Biomedical subjects

Han-Sung Jung

Publications and source records attributed to Han-Sung Jung.

16 recordsLinked to original sources

PubMed2006

Molecular interactions between Tbx3 and Bmp4 and a model for dorsoventral positioning of mammary gland development.

The formation of the dorsoventral (DV) boundary is central to establishing the body plan in embryonic development. Although there is some information about how limbs are positioned along the DV axis and how DV skin color pattern is determined, the way in which mammary glands are positioned is unknown. Here we focus on Bmp4 and Tbx3, a gene associated with ulnar-mammary syndrome, and compare their expression along the DV axis in relation to mammary gland initiation in mouse embryos. Tbx3 is expressed in the mammary gland-forming region with Tbx15, a gene involved in a DV coat color being expressed more dorsally and Bmp4 being expressed more ventrally. When Tbx3 was overexpressed, formation of mammary gland epithelium was extended along the DV axis. In contrast, overexpression of Bmp4 inhibited both Tbx3 and Tbx15 expression. In addition, when BMP signaling was inhibited by NOGGIN, Lef1 expression was lost. Thus, we propose that mutual interactions between Bmp4 and Tbx3 determine the presumptive DV boundary and formation of mammary glands in early mouse embryogenesis. 1,19-Dioctadecyl-3,3,39,39-tetramethyl indocarbocyanine perchloride labeling experiments showed that cells associated with mammary glands originate more dorsally and then move ventrally. This finding, together with previous findings, suggests that the same DV boundary may not only position limbs and determine coat color but also position mammary glands. Furthermore, Bmp signaling appears to be a fundamental feature of DV patterning.

Animals↗

PubMed2006

Modulation of cell proliferation during palatogenesis by the interplay between Tbx3 and Bmp4.

During secondary palate development, two shelves are elevated to a horizontal position above the tongue through a process involving many cellular mechanisms, including proliferation. In particular, the expression patterns of Tbx3 and Bmp4, which are colocalized at embryonic day 13.5 (E13.5) and have unique expression patterns in specific regions at E14.5, have been investigated in early mouse palatogenesis. Tbx3 expression is reported to be associated with Bmp4 signaling during the process of organogenesis in other areas, such as limb development. However, the function of Tbx3 and the relationship between Tbx3 and Bmp4 in palate development have not been determined. We have examined the gene expression pattern and cell proliferation in order to understand the mutual interactions and function of Tbx3 and Bmp4. An electroporation method was used to investigate the altered pattern of these genes after their over-expression in organ cultures. NOGGIN protein-soaked beads were also implanted into the cultured palate to determine the function of Bmp4 in palatogenesis. After electroporation and NOGGIN bead implantation, the number of PCNA-positive cells was counted. The results showed that Tbx3 and Bmp4 strongly up- and down-regulated each other in order to control the proliferation of the palatal shelf. Thus, Tbx3 expression is induced by Bmp4 in the mesenchyme of the anterior palatal shelves, whereas mesenchymal expression of Tbx3 down-regulates Bmp4 expression in the mesenchyme of the palate. The harmonization between Tbx3 and Bmp4 therefore controls cell proliferation to regulate secondary palate development.

Animals↗

PubMed2006

Capacity of dental pulp differentiation after tooth transplantation.

Under pathological conditions, dental pulp elaborates both bone and dentin matrix in which the contribution of periodontal tissue cannot be excluded. This study has aimed to clarify the capability of dental pulp to deposit bone matrix in an auto-graft experiment by using (1) immunohistochemistry for 5-bromo-2'-deoxyuridine (BrdU) and nestin and (2) histochemistry for tartrate-resistant acid phosphatase (TRAP). Following the extraction of the molars of 3-week-old mice, the roots and pulp floor were resected and immediately transplanted into the sublingual region. On Days 5-7, tubular dentin formation commenced next to the pre-existing dentin at the pulp horn in which nestin-positive odontoblast-like cells were arranged. Up until Day 14, bone-like tissue formation occurred in the pulp chamber in which intense TRAP-positive cells appeared. These results suggest that odontoblast- and osteoblast-lineage cells reside in the dental pulp. Overall, specific dental pulp regeneration should provide fundamental knowledge for the realization of human tooth regeneration in the near future.

Acid Phosphatase↗

PubMed2006

Association of Shh and Ptc with keratin localization in the initiation of the formation of circumvallate papilla and von Ebner's gland.

The development of gustatory papillae in mammalian embryos requires the coordination of a series of morphological events, such as proliferation, differentiation and innervation. In mice, the circumvallate papilla (CVP) is a specialized structure that develops in a characteristic spatial and temporal pattern in the posterior region of the tongue dorsal surface. The distinct expression patterns of Shh and Ptc, which play important roles in the development of other epithelial appendages, have been localized in the trench wall that gives rise to von Ebner's gland (VEG). To define the cellular mechanisms responsible for morphogenesis and differentiation during early development of CVP and VEG, the localization patterns of keratins (cytokeratins) K7, K8, K18, K19, K14 and connexin-43, which are dependent on Shh expression in other developmental systems, have been examined in detail. The distinct localization of keratins K7, K8, K18, K19, K14 and connexin-43 in the epithelium giving rise to the CVP and VEG suggests that cytodifferentiation is established prior to morphological changes. Interestingly, the localization of proliferating cell nuclear antigen, a marker for cell proliferation, is similar to that of Shh. An understanding of the regulatory roles of cell-cell interactions and signalling molecules in orchestrating a mutual network will bring us nearer to defining the molecular and cellular mechanisms underlying morphogenesis in mammalian taste bud development.

Animals↗

PubMed2005

Inhibition of connexin 43 alters Shh and Bmp-2 expression patterns in embryonic mouse tongue.

The morphogenesis of fungiform papillae occurs in a stereotyped pattern on the dorsal surface of the mammalian tongue via epithelial-mesenchymal interactions. These interactions are thought to be achieved via intercellular communication. Gap junctions can be observed in many developing tissues and have been suggested to participate in a variety of functions, including the regulation of cell proliferation, differentiation, and apoptosis. Here, we demonstrate that the expression of Connexin 43 (Cx43), a gap junction protein, is correlated significantly with the development of fungiform papillae, which exhibit a pattern formation and morphogenesis similar to the development of other epithelial appendages. Antisense-oligodeoxynucleotide (AS-ODN) against Cx43 was used to assess the developmental functions of Cx43. The expression patterns of the signaling molecules were disrupted by Cx43 inhibition. Interestingly, the expression patterns of Shh, a key molecule in the determination of the spacing patterns of fungiform papillae, were disturbed after treatment with Cx43 AS-ODN. We have also attempted to determine the functions of Bmp-2 by applying NOGGIN protein to tongue cultures. Our results indicate that upstream regulation via Cx43 controls the Shh and Bmp-2 pathways for the morphogenesis and pattern formation of fungiform papillae.

Animals↗

PubMed2005

Characteristic tissue interaction of the diastema region in mice.

Rodents have a toothless diastema between the incisor and the first molar, which may contain rudimentary tooth germs. In the lower diastema region of mice at E13, the rudimentary tooth germs, which developed into the bud stage before its removal by apoptosis, was found. The immunoreactivity to tenascin was observed in the condensed mesenchyme around the normal tooth bud and was detected in only the basement membrane in the diastema bud. This result shows that the relationship between mesenchymal condensation and tooth development. The similar patterns of Msx-1 and Msx-2 expression between the tooth bud and the diastema bud show that the diastema bud may have some other genetic mechanism in the developmental arrest of the rudimentary tooth germs rather than the Msx-1 and Msx-2 expression. Strikingly, the induction of the tooth formation was possible using tissue recombination between the oral epithelium of the diastema bud and the dental mesenchyme of the molar tooth bud, which indicates the potential capability of the diastema in the tooth formation. In conclusion, it is suggested that the condensed mesenchyme may be the key to tooth development.

Animals↗

PubMed2005

Formation of spacing pattern and morphogenesis of chick feather buds is regulated by cytoskeletal structures.

Chick feather buds develop sequentially in a hexagonal array. Each feather bud develops with anterior posterior polarity, which is thought to develop in response to signals derived from specialized regions of mesenchymal condensation and epithelial thickening. These developmental processes are performed by cellular mechanisms, such as cell proliferation and migration, which occur during chick feather bud development. In order to understand the mechanisms regulating the formation of mesenchymal condensation and their role in feather bud development, we explanted chick dorsal skin at stage HH29+ with cytochalasin D, which inhibits cytoskeletal formation. We show that the aggregation of mesenchymal cells can be prevented by cytochalasin D treatment in a concentration-dependent manner. Subsequently, cytochalasin D disrupts the spacing pattern and inhibits feather bud axis formation as well. In addition, expression patterns of Bmp-4 and Msx-2, key molecules for early feather bud development, were disturbed by cytochalasin D treatment. Our results fully indicate that both the cytoskeletal structure and cell activity via gene regulation are of fundamental importance in mesenchymal condensation leading to proper morphogenesis of feather bud and spacing pattern formation.

Animals↗

PubMed2004

Interactions between FGF and Wnt signals and Tbx3 gene expression in mammary gland initiation in mouse embryos.

Interactions between Wnts, Fgfs and Tbx genes are involved in limb initiation and the same gene families have been implicated in mammary gland development. Here we explore how these genes act together in mammary gland initiation. We compared expression of Tbx3, the gene associated with the human condition ulnar-mammary syndrome, expression of the gene encoding the dual-specificity MAPK phosphatase Pyst1/MKP3, which is an early response to FGFR1 signalling (as judged by sensitivity to the SU5402 inhibitor), and expression of Lef1, encoding a transcription factor mediating Wnt signalling and the earliest gene so far known to be expressed in mammary gland development. We found that Tbx3 is expressed earlier than Lef1 and that Pyst1 is also expressed early but only transiently. Patterns of expression of Tbx3, Pyst1 and Lef1 in different glands suggest that the order of mammary gland initiation is 3, 4, 1, 2 and 5. Consistent with expression of Pyst1 in the mammary gland, we detected expression of Fgfr1b, Fgf8 and Fgf9 in both surface ectoderm and mammary bud epithelium, and Fgf4 and Fgf17 in mammary bud epithelium. Beads soaked in FGF-8 applied to the flank of mouse embryos, at a stage just prior to mammary bud initiation, induce expression of Pyst1 and Lef1 and maintain Tbx3 expression in flank tissue surrounding the bead. Grafting beads soaked in the FGFR1 inhibitor, SU5402, abolishes Tbx3, Pyst1 and Lef1 expression, supporting the idea that FGFR1 signalling is required for early mammary gland initiation. We also showed that blocking Wnt signalling abolishes Tbx3 expression but not Pyst1 expression. These data, taken together with previous findings, suggest a model in which Tbx3 expression is induced and maintained in early gland initiation by both Wnt and Fgf signalling through FGFR1.

Animals↗

PubMed2004

Spacing patterns on tongue surface-gustatory papilla.

The dorsal surface of the mammalian tongue is covered with four kinds of papillae, fungiform, circumvallate, foliate and filiform papillae. With the exception of the filiform papillae, these types of papillae contain taste buds and are known as the gustatory papillae. The gustatory papillae are distributed over the tongue surface in a distinct spatial pattern. The circumvallate and foliate papillae are positioned in the central and lateral regions respectively and the fungiform papillae are distributed on the anterior part of the tongue in a stereotyped array. The patterned distribution and developmental processes of the fungiform papillae indicate some similarity between the fungiform papillae and the other epithelial appendages, including the teeth, feathers and hair. This is because 1) prior to the morphological changes, the signaling molecules are expressed in the fungiform papillae forming area with a stereotyped pattern; 2) the morphogenesis of the fungiform papillae showed specific structures in early development, such as epithelial thickening and mesenchymal condensation and 3) the fungiform papillae develop through reciprocal interactions between the epithelium and mesenchymal tissue. These results led us to examine whether or not the early organogenesis of the fungiform papillae is a good model system for understanding both the spacing pattern and the epithelial-mesenchymal interaction during embryogenesis.

Animals↗

PubMed2004

Integument pattern formation involves genetic and epigenetic controls: feather arrays simulated by digital hormone models.

Pattern formation is a fundamental morphogenetic process. Models based on genetic and epigenetic control have been proposed but remain controversial. Here we use feather morphogenesis for further evaluation. Adhesion molecules and/or signaling molecules were first expressed homogenously in feather tracts (restrictive mode, appear earlier) or directly in bud or inter-bud regions ( de novo mode, appear later). They either activate or inhibit bud formation, but paradoxically colocalize in the bud. Using feather bud reconstitution, we showed that completely dissociated cells can reform periodic patterns without reference to previous positional codes. The patterning process has the characteristics of being self-organizing, dynamic and plastic. The final pattern is an equilibrium state reached by competition, and the number and size of buds can be altered based on cell number and activator/inhibitor ratio, respectively. We developed a Digital Hormone Model which consists of (1) competent cells without identity that move randomly in a space, (2) extracellular signaling hormones which diffuse by a reaction-diffusion mechanism and activate or inhibit cell adhesion, and (3) cells which respond with topological stochastic actions manifested as changes in cell adhesion. Based on probability, the results are cell clusters arranged in dots or stripes. Thus genetic control provides combinational molecular information which defines the properties of the cells but not the final pattern. Epigenetic control governs interactions among cells and their environment based on physical-chemical rules (such as those described in the Digital Hormone Model). Complex integument patterning is the sum of these two components of control and that is why integument patterns are usually similar but non-identical. These principles may be shared by other pattern formation processes such as barb ridge formation, fingerprints, pigmentation patterning, etc. The Digital Hormone Model can also be applied to swarming robot navigation, reaching intelligent automata and representing a self-re-configurable type of control rather than a follow-the-instruction type of control.

Animals↗

PubMed2003

A variation of the musculocutaneous nerve absent.

A variation of the brachial plexus, characterized by the absence of the musculocutaneous nerve on the left arm, was found during the dissection of a 28-year old male cadaver. The whole lateral cord was joined to the median nerve, which it met in two points. One was a typical junction of both roots of the median nerve at the level of the coracoid process. The other was a junction of the remaining lateral cord and the median nerve, which was 92 mm away from the typical junction. This case provided some evidence about the absence of the musculocutaneous nerve, rather than a complete fusion of the median and musculocutaneous nerves. As the nerves are named due to their course or innervation, and not from their origin, it is reasonable to assume that the combined nerve was actually the median nerve, and that the musculocutaneous nerve did not exist.

Adult↗

PubMed2003

Morphological evidence of the importance of epithelial tissue during mouse tongue development.

The morphogenesis of fungiform papillae occurs in a stereotyped pattern on the dorsal surface of the tongue in mice from embryonic day 12 (E12) to E17. The histological results and ultrastructural observations showed the development of specific structures in the epithelium into fungiform papillae. Prior to the morphological changes, the Bmp-4 and Shh transcripts are expressed in a restricted area on the dorsal surface. These results suggest that the development of fungiform papillae requires an epithelium and mesenchyme interaction during morphogenesis. In order to obtain direct evidence of the epithelium and mesenchyme interaction during tongue papillae morphogenesis, the formation of fungiform papillae was examined after a recombination assay. In order to confirm the epithelium and mesenchyme interactions during the early development of the mouse tongue, a recombination assay was conducted after the recombination assay at E12.5 and E13.5 for 2 days using an in vitro organ culture. From the recombination assay results, the E13.5 epithelial portion of the fungiform papillae could determine the position of the newly formed fungiform papillae with the epithelial signaling molecules. E13.5 was a critical stage for fungiform papillae morphogenesis. Fungiform papillae can be considered to be small epithelial appendages, which are formed via the epithelium and mesenchyme interactions.

Animals↗

PubMed2003

Study on tooth development, past, present, and future.

For decades, the understanding of craniofacial development has been a central issue in odontology and developmental biology. As a consequence, a significant number of deformities are being studied for their variety of genotype and phenotype. Although there is little doubt about the essential roles of homeobox genes, transcription factors, and growth factors, we now know at least the fundamental strategy of craniofacial biology. The tooth as an organ performs a whole range of functions, each of which is truly indispensable for the maintenance of life. The possession of teeth is, therefore, obviously coupled with the complication of the natural structure of an individual organism. In the following, we shall focus on a brief history of tooth studies and some suggestions for obtaining a full understanding of teeth in the future.

Gene Expression Regulation, Developmental↗

PubMed2003

Analysis of tooth formation by reaggregated dental mesenchyme from mouse embryo.

Tooth morphogenesis is a well-known developmental system related to epithelial-mesenchymal interactions. In mice, the dental epithelium has the potential to induce tooth formation prior to the bud stage, whereas this potential shifts to the dental mesenchyme from the dental epithelium. The reaggregation of mesenchymal tissue leads to previous memories of individual cells being reset, which is useful for studying the predetermination of mesenchyme. Here, the mesenchyme was triturated into single cells after separation of the epithelium and the mesenchyme. These single cells were repelleted and combined with the epithelium. The reaggregated tooth was transplanted into a mice kidney capsule. In order to investigate the essential functions of both the dental epithelium and the dental mesenchyme regarding their mutual interaction, a reaggregation system was introduced using the late bud stage of the mouse first molar. Amelogenin expression was examined to confirm the cytodifferentiation in the reaggregated tooth. The results showed that a new tooth formed after reaggregating the dental mesenchyme. This tooth contained enamel, dentin, dentinal tubules and dental pulp. The inner enamel epithelium of the reaggregated tooth differentiated into ameloblasts. Immunohistochemistry for amelogenin was observed both in the ameloblasts and the enamel. However, the structure of the enamel was different from that of the normal tooth, with the thickness of the predentin becoming wider. These findings suggest that reaggregated dental mesenchyme cells can produce a tooth. The fate of dental epithelium was not affected by reaggregated dental mesenchyme, although the dental mesenchyme appears to lose the information from the dental epithelium.

Amelogenin↗

PubMed2003

Lineage of non-cranial neural crest cell in the dental mesenchyme: using a lacZ reporter gene during early tooth development.

The tooth is one of the ectodermal organs controlled by reciprocal interactions between the epithelium and the mesenchyme. Mesenchymal cells in the developing tooth, so-called dental mesenchymal cells, are derived from two different origins: the cranial neural crest (CNC) and the non-CNC. These CNC-derived cells migrate, proliferate and differentiate into odontoblasts, cementoblasts, fibroblasts, osteoblasts and chondroblasts. Tooth germs of wild-type mice were transplanted into the kidney of adult lacZ-transgenic mice. After 1 week of transplantation, a few lacZ-expressing cells and many red blood cells were found near or inside the blood vessels in the pulp of wild-type tooth germs. This result shows that circulating cells of the adult host could invade the dental pulp during tooth development, through the blood vessels, and be a part of dental pulp tissue. Therefore, it can be suggested that these circulating progenitor cells could be the origin of non-CNC-derived cells in tooth germ and their migration pathways would be the blood vessels invading the dental pulp during tooth development. If variations of this experiment were suitably adjusted, such as the embryonic stage of the tooth germ, duration of transplantation, etc., this transplantation experiment using adult lacZ-transgenic mice could be a good system to reveal the origin and migration pathway of cells in developing organs as well as in dental mesenchymal cells.

Animals↗

PubMed2002

Regulation of hair follicle development by the TNF signal ectodysplasin and its receptor Edar.

X-linked and autosomal forms of anhidrotic ectodermal dysplasia syndromes (HED) are characterized by deficient development of several ectodermal organs, including hair, teeth and exocrine glands. The recent cloning of the genes that underlie these syndromes, ectodysplasin (ED1) and the ectodysplasin A receptor (EDAR), and their identification as a novel TNF ligand-receptor pair suggested a role for TNF signaling in embryonic morphogenesis. In the mouse, the genes of the spontaneous mutations Tabby (Ta) and downless (dl) were identified as homologs of ED1 and EDAR, respectively. To gain insight into the function of this signaling pathway in development of skin and hair follicles, we analyzed the expression and regulation of Eda and Edar in wild type as well as Tabby and Lef1 mutant mouse embryos. We show that Eda and Edar expression is confined to the ectoderm and occurs in a pattern that suggests a role of ectodysplasin/Edar signaling in the interactions between the ectodermal compartments and the formation and function of hair placodes. By using skin explant cultures, we further show that this signaling pathway is intimately associated with interactions between the epithelial and mesenchymal tissues. We also find that Ta mutants lack completely the placodes of the first developing tylotrich hairs, and that they do not show patterned expression of placodal genes, including Bmp4, Lef1, Shh, Ptch and Edar, and the genes for beta-catenin and activin A. Finally, we identified activin as a mesenchymal signal that stimulates Edar expression and WNT as a signal that induces Eda expression, suggesting a hierarchy of distinct signaling pathways in the development of skin and hair follicles. In conclusion, we suggest that Eda and Edar are associated with the onset of ectodermal patterning and that ectodysplasin/edar signaling also regulates the morphogenesis of hair follicles.

Activins↗