Cranial Ectoderm Research Articles

Distribution of the titf2/foxe1 gene product is consistent with an important role in the development of foregut endoderm, palate, and hair.

Titf2/foxe1 is a forkhead domain-containing gene expressed in the foregut, in the thyroid, and in the cranial ectoderm of the developing mouse. Titf2 null mice exhibit cleft palate and either a sublingual or completely absent thyroid gland. In humans, mutations of the gene encoding for thyroid transcription factor-2 (TTF-2) result in the Bamforth syndrome, characterized by thyroid agenesis, cleft palate, spiky hair, and choanal atresia. Here, we report a detailed expression pattern of TTF-2 protein during mouse embryogenesis and show its presence in structures where it has not been described yet. At embryonic day (E) 10.5, TTF-2 is expressed in Rathke's pouch, in thyroid, and in the epithelium of the pharyngeal wall and arches, whereas it is absent in the epithelium of the pharyngeal pouches. According to this expression, at E13.5, TTF-2 is present in endoderm derivatives, such as tongue, palate, epiglottis, pharynx, and oesophagus. Later in embryogenesis, we detect TTF-2 in the choanae and whiskers. This pattern of expression helps to define the complex phenotype displayed by human patients. Finally, we show that TTF-2 is a phosphorylated protein. These results help to characterize the domains of TTF-2 expression, from early embryogenesis throughout organogenesis, providing more detail on the potential role of TTF-2 in the development of endoderm and ectoderm derived structures.

Development of the vertebrate inner ear.

The inner ear, also called the membranous labyrinth, contains the cochlea, which is responsible for the sense of hearing, and the vestibular apparatus, which is necessary for the sense of balance and gravity. The inner ear arises in the embryo from placodes, which are epithelial thickenings of the cranial ectoderm symmetrically located on either side of hindbrain rhombomeres 5 and 6. Placode formation in mice is first visible at the 12-somite stage and is controlled by surrounding tissues, the paraxial mesoderm and neural ectoderm. Diffusible molecules such as growth factors play an important role in this process. The activity of several genes confers the identity to the placodal cells. Subsequent cellular proliferation processes under influences from the adjacent hindbrain cause the inner ear epithelium to invaginate and form a vesicle called the otocyst. Combinatorial expression of several genes and diffusible factors secreted from the vesicle epithelium and hindbrain control specification of distinct inner ear compartments. Transplantation studies and inner ear in vitro cultures show that each of these compartments is already committed to develop unique inner ear structures. Later developmental periods are principally characterized by intrinsic differentiation processes. In particular, sensory patches differentiate into fully functional sensory epithelia, and the semicircular canals along with the cochlear duct are elaborated and ossified.

Competence, specification and commitment in otic placode induction.

The inner ear is induced from cranial ectoderm adjacent to the hindbrain. Despite almost a century of study, the molecular mechanisms of inner ear induction remain obscure. We have identified four genes expressed very early in the anlage of the inner ear, the otic placode. Pax-2, Sox-3, BMP-7 and Notch are all expressed in placodal ectoderm from the 4-5 somite stage (ss) onwards, well before the otic placode becomes morphologically visible at the 12-14ss. We have used these four molecular markers to show that cranial ectoderm becomes specified to form the otic placode at the 4-6ss, and that this ectoderm is committed to a placodal fate by the 10ss. We also demonstrate that much of the embryonic ectoderm is competent to generate an otic placode if taken at a sufficiently early age. We have mapped the location of otic placode-inducing activity along the rostrocaudal axis of the embryo, and have determined that this activity persists at least until the 10ss. Use of the four molecular otic placode markers suggests that induction of the otic placode in birds occurs earlier than previously thought, and proceeds in a series of steps that are independently regulated.

Characterization of avian frizzled genes in cranial placode development

To determine the possible role of Wnt signaling in cranial placode development, we have cloned several chick frizzled genes, a family of putative Wnt receptor molecules, and analyzed their expression during chick embryogenesis. Chick frizzled-2 (cFz-2) and frizzled-7 (cFz-7) are expressed broadly in cranial ectoderm, tissue that is competent to express markers of the trigeminal placode (Stark et al., 1997. Development 124, 4287–4295; Baker et al., 1999. Development 126, 147–156). In addition, cFz-2 and cFz-7 are uniquely expressed in other cranial placodes, including the olfactory, lens, and otic placodes. Chick frizzled-1 (cFz-1) is expressed in the lens, otic placode and, along with cFz-7, in epibranchial placodes. Each frizzled gene expressed in the otic placode displays a unique domain of expression: cFz-1 transcripts are detected in the medial wall of the vesicle, cFz-2 in the rostral rim of the vesicle, and cFz-7 in the lateral half of the vesicle. Other chick frizzled family members cloned that do not show striking expression in cranial placodes include frizzled-4 (cFz-4), frizzled-8 (cFz-8), frizzled-9 (cFz-9), and frizzled-10 (cFz-10). A brief summary of their expression is given, along with a brief summary of non-placodal expression of cFz-1, cFz-2, and cFz-7. In all, frizzled genes show dynamic expression at key times during embryonic development, particularly in the cranial placodes.

Spatiotemporal Expression Pattern of Keratins in Skin of AP-2α-deficient Mice

Transcription factor AP-2alpha has been implicated as being a cell-type-specific regulator of gene expression during vertebrate embryogenesis based on its expression pattern in neural crest cells, ectoderm, and the nervous system in mouse and frog embryos. In mice, AP-2alpha is expressed in surface ectoderm beginning at the single cell layer state around E8.75. AP-2alpha-deficient mice, derived by targeted mutagenesis, display a severe ventral closure defect resulting in cranio-abdominoschisis and a hypoplasia of the cranial ganglia. This study analyzed the effect of a targeted disruption of the AP-2alpha gene on the architecture and the expression of intermediate filaments in skin. We analyzed skin samples from newborn mice and found no difference in either the morphology of the skin or the amount of intermediate filaments expressed. This suggests that despite the results from other analyses, loss of transcription factor AP-2alpha does not affect the expression of intermediate filaments in the skin of newborn animals. We found an altered spatial distribution of intermediate filament expression in the single layered cranial ectoderm during days 9-12 of gestation leading to an evenly distributed expression of keratin 5 and 15 in the mutants. Furthermore, the mutants lack a ring of ectodermal cells highly positive for keratin 15 in the area where lens induction occurs, indicating a defect in the inductive interactions underlying eye formation.

Induction of the epibranchial placodes.

The cranial sensory ganglia, in contrast to those of the trunk, have a dual embryonic origin arising from both neurogenic placodes and neural crest. Neurogenic placodes are focal thickenings of ectoderm, found exclusively in the head of vertebrate embryos. These structures can be split into two groups based on the positions that they occupy within the embryo, dorsolateral and epibranchial. The dorsolateral placodes develop alongside the central nervous system, while the epibranchial placodes are located close to the top of the clefts between the branchial arches. Importantly, previous studies have shown that the neurogenic placodes form under the influence of the surrounding cranial tissues. In this paper, we have analysed the nature of the inductive signal underlying the formation of the epibranchial placodes. We find that epibranchial placodes do not require neural crest for their induction, but rather that it is the pharyngeal endoderm that is the source of the inductive signal. We also find that, while cranial ectoderm is competent to respond to this inductive signal, trunk ectoderm is not. We have further identified the signalling molecule Bmp7 as the mediator of this inductive interaction. This molecule is expressed in a manner consistent with it playing such a role and, when added to ectoderm explants, it will promote the formation of epibranchial neuronal cells. Moreover, the Bmp7 antagonist follstatin will block the ability of pharyngeal endoderm to induce placodal neuronal cells, demonstrating that Bmp7 is required for this inductive interaction. This work answers the long standing question regarding the induction of the epibranchial placodes, and represents the first elucidation of an inductive mechanism, and a molecular effector, underlying the formation of any primary sensory neurons in higher vertebrates.

The differentiation of the olfactory placode in Xenopus laevis: A light and electron microscope study

AbstractThe origin and development of the olfactory neuroepithelium of Xeno‐pus laevis has been investigated with light microscopy and scanning and transmission electron microscopy.The cranial ectoderm of the Xenopus laevis embryo consists of two layers: The inner layer is called the sensory or nervous layer (NL) and the outer layer is called the nonnervous layer of the ectoderm (NNE). Between stages 23 and 26, the presumptive olfactory region consists of two discretely arranged cellular populations: the cells of the olfactory placode proper derived from the NL and the cells of the overlying NNE. After stage 26 the placodal cells begin to move toward the body surface by migrating between the NNE cells. As the placodai cells migrate they sprout an apical process which reaches the body surface at approximately stage 28. The apical process contains free and membrane‐bound ribosomes, elongated mitochondria, and microtubules oriented parallel to the process. Centrioles and basal bodies are present in the apical cytoplasm and microvilli and cilia project from the process's surface. The structure and ultrastructure of the apical process identify it as the dendrite of an olfactory neuron. As the apical process reaches the epithelial free surface, a second process sprouts from the base of the cell. This process pierces the basal lamina at stage 29/30 to reach the underlying telencephalon. The basal process is considerably thinner than the apical one and contains only free ribosomes and mitochondria. Between the stages 29/30 and 32+ it elongates and displays at its distal end the characteristics of a growth cone. The structural and ultrastructural morphology of the basal process identifies it as the axon of the olfactory neuron. Thus the placodai cells which migrate superficially differentiate into the olfactory neurons. Many placodai cells do not migrate, however, butremain at the epi‐thelial base as basal cells.As the placodal cells migrate to the body surface the NNE cells, originally confined to the surface layer (NNE), elongate and project processes toward the basal lamina. Beginning at stage 27 the apical surface of these cells becomes domed; the dome is filled with dense‐core vesicles and is covered with microvilli and globular material. The supranuclear cytoplasm of these cells contains free and membrane‐bound ribosomes, microtubules, and bundles of filaments. These cells can be characterized as the supporting cells of the olfactory neuroepithelium.The results indicate that (1) the olfactory neurons and basal cells are derived from the cells of the nervous layer, (2) the supporting cells are derived from the cells of the nonnervous ectoderm, (3) the olfactory nerve emerges from the olfactory neuroepithelium at stage 29/30, and (4) the placodai cells begin to display the morphological features of the mature olfactory neurons before their axons reach their target, the olfactory bulb.

The role of tissue interactions in the skeletogenic differentiation of avian neural crest cells

In the avian embryo, cranial neural crest (NC) cells migrate extensively throughout the head region and give rise to most of the cranial skeleton (Le Lievre, C. S. (1978). J. Embryol. Exp. Morphol. 47, 17–37). To investigate the skeletogenic differentiation of these cells, NC explants from the mesencephalic level of st. 9+ embryos were grown in standard organ culture on Millipore filter substrates either in isolation or in combination with those tissues with which the cells normally associate during their in vivo migration and at their final tissue sites. The results demonstrate that interaction between premigratory NC and cranial ectoderm leads to chondrogenic differentiation of NC cells. Combination of premigratory NC with presumptive site tissues led to a pattern of NC cell differentiation normally expressed after in vivo migration: Combinations of NC with retinal pigmented epithelia gave cartilage, whereas NC with maxillary ectoderm formed cartilage and membrane bone. Both resulting skeletal tissues possessed their characteristic collagen types (II in cartilage and I in bone) as shown by indirect immunofluorescence using antibodies raised against specific types of collagen. It is concluded that avian cephalic NC cells require tissue interactions if chondrogenic and osteogenic differentiation is to ensue, but that migration per se is not an absolute prerequisite for these types of differentiation. The degree of specificity underlying such interactions is discussed.