Organ function depends on tissues adopting the correct architecture. However, insights into organ architecture are currently hampered by an absence of standardized quantitative 3D analysis. We aimed to develop a robust technology to visualize, digitalize, and segment the architecture of two tubular systems in 3D: double resin casting micro computed tomography (DUCT). As proof of principle, we applied DUCT to a mouse model for Alagille syndrome (Jag1Ndr/Ndr mice), characterized by intrahepatic bile duct paucity, that can spontaneously generate a biliary system in adulthood. DUCT identified increased central biliary branching and peripheral bile duct tortuosity as two compensatory processes occurring in distinct regions of Jag1Ndr/Ndr liver, leading to full reconstitution of wild-type biliary volume and phenotypic recovery. DUCT is thus a powerful new technology for 3D analysis, which can reveal novel phenotypes and provide a standardized method of defining liver architecture in mouse models.
Many essential parts of the body contain tubes: the liver for example, contains bile ducts and blood vessels. These tubes develop right next to each other, like entwined trees. To do their jobs, these ducts must communicate and collaborate, but they do not always grow properly. For example, babies with Alagille syndrome are born with few or no bile ducts, resulting in serious liver disease. Understanding the architecture of the tubes in their livers could explain why some children with this syndrome improve with time, but many others need a liver transplant. Visualising biological tubes in three dimensions is challenging. One major roadblock is the difficulty in seeing several tubular structures at once. Traditional microscopic imaging of anatomy is in two dimensions, using slices of tissue. This approach shows the cross-sections of tubes, but not how the ducts connect and interact. An alternative is to use micro computed tomography scans, which use X-rays to examine structures in three dimensions. The challenge with this approach is that soft tissues, which tubes in the body are made of, do not show up well on X-ray. One way to solve this is to fill the ducts with X-ray absorbing resins, making a cast of the entire tree structure. The question is, can two closely connected tree structures be distinguished if they are cast at the same time? To address this question, Hankeova, Salplachta et al. developed a technique called double resin casting micro computed tomography, or DUCT for short. The approach involved making casts of tube systems using two types of resin that show up differently under X-rays. The new technique was tested on a mouse model of Alagille syndrome. One resin was injected into the bile ducts, and another into the blood vessels. This allowed Hankeova, Salplachta et al. to reconstruction both trees digitally, revealing their length, volume, branching, and interactions. In healthy mice, the bile ducts were straight with uniform branches, but in mice with Alagille syndrome ducts were wiggly, and had extra branches in the centre of the liver. This new imaging technique could improve the understanding of tube systems in animal models of diseases, both in the liver and in other organs with tubes, such as the lungs or the kidneys. Hankeova, Salplachta et al. also lay a foundation for a deeper understanding of bile duct recovery in Alagille syndrome. In the future, DUCT could help researchers to see how mouse bile ducts change in response to experimental therapies.
- Klíčová slova
- Alagille syndrome, MicroCT, cholangiopathy, human, mouse, physics of living systems, regenerative medicine, resin, stem cells, vasculature,
- MeSH
- Alagillův syndrom patofyziologie MeSH
- modely nemocí na zvířatech MeSH
- myši transgenní MeSH
- myši MeSH
- rentgenová mikrotomografie klasifikace metody MeSH
- žlučové cesty růst a vývoj patofyziologie MeSH
- zvířata MeSH
- Check Tag
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Plants as non-mobile organisms constantly integrate varying environmental signals to flexibly adapt their growth and development. Local fluctuations in water and nutrient availability, sudden changes in temperature or other abiotic and biotic stresses can trigger changes in the growth of plant organs. Multiple mutually interconnected hormonal signaling cascades act as essential endogenous translators of these exogenous signals in the adaptive responses of plants. Although the molecular backbones of hormone transduction pathways have been identified, the mechanisms underlying their interactions are largely unknown. Here, using genome wide transcriptome profiling we identify an auxin and cytokinin cross-talk component; SYNERGISTIC ON AUXIN AND CYTOKININ 1 (SYAC1), whose expression in roots is strictly dependent on both of these hormonal pathways. We show that SYAC1 is a regulator of secretory pathway, whose enhanced activity interferes with deposition of cell wall components and can fine-tune organ growth and sensitivity to soil pathogens.
- MeSH
- Arabidopsis genetika růst a vývoj metabolismus MeSH
- buněčná stěna chemie metabolismus MeSH
- cytokininy metabolismus MeSH
- endozomy metabolismus MeSH
- geneticky modifikované rostliny metabolismus MeSH
- Golgiho aparát metabolismus MeSH
- kořeny rostlin metabolismus mikrobiologie MeSH
- kyseliny indoloctové metabolismus MeSH
- membránové proteiny genetika metabolismus MeSH
- odolnost vůči nemocem genetika MeSH
- Plasmodiophorida patogenita MeSH
- proteiny huseníčku genetika metabolismus MeSH
- půda MeSH
- regulace genové exprese u rostlin genetika MeSH
- regulátory růstu rostlin metabolismus MeSH
- sekreční dráha genetika MeSH
- stanovení celkové genové exprese MeSH
- transkriptom genetika MeSH
- vezikulární transportní proteiny metabolismus MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- AT2G18840 protein, Arabidopsis MeSH Prohlížeč
- AT4G30260 protein, Arabidopsis MeSH Prohlížeč
- cytokininy MeSH
- ECHIDNA protein, Arabidopsis MeSH Prohlížeč
- kyseliny indoloctové MeSH
- membránové proteiny MeSH
- proteiny huseníčku MeSH
- půda MeSH
- regulátory růstu rostlin MeSH
- SYAC1 protein, Arabidopsis MeSH Prohlížeč
- vezikulární transportní proteiny MeSH
Auxin is unique among plant hormones due to its directional transport that is mediated by the polarly distributed PIN auxin transporters at the plasma membrane. The canalization hypothesis proposes that the auxin feedback on its polar flow is a crucial, plant-specific mechanism mediating multiple self-organizing developmental processes. Here, we used the auxin effect on the PIN polar localization in Arabidopsis thaliana roots as a proxy for the auxin feedback on the PIN polarity during canalization. We performed microarray experiments to find regulators of this process that act downstream of auxin. We identified genes that were transcriptionally regulated by auxin in an AXR3/IAA17- and ARF7/ARF19-dependent manner. Besides the known components of the PIN polarity, such as PID and PIP5K kinases, a number of potential new regulators were detected, among which the WRKY23 transcription factor, which was characterized in more detail. Gain- and loss-of-function mutants confirmed a role for WRKY23 in mediating the auxin effect on the PIN polarity. Accordingly, processes requiring auxin-mediated PIN polarity rearrangements, such as vascular tissue development during leaf venation, showed a higher WRKY23 expression and required the WRKY23 activity. Our results provide initial insights into the auxin transcriptional network acting upstream of PIN polarization and, potentially, canalization-mediated plant development.
- MeSH
- Arabidopsis genetika růst a vývoj MeSH
- geneticky modifikované rostliny MeSH
- genové regulační sítě * účinky léků MeSH
- kořeny rostlin účinky léků genetika růst a vývoj metabolismus MeSH
- kyseliny indoloctové metabolismus farmakologie MeSH
- membránové transportní proteiny genetika metabolismus MeSH
- mikročipová analýza MeSH
- polarita buněk * genetika MeSH
- proteiny huseníčku genetika metabolismus fyziologie MeSH
- regulace genové exprese u rostlin účinky léků MeSH
- stanovení celkové genové exprese MeSH
- transkripční faktory fyziologie MeSH
- zpětná vazba fyziologická účinky léků MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- kyseliny indoloctové MeSH
- membránové transportní proteiny MeSH
- PIN1 protein, Arabidopsis MeSH Prohlížeč
- proteiny huseníčku MeSH
- transkripční faktory MeSH
- WRKY23 protein, Arabidopsis MeSH Prohlížeč
