Foxc1a regulates zebrafish vascular integrity and brain vascular development through targeting amotl2a and ctnnb1
Xuchu Duan1, Yuanyuan Shi2, Shu Zhao1
1School of Life Science, Nantong Laboratory of Development and Diseases, Department of Endocrine, Affiliated Hospital, Key Laboratory of Neuroregeneration of Jiangsu and Ministry of Education, Co-innovation Center of Neuroregeneration, Medical School, Nantong University, Nantong, China.
Abstract:
Accumulating evidences have pointed that foxc1a is essential for vascular development and integrity maintenance through regulating the expression of downstream genes and interacting with signaling pathways. However, the underling cellular and molecular mechanisms of foxc1a in regulating vascular development remain undetermined. Based on two different foxc1a mutant zebrafish lines (foxc1anju18 and foxc1anju19 which generated predicted truncated foxc1a proteins with 50aa and 315aa respectively), we found that around 30 % of foxc1anju18 zebrafish exhibited severe vascular developmental defects with obvious hemorrhage in hindbrain and trunk at embryonic stages. Confocal imaging analysis revealed that the formation of middle cerebral vein (MCeV), intra-cerebral central arteries (CtAs) and dorsal longitudinal vein (DLV) of brain vessels was significantly blocked in foxc1anju18enbryos. Injection of exogenous full length and foxc1anju19 truncated foxc1a mRNA both rescued the deficiency of foxc1anju18 embryos. Transcriptome analysis revealed 186 DEGs in foxc1anju18 zebrafish among which amotl2a and ctnnb1 expression were reduced and functionally associated with adherens junctions. Dual-Luciferase assays validated amotl2a and ctnnb1 were both directly transactivated by foxc1a. Rescue experiments demonstrated that amotl2a was mainly responsible for the vascular integrity caused by foxc1a mutation and also coordinated with ctnnb1 to regulate brain vascular development. Our data point to a novel clue that foxc1a regulates vascular integrity and brain vascular development through targeting amotl2a and ctnnb1.
Insights
Forkhead box C1a (foxc1a) is crucial for vascular development. This study reveals foxc1a regulates vascular integrity and brain vessel formation by targeting amotl2a and ctnnb1 genes.
Area of Science:
- Developmental biology
- Vascular biology
- Genetics
Background:
- Foxc1a is known to be essential for vascular development and integrity.
- The precise cellular and molecular mechanisms by which foxc1a regulates vascular development are not fully understood.
Purpose of the Study:
- To elucidate the underlying cellular and molecular mechanisms of foxc1a in regulating vascular development and integrity.
- To identify downstream targets of foxc1a involved in vascular development.
Main Methods:
- Utilized two foxc1a mutant zebrafish lines (foxc1a nju18 and foxc1a nju19) to study vascular defects.
- Employed confocal imaging, mRNA injection for rescue experiments, transcriptome analysis, and Dual-Luciferase assays.
Main Results:
- foxc1a nju18 mutants exhibited severe vascular defects, including hemorrhage and blocked brain vessel formation.
- Transcriptome analysis identified amotl2a and ctnnb1 as downregulated genes in foxc1a nju18 mutants.
- Amotl2a and ctnnb1 were validated as direct targets of foxc1a, with amotl2a playing a key role in vascular integrity.
Conclusions:
- Foxc1a regulates vascular integrity and brain vascular development.
- Foxc1a targets amotl2a and ctnnb1 to control vascular development and integrity.
- Amotl2a is critical for foxc1a-mediated vascular integrity and functions in coordination with ctnnb1.
