percentage of coverage of neurovasculature by pericytes and glia may be lower in zebrafish. In mammals, microglia are known to play important roles in the control of the BBB and neurovascular unit (Mayer and Fischer, 2024). Zebrafish have a microglial/macrophage population, but it is yet to be explored in detail in the control of the BBB (Wu et al., 2018; Xu et al., 2015; Herbomel et al., 2001) and differences in other innate immune cells in the CNS has been recently reported (Gaudi et al., 2026). Overall, while there are some areas remaining to be fully explored, the development, cellular composition and the central role of ECs in the control of the zebrafish BBB is very similar to mammals. As such, zebrafish are accepted as a highly accessible model to study BBB function, as well as mural cell control of the BBB, relevant to mammalian biology (Quiñonez-Silvero et al., 2020).

Here, we explored mural cell control of BBB function in zebrafish. We used pdgfrb mutants with loss of both pericytes and vSMCs, coupled with carefully optimized intravascular tracer dye injections. We analyzed barrier function from developmental to adult stages and found that at larval stages up to 14 dpf, well after the establishment of the BBB, loss of mural cells does not lead to measurable BBB leakage or loss of BBB integrity. In adult animals, we observed loss of BBB integrity that was consistent with leakage at hotspots, found to be aneurysm associated hemorrhages. These hotspots were also observed in juvenile animals (1 month old) and appeared to increase in number as animals aged. Our observations provide no definitive evidence of BBB leakage at capillary networks that might be consistent with a broad increase in EC transcytosis or loss of barrier integrity independent of focal hemorrhages. This work demonstrates that young animals can have a functional BBB in the absence of mural cells and highlights the central role for the EC in control of barrier function across the zebrafish neurovasculature.

To investigate the influence of mural cells upon the neurovasculature, we generated a pdgfrb mutant (pdgfrb^uq30bh, hereafter pdgfrb^−/−), which possesses an early deletion leading to a frameshift, predicted to cause a premature stop codon and a putative null allele (Figure 1—figure supplement 1). Using both the TgBAC(pdgfrb:EGFP)^uq15bh and TgBAC(abcc9:abcc9-T2A-mCherry)^uom139 transgenic marker strains, we observed that pericytes were lost in the cerebral central arteries of pdgfrb mutants (Figure 1a, Figure 1—figure supplement 1), as observed in previous studies (Ando et al., 2021b; Ando et al., 2016). These mutants showed indistinguishable development from their wild-type siblings up to 30 dpf with progressively reduced survival and craniofacial defects thereafter (Figure 1—figure supplement 1), consistent with previous studies of pdgfrb null loss-of-function mutants (Ando et al., 2021b; Ando et al., 2021a). To assess how brain vasculature develops without pericytes, we imaged, traced and quantified central arteries in the midbrain (Figure 1a–c). At 7 dpf, mutants displayed significantly reduced vascular complexity compared to siblings, with reduced vessel length and branch points (Figure 1d, e). This was more severe at 14 dpf, demonstrating the phenotype is progressive (Figure 1d, e). Timecourse imaging revealed the reduction in vascular complexity was apparent from 5 dpf (Figure 1f, g). These results suggest that the pericytes (as vSMCs mature after 5 dpf; Ando et al., 2019) regulate cerebral angiogenesis or remodeling from early in larval development.

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We next tested BBB function in mutants at 7 and 14 dpf. We performed intravenous injection of fluorescently conjugated tracers of various molecular weights (1-kDa NHS, 10- and 70-kDa Dextran), and live imaged fish at 2 hours post injection (hpi), the same timing used in O’Brown et al., 2019; Figure 2—figure supplement 1. 2000-kDa Dextran was co-injected to normalize for the amount of vascular tracer delivered in each animal, and we measured intravascular vs extravascular tracer intensity to assess leakage (see methods for full details). In control animals, quantification confirmed that 2000-kDa tracer remained stably within the vasculature, while 1 and 10 kDa were more readily detected in the brain parenchyma and 70 kDa provided intermediate sensitivity (Figure 2—figure supplement 1). This validated the use of 2000-kDa tracer to normalize for amount of tracer injected, as reported elsewhere (Lim et al., 2026). To provide additional confidence in our measurements, we also normalized the extravasated tracer intensity to the EC marker kdrl (Tg(kdrl:EGFP)^s843 or Tg(kdrl:Hsa.HRAS-mCherry)^s916). We found no evidence of elevated 10- or 70-kDa Dextran tracer extravasation into the brain parenchyma of pdgfrb mutants compared to siblings at either timepoint (7 or 14 dpf) using any of our normalization methods (Figure 2a–d, Figure 2—figure supplement 2). To confirm that our quantification methods were sufficiently sensitive to detect BBB leakage, we assessed 10- and 70-kDa tracer leakage before and after the establishment of BBB function (at 3 and 7 dpf; O’Brown et al., 2019). With both 10- and 70-kDa tracers, we could detect an open BBB at 3 dpf that was closed at 7 dpf (Figure 2e, f). Together, these results show that we can accurately measure BBB leakage and integrity in zebrafish and that based on these measures neither pericytes nor vSMCs are necessary for BBB integrity during larval stages.

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To investigate how mural cells control cerebral vasculature at later stages of life, we tissue-cleared brains of adult stage pdgfrb mutants and siblings (Matsumoto et al., 2019) and performed whole brain imaging. We confirmed loss of pericytes and vSMCs at this stage using TgBAC(pdgfrb:EGFP)^uq15bh and TgBAC(acta2:EGFP)^uq17bh (Figure 3a, b). We examined a consistent region of the left optic tectum (midbrain) and detected decreased branching of capillaries and increased diameter of major vessels (Figure 3d–f). The observed vessel dilation was restricted to larger caliber arteries that would normally be invested with vSMCs (Figure 3—figure supplement 1) and was not seen in capillaries in this region of the brain (Figure 3e, f). The loss of vSMCs was observable from the earliest stages of vSMC development and artery dilation was observed by 21 dpf, consistent with previous studies (Ando et al., 2021b; Figure 3c, Figure 3—figure supplement 1). This suggests that major vessel dilation seen in pdgfrb mutants is due to loss of vSMCs and that reduced branching of capillaries is due to loss of pericytes (given the timing observed in Figure 1).

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To investigate BBB integrity at later stages, we injected 3-month-old adult zebrafish with 10-kDa Dextran. Brightfield imaging of vibratome-sectioned brains revealed severe aneurysms across large caliber vessels (Figure 4a) with blood pooling that was potentially consistent with vessel ruptures (Figure 4c) as previously reported (Ando et al., 2021a). Furthermore, we detected parenchymal accumulation of tracer dye closely associated with large caliber vessel aneurysms (Figure 4b, c). To quantify the distribution of parenchymal tracer dye from deep (around major artery aneurysms) to superficial (capillary beds) brain regions, we measured fluorescence intensity in 10 brain regions approximately 100 µm apart from medial to lateral locations. This demonstrated accumulation of tracer dye in medial regions associated with aneurysm in mutants (Figure 4d, e). These animals showed little evidence of leakage in capillary beds, with marginally higher tracer intensity in mutants than controls in lateral locations that could be due to diffusion from medial locations with high concentrations of tracer (Figure 4e). To better understand this extravasated tracer accumulation, we conducted whole brain imaging after tracer injection in 5-month-old adults. Interestingly, we observed highly localized 70-kDa tracer accumulation in regions that could be considered leakage ‘hotspots’ at large caliber vessel aneurysms (Figure 5a; Video 1 and Video 2).

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As 10-kDa tracer showed higher sensitivity than 70-kDa tracer in our control experiments (Figure 2—figure supplement 1), we examined 10-kDa tracer injections at earlier stages and found that hotspots were readily detectable from as early as 1 month using this lower molecular weight tracer (Figure 5b). The hotspots were not readily detectable using 70-kDa Dextran in 2-month-old zebrafish, consistent with increased sensitivity of 10-kDa tracers (Figure 5—figure supplement 1). Interestingly, the quantification of hotspots in whole cleared brains at 1 and 5 months of age showed that the number increased with aging, suggesting a progressive loss of BBB integrity at focal sites of leakage (Figure 5c). To further understand the nature of leakage hotspots, we examined red blood cells in pdgfrb loss-of-function adults. We used pdgfrb Crispant knockouts, which we validated by finding that F0 CRISPR consistently reproduced the adult mutant phenotype (Figure 5—figure supplement 1). Using the Tg(gata1:DsRed)^sd2 transgenic line to label erythrocytes, together with tracer injection, we found that knockout animals showed tracer accumulation concomitant with extravasated red blood cells at hotspots (Figure 5d). This identified hotspots as sites of focal hemorrhage. Notably, these sites were not detected at capillaries (Figure 5e). Taken together, this shows that as mutants age, loss of BBB integrity is associated with an increasing number of hotspot sites of focal hemorrhage, that are found along aneurysmal vessels.

The quantification of tracer dye intensity at 2 hpi in adult mutants from medial (major vessels) to lateral (capillaries) locations indicated leakage most closely associated with hotspot focal hemorrhages. However, the intensity of tracer dye found in the lateral capillary regions was measurably higher than in control animals (Figure 4e). To test if this was likely due to progressive diffusion of tracers from medial to lateral locations or if there may be some local leakage at capillaries, we examined 10-kDa tracer leakage at 0.5 and 6 hpi. While hotspots could be detected at 0.5 hpi, we detected little tracer accumulation in medial locations and no evidence of parenchymal tracer leakage at lateral capillary locations (Figure 5f, h, Figure 5—figure supplement 2). However, after 6 hr of diffusion of tracer within the brain, we clearly observed tracer at both medial and lateral locations, with higher intensity in medial locations (Figure 5g, i; Figure 5—figure supplement 2). While we cannot rule out the possibility of slow leakage of tracer at capillaries occurring concurrently with leakage at hemorrhages, these observations seem consistent with a primary source of leakage at focal hemorrhages, followed by diffusion of tracer throughout the parenchyma.

Pericytes are proposed to regulate the BBB by suppressing transcytosis through brain capillary ECs (Armulik et al., 2010; Daneman et al., 2010). This model is built on the observation by electron microscopy of the induction of increased numbers of vesicular structures and caveolae in leaky pericyte-deficient vessels (Armulik et al., 2010; Daneman et al., 2010). To investigate if the loss of BBB integrity is associated with ultrastructural changes, we performed electron microscopy. We imaged large caliber vessels and capillaries (Figure 6a, b, Figure 6—figure supplement 1) and examined cellular junctions, basement membrane and caveolae (defined as uncoated vesicular profiles of <100 nm diameter). While tight junctions between ECs remained intact, caveolae numbers along large vessel aneurysms in pdgfrb mutants were significantly increased (Figure 6a′′–c). This increase was almost exclusive to the abluminal side of the ECs (Figure 6c). This abluminal accumulation of caveolae may be more associated with changes in the mechanical state of ECs (as caveolae are highly regulated by membrane tension Sinha et al., 2011; Del Pozo et al., 2021) than changes in intracellular transport. Furthermore, we detected thickening of basement membrane (BM) (Figure 6d), gaps in the BM and fluid accumulation outside the vessel wall at aneurysms in mutants. We found no obvious difference in the caveolae density in capillary ECs of pdgfrb mutants compared with siblings (Figure 6—figure supplement 1). Thus, leakage hotspots at aneurysms are associated with a loss of the normal flattened morphology of ECs, caveola induction and disruption of the BM, all factors that could contribute to or be observed concurrent with vessel wall rupture.

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Brain vasculature is endowed with mural cells that control vascular development and stability (Levéen et al., 1994; Lindahl et al., 1997; Ando et al., 2021b; Rajan et al., 2020). Studies in rodents have shown that pericytes regulate BBB integrity, with knockout of Pdgfb or Pdgfrb leading to vessel dilations and brain hemorrhages from early in development (Levéen et al., 1994; Lindahl et al., 1997). However, hypomorphic mutant alleles impacting Pdgfb or Pdgfrb show increased leakage of the BBB, with the severity of the leakage phenotype correlating with the expressivity of the pericyte phenotype (Armulik et al., 2010; Daneman et al., 2010). BBB leakage is associated with increased vesicular structures in ECs and the ‘open’ BBB state in the absence of mural cells is thought to be caused by hotspots of increased EC transcytosis (Armulik et al., 2011; Tjakra et al., 2019). Other potentially relevant pathways in ECs that could be altered by pericytes include altered cell–cell adhesion (e.g. Cldn5; Mäe et al., 2021), control of Mfsd2a (O’Brown et al., 2019; Andreone et al., 2017), Netrin1-Unc5B signaling (Boyé et al., 2022), or vitronectin regulation of integrin receptors (Ayloo et al., 2022), but direct links are yet to be established. Some studies using other models to deplete pericytes in mice have reported loss of pericytes without vessel leakage. For example, optical or chemical ablation of pericytes in the brain or retina of mice can be achieved without compromising the integrity of BBB or blood–retinal barrier (Berthiaume et al., 2022; Park et al., 2017), however these methods may induce inflammatory or compensatory mechanisms that also influence local vasculature. Taken together, these studies show that mural cells and in particular pericytes control the BBB in mammals, yet the precise downstream mechanisms by which pericytes regulate EC function require further study.

We show using zebrafish that mural cells are involved in cerebrovascular patterning in larval and juvenile animals, but these vessels maintain BBB function in the absence of mural cells. While the zebrafish BBB has been shown to be established between 3 and 5 dpf, our leakage assays at 7 and 14 dpf in mural cell-deficient pdgfrb mutants indicate intact BBB function and vessel integrity. This is unlikely to be due to technical issues with detecting leakage, as multiple measures and methods of tracer normalization produced consistent results. Furthermore, we can readily detect the ‘open’ BBB before it closes during development (Figure 2e, f). This demonstrates that a functional vertebrate BBB can exist in the absence of pericytes. Importantly, while other species have a glial cell BBB (e.g. Drosophila, some sharks and sturgeon), the zebrafish is well established to have an EC barrier and the glial cell coverage at the stages analyzed here is far from complete (Gall et al., 2025), making glial cell compensation for a loss of pericytes unlikely. There are other evolutionary differences (e.g. developmental timing, innate immune differences Gaudi et al., 2026; Renshaw and Trede, 2012) between zebrafish and mammals and our observation of intact BBB function without mural cells could represent an additional divergence. However, the high level of conservation of developmental pathways in BBB ECs and the very similar transcriptional signatures of both neurovascular ECs and mural cells between species (Quiñonez-Silvero et al., 2020; Shih et al., 2021; Ando et al., 2019; Vanlandewijck et al., 2018) may make this surprising. Once additional mechanisms of pericyte–EC crosstalk are established in mice and zebrafish, deeper mechanistic comparative studies will help to better understand if our observations identify major differences in BBB control between these organisms or not.

Ultimately, in older mutants from 1 month of age onwards, BBB integrity defects are observable with major arteries progressively harboring aneurysms and focal hemorrhages (leakage hotspots). These aneurysms are likely due to the loss of vSMCs seen in our pdgfrb mutants and other zebrafish pdgfrb mutant models (Ando et al., 2021b; Ando et al., 2021a). Adult mutant hotspots show extravascular erythrocytes and tracer dye, demonstrating that focal hemorrhages are a cause of vessel leakage. Similar to this, in Pdgfb or Pdgfrb mutant mice with strong phenotypic expressivity, hemorrhages are also reported (although not in mild or hypomorphic mutants used in BBB studies) (Levéen et al., 1994; Lindahl et al., 1997; Soriano, 1994). These observations suggest a highly conserved role for mural cells in the regulation of aneurysm and hemorrhage in mice and zebrafish. In our zebrafish model, BBB leakage in juvenile and adult animals appears to only be observed once hemorrhages or hotspots are present. Coupled with the fact that larval zebrafish can have no mural cells, but intact barrier function, we suggest that any roles for mural cells in BBB integrity beyond control of aneurysm and hemorrhage are likely to be minor roles in this setting. It remains possible that there is slow undetectable capillary leakage, but future models that specifically lack capillary pericytes and not vSMCs will be needed to test this. Understanding the nature of hotspot leakage more broadly in various mammalian and non-mammalian models of mural cell deficiency would seem likely to uncover further insights into BBB regulation. Deeper understanding of the fundamental biology of the BBB is still needed and will pave the way for increasingly sophisticated efforts to target the BBB in disease in the future.

Source data 1 contains the numerical data used to generate the figures.

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Author details

1. Oguzhan F Baltaci

   1. Organogenesis and Cancer Program, Peter MacCallum Cancer Centre, Melbourne, Australia
   2. Sir Peter MacCallum Department of Oncology, University of Melbourne, Melbourne, Australia

   Contribution

   Conceptualization, Data curation, Formal analysis, Validation, Visualization, Methodology, Writing – original draft, Writing – review and editing

   For correspondence

   oguzhan.baltaci@petermac.org

   Competing interests

   No competing interests declared

   "This ORCID iD identifies the author of this article:" 0009-0001-5651-1331

2. Andrea Usseglio Gaudi

   1. Organogenesis and Cancer Program, Peter MacCallum Cancer Centre, Melbourne, Australia
   2. Sir Peter MacCallum Department of Oncology, University of Melbourne, Melbourne, Australia

   Contribution

   Investigation, Methodology

   Competing interests

   No competing interests declared

3. Stefanie Dudczig

   1. Organogenesis and Cancer Program, Peter MacCallum Cancer Centre, Melbourne, Australia
   2. Sir Peter MacCallum Department of Oncology, University of Melbourne, Melbourne, Australia

   Contribution

   Investigation, Methodology

   Competing interests

   No competing interests declared

4. Weili Wang

   Institute for Molecular Bioscience, The University of Queensland, St Lucia, Australia

   Contribution

   Investigation, Methodology

   Competing interests

   No competing interests declared

5. Scott Paterson

   1. Organogenesis and Cancer Program, Peter MacCallum Cancer Centre, Melbourne, Australia
   2. Sir Peter MacCallum Department of Oncology, University of Melbourne, Melbourne, Australia

   Contribution

   Investigation, Methodology

   Competing interests

   No competing interests declared

6. Maria Cristina Rondon-Galeano

   1. Organogenesis and Cancer Program, Peter MacCallum Cancer Centre, Melbourne, Australia
   2. Sir Peter MacCallum Department of Oncology, University of Melbourne, Melbourne, Australia

   Contribution

   Investigation, Methodology

   Competing interests

   No competing interests declared

7. Ye-Wheen Lim

   Institute for Molecular Bioscience, The University of Queensland, St Lucia, Australia

   Contribution

   Investigation, Methodology

   Competing interests

   No competing interests declared

8. James Rae

   Institute for Molecular Bioscience, The University of Queensland, St Lucia, Australia

   Contribution

   Investigation, Methodology

   Competing interests

   No competing interests declared

9. Anne Lagendijk

   Institute for Molecular Bioscience, The University of Queensland, St Lucia, Australia

   Contribution

   Conceptualization, Supervision

   Competing interests

   No competing interests declared

10. Robert G Parton

    1. Institute for Molecular Bioscience, The University of Queensland, St Lucia, Australia
    2. Centre for Microscopy and Microanalysis, The University of Queensland, St Lucia, Australia

    Contribution

    Formal analysis, Investigation, Methodology

    Competing interests

    No competing interests declared

    "This ORCID iD identifies the author of this article:" 0000-0002-7494-5248

11. Alison Farley

    1. Organogenesis and Cancer Program, Peter MacCallum Cancer Centre, Melbourne, Australia
    2. Sir Peter MacCallum Department of Oncology, University of Melbourne, Melbourne, Australia
    3. Walter and Eliza Hall Institute of Medical Research, Melbourne, Victoria, Australia

    Contribution

    Investigation, Methodology

    Competing interests

    No competing interests declared

12. Benjamin M Hogan

    1. Organogenesis and Cancer Program, Peter MacCallum Cancer Centre, Melbourne, Australia
    2. Sir Peter MacCallum Department of Oncology, University of Melbourne, Melbourne, Australia
    3. Department of Anatomy and Physiology, University of Melbourne, Melbourne, Australia

    Contribution

    Conceptualization, Formal analysis, Supervision, Funding acquisition, Writing – original draft, Writing – review and editing

    For correspondence

    ben.hogan@petermac.org

    Competing interests

    Reviewing editor, eLife

    "This ORCID iD identifies the author of this article:" 0000-0002-0651-7065

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