Three-dimensional reconstruction of systematic histological sections: application to observations on palatal shelf elevation

Liu, Weilong; Wang, Xiaoming; Wang, Yinuo; Wang, Yahong; Zhang, Jing; Shi, Bing; Li, Chenghao

doi:10.1038/s41368-021-00122-8

Cited by 7 publications

(7 citation statements)

References 33 publications

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“…Previous studies have highlighted anteroposterior differences in PS elevation (Bush and Jiang 2012; Chiquet et al 2016; Liu et al 2021; Walker and Fraser 1956b; Yu and Ornitz 2011). Given our ability to consistently obtain embryos in the process of PS elevation, we were able to collect and scan several of them using magnetic resonance imaging (MRI).…”

Section: Resultsmentioning

confidence: 99%

“…This would imply that one PS elevates above the tongue prior to the other shelf and that the tongue goes through a rocker like motion. There have been a few subsequent studies where unilaterally elevated PS have been captured in normal embryos (Bush and Jiang 2012; Liu et al 2021; Yu and Ornitz 2011), and several studies where unilateral elevation was captured in mutant mouse models of cleft palate, including those with Specc1l deficiency (Butali et al 2019; Goodwin et al 2020; Hall et al 2020; Hill et al 2015; Liu et al 2008). We were keen to see if we would routinely capture unilaterally elevated PS in our study.…”

Section: Discussionmentioning

confidence: 99%

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Novel insights into the fundamentals of palatal shelf elevation dynamics in normal mouse embryos

Goering

Moedritzer

Stetsiv

et al. 2022

Preprint

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Embryonic palate development involves bilateral vertical growth of palatal shelves - extensions from the maxillary processes - next to the tongue until embryonic day (E) 13.5. Following vertical growth, palatal shelves elevate and adhere above the tongue by E14.5. Current models indicate that this process of elevation involves a complex vertical to horizontal reorienting of the palatal shelves. While earlier studies have implied that this is a rapid process, the precise timing has not been resolved. To understand the dynamics of palatal shelf elevation, we employed time-restricted pregnancies with a one-hour resolution and magnetic resonance imaging of intermediate stages. Our data showed that in almost all C57BL/6J embryos, palatal shelves have not yet elevated by E14.0. However, six hours later at E14.25, palatal shelves have completed elevation in 80% of embryos. Interestingly, all E14.25 embryos with unelevated palatal shelves (20%) were female, suggesting a delay in female embryos. In FVB/NJ embryos, the elevation window started earlier (E13.875-E14.25) without any noticeable sex differences. We frequently captured an intermediate stage with unilateral elevation of either right or left palatal shelf. Magnetic resonance imaging of various stages showed that palatal shelf elevation began with the formation of bilateral bulges in the posterior. These bulges progressed laterally and anteriorly over time. During elevation, we observed increased cell proliferation in the lingual region of the palatal shelf. Within the bulge, cell orientation was acutely tilted towards the tongue and actomyosin activity was increased, which together may participate in the projection of the bulge in the horizontal direction. Thus, our data reveal novel insights into the rapid dynamic changes in palatal shelf elevation that lay the foundation for future studies of normal and abnormal palatogenesis.

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Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Novel insights into the fundamentals of palatal shelf elevation dynamics in normal mouse embryos

Goering

Moedritzer

Stetsiv

et al. 2022

Preprint

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“…The entire palatal shelf was lingually curved (Figure 3), and the morphological changes at the base of the shelf occurred almost simultaneously on both the lingual and buccal sides (Figure 4). In the “flow” model of elevation, the protrusion is formed on the lingual wall of the palatal shelf, and morphological changes in the base of the lingual side occur prior to those on the buccal side (Brinkley & Vickerman, 1979; Jin et al, 2010; Liu et al, 2021). Such changes in shape are not observed in the “flip‐up” model of elevation (Bush & Jiang, 2012; Liu et al, 2021) and were not observed in this study.…”

Section: Resultsmentioning

confidence: 99%

“…In the “flow” model of elevation, the protrusion is formed on the lingual wall of the palatal shelf, and morphological changes in the base of the lingual side occur prior to those on the buccal side (Brinkley & Vickerman, 1979; Jin et al, 2010; Liu et al, 2021). Such changes in shape are not observed in the “flip‐up” model of elevation (Bush & Jiang, 2012; Liu et al, 2021) and were not observed in this study. Hence, the morphological changes observed in the present study were consistent with the “flip‐up” model of palatal shelf elevation.…”

Section: Resultsmentioning

confidence: 99%

Live imaging observation of elevation of the anterior palatal shelf in mouse embryos

et al. 2023

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The mammalian secondary palate develops through complex processes including palatal shelf growth, elevation, and fusion. Palatal shelf elevation is a process accompanied by large-scale morphological changes over a short period. The elevation pattern changes along the anterior-posterior axis; the anterior region elevates by the "flip-up" model, and the middle and posterior regions reorient through the "flow" model. However, the mechanisms of both models are unclear because of the rapid progression of the elevation in utero. To observe palatal elevation in real time in detail, we aimed to establish a live imaging method using explants of the anterior region of the palatal shelf in mouse embryos before the beginning of elevation.Changes in the degree of shelf orientation were measured, which showed that the palatal shelf continuously changed shape toward the lingual side. The changes in the angle between the lingual and buccal bases of the palatal shelf were different; the morphological change at the lingual side resulted in a more acute angle, and the change at the buccal side resulted in a more obtuse angle. The morphological changes of the lingual and buccal sides occurred nearly simultaneously, suggesting that the anterior region of the palatal shelf in vitro elevated according to the "flip-up" model. This live imaging method enables the continuous observation of palatal shelf elevation and provides new insights into palatogenesis.

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“…Palatal development in both mice and humans typically occurs in three stages: palatal expansion and elongation before embryonic day 13.5 (E13.5), elevation and contact before E14.5, and fusion before E15.5 [ 9 ]. Palatal elevation is an essential process in palatogenesis led by the intrinsic force produced by HA accumulation [ 10 , 11 ]. HA has unique biophysical properties to bind up to 10 times its own weight with water and to enmesh other extracellular molecules [ 12 , 13 ].…”

Section: Introductionmentioning

confidence: 99%

FGF9 Promotes Expression of HAS2 in Palatal Elevation via the Wnt/β-Catenin/TCF7L2 Pathway

Sun

Ying

et al. 2022

Biomolecules

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Background: Fgf9 mutation was found in cleft palate patients. Our previous study indicated that Fgf9 promotes timely elevation of palate by regulating hyaluronic acid (HA) accumulation at embryonic day 13.5 (E13.5). HA is synthesized by hyaluronic acid synthases (HAS) isoforms 1, 2, or 3. However, how FGF9 regulates HA in palatogenesis is still unclear. Methods: Using Ddx4-Cre mice, we generated the Fgf9−/− mouse model (with exon 2 deletion). Immunohistochemistry was used to detect the location and expression of HAS2 in WT and the Fgf9−/− palate at E13.5. We also predicted the association between Fgf9 and Has2 within the developing palate by performing a bioinformatics analysis. The expression of β-catenin, HAS2, and TCF7L2 were verified by Western blotting after knockout of Fgf9. Rescue experiments were performed by ELISA in vitro. Results: Fgf9−/− mice exhibited 100% penetrance of the cleft palate. A knockout of Fgf9 confirmed that HAS2 and TCF7L2 expression was positively correlated with FGF9. TCF7L2 binds to the Has2 promoter, exhibiting the high specificity predicted by JASPAR. Additionally, increased HA expression by BML-284, TCF-dependent agonist, was blocked in Fgf9−/− palate because of the significant decline in TCF7L2 expression. Conclusions: FGF9 promotes HAS2 expression via Wnt/β-catenin/TCF7L2 pathway with TCF7L2 activating transcription of Has2 in the palate.

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Three-dimensional reconstruction of systematic histological sections: application to observations on palatal shelf elevation

Cited by 7 publications

References 33 publications

Novel insights into the fundamentals of palatal shelf elevation dynamics in normal mouse embryos

Novel insights into the fundamentals of palatal shelf elevation dynamics in normal mouse embryos

Live imaging observation of elevation of the anterior palatal shelf in mouse embryos

FGF9 Promotes Expression of HAS2 in Palatal Elevation via the Wnt/β-Catenin/TCF7L2 Pathway

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