Transient connection or origin of the inferior pharyngeal constrictor during fetal development: A study using human fetal sagittal sections.

The inferior pharyngeal constrictor (IPC) originates from the thyroid and cricoid cartilages and inserts to the pharyngeal raphe. In serial sagittal sections of 37 embryos and fetuses at 6-15 weeks (crown rump length 15-115mm), we found (1) the IPC connecting to the sternothyroideus and thyrohyoideus muscles (16 fetuses at 6-11 weeks) or (2) the cricothyroideus muscle (6 fetuses at 12-15 weeks) in addition to the usual cricoid origin. These aberrant connections were most likely to be transient origins of the IPC not from a hard tissue but nearby striated muscles. In four of the latter six specimens, a tendinous band from the IPC inferior end connected to the cricothyroideus muscle to provide a digastric muscle-like appearance. These aberrant connections with nearby muscles seemed to become separated by a growing protrusion of the thyroid cartilage. Therefore, these aberrant origins were, even if developed, most likely to be "corrected" to the adult morphology during midterm or late prenatal period. The aberrant or transient origin of the IPC seemed to result from a discrepancy in growth of the cartilage and muscles. Such a discrepancy in growth seems to resemble the IPC wrapping around the superior cornu of thyroid cartilage. In addition, a final or adult-like morphology was found in two of the present 37 fetal specimens. It seemed to suggest a significant redundancy in growth rate of the laryngeal structures.

Regulation of Sox9 by Sonic Hedgehog (Shh) is essential for patterning and formation of tracheal cartilage.

We report that Sonic Hedgehog (Shh) regulates both formation and patterning of tracheal cartilage by controlling the expression pattern and level of the chondrogenic gene, Sox9. In Shh(-/-) tracheo-esophageal tubes, Sox9 expression is transient and not restricted ventrally to the site of chondrogenesis, and is absent at the time of chondrogenesis, resulting in the failure of tracheal cartilage formation. Inhibition of Hedgehog signalling with cyclopamine in tracheal cultures prevents tracheal cartilage formation, while treatment of Shh(-/-) tracheal explant with exogenous Shh peptide rescues cartilage formation. Both exogenous Bmp4 and Noggin rescue cartilage phenotype in Shh(-/-) tracheal culture, while promoting excessive cartilage development in wild-type trachea through induction of Sox9 expression. The ventral and segmented expression of Sox9 in tracheal primordia under Shh modulated by Bmp4 and Noggin thus determine where and when tracheal cartilage develops. These results indicate that Shh signalling is a critical determinant in tracheal cartilage development.

Fibroblast growth factor 18 gives growth and directional cues to airway cartilage.

OBJECTIVES/HYPOTHESIS: The majority of congenital airway anomalies arise from deficits in the respiratory tract cartilage, emphasizing the importance of this cartilage to the form and function of the upper airway. The primary objective of this study was to characterize molecular mechanisms that regulate rate and direction of chondrocyte growth in the larynx and trachea. Our hypothesis for this study was that fibroblast growth factor 18 (FGF18) provides proliferative and directional cues to the developing laryngeal and tracheal cartilage in the mouse by up-regulating the cartilage-specifying gene, Sox9. STUDY DESIGN: Molecular genetic and histological analyses of gene expression and cartilage growth in a mouse model. METHODS: Controlled mating of wild-type FVB/N (Friend Virus B-type/NIH mouse) mice and FGF18 overexpressing mice were carried out, and embryos ranging from embryonic (E) day 10.5 to E18.5 were obtained. The respiratory tract, including the larynx, trachea, and lung, was removed through meticulous dissection, and subjected to whole-mount in situ hybridization with RNA probes, or was sectioned and subjected to immunohistochemistry. Respiratory tracts from FVB/N mice were grown in culture in the presence of exogenous FGF18 or known inhibitors of the FGF pathway, and then subjected to quantitative reverse transcriptase polymerase chain reaction to measure the expression of cartilage-specific genes. RESULTS: The upper respiratory tract begins as a simple out-pouching from the ventral foregut endoderm at E10.5. The chondrocytes that form the cartilaginous structures of the upper respiratory tract are located at the junction of the respiratory tract out-pouching and the ventral foregut endoderm. This population of chondrocytes then undergoes directional proliferation to eventually assume the mature three-dimensional configuration of the upper respiratory tract cartilaginous framework. Immunohistochemical localization of extracellular signal-regulated kinases, a known modulator of FGF signaling, demonstrated the presence of this enzyme at the periphery of growing cartilage. Explants of larynx-trachea-lung grown in culture with exogenous FGF18 demonstrated hyperplastic growth and directed growth towards the FGF18 source. Finally, both FGF18 overexpressing tracheas and tracheas cultured with exogenous FGF18 demonstrated increased expression of the cartilage-specifying gene, Sox9. CONCLUSIONS: FGF18 provided both directional and proliferative cues to chondrocytes in the developing upper respiratory tract. FGF18 exerted this effect on developing chondrocytes by up-regulating Sox9 expression. Laryngoscope, 2009.

Prenatal and early postnatal morphogenesis and growth of human laryngotracheal structures.

Advances in neonatal medicine have resulted in increased care of fetal and neonatal airways. These advances have required an exhaustive knowledge of fetal airway anatomy and development. The aim of this study was to determine the anatomical development of laryngotracheal structures during the fetal and immediate postnatal period and to correlate these observations with other fetal biometric parameters to estimate developmental particularities of the fetal airway. An anatomical prospective study was based on examination of larynx and trachea from 300 routine autopsies of fetuses and infants, free of malformation and never intubated. Anatomical measurements of cricoid cartilage, thyroid cartilage, glottis, arytenoid cartilage and trachea were performed using a precision calliper and precision divider. Statistical analysis was performed to represent the growth of anatomical structures and to evaluate the correlation with biometric data. Raw data and 10th and 90th percentile curves were fitted satisfactorily with a linear model for gestational age. A linear relationship between laryngotracheal measurement and body weight and height was observed except for glottis length, interarytenoid distance and anterior cricoid height. The diameter of the cricoid lumen was significantly less than that of the trachea and glottis lumen. A sexual dysmorphism was noted for thyroid cartilage measurements and interarytenoid distance, with measurements significantly smaller in females. This study reports the anatomical development of normal laryngotracheal structures during the fetal period. Despite the fact that this study was performed during postmortem examination, these observations can be useful to develop criteria, materials and surgical procedures adapted to fetal and neonatal airways as well as for the purposes of early diagnosis and management of laryngotracheal malformations.

[The cartilaginous differentiation of the second arch in the human. From the traditional to the actual theory. Personal contribution]. / La diferenciación cartilaginosa del segundo arco en el hombre. De la teoría tradicional a la actual. Aportación personal.

Classically, the cartilaginous formation of the second pharyngeal arch has been described as a continuous structure wich will be the primary skeleton of the arch. Actually this theory has experimented a deep change Rodríguez Vázquez, 2005, and Rodríguez Vázquez et al. 2006, have a new cartilaginous differentiation model in the second pharyngeal arch and thus of its derivates in the human craniofacial development. The stapes and Reichert's cartilage have been formed by independent anlages. The cartilaginous differentiation model of the second arch, has allowed us to know and interpret the variations and classify them.

Androgen-induced myogenesis and chondrogenesis in the larynx of Xenopus laevis.

We investigated a possible role for testosterone-induced cell proliferation in the development of sexual dimorphism in the larynx of South African clawed frogs, Xenopus laevis. Androgen-induced cell proliferation was studied using [3H]thymidine autoradiography. Nuclei of cartilage, perichondrium, and muscle were labeled in the larynx of sexually immature frogs of both sexes but not in adults. Cell proliferation did not occur with estradiol treatment nor was it seen in nonlaryngeal muscle or cartilage. Electron microscopic/autoradiographic studies of laryngeal muscle indicate that testosterone stimulates satellite cell division which later results in formation of myonuclei. We conclude that testosterone induces both chondrogenesis and myogenesis in juvenile larynx and that this process may contribute to the pronounced sexual dimorphism of the adult vocal organ.

Local immune system in the developing fetal larynx.

Using histopathological and immunofluorescent techniques, we investigated the development of the glandular system of the larynx from 12 human fetuses. Glandular buds were first observed at the fourth month of gestation and glandular distribution became evident from the fifth month in the larynx. Ciliated or nonciliated epithelium covered the cavity of the larynx, with the exception of the true vocal cord. Immunofluorescent examination revealed the presence of IgG in the fetal larynx, but there was little fluorescence for IgA, IgM and IgE. Secretory component (SC) synthesis, on the other hand, was found in each serous-type glandular epithelium or acinus from the fourth month on. Our observations suggest that the local immune system by SC, which binds to IgA as SIgA in human laryngeal mucosa, is inherently acquired in the fetal larynx.

The human chondrocranium at the end of the embryonic period, proper, with particular reference to the nervous system.

The chondrocranium at stage 23 (27-32 mm C.-R., 8 postovulatory weeks) was studied in detail to fill the descriptive gap between 7 and 9 weeks. Serial sections of ten embryos, all belonging to the same developmental stage, were investigated and compared. Graphic and Perspektomat reconstructions were made from three embryos. The chondrocranium forms a continuous mass of well-formed cartilage, which blends with early cartilage in the nasal, orbital, and otic regions, around the future foramen rotundum, and at the site of the parietal lamina. The chondrocranium and "membrane" bones cover a relatively small portion of the brain, namely 1) the medulla and pons, 2) the caudal half of the lateral portion of the cerebellar primordium, and 3) the rostroventral part of the cerebral hemispheres. The relationships of the cranial nerves were are also investigated. The degree of cranial development is intermediate between that at stage 20 (21 mm) studied by Lewis (the only other skull so far studied adequately in a staged embryo) and that of fetuses of 40 and 43 mm investigated by Macklin. On the whole, the resemblances to the fetal skulls are greater: ossification is beginning in the tectum posterius (to form the supraoccipital), and most of the "membrane" bones (except the parietal, tympanic, lacrimal, and nasal) show osseous areas. The foramen rotundum is present. The lower angle between the chordal and prechordal parts of the basal stem ranges from 129 degrees (which resembles that at stage 20, namely 122 degrees) to 115 degrees (which is the same as that at 40-43 mm). An orbito-parietal commissure is not yet present. The lower border of the larynx is at the level of the lower border of the fourth cervical vertebra. In the larynx, the thyroid laminae have usually not fused, the hyoid cartilage possesses greater and lesser cornua, the cricoid cartilage displays its signet-ring form, and the arytenoids are identifiable as condensed mesenchyme with some early cartilage. The cartilage of the first pharyngeal arches is covered in its ventral three-fourths by an osseous mandibular plate. Morphological variations within stage 23 are minor and appeared not to be related closely to each other, so that it was not possible to arrange the embryos in a linear developmental series based on their level of cranial morphogenesis.

Ultrastructure of human foetal and neonatal hyaline cartilage.

Ultrastructural studies showed that human foetal and neonatal hyaline cartilage contained at least two types of chondrocytes. The predominant chondrocytes of chief cells were characterised by scanty, flat endoplasmic reticulum, few mitochondria, a rudimentary Golgi zone, and by the presence of clear cytoplasmic vacuoles which contained electron-dense granules and beaded filaments. The second type of cells had dark or electron-dense cytoplasm which contained numerous dilated endoplasmic reticulum and prominent Golgi zones; these cells gave rise to slender cytoplasmic processes from which the matrix vesicles were probably derived. The lacunar matrices surrounding both cell types were morphologically different reflecting some functional differences between these cells. Both types of cells participated in the formation of costochondral growth zones. Condrocytes with morphology of the chief cells proliferated, columnised, underwent hypertrophy, and differentiated either into a compact hypertrophic cell or a swollen pale cell. The cells also became hypertrophic and gave rise to increasing numbers of dense vesicles. Calcification was first noted in the longitudinal septa inside or adjacent to matrix vesicles. The recognition of the heterogeneity of chondrocytes in human hyaline cartilage may be helpful in understanding the pathology of the chondrodysplasias.

Laryngotracheoesophageal cleft. Diagnosis, management, and presentation of a new diagnostic device.

Congenital posterior clefts between the laryngotrachea and the esophagus have rarely been encountered as evidenced by only 30 cases reported in the world literature. Of these, patients in nine have survived. We present another patient with this condition who was successfully treated. Important aspects in evaluation, including the need for an experienced endoscopist and cineradiography, are discussed. Postoperative management using a continuous infusion pump to administer formula per gastrostomy prevents gastric distension and emesis that lead to stress on the closure. This should be a basic consideration in the management of these patients.

Infant primate larynx: developmental histology.

Laryngeal epithelium of monkeys (M. mulatta, M. nemestrina) ranging in age from late fetal to adult was found to undergo the following developmental changes: (1) progressive elaboration of commissural epithelium prenatally, reaching the adult form of pseudostratified columnar ciliated epithelium early in the postnatal period;(2) vocal process epithelium changing from cuboid to stratified squamous tissue during the late prenatal period, and (3) progressive increments in overall epithelial thicknees during development, reaching a maximum in the early postnatal period. Chondrocyte density of the cricoid and thyroid cartilage decreased during the late prenatal period, reaching the adult form at the neonatal stage. The evidence suggests that mechanical or metaplastic stresses do not determine the observed tissue changes.

Some aspects of fetal laryngeal development.

Some aspects of morphological and physiological development of the fetal larynx are presented. Histochemical confirmation and determinations of fetal glandular activity are needed. Anomalous development of the larynx is still conjecture, but congenital cystic formation with glandular development appears possible after the fifth month of human fetal life.