Transgelin-expressing myofibroblasts orchestrate ventral midline closure through TGFβ signalling

Aldeiri, Bashar; Roostalu, Urmas; Albertini, Alessandra; Wong, Jason; Morabito, Antonino; Cossu, Giulio

doi:10.1242/dev.152843

Cited by 16 publications

(22 citation statements)

References 54 publications

(72 reference statements)

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“…Clusters with similar marker gene expressions were merged manually into one. Neuronal cells and progenitors were identified according to their expressions of corresponding marker genes: progenitor markers (SOX2, VIM, ID3, MKI67, TOP2A, CENPF) [46], pan-neuron markers (MAP2, SYT4, SYT1, STMN2, GAP43, DCX, NEFM, VGLUT2) ) [47,48], sensory neuron genes (PRPH, ISL1, BRN3A, PHOX2B, TAC1, GAL, SSTR2, PIEZO2, FGF3, SCN9A, NTRK2) [28,29] and fibroblast cluster markers (COL1A1, COL1A2, MYL9, ACTA2, TAGLN) [49]. 24 Differentially expressed genes (DEGs) between neuronal cells and neural progenitors of each species at each time point were identified by using the "FindMarkers" function in the Seurat R package with the cut-offs of adjusted p value less than 0.05 and log2 transformed fold change larger than 0.25.…”

Section: Identification Of Neuronal Cells and Differentially Expressementioning

confidence: 99%

Comparison of induced neurons reveals slower structural and functional maturation in humans than in apes

Schoernig

Fast

et al. 2020

Preprint

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We generated induced excitatory sensory neurons (iNeurons, iNs) from chimpanzee, bonobo and human stem cells by expressing the transcription factor neurogenin-2 (NGN2). Single cell-RNA sequencing showed that genes involved in dendrite and synapse development are expressed earlier during iNs maturation in the chimpanzee than the human cells. In accordance, during the first two weeks of differentiation, chimpanzee and bonobo iNs showed repetitive action potentials and more spontaneous excitatory activity than human iNs, and extended neurites of higher total length. However, the axons of human iNs were slightly longer at 5 weeks of differentiation. The timing of the establishment of neuronal polarity did not differ between the species. Chimpanzee, bonobo and human neurites eventually reached the same level of structural complexity.Thus, human iNs develop slower than chimpanzee and bonobo iNs and this difference in timing likely depends on functions downstream of NGN2. 4Here, we use a direct conversion system in which chimpanzee, bonobo and human iPSCs are converted into neurons using an NGN2 overexpression system [23,24].Unlike what has been previously reported, we obtain a neuronal cell population containing a majority of sensory neurons. We describe their temporal differentiation in terms of gene expression, electrophysiology and dendritic arborization. 5 RESULTS Maturation of human and ape induced neurons in vitroWe generated iNeurons from three chimpanzee (SandraA, JoC, ciPS01), one bonobo (BmRNA) and three human (409B2, SC102A, HmRNA) iPSC lines and from one human ESC line (H9) using forced expression of the pan-neurogenic transcription factor neurogenin 2 (NGN 2; Figure 1A) [23,24]. We followed their differentiation using molecular, electrophysiological and morphological approaches for up to 8 weeks, as indicated in the ( Figure 1A, B). Single cell RNA sequencing (scRNAseq) was performed for one chimpanzee (SandraA) and three human (409B2, SC102A1 and H9) cell lines, electrophysiology was performed using all eight cell lines and morphological analyses were done with two chimpanzee (SandraA and JoC), the bonobo (BmRNA) and three human (409B2, SC102A1 and H9) cell lines. We will refer to ape iNs for results where we combined chimpanzee and bonobo iNs for analysis.Differentiation was characterized by a downregulation of the stem cell markers NANOG, OCT4 and SOX2 and of the neural progenitor's markers ID1 and ID3 in both chimpanzee and human cells ( Supplementary Figure 1), by a change in cellular morphology and by the extension of neurites ( Figure 1C). Chimpanzee, bonobo and human iNs showed a neuron-like morphology at day 7 (d7) of differentiation and formed a dense network by d14. Neurites were positive for TUJI (beta-III-tubulin, a neuronal marker) starting from d3 of differentiation in apes and humans ( Supplementary Figure 2).By the end of the differentiation at d35, both ape and human cells formed networks that were positive for MAP2 (microtubule associated protein-2, marker for mature

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Section: Identification Of Neuronal Cells and Differentially Expressementioning

confidence: 99%

Comparison of induced neurons reveals slower structural and functional maturation in humans than in apes

Schoernig

Fast

et al. 2020

Preprint

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“…The Tagln -Cre: Tgfbr2 flx/flx model exhibits three main categories of congenital anomalies. The first striking anomaly is the complete failure of ventral body wall closure as we have recently shown 15 . We used here micro-CT scanning to characterise the extent of the defect and to delineate its 3D topography.…”

Section: Resultsmentioning

confidence: 89%

“…At embryonic stages TAGLN is not a specific VSMC marker and is widely expressed by non-vascular mesenchymal tissues 24 . Moreover, Tagln was recently found to label a migratory myofibroblasts cell population that respond to TGFβ signalling 15 . TGFβ is also known to induce Transgelin ( Tagln ) in vitro and in vivo 25 – 27 through SMAD binding to the Tagln promoter 28 .…”

Section: Introductionmentioning

confidence: 99%

“…TGFβ is also known to induce Transgelin ( Tagln ) in vitro and in vivo 25 – 27 through SMAD binding to the Tagln promoter 28 . Furthermore, TGFβ signalling inactivation in Tagln expressing cells leads to ventral body wall closure defect, cardiac and great vessels anomalies 15 , 17 , 29 .…”

Section: Introductionmentioning

confidence: 99%

“…We have recently demonstrated that selective removal of Tgfbr2 from TAGLN expressing cells results in exomphalos and ectopia cordis 15 . In addition, previous reports of the Tagln -Cre: Tgfbr2 flx/flx have demonstrated cardiac and major vessels defect that overlap with the spectrum of cardiac anomalies seen in PC 17 , 29 .…”

Section: Introductionmentioning

confidence: 99%

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Abrogation of TGF-beta signalling in TAGLN expressing cells recapitulates Pentalogy of Cantrell in the mouse

Aldeiri

Roostalu

Albertini

et al. 2018

Sci Rep

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Pentalogy of Cantrell (PC) is a rare multi-organ congenital anomaly that impedes ventral body wall closure and results in diaphragmatic hernia, intra- and pericardial defects. The underlying cellular and molecular changes that lead to these severe developmental defects have remained unknown largely due to the lack of representative animal models. Here we provide in depth characterization of a mouse model with conditional ablation of TGFβRII in Transgelin (Tagln) expressing cells. We show that Tagln is transiently expressed in a variety of cells that participate in the embryonic development and patterning of ventral structures. Genetic ablation of TGFβRII in these cells leads to ventral midline closure defect, diaphragmatic hernia, dilated cardiac outflow tract and aberrant cardiac septation, providing a reliable model to study the morphological changes leading to PC. We show that myogenisis in the diaphragm is independent of TGFβ and the diaphragmatic hernia arises from fibroblast-specific migration defect. In the dorsal body wall Tagln expression is initiated after the closure process, revealing a remarkable difference between ventral and dorsal body walls development. Our study demonstrates the use of micro-CT scanning to obtain a 3-dimensional high-resolution overview of embryonic anomalies and provides the first mechanistic insight into the development of PC.

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Single-cell atlas of craniogenesis uncovers SOXC-dependent, highly proliferative, and myofibroblast-like osteodermal progenitors

et al. 2022

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Transgelin-expressing myofibroblasts orchestrate ventral midline closure through TGFβ signalling

Cited by 16 publications

References 54 publications

Comparison of induced neurons reveals slower structural and functional maturation in humans than in apes

Comparison of induced neurons reveals slower structural and functional maturation in humans than in apes

Abrogation of TGF-beta signalling in TAGLN expressing cells recapitulates Pentalogy of Cantrell in the mouse

Single-cell atlas of craniogenesis uncovers SOXC-dependent, highly proliferative, and myofibroblast-like osteodermal progenitors

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