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Embryonic development of the head and neck: part 2, the branchial region.

The embryology of the branchial (pharyngeal) region is reviewed in terms of the embryonic pharynx, branchial apparatus and derivatives, tongue, larynx, and trachea. Derivatives of the branchial pouches, grooves (clefts), and arches are analyzed in relation to each other and to the first arch syndrome. Other common head and neck defects attributable to abnormal development of the branchial region are also discussed, particularly those defects involving the incomplete degeneration or the abnormal migration of branchial tissues.

Branchial Region↗

A distinct Hox code for the branchial region of the vertebrate head.

The branchial region of the vertebrate head forms through complex interactions involving rhombomeric segments, neural crest and branchial arches. It is though that aspects of their patterning mechanisms are linked and involve Hox-2 genes, whose overlapping and spatially restricted expression domains represent a combinatorial code for generating regional diversity. Vertebrates possess four Hox clusters of Antennapedia class homeobox genes, related to each other by duplication and divergence from a common ancestral complex. In consequence, at equivalent positions in different clusters there are highly related genes known as subfamilies or paralogous groups. As Hox-2 genes cannot fully account for patterning individual rhombomeres, we investigated whether offsets in expression limits of paralogous genes could account for the generation of regional diversity. We report here that, with the exception of the labial subfamily, paralogues show identical expression limits in rhombomeres, cranial ganglia and branchial arches, providing a combinatorial Hox code for the branchial region that seems to be different in organization to that of the trunk.

Alleles↗

Sites of aspecific ATP-phosphohydrolase activity in the branchial region of chick embryos, with particular emphasis on the myogenic cell population.

The distribution pattern of unspecific ATP-phosphohydrolase (ATP-PH) activity was studied in serial sections through the branchial region of chick embryos from HH stage 14 to 27. A selective reaction was found in the prospective myogenic mesoderm of voluntary muscles of the first three branchial arches. This property initially characterized uncondensed myogenic cell populations in the mandibular and hyoid arches and appeared later on in the third arch. In each case, the positive premuscular mesoderm was closely apposed to the lateral wall of the corresponding aortic arch. Myogenic primordia retained a strong ATP-PH activity up to stage 27 when they began to segregate into smaller premuscular units. Discrete changes observed in the mandibular and hyoid blastemas suggested that non reacting cells probably of neural crest origin invade their proximal core and contribute to the subsequent pathfinding of muscular nerves. Other selective sites of ATP-PH reactivity were detected in the odd-numbered rhombomeres, rhombencephalic floorplate, notochord, arterial endothelial cells, prospective dermis and perichondrium.

Adenosine Triphosphate↗

A homeotic transformation is generated in the rostral branchial region of the head by disruption of Hoxa-2, which acts as a selector gene.

The Hoxa-2 gene was disrupted by homologous recombination. Homozygous mutant mice died at birth. Defects were found in the branchial region of the head, which corresponds to the Hoxa-2 rostral expression domain. While rhombomeric and neural crest cell (NCC) segmentation was not affected, mesenchymal NCC derivatives of the second arch were lacking, and second arch mesenchymal NCC identity was changed to first arch identity, resulting in homeotic transformation of second to first arch skeletal elements. These results reveal the existence of a skeletogenic ground pattern program common to at least the mesenchymal NCC that originated from rhombomeres 2 and 4. The appearance of an atavistic reptilian pterygoquadrate element in Hoxa-2 mutants suggests that this ground pattern is intermediate between reptiles and mammals. The ground pattern program appears to be modified in the mouse first arch by a Hox-independent process, whereas Hoxa-2 acts as a selector gene in the second arch.

Animals↗

Segmentation and specification in the branchial region of the head: the role of the Hox selector genes.

Hox genes are segmentally expressed in the developing vertebrate hindbrain, neural crest cells and pharyngeal arches suggesting an important role in patterning these structures. Here we discuss the cellular and molecular mechanisms controlling segmentation and specification in the branchial region of the head. In addition, based on the recent phenotypical and molecular analysis of loss-of-function mutants in the mouse, we speculate that Hox genes may act like Drosophila selector genes in this system.

Animals↗

[The branchial region and its proper role in the morphogenesis of the neck].

The branchial apparatus is a keystone in the embryonic arrangement of the neck region. However the developing cervical region is also under the influence of the fundamental metamerism of the body--the formation of sonites-, which gives origin to the cervical spine, the prechordal space and the anterior cervical area. The developmental history of the cranial portion depends on the connection between these two distinct systems of segmentation.

Branchial Region↗

Nkx2.6 expression is transiently and specifically restricted to the branchial region of pharyngeal-stage mouse embryos.

The Nkx2.6 gene belongs to the NK superfamily of homeobox genes (Harvey, 1996). We report here the expression pattern of the murine Nkx2.6 gene during early mouse development, which is unique among the NK family of homeobox genes in that its expression is restricted to the very narrow development period between stages E8.5 and E10.5 of embryogenesis. The distribution of Nkx2.6 transcripts is also quite restricted spatially, with expression detected uniquely within the caudal branchial arches. Nkx2.6 is expressed in all three layers comprising the caudal branchial arches (ectoderm, mesectoderm and endoderm) with the strongest expression being detected in the surface ectoderm.

Animals↗

Correlation between loss of middle ear bones and altered goosecoid gene expression in the branchial region following retinoic acid treatment of mouse embryos in vivo.

The homeobox gene goosecoid marks the Spemann organizer in vertebrate gastrula embryos, and is expressed in the craniofacial region, body wall and limbs during organogenesis. Mouse mutants of goosecoid displayed a variety of phenotypes related to the expression pattern at mid-embryogenesis. These defects included loss of the tympanic ring and malformation of the malleus, phenotypes which were reminiscent of the teratogenic effects of retinoic acid (RA). Here we investigated the correlation of goosecoid gene expression and RA-teratogenicity following treatment of mouse embryos in vivo at embryonic day (E) 8 + 5 h. We found that goosecoid was specifically affected at E10.5 in branchial arches I and II. Expression was either reduced to background levels or restricted to the branchial cleft region. This change in goosecoid gene expression correlated with a loss of middle ear ossicles and a partial or complete deletion of the tympanic ring, suggesting a role for goosecoid in executing the RA teratogenic effects.

Animals↗

Apoptosis of premigratory neural crest cells in rhombomeres 3 and 5: consequences for patterning of the branchial region.

In the avian hindbrain, premigratory neural crest cells undergo programmed cell death (apoptosis) in rhombomeres 3 and 5 (r3, r5). Here, we have attempted to analyze the significance of the loss of neural crest cells from these odd-numbered rhombomeres. When apoptosis is prevented in r3 and r5, r3 crest migrate into the first arch and r5 into the third arch. Interestingly, these extra neural crest cells contributed to the formation of ectopic muscle attachment sites that are also found in those species in which r3 and r5 neural crest cells do not undergo apoptosis. Thus, apoptosis in the odd-numbered rhombomeres appears to be an evolutionarily derived mechanism that is required to eliminate r3 and r5 crest migration into first and third arches and thereby remove these muscle attachment sites.

Animals↗

Hox genes and pattern formation in the branchial region of the vertebrate head.

The conservation of molecular mechanisms in evolution has stimulated interest in comparative embryology, and the Hox/HOM-C homeobox complexes are among the best examples of the remarkable conservation between species. An important region associated with Hox patterning is the head, where the evolutionary and molecular studies have reawakened interest in the cellular processes and complex interactions required to regulate craniofacial morphogenesis.

Animals↗

The branchial arches and HGF are growth-promoting and chemoattractant for cranial motor axons.

During development, cranial motor neurons extend their axons along distinct pathways into the periphery. For example, branchiomotor axons extend dorsally to leave the hindbrain via large dorsal exit points. They then grow in association with sensory ganglia, to their targets, the muscles of the branchial arches. We have investigated the possibility that pathway tissues might secrete diffusible chemorepellents or chemoattractants that guide cranial motor axons, using co-cultures in collagen gels. We found that explants of dorsal neural tube or hindbrain roof plate chemorepelled cranial motor axons, while explants of cranial sensory ganglia were weakly chemoattractive. Explants of branchial arch mesenchyme were strongly growth-promoting and chemoattractive for cranial motor axons. Enhanced and oriented axon outgrowth was also elicited by beads loaded with Hepatocyte Growth Factor (HGF); antibodies to this protein largely blocked the outgrowth and orientation effects of the branchial arch on motor axons. HGF was expressed in the branchial arches, whilst Met, which encodes an HGF receptor, was expressed by subpopulations of cranial motor neurons. Mice with targetted disruptions of HGF or Met showed defects in the navigation of hypoglossal motor axons into the branchial region. Branchial arch tissue may thus act as a target-derived factor that guides motor axons during development. This influence is likely to be mediated partly by Hepatocyte Growth Factor, although a component of branchial arch-mediated growth promotion and chemoattraction was not blocked by anti-HGF antibodies.

Animals↗

Gene expression analysis of Gata3-/- mice by using cDNA microarray technology.

Transcription factor Gata3 is implicated in the formation of autosomal dominant hypoparathyroidism, sensorineural deafness, and renal anomaly (HDR) syndrome. We pursued to identify the potential Gata3 target genes by profiling the gene expression pattern in E9.5 Gata3-/- mouse embryos. Altogether four independent microarray hybridizations were carried out on NIA Mouse15K cDNA arrays. We discovered two hundred and sixty one genes that are downregulated in Gata3 mutant embryos at E9.5 (with a minimal 2.0-fold change). The majority of the differentially expressed genes belong to two functional groups--genes involved in transcription regulation and cellular signaling. One of the genes discovered to be downregulated in Gata3 mutant embryos was tumor suppressor gene Disabled 2. The validity of this finding was checked by using the whole mount in situ hybridization technology. This study revealed that the sites, where Dab2 is downregulated in the mutant embryos partly overlap with the Gata3 expression domains, including the mid-embryo region, branchial arches and facio-acoustic (VII-VIII) neural crest complex. This is the first time when tumor supressor gene Dab2 is shown to be implicated in the defective phenotype of Gata3 mutant mice.

Adaptor Proteins, Signal Transducing↗

Receptive fields and properties of a new cluster of mechanoreceptor neurons innervating the mantle region and the branchial cavity of the marine mollusk Aplysia californica.

The rostral LE cluster (rLE) is a new set of mechanoreceptor neurons of the abdominal ganglion innervating the mantle area, the branchial cavity, the gill and the siphon of the marine mollusk Aplysia californica Cooper. We have compared the organization of rLE cell receptive fields with that of three other clusters of sensory neurons in the abdominal ganglion (LE, RE and RF) that we have reanalysed. There is extensive overlap of receptive fields from the four populations of sensory cells, and the most exposed areas of the mantle are the most densely innervated. The sensory threshold is similar for all groups. The action potentials of the LE, rLE and RE neurons are broadened by serotonin and the peptide SCPB and narrowed by dopamine and FMRFamide. The RF group does not show the same kind of sensitivity to these neuromodulators. The synaptic outputs of the LE and rLE neurons undergo similar synaptic depression and homosynaptic and heterosynaptic facilitation. We estimate that 100 mechanoreceptor neurons innervate the entire mantle and siphon skin, gill and branchial cavity of Aplysia. The degree of their convergence onto various interneurons and motor neurons mediating the gill- and siphon-withdrawal reflex and other reflexes is under investigation.

Animals↗

Morphometric discrimination of early life stage Lampetra tridentata and L. richardsoni (Petromyzonidae) from the Columbia River Basin.

The effectiveness of morphometric and meristic characteristics for taxonomic discrimination of Lampetra tridentata and L. richardsoni (Petromyzonidae) during embryological, prolarval, and early larval stages (i.e., age class 1) were examined. Mean chorion diameter increased with time from fertilization to hatch and was significantly greater for L. tridentata than for L. richardsoni at 1, 8, and 15 days postfertilization. Lampetra tridentata larvae had significantly more trunk myomeres than L. richardsoni; however, trunk myomere numbers were highly variable within species and deviated from previously published data. Multivariate examinations of prolarval and larval L. tridentata (7.2-11.0 mm; standard length) and L. richardsoni (6.6-10.8 mm) were conducted based on standard length and truss element lengths established from eight homologous landmarks. Principal components analysis indicated allometric relationships among the morphometric characteristics examined. Changes in body shape were indicated by groupings of morphometric characteristics associated with body regions (e.g., oral hood, branchial region, trunk region, and tail region). Discriminant function analysis using morphometric characteristics was successful in classifying a large proportion (> 94.7%) of the lampreys sampled.

Animals↗

A novel MAP kinase phosphatase is localised in the branchial arch region and tail tip of Xenopus embryos and is inducible by retinoic acid.

Using a differential display strategy, we have isolated a cDNA corresponding to a mRNA which is induced by retinoic acid treatment of late gastrula Xenopus embryos, and much more strongly induced by retinoic acid and cycloheximide. The cDNA, designated X17C, encodes a novel mitogen-activated protein (MAP) kinase phosphatase of 378 amino acid residues which is only distantly related to other known MAP kinase phosphatases. In normal embryogenesis, the X17C mRNA is expressed after the midblastula transition and accumulates during gastrulation. In neurula and tailbud stage embryos the mRNA is localised in two domains, one in the anterior region of the embryo, and one at the tail tip. When expressed from synthetic mRNA injected into oocytes, the X17C protein is found within the cytosolic fraction and not in the nucleus. The X17C protein dephosphorylates and inactivates Xenopus MAP kinase in oocytes stimulated to undergo maturation by progesterone. We indicate the application of X17C as a tool for interfering with MAP kinase signaling in somatic cells of embryos, using FGF receptor-mediated MAP kinase activation in animal cap explants.

Amino Acid Sequence↗