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Gene expression during Drosophila wing morphogenesis and differentiation.

The simple cellular composition and array of distally pointing hairs has made the Drosophila wing a favored system for studying planar polarity and the coordination of cellular and tissue level morphogenesis. We carried out a gene expression screen to identify candidate genes that functioned in wing and wing hair morphogenesis. Pupal wing RNA was isolated from tissue prior to, during, and after hair growth and used to probe Affymetrix Drosophila gene chips. We identified 435 genes whose expression changed at least fivefold during this period and 1335 whose expression changed at least twofold. As a functional validation we chose 10 genes where genetic reagents existed but where there was little or no evidence for a wing phenotype. New phenotypes were found for 9 of these genes, providing functional validation for the collection of identified genes. Among the phenotypes seen were a delay in hair initiation, defects in hair maturation, defects in cuticle formation and pigmentation, and abnormal wing hair polarity. The collection of identified genes should be a valuable data set for future studies on hair and bristle morphogenesis, cuticle synthesis, and planar polarity.

Animals↗

[The action of insulin in the morphogenesis of Gallus gallus domesticus embryos].

Aspects concerned with morphogenesis of Gallus gallus domesticus, avail studies related to the action of the insulin in the topography and embryonic structures. At the temperature of 37.5 degrees C, eggs were incubated during 24 h, injected with 5 ml of swine insulin in three concentrations and reincubated for more 72 h. The morphological characteristics of 80 embryos were evaluated and, according to the presented organization, classified in 5 morphogenetic levels. It was registered generalized dysmorphism (4th level) in 21 embryos that went through the tests with insulin. Standard morphogenesis (1st level) and located dysmorphism (3rd level) were verified among those from the control experiments. Those individuals concerned with the 4th level, showed reduced dimension of the body and were characterized by anterior-dorsal limits organized in a cephalic projection, and also presented alterations in the posterior-ventral region. These features evidence a pattern of abnormality in the determination of the cephalic-caudal axis and indicate a specific action of the insulin in the embryonic morphogenesis, in the period of 96 hours of incubation.

Animals↗

Effects of fetus weight, dam strain, dam weight, and litter size on the craniofacial morphogenesis of CL/Fr mouse fetuses affected with cleft lip and palate.

OBJECTIVE: This study examined the factors related to the morphogenesis of the craniofacial complex of the CL/Fr mouse fetus affected with CLP based on the findings of a lateral cephalogram. DESIGN: Embryo transfer experiments were performed to determine the effect of the fetus weight, dam strain, dam weight, and litter size on the Intra-uterine craniofacial morphogenesis of CL/Fr mouse fetuses. On the 18th gestational day, each pregnant dam that had received CL/Fr mouse embryos was laparotomized to remove the transferred fetuses that had developed in the uteri of the cleft lip and palate (CLP)-susceptible CL/Fr strain dam and the CLP-resistant C57BL strain dam. A cephalometric observation of the craniofacial morphology of each fetus was subsequently performed. RESULTS: Based on a multiple regression analysis, the standardized partial regression coefficients of the affected fetus weight, the dam weight, and the litter size on the maxillary size of the affected CL/Fr fetus were 0.71 (p < .01), 0.03, and -0.07. According to a least-squares analysis of variance, the dam strain effect in addition to the effect of the affected fetus weight on the maxillary size and the cranial size of the affected fetuses was significant (p < .01 for cranial size, p < .05 for maxillary size) and close to a significant level (p = .09) for the mandibular size of the affected fetuses. The adjusted maxillary size and cranial size after statistically eliminating the effects of the affected fetus weight, dam weight, and litter size on each original craniofacial size of the affected fetuses that had developed in the CL/Fr dam strain were also significantly smaller than those of the affected fetuses that had developed in the C57BL dam strain. CONCLUSIONS: The present results indicate that the craniofacial growth of the CL/Fr mouse fetus affected with CLP increased in proportion to the fetus weight. The dam strain effect, in addition to the effect of the affected fetus weight, could thus not be ignored when the etiology of the spontaneous CLP was examined, while the uterine environment, provided by the CL/Fr strain dam, retarded the intra-uterine craniofacial growth of the affected fetuses. It was therefore concluded that the dam strain effect, as well as the effect of the affected fetus weight, both play an important role on the craniofacial morphogenesis of the CL/Fr strain of the affected fetuses that developed in both strain dams.

Analysis of Variance↗

Dystroglycan and laminins: glycoconjugates involved in branching epithelial morphogenesis.

Branching epithelial morphogenesis is crucial for the development of several organs, such as lung, submandibular gland, mammary gland, tooth, pancreas, and kidney. During early embryogenesis, these organs are composed of a small epithelial rudiment surrounded by mesenchymal cells. Interactions between the two tissue compartments induce growth and branching of the epithelium into the mesenchyme. In each tissue, the epithelial branching has tissue-specific features, but there are many similarities both at the morphological and molecular level. Basement membrane components such as laminin have been implicated in the regulation of epithelial morphogenesis. Here data are reviewed that suggest that interactions between laminin-1 and other basement membrane components and the cell surface are important for epithelial morphogenesis in the kidney, lung, and salivary gland.

Animals↗

Tooth crown morphogenesis and cytodifferentiations: candid questions and critical comments.

Teeth are probably meristic units and crown morphogenesis leads to tooth specific distribution of functional cells. Since heterodonty is derived from homodonty, one way to understand tooth morphogenesis would be to unravel the involved phenomena in homodont species and then to characterize the "put up job" of evolution leading to species specific dentitions with particular functional abilities. Interaction of paleontologists and developmental biologists should be initiated. My naive "developmental" point of view will illustrate only one facet of mouse tooth morphogenesis and cytodifferentiation. The main concern will be to try to discriminate between known facts and speculations, between hypotheses and anticipated or deduced certitudes, to call attention to conflicting data, to suggest some further investigations and to advocate the point of view that molecular interpretations should be founded on indisputable morphological data.

Animals↗

[Cytomechanical control of morphogenesis].

The role of mechanically strained state of cells and multicellular structures in morphogenesis regulating in vertebrate embryos is discussed. Regular changes in patterns of mechanical strain during embryonic development are described. Artificial relaxation of mechanical strain performed on definite developmental stages and retension of embryonic tissues in arbitrary directions considerably affects morphogenesis and cell differentiation patterns. Cytomechanical models of morphogenesis are reviewed and a concept of hyperrestoration of mechanical strain as a possible driving force of morphogeneiss is suggested.

Animals↗

Epithelial-mesenchymal interactions and lung branching morphogenesis. Role of polyamines and transforming growth factor beta1.

Lung branching morphogenesis is a result of epithelial-mesenchymal interactions, which are in turn dependent on extracellular matrix composition and cytokine regulation. Polyamines have recently been demonstrated as able to modify chick embryo skin differentiation. In this work we have examined the effects of putrescine and spermidine during chick embryo lung morphogenesis in organotypic cultures by morphological, histochemical and biochemical examination. To verify the role of polyamines, we used specific inhibitors, such as bis-cyclohexylammonium sulphate and alfa-difluoromethylornithine, and transforming growth factor beta1, an ornithine decarboxylase and polyamine stimulator. Our data show that lung morphogenesis is significantly altered following the induced mesenchymal glycosaminoglycan changes. The increase of mesenchymal glycosaminoglycans is correlated with a stimulation of lung development in the presence of polyamines, and with its inhibition when transforming growth factor beta1 is added to the culture medium. The morphometric data show a uniform increase of both the mesenchyme and epithelial branching with spermidine and putrescine stimulus, whereas the mesenchymal substance alone is significantly increased in apical-median lung sections with transforming growth factor beta1 and transforming growth factor beta1 + spermidine lung cultures. Transforming growth factor beta1 and transforming growth factor beta1 + spermidine confirm the blocking of epithelial branching formations and fibroblast activation, and show that polyamines are unable to prevent the blocking of epithelial cells due to the inhibitory effect of transforming growth factor beta1.

Animals↗

[The effect of retinoic acid on crown morphogenesis of mouse tooth germs in vitro].

OBJECTIVE: To study the effects of retinoic acid on crown morphogenesis of early tooth germs. METHODS: Mouse tooth organ culture model was used and the functions of different retinoic acid concentration on the first mandibular molar germs of 14-day old mouse embryo were examined. RESULTS: The first mandibular molar germ of 14-day old mouse was at the bud stage. After 6 days culture, tooth germs in culture medium (not containing retinoic acid) developed from bud stage to bell stage and formed normal cusps. The morphogenesis of tooth germs in medium containing 1 x 10(-7) mol/L retinoic acid was similar to molar germs in the medium without retinoic acid. When the concentration of retinoic acid increased to 1 x 10(-6) mol/L, the mouse tooth germs still developed from bud stage to bell stage, but normal cusps was not formed. CONCLUSION: The results suggest that high concentration of retinoic acid alter crown morphogenesis of tooth germs.

Animals↗

[Expression of transforming growth factor-beta 1 mRNA during mouse molar morphogenesis].

OBJECTIVE: To investigate the temporal and spatial expression and localization of transforming growth factor-beta 1 mRNA during mouse molar morphogenesis. METHODS: The distribution pattern of TGF-beta 1 mRNA was analysed on 5 microns serial sections of paraformaldehyde-fixed and paraffin-embedded embryonic mouse heads or mandibles using in situ hybridization. RESULTS: The results show that TGF-beta 1 mRNA was expressed during mouse molar morphogenesis in a temporally and spatially regulated fashion. Local expression of TGF-beta 1 mRNA in the dental epithelium at bud-staged (E13) and cap-staged (E15) teeth were observed. During bell stage (E16-18), TGF-beta 1 mRNA was very abundant in the ameloblast layer and dental papilla cells. The expression of TGF-beta 1 was increased in the layer of odontoblast and ameloblast with the differentiation of these cells. CONCLUSIONS: These results suggest that TGF-beta 1 may have an important role in tooth morphogenesis, and TGF-beta 1 acts as a paracrine and autoinducing factor.

Animals↗

[Morphogenesis of Okhotsky virus from the genus orbisvirus].

Morphogenesis of Okhotsky virus isolated in the USSR was studied. Virions of 50-65 nm in diameter were found in the peripheral zone of cytoplasmic matrices. Crystalline-like arrays of virions were observed not reported by other investigators studying morphogenesis of orbisviruses. Besides, not directly associated with matrices, crystalline arrays of some proteins were found in infected cells for the first time with orbisviruses. Nevertheless the main stages of morphogenesis of Okhotsky virus were similar to those of other members of the orbisvirus group.

Animals↗

A chondroitin sulfate proteoglycan PTPzeta /RPTPbeta regulates the morphogenesis of Purkinje cell dendrites in the developing cerebellum.

PTPzeta/RPTPbeta, a receptor-type protein tyrosine phosphatase synthesized as a chondroitin sulfate (CS) proteoglycan, uses a heparin-binding growth factor pleiotrophin (PTN) as a ligand, in which the CS portion plays an essential role in ligand binding. Using an organotypic slice culture system, we tested the hypothesis that PTN-PTPzeta signaling is involved in the morphogenesis of Purkinje cell dendrites. An aberrant morphology of Purkinje cell dendrites such as multiple and disoriented primary dendrites was induced in slice cultures by (1) addition of a polyclonal antibody against the extracellular domain of PTPzeta, (2) inhibition of protein tyrosine phosphatase activity, (3) enzymatic removal of the CS chains, (4) addition of exogenous CS chains, and (5) addition of exogenous PTN, all of which disturb PTN-PTPzeta signaling. These treatments also reduced the immunoreactivity to GLAST, a glial glutamate transporter, on Bergmann glial processes. Furthermore, a glutamate transporter inhibitor also induced the abnormal morphogenesis of Purkinje cell dendrites. Altogether, these findings suggest that PTN-PTPzeta signaling regulates the morphogenesis of Purkinje cell dendrites and that the mechanisms underlying that regulation involve the GLAST activity in Bergmann glial processes.

Amino Acid Transport System X-AG↗

Endogenous epidermal growth factor regulates the timing and pattern of embryonic mouse molar tooth morphogenesis.

The tooth organ provides a model for discrete patterns of morphogenesis over short periods of developmental time. Studies were designed to test the hypothesis that endogenous epidermal growth factor (EGF) functions to regulate multiple cusp molar tooth morphogenesis during embryonic mouse development. The relative levels of endogenous EGF and EGF receptor (EGFR) transcripts were determined in both enamel organ epithelia and dental ectomesenchyme by reverse transcription-polymerase chain reaction (RT-PCR) assays. EGF and EGFR were localized by immunohistochemistry; both antigenic determinants were demonstrated on the same odontogenic cells in cultured tooth explants. To examine EGF-mediated signal transduction, cap stage mouse molar tooth organs (E16) were cultured in serumless, chemically-defined medium as either (i) controls, or supplemented with (ii) tryphostin (an EGF receptor kinase inhibitor), (iii) tyrphostin plus exogenous EGF, and (iv) exogenous EGF. Antisense oligodeoxynucleotide (ODN) strategy was used to investigate the functions of endogenous EGF employing (i) non-treated control, (ii) sense ODN control, (iii) antisense ODN, (iv) exogenous EGF, (v) sense ODN with exogenous EGF, and (vi) antisense ODN with exogenous EGF. Tyrphostin inhibited DNA synthesis and produced a significant decrease in the volume of the explants. These effects were recovered by addition of exogenous EGF. Antisense ODN inhibition resulted in abnormal cusp formations, decreased DNA synthesis, total DNA, RNA and protein content, and decreased stellate reticulum and tooth explant volumes. The decreased tooth size was not uniform, the most pronounced effect was in the stellate reticulum. This pattern of changes was not seen when antisense ODN treatment was supplemented with exogenous EGF. These results suggest that during cap stage of odontogenesis endogenous EGF acts to stimulate DNA synthesis, which increases the cell number of specific phenotypes within the enamel organ epithelia, and thereby regulates molar tooth morphogenesis.

Animals↗

Hydrocortisone perturbs the cell proliferation pattern during feather morphogenesis: evidence for disturbance of cephalocaudal orientation.

In this study, we have monitored the spatial distribution of S-phase cells during successive stages of normal feather morphogenesis using the specific marker BrdU. We also disturbed the development program by administration of hydrocortisone on the chorioallantoic membrane of 6.5-day chick embryos and examined the resulting pattern of BrdU incorporation. Our results show that a specific spatio-temporal pattern of cell proliferation occurs during successive stages of feather development and that this pattern accounts for the growth of feather buds according to the cephalocaudal orientation. Our experimental analysis showed that the stage-dependent alteration of feather morphogenesis (as shown by Züst, Ann. Embryol. Morphogen. 4, 1971 and confirmed by Démarchez et al., Dev. Biol. 106, 1984), is based on a stage-dependent alteration of the proliferation pattern in the epidermis. Forty-eight hours after treatment, non-induced epidermis ceases DNA synthesis and is unable to form placodes. Induced epidermis at the placodal and dermal condensation stages fails to produce the cohorts of S-phase cells responsible for the caudal outgrowth and the slanting shape of the buds. These young buds display anarchic proliferation in the whole epidermis possibly resulting in the appearance of "curly" feathers. Together, these results show the importance of the spatial pattern of ectodermal and mesodermal cell proliferation during the normal feather morphogenesis. Moreover, they corroborate the particular role of epidermis both in the establishment of feather rudiments and in the cephalocaudal orientation of the feathers.

Animals↗

MEK kinase 1 regulates c-Jun phosphorylation in the control of corneal morphogenesis.

PURPOSE: To study the in vivo role of MEK kinase 1 (MEKK1) in corneal development. METHODS: Wild type and Mekk1DeltaKD/DeltaKD mice eye tissues were examined by staining with hematoxylin and eosin for morphogenesis and Masson's trichrome for extracellular matrix (ECM) deposition. The cells expressing ECM gene transcripts of Collagen I, Keratocan, and Lumican in corneal stroma were identified by in situ hybridization and the level of Collagen I mRNA in the developing cornea was quantified by real-time RT-PCR. Immunohistochemistry staining was employed to study the expression and N-terminal phosphorylation of c-Jun and the expression of epithelium differentiation markers and intercellular structural proteins of the corneal epithelium. RESULTS: Mekk1DeltaKD/DeltaKD mice exhibited the "eye open at birth" phenotype (EOB), and developed eye defects and severe pathology secondary to impaired eyelid formation. The corneal stroma of Mekk1DeltaKD/DeltaKD fetuses, although exhibiting normal morphology, thickness, and keratocyte proliferation, showed reduced extracellular matrix (ECM) accumulation, corresponding to a decrease in transcription of Lumican, Keratocan, and Collagen I. Immunohistochemistry studies demonstrated that MEKK1 ablation caused a remarkable reduction in the expression of occludin and zonula occluden protein-1 (ZO-1), components of tight junction, but had no effect on the expression of E-cadherin and beta-catenin for adherens junctions, desmoplakin and desmoglein for desmosomes and cytokeratins 12 and 14 for cornea-type epithelial differentiation in the developing cornea. c-Jun was abundantly expressed in the developing corneal epithelium and its phosphorylation was considerably reduced in Mekk1DeltaKD/DeltaKD fetuses. CONCLUSIONS: In addition to its role in eyelid morphogenesis, MEKK1 is crucial for corneal development such that its ablation caused a reduction of ECM deposition in corneal stroma and disturbance of tight junctions in corneal epithelium. c-Jun phosphorylation in corneal epithelium is a downstream event of the MEKK1 pathway, likely contributing to corneal development and function. Altogether, MEKK1 plays a major role in ocular surface morphogenesis and its ablation leads to damage and various eye manifestations at postnatal stages.

Animals↗

Inhibition of branching morphogenesis of mouse fetal submandibular gland by sodium fluoride--protection by epidermal growth factor.

As an initial step to study the effect of sodium fluoride on the oral environment, we investigated how sodium fluoride (NaF) affects the branching morphogenesis of fetal mouse submandibular gland (SMG). When mouse SMG was prepared from the embryo at 13 days post prenatal stage and cultured, gradual development of branching morphogenesis was observed. Addition of NaF affected this morphological change in bimodal fashions. At a lower concentration of NaF (< 2 microM), the branching morphogenesis was slightly enhanced, whereas at a higher concentration of NaF (4 - 8 microM), it was almost completely inhibited. The inhibitory effect of NaF at the higher concentration was abrogated by stimultaneous addition of epidermal growth factor (EGF), but not by 5alpha-dihydrotestosterone (DHT) or insulin-like growth factor (IGF). These data demonstrate that EGF can effectively reduce the cytotoxic activity of NaF at micromolar concentration.

Animals↗

The distribution of syndecan during murine secondary palate morphogenesis.

The distribution of syndecan, an integral membrane proteoglycan, has been immunohistochemically mapped during the course of murine secondary palate morphogenesis, gestational days 12-15. Syndecan has been shown to mediate cell adhesion and shape change and to be involved in epithelial-mesenchymal interactions during the morphogenesis of several structures. Changes in epithelial cell architecture accompany and may serve to direct the reorientation of the murine secondary palatal shelves from a vertical position on either side of the tongue to a horizontal and adhering position above it. Using a monoclonal antibody made to the core protein of the ectodomain of syndecan, staining was observed to correlate with epithelial cell shape, packing and degree of differentiation. Staining of condensing mesenchyme was also observed. Syndecan may be involved in modulating epithelial cell shape, architecture and fates during both major phases of secondary palate morphogenesis: shelf reorientation and midline epithelial seam dissolution.

Animals↗

[Mechanics of growth pulsations as the basis of growth and morphogenesis in colonial hydroids].

Growth and shaping in colonial hydroids (Hydrozoa, Cnidaria) are realized due to the functioning of special colony elements, growing tips located at the terminuses of branched colony body. Unlike in plants, the growing tips of colonial hydroids are sites of active cell movements related to morphogenesis and lacking proliferation. The activity of hydroid growing tips is expressed as growth pulsations: cyclic repetitions of their apex extensions and retractions. The parameters of growth pulsations are species specific and related to the shape of a forming element. Here, the succession of cell movements and changes in mutual arrangement within the growing tip are described in detail at all pulsation phases. The role of the inner cell layer in the tip activity was demonstrated for the first time. Relationships between the growing tip parameters, length and diameter, and pulsations are discussed. A scheme is proposed for cyclic processes in both epithelial layers. An explanation is provided for the two-step mode of growth pulsations with relative independence of the main phases. It was proposed that successive activities of the tip ecto- and endoderm serve as driving forces provided there is a hard outer skeleton. This scheme makes it possible to explain some patterns of growth and morphogenesis in colonial hydroids, such as gradually increasing growth rate of a new tip and its maximum growth rate, differences in the parameters of growth pulsations between shoot and stolon tips, shoot base inclination towards the stolon tip, etc., and provides a basis for further improvement of the model of morphogenesis in hydroids.

Animals↗

Recent progress in vertebrate limb morphogenesis.

Morphogenesis of vertebrate limb, specifically that of the chick wing, has been recognized as a suitable model to study the cellular and molecular mechanisms of pattern formation. The importance of cellular inductive phenomena and the relevance of the processes such as cell division and cell death in the above model are discussed. These studies have revealed the retinoic acid (RA) and retinols as convincing candidates for vertebrate morphogens. The recent discovery that the RA receptors belong to the steroid hormone receptor superfamily might indicate the universality of the RA morphogen and might enlighten the possible mode of its action. Identification and characterization of the 1d locus genes associated with the mouse limb morphogenesis and the possible involvement of the homeobox proteins in chick wing development have opened new prospects in understanding the molecular mechanisms of vertebrate morphogenesis.

Animals↗