Cleft lip and palate are the most common congenital malformations in humans. Using 43 staged human embryos and early fetuses ranging from the 4th to 12th week of development, we investigated the development of the lip and palate in order to provide the basic developmental concepts required for managing these anomalies. The lower lip appeared as bilateral mandibular arches at Carnegie stage 11, and these were completely merged at stage 15. The components of the upper lip, medial nasal prominence and maxillary process, appeared at stage 16, and completely merged at stage 20. The median palatine process appeared at stage 16, and the lateral palatine process, at stage 17. The palatine processes and the nasal septum started to fuse abruptly at stage 23, and from external observation seemed to be fused at the 9th week. However, complete fusion did not take place until the 12th week of development. The tongue was prominent at stage 16, showed differentiation of the muscular tissue at stage 21, and was located superior to the lateral palatine process before stage 23. These results may be used in understanding the different mechanisms present in the formation of various congenital anomalies in this region.
Female ; Gestational Age ; Human ; Lip/embryology* ; Palate/embryology* ; Pregnancy

OBJECTIVETo quantitatively compare the relationship between the congenital cleft palate and development of the palatal shelf.

METHODSFifty two pairs palatal shelves were macroscopic measured, and 60 series coronal sections were microscopically measured, which were precisely orientated in the coronal plane and serially sectioned at 7 micro m thickness. With the aid of computer imaging analysis system the widths and areas of the palatal shelves in vertical and coronal direction, the maximal areas of the palatal bone and palatal process and alveolar process were measured and compared quantitatively between the cleft group and non-cleft group.

RESULTSThe widths and areas of palatal shelves in cleft foetuses showed significant reduction macroscopically and microscopically as well as the maximal areas of the palatal bone, in addition, both of two processes of the maxilla showed significant developmental deficiency.

CONCLUSIONSThe palatal shelves show significant developmental hypoplasia in three dimension directions, which have significant correlation between palatal cleft and trisomic condition.

Animals ; Cleft Palate ; embryology ; genetics ; Female ; Image Processing, Computer-Assisted ; Mice ; Palate ; anatomy & histology ; embryology ; Trisomy

INTRODUCTION: Congenital anophthalmos is a very rare malformation. In terms of embryology, congenital anophthalmos is caused by the lack of developing of eye bulb that leads to the absence of vital component of eyeball. In Korea, about 5 cases have been reported. CASE: It was a female newborn who had congenital anophthalmos in the left eye with cleft palate associated with the marks of an eyelid with stricture and sinking in left orbit. We have observed conduction defects on visual evoked potential(VEP) pathways while the right eye responded properly on the photosensitivity test, and showed normal state on the chromosome test. RESULT: We experienced a case of congenital anophthalmos with cleft palate. But we have not found the cause through systemic laboratory or chromosomal study results, the parent's past medical history.
Anophthalmos* ; Cleft Palate* ; Constriction, Pathologic ; Embryology ; Eyelids ; Female ; Humans ; Infant, Newborn ; Korea ; Orbit

OBJECTIVETo investigate the morphologic changes of embryonic palatal development exposed to retinoic acid (RA) in mouse, and to detect the significance of the expression of TGFbeta1, TGFbeta3, EGF and BCL2.

METHODSThe stage of palatal development was examined by light microscopy. S-P immunohistochemistry and in-situ hybridization was used to detect spatio-temporal patterns of expression of TGFbeta1, TGFbeta3, EGF and BCL2 in embryonic palate.

RESULTSThe fetus exposed to RA resulted in formation of small palatal shelves without contact and fusion of each other to form and intact palate. RA can regulate the embryonic palatal expression of genes involved in RA-induced cleft palate.

CONCLUSIONSRA can inhibit the proliferation of MEPM cell to form small palatal shelves and induce abnormal differentiation of MEE cell causing the bi-palatal shelves no contact and fuse with each other, then induce the formation of cleft palate. RA can regulate the spatio-temporal patterns of expression of TGFbeta1, TGFbeta3 and EGF in embryonic palatal processes and the change of special expression of these genes in embryonic palatal processes are involved in RA-induced cleft palate.

Abnormalities, Drug-Induced ; etiology ; Animals ; Cleft Palate ; chemically induced ; embryology ; Embryo, Mammalian ; Epidermal Growth Factor ; biosynthesis ; Female ; Mice ; Mice, Inbred C57BL ; Palate ; embryology ; metabolism ; Transforming Growth Factor beta ; biosynthesis ; Tretinoin ; toxicity

Cleft lip and/or palate (CL/P) is a common congenital malformation with a complex etiology which is not fully elucidated yet. Epidemiological studies point to different etiologies in the cleft lip and palate subgroups, isolated cleft lip (CL), isolated cleft palate (CP) and combined cleft lip and palate (CLP). In order to understand the biological basis in these cleft lip and palate subgroups better we studied the expression profiles in human tissue from patients with CL/P. In each of the CL/P subgroups, samples were obtained from three patients and gene expression analysis was performed. Moreover, selected differentially expressed genes were analyzed by quantitative RT-PCR, and by immunohistochemical staining of craniofacial tissue from human embryos. Osteopontin (SPP1) and other immune related genes were significantly higher expressed in palate tissue from patients with CLP compared to CP and immunostaining in palatal shelves against SPP1, chemokine receptor 4 (CXCR4) and serglycin (PRG1) in human embryonic craniofacial tissue were positive, supporting a role for these genes in palatal development. However, gene expression profiles are subject to variations during growth and therefore we recommend that future gene expression in CL/P studies should use tissue from the correct embryonic time and place if possible, to overcome the biases in the presented study.
Cleft Lip/*genetics/*immunology ; Cleft Palate/embryology/*genetics/*immunology ; Gene Expression Profiling ; Humans ; Immunohistochemistry ; Infant ; Oligonucleotide Array Sequence Analysis ; Osteopontin/*genetics/immunology ; Reverse Transcriptase Polymerase Chain Reaction

OBJECTIVEThe glucocorticoid dexamethasone (DEX) can induce palatal cleft; however, the mechanism involved remains unclear. E-cadherin is an important cell adhesion molecule, and it can significantly affect cell fate and embryonic development. Recent studies have indicated that E-cadherin expression in palatal epithelial cells is suppressed in normal palate fusion. This study aimed to determine whether the change in E-cadherin expression is related to the incidence of cleft palate in DEX-induced mice.

METHODSMice were divided into the experimental group and the control group. Pregnant mice were injected with DEX on E10.0-E12.0, whereas mice in the control group were injected with normal saline. Hematoxylin and eosin (HE) staining, immunohistochemistry, and real-time quantitative polymerase chain reaction were employed to evaluate the effect of DEX on fetal mouse palatal processes, particularly the changes in E-cadherin and β-catenin expression levels in the phases of the experimental and control groups.

RESULTSData indicated that the incidence of cleft palate in the DEX group was 43.59% (17/39), whereas that in the control group was only 3.03% (1/33). The results of HE staining showed that the obviously shortened palatal processes could not contact and fuse with one another in the DEX-treated mice model compared with those in the control group. The ectopic expression of E-cadherin in embryonic palatal mesenchymal cells was also analyzed. The expression levels of E-cadherin and β-catenin in the experimental group were higher than those in the control group.

CONCLUSIONThese findings indicated that DEX could induce E-cadherin gene upregulation and ectopic expression, as well as high β-catenin expression, thereby inhibiting the growth of mesenchyme cells and cleft palate.

Animals ; Cadherins ; genetics ; metabolism ; Cleft Palate ; chemically induced ; embryology ; Dexamethasone ; adverse effects ; Disease Models, Animal ; Epithelial Cells ; Female ; Glucocorticoids ; Immunohistochemistry ; Mice ; Pregnancy ; beta Catenin ; metabolism

OBJECTIVETo test the changing expression level of Annexin I, cPLA(2) and PCNA in the palatine process of cleft-palate mice A/J and C57B/6j induced by dexamethasone. To discuss the developing mechanism of cleft-palate and the corresponding preventive methods.

METHODs Pregnant mice A/J and C57BL/6j were randomly divided into normal group as blank control, group with deformity induced by dexamethasone, group given VitB(12) as antagonist to deforming factor, group given only VitB(12). The relative quantity of Annexin I, Cpla2 and PCNA were detected by immunoblotting.

RESULTSAmong mice A/J, with the development of palatine process, Annexin I's expression level was increased in the normal group, and other groups showed the similar changes. Annexin I's expression level was significantly higher in group DEX and group DEX + VitB(12) than in normal group and VitB(12) group of, while there was no significant difference between normal group and VitB(12) group. But the changes of cPLA(2) and PCNA expression level was in an opposite direction, with development it decreased in the normal mice's palatine process. In mice C57B/6j there was no significant difference between normal group and group DEX on the measured quantity of Annexin I, cPLA(2) and

CONCLUSIONSAnnexin I and cPLA(2) introduce glucocorticoid to induce cleft-palate. VitB(12) can not inhibit DEX's enhancing effect on the expression level of Annexin I, but it can antagonize DEX's inhibiting effect on expression level of cPLA(2), which is probably one of the mechanisms why VitB(12) antagonize glucocorticoid's deforming effect.

Abnormalities, Drug-Induced ; etiology ; Animals ; Annexin A1 ; biosynthesis ; Cleft Palate ; chemically induced ; Dexamethasone ; toxicity ; Female ; Male ; Mice ; Mice, Inbred C57BL ; Palate ; embryology ; metabolism ; Phospholipases A ; biosynthesis ; Pregnancy ; Proliferating Cell Nuclear Antigen ; biosynthesis ; Random Allocation

OBJECTIVETo study the effect of dexamethasone on the differentiation and proliferation of type A mouse palatal medial edge epithelial cells when there is type A mouse embryonic palatal mesenchymal cells.

METHODSThe mouse palatal shelves were harvested from a female mouse of gestation day 14 by microsurgical dissection and cultured in vitro. The differentiation was investigated through microscope and transmission electron microscope under condition of the palatal shelves fusion.

RESULTSDexamethasone promoted the palatal medial edge epithelium differentiated into squamause epithelium and affected normal development and obstructed the fusion of mouse palatal shelves.

CONCLUSIONThe results of histological observation indicate that dexamethasone promotes the proliferation of palatal meseuchymal cells and inhibits the normal differentiation of palatal medial edge epithelial cells, which results in cleft palate.

Animals ; Cell Differentiation ; drug effects ; Cell Proliferation ; drug effects ; Cleft Palate ; chemically induced ; Dexamethasone ; adverse effects ; pharmacology ; Epithelial Cells ; Epithelium ; Female ; Glucocorticoids ; adverse effects ; pharmacology ; Mice ; Organ Culture Techniques ; Palate, Hard ; drug effects ; embryology

Excessive dexamethasone (Dex) administrated into pregnant mice during critical periods of palatal development can produce a high incidence of cleft palate. Its mechanisms remain unknown. Vitamin B12 has been shown to antagonize the teratogenic effects of Dex, which, however, remains controversial. In this study, we investigated the effects of Dex and vitamin B12 on murine embryonic palatal shelf fusion using organ culture of murine embryonic shelves. The explanted palatal shelves on embryonic day 14 (E14) were cultured for 24, 48, 72 or 96 h in different concentrations of Dex and/or vitamin B12. The palatal shelves were examined histologically for the morphological alterations on the medial edge epithelium (MEE) and fusion rates among different groups. It was found that the palatal shelves were not fused at 72 h or less of culture in Dex group, while they were completely fused in the control and vitamin B12-treated groups at 72 and 96 h, respectively. The MEE still existed and proliferated. In Dex+vitamin B12 group the palatal shelves were fused at each time point in a similar rate to controls. These results may suggest that Dex causes teratogenesis of murine embryonic palatal shelves and vitamin B12 prevents the teratogenic effect of Dex on palatogenesis on murine embryos in vitro.
Animals ; Cell Proliferation ; Cleft Palate ; embryology ; metabolism ; prevention & control ; Dexamethasone ; toxicity ; Epithelial Cells ; cytology ; Female ; Glucocorticoids ; pharmacology ; Male ; Mice ; Mice, Inbred C57BL ; Microscopy, Electron, Scanning ; Organ Culture Techniques ; methods ; Teratogens ; Time Factors ; Vitamin B 12 ; metabolism

We used Smo siRNA to inhibit hedgehog signaling pathway in embryonic day (E) 13 palatal shelves in organ culture. SiRNA 4 was chosen as the most efficient from four synthesized Smo siRNAs. Palatal shelf fusion rate of 4 μg/mL cyclopamine group was the lowest and significantly lower than that of blank control group (P<0.05), and that of siRNA 4 group was also lower than that of blank control group (P=0.183). At 48 h after transfection, Smo protein level of siRNA 4 group was 64.8% lower than that of blank control group (P<0.05), and Gli1 protein level of 4 μg/mL cyclopamine group was 68.9% lower than that of blank control group (P<0.05). Hedgehog signaling pathway inhibition decreased palatal fusion in organ culture, probably owing to downregulation of Smo and Gli1 proteins.
Animals ; Hedgehog Proteins ; genetics ; metabolism ; Kruppel-Like Transcription Factors ; genetics ; metabolism ; Mice ; Nerve Tissue Proteins ; genetics ; metabolism ; Palate ; embryology ; metabolism ; RNA, Small Interfering ; genetics ; metabolism ; Signal Transduction ; Zinc Finger Protein Gli2 ; Zinc Finger Protein Gli3