Characterization of the Effect of Tongue on Palatal Shelf Elevation Patterns in a Mouse Model of Palatogenesis

ZHANG Yu, YANG Hui, ZHANG Demao, LI Dou, WANG Xiaoming

Abstract

Objective 

To investigate the mechanisms underlying regional heterogeneity in the elevating patterns of palatal shelf during mammalian craniofacial development.

Methods 

Using a mouse model of embryonic palatal development, we acquired coronal multi-plane slices of the palatal shelves before elevation (early E13.5), during elevation (late E13.5), and after elevation (early E14.5). Hematoxylin and eosin (HE) staining was performed to compare the morphological changes and spatial correlations between the palate and tongue. Immunofluorescence staining of myosin heavy chain 1 (MYH1), a marker found in slow muscle fibers and responsible for muscle contraction and movement, was performed to observe the tongue muscle development characteristics at different stages. We also observed changes in the palatal shelf elevating patterns at early E13.5 in the absence of the tongue through HE-stained in vitro palate organ culture. Further immunofluorescence staining of tenascin-C, an extracellular matrix protein, was performed to evaluate the effect of the tongue on the elevating pattern of the palatal shelf along the anterior-posterior axis.

Results 

HE staining results of the coronal multi-plane slices showed that during the elevation period, from the posterior toward anterior, the coronal height of the tongue decreased, lateral inclination and flattening increased, but the sagittal length of the tongue increased. The elevating pattern of the palatal shelf changed from slow remodeling to rapid flipping, and MYH1 was abundantly expressed in both the internal and external muscle bundles of the tongue during this period. According to findings from in vitro cultivation of palatal organs, the posterior part of the palatal shelf elevated without forming new lateral lingual protrusions in the absence of the tongue. The regional expression pattern of tenascin-C was consistent with that observed before elevation. The posterior palate exhibited an elevation pattern similar to that of the anterior region.

Conclusion 

The tongue may play a crucial role in shaping the posterior morphological remodeling and distinct elevation patterns of the palatal shelf.

 

Keywords: Cleft lip and palate, Palatal shelf elevation, Extracellular matrix remodeling, Tongue

 

Full Text:

PDF


References


WANG X, LIU W, LUO X, et al. Mesenchymal β-catenin signaling affects palatogenesis by regulating α-actinin-4 and F-actin. Oral Dis, 2023, 29(8): 3493-3502. doi: 10.1111/odi.14408.

LI C, LAN Y, JIANG R. Molecular and cellular mechanisms of palate development. J Dent Res, 2017, 96(11): 1184-1191. doi: 10.1177/ 0022034517703580.

BUSH J O, JIANG R. Palatogenesis: morphogenetic and molecular mechanisms of secondary palate development. Development, 2012, 139: 231-243. doi: 10.1242/dev.067082.

LAN Y, QIN C, JIANG R. Requirement of hyaluronan synthase-2 in craniofacial and palate development. J Dent Res, 2019, 98(12): 1367-1375. doi: 10.1177/0022034519872478.

MATTHIAS C, SUSAN B, MANUELA A, et al. Mesenchymal remodeling during palatal shelf elevation revealed by extracellular matrix and F-actin expression patterns. Front Physiol, 2016, 7: 392. doi: 10.3389/fphys.2016.00392.

PARADA C, HAN D, GRIMALDI A, et al. Disruption of the ERK/MAPK pathway in neural crest cells as a potential cause of Pierre Robin sequence. Development, 2015, 142: 3734-3745. doi: 10.1242/dev.125328.

PANG X, WANG X, WANG Y, et al. Sox9CreER-mediated deletion of β-catenin in palatal mesenchyme results in delayed palatal elevation accompanied with repressed canonical Wnt signaling and reduced actin polymerization. Genesis, 2021, 59: e23441. doi: 10.1002/dvg.23441.

LIU W, WANG X, WANG Y, et al. Three-dimensional reconstruction of systematic histological sections: application to observations on palatal shelf elevation. Int J Oral Sci, 2021, 13: 17. doi: 10.1038/s41368-021-00122-8.

NAGASAKA A, BANDO Y, TODA-FUJII M, et al. Differences in palatal shelf epithelial stiffness between the lingual/nasal and buccal/oral surfaces during palatal shelf elevation in developing mice. Dev Dyn, 2025. doi:10.1002/dvdy.70044.

NAGASAKA A, SAKIYAMA K, BANDO Y, et al. Live imaging observation of elevation of the anterior palatal shelf in mouse embryos. Dev Growth Differ, 2023, 65: 224-229. doi: 10.1111/dgd.12851. SNYDER-

WARWICK A K, PERLYN C A, PAN J, et al. Analysis of a gain-of-function FGFR2 Crouzon mutation provides evidence of loss of function activity in the etiology of cleft palate. Proc Natl Acad Sci U S A, 2010, 107: 2515-2520. doi: 10.1073/pnas.0913985107.

WANG X M, LIU W L, CHEN Y, et al. Lithium-induced overexpression of β-catenin delays murine palatal shelf elevation by Cdc-42 mediated F-actin remodeling in mesenchymal cells. Birth Defects Res, 2021, 113: 427-438. doi: 10.1002/bdr2.1853.

MA Y Q, ZHANG X Y, ZHAO S W, et al. Retinoic acid delays murine palatal shelf elevation by inhibiting Wnt5a-mediated noncanonical Wnt signaling and downstream Cdc-42/F-actin remodeling in mesenchymal cells. Birth Defects Res, 2023, 115: 1658-1673. doi: 10.1002/bdr2.2244.

CHEN X, LI N, HU P, et al. Deficiency of Fam20b-catalyzed glycosaminoglycan Chain synthesis in neural crest leads to cleft palate. Int J Mol Sci, 2023, 24(11): 9634. doi: 10.3390/ijms24119634.

XU J, LIU H, LAN Y, et al. The transcription factors Foxf1 and Foxf2 integrate the SHH, HGF and TGFβ signaling pathways to drive tongue organogenesis. Development, 2022, 149(21): dev200667. doi: 10.1242/dev. 200667.

PARADA C, HAN D, CHAI Y. Molecular and cellular regulatory mechanisms of tongue myogenesis. J Dent Res, 2012, 91: 528-535. doi: 10. 1177/0022034511434055.

LLOYD R A, DISSANAYAKE E, JUGÉ L, et al. How mandibular and hyoid morphology alters tongue muscle architecture in healthy adults: an anatomical atlas and statistical shape model of the tongue. Comput Biol Med, 2025, 189: 110006. doi: 10.1016/j.compbiomed.2025.110006.

MAY C A. Aponeurosis linguae-myocutaneous or myotendinous junctions of skeletal muscle fibres in the human tongue? J Anat, 2022, 241: 168-172. doi: 10.1111/joa.13637.

FRIEDL R M, RAJA S, METZLER M A, et al. RDH10 function is necessary for spontaneous fetal mouth movement that facilitates palate shelf elevation. Dis Model Mech, 2019, 12(7): dmm039073. doi: 10.1242/dmm.039073.

ZHANG X Y, CAI M Q, MA Y Q, et al. Teratogenic effect of TCDD on bone of facial cranium development in mice evaluated by three-dimensional CT reconstruction and genetic testing. Journal of Lanzhou University (Medical Sciences), 2023, 49(9): 9-15. doi: 10.27204/d.cnki. glzhu.2024.002793.

SHI J X, WANG C Y, LI J T. Research progress on cleft palate repair among patients with Pierre Robin sequence. International Journal of Stomatology, 2023, 50(2): 237-242. doi: 10.7518/gjkq.2023002.

XU J, LIU H, LAN Y, et al. Hedgehog signaling patterns the oral-aboral axis of the mandibular arch. Elife, 2019, 8: e40315. doi: 10.7554/eLife.40315.

OKA K, HONDA M J, TSURUGA E, et al. Roles of collagen and periostin expression by cranial neural crest cells during soft palate development. J Histochem Cytochem, 2012, 60: 57-68. doi: 10.1369/0022155411427059.

OHKI S, OKA K, OGATA K, et al. Transforming growth factor-beta and sonic hedgehog signaling in palatal epithelium regulate tenascin-C expression in palatal mesenchyme during soft palate development. Front Physiol, 2020, 11: 532. doi: 10.3389/fphys.2020.00532.


Refbacks

  • There are currently no refbacks.