Biology subjects

Farmer, D.

Publications and source records attributed to Farmer, D..

4 recordsLinked to original sources

bioRxiv2026

BMP antagonism is required for mandible outgrowth in zebrafish

The Meckels cartilage (MC) is a fundamental component of mandibular development across vertebrates. In mammals, MC is transient and functions primarily as an early template for mandibular ossification, whereas other vertebrates, including zebrafish, retain MC within the mandible throughout life. Despite its importance, the requirements for MC in sustaining mandibular growth and how signaling pathways implicated in MC development contribute to this process remain unclear. Here, we investigated the dosage-dependent roles of BMP antagonists during zebrafish MC development using mutant alleles of grem1a, nog2, and nog3. Compound mutant adults exhibited fully penetrant mandibular truncation. MC shortening emerged after early larval stages, indicating a requirement for BMP antagonism to sustain cartilage growth. Chondrocyte number remained unchanged as phenotypes developed, but mutants displayed disorganized cartilage morphology and increased chondrocyte volume. Molecular analyses revealed reduced col2a1a domains and expanded ihha and col10a1a expression, consistent with ectopic hypertrophic-like differentiation. Constitutive activation of BMP receptor signaling in chondrocytes recapitulated these phenotypes. Although osteogenesis appeared unaffected by 14 dpf, loss of a tnn skeletal mesenchyme population was observed. Together, these findings demonstrate that BMP antagonists sustain MC growth by regulating chondrocyte differentiation and cartilage organization to support mandibular growth in non-mammalian vertebrates. Summary StatementThis study leverages zebrafish to define the cellular and molecular mechanisms by which BMP antagonism sustains mandibular growth.

developmental biology↗

bioRxiv2026

Fibronectin 1 is required for suture patency and dysregulated across craniosynostosis models in the mouse

The mammalian skull roof is comprised of calvarial bones, connected through fibrous sutures, that protect the brain and allow for growth. Premature suture closure, or craniosynostosis (CS), impedes expansion, impacting 1 in every 2500 newborns. Despite its genetic heterogeneity, with nearly 80 associated genes, CS manifests as a common phenotype, hinting at a convergent etiological mechanism. Recently, we described how graded expression of extracellular matrix protein Fibronectin (FN1) is required for apical expansion of calvaria and coronal suture patency. Dysregulated FN1 expression has been identified in two human CS syndromes, suggesting its potential as a convergent mechanism of CS. Here, we further investigate the cellular basis for the CS phenotype in the Fn1 mutant mouse. Graded expression of FN1 baso-apically in the cranial mesenchyme was variably dysregulated across mouse models of syndromic and non-syndromic CS and accompanied by diminished apical expansion of frontal bone primordia. In parallel, at later developmental stages we find ectopic osteogenic induction of Six2+ patent suture mesenchyme in the Fn1 mutant. These findings pinpoint FN1 as a crucial regulator of suture patency by modulating calvarial growth and driving cell identity, and differentiation, thus providing a potential target for matrix-mediated treatments.

developmental biology↗

bioRxiv2022

Cryo-plasma FIB/SEM volume imaging of biological specimens

Serial focussed ion beam scanning electron microscopy (FIB/SEM) enables imaging and assessment of sub-cellular structures on the mesoscale (10 nm to 10 m). When applied to vitrified samples, serial FIB/SEM is also a means to target specific structures in cells and tissues while maintaining constituents hydration shells for in-situ structural biology downstream. However, the application of serial FIB/SEM imaging of non-stained cryogenic biological samples is limited due to low contrast, curtaining and charging artefacts. We address these challenges using a cryogenic plasma FIB/SEM (cryo-pFIB/SEM). We evaluated the choice of plasma ion source and imaging regimes to produce high quality SEM images of a range of different biological samples. Using an automated workflow we produced three dimensional volumes of bacteria, human cells, and tissue, and calculated estimates for their resolution, typically achieving 20 to 50 nm. Additionally, a tag-free tool is needed to drive the application of in situ structural biology towards tissue. The combination of serial FIB/SEM with plasmabased ion sources promises a framework for targeting specific features in bulk-frozen samples (>100 m) to produce lamella for cryogenic electron tomography.

cell biology↗

bioRxiv2021

Embryonic requirements for Tcf12 in the development of the mouse coronal suture

A major feature of Saethre-Chotzen syndrome is coronal craniosynostosis, the fusion of the frontal and parietal bones at the coronal suture. It is caused by heterozygous loss-of-function mutations in the basic HLH transcription factors TWIST1 and TCF12. While compound heterozygous Tcf12; Twist1 mice display severe coronal synostosis, the individual role of Tcf12 has remained unexplored. Here we show that Tcf12 controls several key processes in calvarial development, including the rate of frontal and parietal bone growth, and the boundary between sutural and osteogenic cells. Genetic analysis supports an embryonic requirement for Tcf12 in suture formation, as combined deletion of Tcf12 in the embryonic neural crest and mesoderm, but not in the postnatal suture mesenchyme, disrupts the coronal suture. We also detect asymmetric distribution of Grem1 + mesenchymal cells on opposing sides of the wild-type frontal and parietal bones, which prefigures later bone overlap at the sutures. In Tcf12 mutants, reduced asymmetry correlates with lack of bone overlap. Our results indicate a largely embryonic function of Tcf12 in controlling the rate and asymmetrical growth of calvarial bones and establishment of suture boundaries, which together ensure the proper formation of the overlapping coronal suture.

developmental biology↗