scholarly journals SPECC1L-deficient primary mouse embryonic palatal mesenchyme cells show speed and directionality defects

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Jeremy P. Goering ◽  
Dona G. Isai ◽  
Everett G. Hall ◽  
Nathan R. Wilson ◽  
Edina Kosa ◽  
...  
Keyword(s):  
Wound Repair ◽  
Cleft Lip ◽  
Cell Movement ◽  
Live Imaging ◽  
Primary Role ◽  
Palatal Shelf ◽  
Collective Movement ◽  
Primary Mouse ◽  
Mutant Mouse Embryos ◽  
Mesenchyme Cells

AbstractCleft lip and/or palate (CL/P) are common anomalies occurring in 1/800 live-births. Pathogenic SPECC1L variants have been identified in patients with CL/P, which signifies a primary role for SPECC1L in craniofacial development. Specc1l mutant mouse embryos exhibit delayed palatal shelf elevation accompanied by epithelial defects. We now posit that the process of palate elevation is itself abnormal in Specc1l mutants, due to defective remodeling of palatal mesenchyme. To characterize the underlying cellular defect, we studied the movement of primary mouse embryonic palatal mesenchyme (MEPM) cells using live-imaging of wound-repair assays. SPECC1L-deficient MEPM cells exhibited delayed wound-repair, however, reduced cell speed only partially accounted for this delay. Interestingly, mutant MEPM cells were also defective in coordinated cell movement. Therefore, we used open-field 2D cultures of wildtype MEPM cells to show that they indeed formed cell streams at high density, which is an important attribute of collective movement. Furthermore, activation of the PI3K-AKT pathway rescued both cell speed and guidance defects in Specc1l mutant MEPM cells. Thus, we show that live-imaging of primary MEPM cells can be used to assess mesenchymal remodeling defects during palatal shelf elevation, and identify a novel role for SPECC1L in collective movement through modulation of PI3K-AKT signaling.

scholarly journals SPECC1L-deficient palate mesenchyme cells show speed and directionality defect

2019 ◽  
Author(s):  
Jeremy P. Goering ◽  
Dona Greta Isai ◽  
Everett G. Hall ◽  
Nathan R. Wilson ◽  
Edina Kosa ◽  
...  
Keyword(s):  
Wound Repair ◽  
Cell Tracking ◽  
Time Lapse ◽  
Primary Role ◽  
Palatal Shelf ◽  
Collective Movement ◽  
Primary Mouse ◽  
Mesenchyme Cells ◽  
Delayed Closure

AbstractClefts of the lip and/or palate (CL/P) are common anomalies that occur in 1/800 live births. Pathogenic SPECC1L variants identified in patients with rare atypical clefts and syndromic CL/P suggest the gene plays a primary role in face and palate development. We have generated Specc1l gene-trap (Specc1lcGT) and truncation (Specc1lΔC510) alleles that cause embryonic or perinatal lethality, respectively. Specc1lcGT/ΔC510 compound mutants show delayed and abnormal palatal shelf elevation at E14.5. By E15.5, the mutant shelves do elevate and fuse, however, the palatal rugae form abnormally. Palatogenesis requires extensive mesenchymal remodeling, especially during palatal shelf elevation. We posit that this remodeling involves collective movement of neural crest-derived palatal mesenchyme cells. Live time-lapse microscopy was performed to visualize in vitro wound-repair assays with wildtype and SPECC1L-deficient primary mouse embryonic palatal mesenchyme (MEPM) cells. SPECC1L-deficient MEPM cells consistently showed delayed closure in wound-repair assays. To evaluate which features of cellular movement were responsible, we performed automated particle image velocimetry (PIV) and manual cell tracking. The analyses revealed that both cell speed and directionality are disrupted in SPECC1L-deficient cells compared to controls. To determine if primary MEPM cells can move collectively, we assayed stream formation, which is a hallmark of collective movement. Indeed, MEPM cultures displayed correlated movement of neighboring cells. Importantly, correlation length was reduced in SPECC1L-deficient cultures, consistent with a role for SPECC1L in collective migration. Furthermore, we demonstrated that activation of the PI3K-AKT pathway with the 740Y-P small molecule can rescue the wound-closure delay in SPECC1L-deficient MEPM cells. Cell tracking analyses showed that this rescue was due to both increased speed and improved directionality. Altogether, our data showed a novel role for SPECC1L in guided movement through modulation of PI3K-AKT signaling.

Scanning electron microscopy of gastrulation in a sea urchin (Anthocidaris crassispina)

Development ◽  
1982 ◽  
Vol 67 (1) ◽  
pp. 27-35
Author(s):  
Shonan Amemiya ◽  
Koji Akasaka ◽  
Hiroshi Terayama
Keyword(s):  
Electron Microscopy ◽  
Scanning Electron Microscopy ◽  
Cell Formation ◽  
Cell Movement ◽  
Animal Pole ◽  
Vegetal Pole ◽  
Mesenchyme Cell ◽  
Major Motive ◽  
Mesenchyme Cells ◽  
Scanning Electron

Gastrulation in Anthocidaris was investigated by observing the inside and the outside of embryos by scanning electron microscopy. Furrows which possibly rėflect changes in intercellular interactions were observed on the outer surface (hyaline layer side) of embryos twice in development: firstly at the time of primary mesenchyme cell formation, and secondly at the time of vegetal plate indentation. In the latter case, the cells within and surrounding the vegetal plate appeared to change their shapes differently; the former (within the plate) having broader surfaces on the blastocoel side whereas the latter (surrounding the plate) having broader surfaces on the hyaline layer side. This suggests that the first phase of indentation may be mediated by the autonomous change of cell shape and intercellular adhesiveness, accompanied by an autonomous cell movement in the vegetal pole region. Although some pseudopodial linkages were observed between secondary mesenchyme cells on the top of the invaginating archenteron and the animal pole in the mid-gastrula and later stage embryos, they were thinner and smaller in number as compared to those in the Pseudocentrotus embryos. The rate of invagination appeared rather constant throughout gastrulation in contrast to the accelerated invagination in other embryos with larger blastocoel cavities. Moreover, the number of columnar cells on the dissected surface of embryos remained unaltered. These findings suggest that the secondary mesenchyme cells may act as a linker between the archenteron tip and the animal pole, but they may not generate major motive forces for archenteron invagination at least in the Anthocidaris embryos.

scholarly journals Epithelial invagination by vertical telescoping

2019 ◽  
Author(s):  
Jingjing Li ◽  
Andrew D. Economou ◽  
Jeremy B.A. Green
Keyword(s):  
Cell Migration ◽  
Salivary Glands ◽  
Shape Change ◽  
Cell Movement ◽  
Leading Edge ◽  
Apical Constriction ◽  
Extrinsic Compression ◽  
Fundamental Process ◽  
Mesenchyme Cells ◽  
Vertical Cell

AbstractEpithelial bending is a fundamental process that shapes organs during development. All currently known mechanisms involve cells locally changing shape from columnar to wedge-shaped. Often this shape change occurs by cytoskeletal contraction at cell apices (“apical constriction”) but mechanisms such as basal nuclear positioning (“basal wedging”) or extrinsic compression are also known. Here we demonstrate a completely different mechanism which occurs without cell wedging. In mammalian salivary glands and teeth, we show that initial invagination occurs through coordinated vertical cell movement. Specifically, we show that cells towards the periphery of the placode move vertically upwards while their more central neighbours move downwards to create the invagination. We further show that this occurs by active cell-on-cell migration: outer cells migrate with an apical leading edge protrusion, depressing the central cells to “telescope” the epithelium downwards into the underlaying mesenchyme. Cells remain basally attached to the underlying lamina while their apical protrusions are dynamic and planar polarised centripetally. These protrusions depend on the actin cytoskeleton, and inhibition of the branching molecule Arp2/3 inhibits them and the invagination. FGF and Hedgehog morphogen signals are also required, with FGF providing a directional cue. These findings show that epithelial bending can be achieved by novel morphogenetic mechanism of coordinated cell rearrangement quite distinct from previously recognised invagination processes.

scholarly journals Dexamethasone Suppresses Palatal Cell Proliferation through miR-130a-3p

2021 ◽  
Vol 22 (22) ◽  
pp. 12453
Author(s):  
Hiroki Yoshioka ◽  
Goo Jun ◽  
Akiko Suzuki ◽  
Junichi Iwata
Keyword(s):  
Cell Proliferation ◽  
Cleft Palate ◽  
Birth Defects ◽  
Cleft Lip ◽  
Neural Crest Cell ◽  
Congenital Birth Defects ◽  
Cranial Neural Crest Cell ◽  
Primary Mouse ◽  
Mirna Sequencing ◽  
Crest Cell

Cleft lip with or without cleft palate (CL/P) is one of the most common congenital birth defects. This study aims to identify novel pathogenic microRNAs associated with cleft palate (CP). Through data analyses of miRNA-sequencing for developing palatal shelves of C57BL/6J mice, we found that miR-449a-3p, miR-449a-5p, miR-449b, miR-449c-3p, and miR-449c-5p were significantly upregulated, and that miR-19a-3p, miR-130a-3p, miR-301a-3p, and miR-486b-5p were significantly downregulated, at embryonic day E14.5 compared to E13.5. Among them, overexpression of the miR-449 family (miR-449a-3p, miR-449a-5p, miR-449b, miR-449c-3p, and miR-449c-5p) and miR-486b-5p resulted in reduced cell proliferation in primary mouse embryonic palatal mesenchymal (MEPM) cells and mouse cranial neural crest cell line O9-1. On the other hand, inhibitors of miR-130a-3p and miR-301a-3p significantly reduced cell proliferation in MEPM and O9-1 cells. Notably, we found that treatment with dexamethasone, a glucocorticoid known to induce CP in mice, suppressed miR-130a-3p expression in both MEPM and O9-1 cells. Moreover, a miR-130a-3p mimic could ameliorate the cell proliferation defect induced by dexamethasone through normalization of Slc24a2 expression. Taken together, our results suggest that miR-130-3p plays a crucial role in dexamethasone-induced CP in mice.

A role for fibronectin in cell sorting

1984 ◽  
Vol 69 (1) ◽  
pp. 179-197
Author(s):  
P.B. Armstrong ◽  
M.T. Armstrong
Keyword(s):  
Organ Culture ◽  
Control Cell ◽  
Cell Movement ◽  
Mesenchymal Cell ◽  
Surface Zone ◽  
The Matrix ◽  
Chick Heart ◽  
Embryonic Chick Heart ◽  
Embryonic Morphogenesis ◽  
Mesenchyme Cells

A useful approach to the investigation of embryonic morphogenesis is the study of the factors that control cell movement in cell aggregates in organ culture. Previous studies, in which aggregates of embryonic chick heart ventricle tissue were paired in organ culture, supported the hypothesis that the associative behaviour is dominated by the mesenchymal cell (at the stages used the ventricle is composed of approximately 25% mesenchyme (Mes) and 75% myocyte tissue (My)) by virtue of this cell's ability to establish a pericellular matrix rich in fibronectin. In aggregate pairs, the aggregate types that develop a fibronectin-rich matrix rapidly are spread over by the aggregate types that are less able to deposit fibronectin in the matrix. In sorting conditions, Mes sorts to the surface of My. This is explained as a consequence of a requirement that Mes have access to a component in the serum fraction of the culture medium for deposition of fibronectin in the matrix. It is proposed that the factor penetrates to a shallow depth in aggregates, limiting the establishment of a fibronectin-rich matrix to superficially located Mes. As fibronectin appears in the matrix, Mes becomes more cohesive than My, allowing it to exclude myocytes and establish itself as a pure tissue that increases in volume as mesenchyme cells migrating within the interior contact the surface zone, becoming immobilized and also activated to secrete fibronectin. The analysis presented includes an experimental investigation of the different elements of this hypothesis and also explores some of the predictions of the hypothesis.

scholarly journals Sri Lankan Cleft Lip and Palate Project: A Preliminary Report

1988 ◽  
Vol 81 (12) ◽  
pp. 705-709 ◽  
Author(s):  
S P Lambadusuriya ◽  
M Mars ◽  
C M Ward
Keyword(s):  
Sri Lanka ◽  
Older Patients ◽  
Preliminary Report ◽  
Cleft Lip ◽  
Cleft Lip And Palate ◽  
Adult Life ◽  
Psychosocial Impact ◽  
Sri Lankan ◽  
Facial Growth ◽  
Palatal Shelf

By means of a surgical expedition involving an independently financed team of surgeons, anaesthetists, nurses, orthodontists and speech therapists, 195 patients in Sri Lanka with cleft lip and palate were treated over a period of 4 weeks while over 300 patients were examined in detail to assess faciomaxillary growth, components of speech and the psychosocial impact of the untreated deformity in childhood and adult life. Lip surgery proved to be simple and safe but in certain older patients palatal closure was complicated by wide palatal shelf displacement, mucosal fibrosis and heavy bleeding. This is a preliminary report and much data has yet to be analysed but there is little doubt that impaired facial growth following palatal repair is predominantly an iatrogenic deformity.

Atrial Septal Defect in Cyanotic CL/Fr Mice

1998 ◽  
Vol 35 (1) ◽  
pp. 58-64
Author(s):  
Shingo Kadowaki ◽  
Michiko Sakamoto ◽  
Hiroshi Kamiishi ◽  
Takashi Tanimura
Keyword(s):  
Atrial Septal Defect ◽  
Airway Obstruction ◽  
Cleft Lip ◽  
Septal Defect ◽  
Cleft Lip And Palate ◽  
Atrial Septum ◽  
Palatal Shelf ◽  
Icr Mice ◽  
Septum Primum ◽  
Septal Development

Objective It is widely known that some newborn CL/Fr mice with cleft lip and palate (CLP) also have cyanotic symptoms, which have been thought to depend on an atrial septal defect (ASD). In a previous study, we found that cyanotic mice tended to have more severe types of CLP. We hypothesize that the mechanical airway obstruction due to a poorly developed palatal shelf and unmoved tongue in CLP(+) mice might be related to the occurrence of cyanosis. The purpose of this study was to examine the relationships between ASD and cyanosis in CLP(+) newborns. Method The newborn hearts from CLP(-), noncyanotic CLP(+), cyanotic CLP(+), CL/Fr mice and ICR mice were examined histologically, and the incidence and size of ASD was determined on neonatal day (ND) 0. In CLP(-) newborns, similar procedures were performed from ND 1 to ND 4. Furthermore, in CLP(+) newborns, development of the palatal shelf was examined. Results While all the ICR mice had a well-developed atrial septum, and the incidence of ASD was 0%, about 80% of CL/Fr mice had ASD, irrespective of the presence or absence of CLP and cyanosis. On ND 0, the septum primum was significantly shorter in cyanotic CLP(+) mice than in CLP(-) mice. It also tended to be shorter in CLP(+) mice than in CLP(-) mice. Between the cyanotics and noncyanotics, there were no significant differences in the incidences of ASD and the rate of septal development. In CLP(-) mice, the septum primum developed well later and no ASD was observed on ND 4. Cyanotic newborns had significantly less developed palatal shelves than did noncyanotics. Conclusions Cyanosis may not be related to ASD and the rate of septal development, but may be related to the occurrence of CLP in this strain. Furthermore, we confirmed that some relationship exists between the development of the palatal shelf and cyanosis. The present study supports our hypothesis concerning the cause of cyanosis in CL/Fr mice.

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