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PubMed1994

Fine autoradiographical study on scale morphogenesis in the regenerating tail of lizards.

Regenerating scales in lizards originate as pockets in the epidermis instead of epidermal elevations as during embryo development. The morphogenesis of scales in the regenerating tail of the lizards. Anolis and Lamphropholis was studied after peritoneal injection of 3H-thymidine. The tracer was localized in the forming epidermis after progressive post-injection times, by means of autoradiography on plastic sections. After 4-5 hours post-injection of 3H-thymidine, the radioactivity was localized in the basal layer. After 2 to 4 days post-injection labelled cells were seen in the basal and intermediate spinosus layers but not in the uppermost keratinizing layers. Labelled cells were seen in the differentiating cornifying layers (pre-beta and pre-alpha) 6-8 days post-injection. At 12-14 days post-injection almost no radioactivity was seen in the basal layer or in the living part of the epidermis. A few labelled cells were present in the dense keratinizing layers of the sloughing wound and interscale lacunar layers. This study shows that scale formation and morphogenesis in the regenerating tail is brought about by a localized cell proliferation along the regenerating epidermis. In the forming scales the percentage of labelled cells in the distal side (future dorsal part of the new scale) is much higher than in the proximal side (the future ventral side of the scale), so that overlapped scales are generated.

Animals↗
PubMed1993

[Genetic expression and morphogenesis of the skin in vertebrates].

Retinoic acid is an exception amongst teratogenic agents: indeed, it not only interferes with morphogenesis, but it also alters positional values, which leads to the formation of additional structures. The latter effect, similar to that of homeotic mutations, which have been characterized in drosophila, is the most instructive way to study morphogenetic processes. Intramniotic injection of retinoic acid into 10-day-old chick embryos causes feather formation on the scales of the anterior side of tarsometatarsus. Likewise, when the upper-lip skin of 13.5-day mouse embryos is treated in vitro with retinoic acid, there is a development of mouse vibrissa hair buds into glands. The expression of retinoic acid nuclear receptors (RARs) has been studied by in situ hybridization during normal morphogenesis of hair vibrissa follicles as well as during retinoic acid-induced glandular metaplasia. During the normal development of hair follicle, only RAR alpha and RAR gamma genes are transcribed, whereas RAR beta gene expression remains undetected by the techniques used. Retinoic acid skin treatment brings about RAR beta expression in the dermis, leading to the formation of glomerular glands. Now, hair, gland, scale or feather differentiation implies dermal-epidermal interactions which have been extensively studied. However, little is known about the language of communication used by the two tissues. Even if the inductive role of the dermis seems dominant, the competence of the epidermis must not be neglected: the latter determines in some cases the type of cutaneous appendage which is formed.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMedDate not supplied

Alterations in cranial morphogenesis in the Lp mutant mouse.

The effects of exencephaly on cranial morphogenesis were studied at 10 to 12 days of gestation in the loop-tail (Lp) mutant mouse in which the hindbrain and spinal cord fail to close. At the level of the hindbrain, the otocysts became displaced ventrally in abnormal (Lp/Lp) embryos, and the everted neuroepithelium showed a diminished "luminal" reaction to the lectins WGA and Con A, as compared with normal embryos. Also, occasional clusters of rounded cells that resembled presumptive neural crest cells and that labeled with WGA, Con A, and anti-N-CAM were observed at the everted tips of the open neural folds. By 12 days' gestation, there was a loss of integrity in some areas of the neuroepithelium. However, despite the topographic and neuroepithelial distortions, normal differentiation of a roof plate-like neuroepithelium occurred at the ends of the everted neural folds. In addition, the mesenchyme showed normal condensations that labeled with WGA, Con A, and anti-N-CAM in the perinotochordal basicranium and periotic regions. Thus, in this mutant mouse model of neural dysraphism, some features of cranial morphogenesis and differentiation appear to be dependent on timely and proper closure of the cranial neural tube, whereas other aspects may proceed independently of neural closure.

Animals↗
PubMed1996

The DCL gene of tomato is required for chloroplast development and palisade cell morphogenesis in leaves.

The defective chloroplasts and leaves-mutable (dcl-m) mutation of tomato was identified in a Ds mutagenesis screen. This unstable mutation affects both chloroplast development and palisade cell morphogenesis in leaves. Mutant plants are clonally variegated as a result of somatic excision of Ds and have albino leaves with green sectors. Leaf midribs and stems are light green with sectors of dark green tissue but fruit and petals are wild-type in appearance. Within dark green sectors of dcl-m leaves, palisade cells are normal, whereas in albino areas of dcl-m leaves, palisade cells do not expand to become their characteristic columnar shape. The development of chloroplasts from proplastids in albino areas is apparently blocked at an early stage. DCL was cloned using Ds as a tag and encodes a novel protein of approximately 25 kDa, containing a chloroplast transit peptide and an acidic alpha-helical region. DCL protein was imported into chloroplasts in vitro and processed to a mature form. Because of the ubiquitous expression of DCL and the proplastid-like appearance of dcl-affected plastids, the DCL protein may regulate a basic and universal function of the plastid. The novel dcl-m phenotype suggests that chloroplast development is required for correct palisade cell morphogenesis during leaf development.

Amino Acid Sequence↗
PubMed1996

Mouse molar morphogenesis revisited by three-dimensional reconstruction. II. Spatial distribution of mitoses and apoptosis in cap to bell staged first and second upper molar teeth.

Tooth morphogenesis is a complex multifactorial process in which differential mitotic activities and cell death play important roles. Upper first (m1) and second (m2) molars from mouse embryos were investigated from early cap to bell stage. m2 differed from m1 by delayed origin of the enamel grooves delimiting the protrusion of the cap bottom towards the dental papilla, and retardation of the enamel knot formation. The width of the m2 enamel organ was conspicuously smaller during cap formation and length remained smaller throughout the period of observation. Formation of the cap depression was comparable in m1 and m2, however margins delimiting the enamel organ cavity arose in m1 and m2 as mirror images. Attempts were made to correlate changes in the distribution of apoptotic cells and bodies and/or mitoses with morphogenesis. These cellular activities were recorded from histological sections and represented in space using computer-assisted three-dimensional reconstructions. Mitoses in the epithelial compartment were associated with the development of the cervical loop. In the mesenchyme of m1 at early bell stage, a postero-anterior increasing gradient of mitoses was observed which might be correlated with the anterior growth of the molar. Cells in the enamel knot demonstrated a high level of apoptosis, retarded in m2, but absolutely no division. Apoptotic processes were also involved in the anterior delimitation of the m1 epithelium. Apoptosis might correspond to the programmed destruction of cells whose function had to be suppressed or whose potential activity had to be avoided.

Animals↗
PubMed1997

Hydrocortisone modulates the expression of c-ets-1 and 72 kDa type IV collagenase in chicken dermis during early feather morphogenesis.

At the onset of chicken feather morphogenesis, dermal cells migrate along bundles of collagen fibers to colonize areas where bud outgrowth takes place. Chicken embryos treated with hydrocortisone during the critical phase of dermal rearrangement show featherless skin areas in which the dermis exhibits an increase of interstitial collagen. We had previously demonstrated that c-ets-1 is a nuclear transcription factor expressed in the dermis at the beginning of feather morphogenesis. Here we study, by in situ mRNA hybridization, the expression of c-ets-1 in the dermis of chicken embryos treated with hydrocortisone. We found that, among the two distinct products (p54 and p68) encoded by the chicken c-ets-1, the expression of the p68 product increased while expression of p54 decreased after hydrocortisone treatment. Since Ets-1 regulates matrix-metalloproteinases genes, we analyzed the expression of the 72 kDa type IV collagenase in both normal and hydrocortisone-treated embryos. We demonstrated that 72 kDa type IV collagenase mRNA expression decreased in the dermis after hydrocortisone treatment and that its expression correlated with that of p54c-ets-1. Taken together, these results indicate that hydrocortisone modulates c-ets-1 expression. In addition, they raise the interesting possibility that c-ets-1 might be involved in an altered pattern of feather development mediated by the accumulation of collagen due to a decrease in collagenase activities.

Animals↗
PubMed1998

Regulatory control of signal transduction during morphogenesis in Drosophila.

Morphogenesis shapes pattern and size during development. The initiation and propagation of morphogenetic processes is led by the integrated activation of signaling cascades. Much is known about regulatory control of signaling cascades in cell culture systems. However, how this regulatory elements act when cells need to behave coordinately is still unknown territory. The morphogenetic process of dorsal closure proceeds through changes in cell shape and polarity under the control of JNK signaling. Amongst other regulatory elements, Puckered, a Drosophila MAPK phosphatase, is involved in a negative feedback loop that controls JNK signaling activity. puckered is expressed in many other tissues, could influence other developmental events and might regulate different signaling cascades. The negative regulatory control of signal transduction pathways could be a general mechanism regulating differentiation and morphogenesis.

Animals↗
PubMed1999

Progressive association of a "soluble" glycolytic enzyme with the detergent-insoluble cytoskeleton during in vitro morphogenesis of MDCK epithelial cells.

In MDCK epithelial cells, cell contact at confluency initiates a protracted process of morphogenesis during which several proteins known to bind the cytoskeleton become progressively associated with the detergent-resistant cell fraction and distributed to their characteristic polarized domains. Using extraction protocols that identify this tight cytoskeletal linkage, here we show a similar but slower, time-dependent enrichment in the detergent resistant fraction of glyceraldehyde-3-phosphate dehydrogenase (GAPDH), a highly abundant glycolytic enzyme that is traditionally considered soluble. Similar enrichment did not occur for two other glycolytic enzymes, phosphoglycerate mutase or lactate dehydrogenase. Insoluble GAPDH was not homogeneously distributed in the cytoplasm but rather displayed several discrete patterns that varied within and among MDCK cells. It also localized prominently to a few nuclei in the phenotypically heterogeneous cells of late confluency cultures. Disruptors of cytoskeletal filaments were relatively ineffective in the postconfluent epithelial monolayers, although use of disrupting agents implicated actin as the cytoplasmic filament that tethers insoluble GAPDH. Catalytic activity could be demonstrated in the insoluble fraction of GAPDH from postconfluent cultures, but only after release by mechanical disruption of insoluble extracts. Treatment of postconfluent cells with agents that deplete ATP diminished the fraction of cytoskeletally associated GAPDH, and levels of insoluble GAPDH were restored with ATP repletion, suggesting that ATP levels may regulate cytoskeletal linkage and thereby local enzyme activity. We conclude that the highly abundant and ubiquitous enzyme GAPDH becomes progressively enriched in detergent stable subcellular compartments during the process of epithelial morphogenesis. The process that produces GAPDH compartments is slow, suggesting that epithelial cells just at confluency, when they are typically analyzed, have not yet maximized the organizational state that can be attained in monolayer culture.

Acrylamide↗
PubMed1996

Selective regulation of cardiomyocyte gene expression and cardiac morphogenesis by retinoic acid.

Early heart development is known to be sensitive to retinoid concentrations; a specific pattern of malformations is observed in both vitamin A-deficiency and retinoid-toxicity states. While the influence of retinoids on early cardiac morphogenesis has been described previously, the effect of retinoids upon cardiomyocyte differentiation and gene expression is largely uncharacterized. We have established an in ovo chick embryo model in which slow-release retinoic acid (RA) induces four distinct cardiac malformations in a dose-dependent fashion. Late primitive streak-stage chick embryos were treated with all-trans-retinoic acid released from anion exchange beads placed on the embryo's left side and then allowed to develop further for 20-24 hr. At low doses (10 and 25 micrograms/ml RA) an abnormal loop structure was observed. At higher doses (50 and 100 micrograms/ml RA) cardia bifida and clustered heart tissue were noted. Situs inversus only occurred after treatment with 100 micrograms/ml RA. RA-treated embryos were subsequently analyzed for appropriate cardiac myocyte differentiation using antibody staining and ELISA analysis to detect sarcomeric myosin heavy chain, tropomyosin, titin, and alpha-actinin protein expression. Alpha-actinin expression was significantly decreased in RA-treated embryos, as compared to DMSO-treated controls. Also, heart contraction rate was depressed after RA exposure. RA exposure did not alter the protein expression levels of sarcomeric myosin heavy chain or tropomyosin. The observed alterations are consistent with suggestions that retinoids may affect both morphogenesis and myofibril formation in the developing heart.

Animals↗
PubMed1999

Nerve growth factor (NGF) supports tooth morphogenesis in mouse first branchial arch explants.

Posterior midbrain and anterior hindbrain neuroectoderm trans-differentiate into cranial neural crest cells (CNCC), emigrate from the neural folds, and become crest-derived ectomesenchyme within the mandibular and maxillary processes. To investigate the growth factor requirement specific for the initiation of tooth morphogenesis, we designed studies to test whether nerve growth factor (NGF) can support odontogenesis in a first branchial arch (FBA) explant culture system. FBA explants containing neural-fold tissues before CNCC emigration and the anlagen of the FBA were microdissected from embryonic day 8 (E8) mouse embryos, and cultured for 8 days in medium supplemented with 10% fetal calf serum only, or serum-containing medium further supplemented with either NGF or epidermal growth factor (EGF) at three different concentrations: 50, 100, or 200 ng/ml. Morphological, morphometric, and total protein analyses indicated that growth and development in all groups were comparable. Meckel's cartilage and tongue formation were also observed in all groups. However, odontogenesis was only detected in explants cultured in the presence of exogenous NGF. NGF-supplemented cultures were permissive for bud stage (50 ng/ml) as well as cap stage of tooth morphogenesis (100 and 200 ng/ml). Morphometric analyses of the volume of tooth organs showed a significant dose-dependent increase in tooth volume as the concentration of NGF increased. Whole-mount in situ hybridization and semiquantitative reverse transcription-polymerase chain reaction for Pax9, a molecular marker of dental mesenchyme, further supported and confirmed the morphological data of the specificity and dose dependency of NGF on odontogenesis. We conclude that (1) E8 FBA explants contain premigratory CNCC that are capable of emigration, proliferation, and differentiation in vitro; (2) serum-supplemented medium is permissive for CNCC differentiation into tongue myoblasts and chondrocytes in FBA explants; and (3) NGF controls CNCC cell fate specification and differentiation into tooth organs.

Animals↗
PubMed1996

Differential expression of extracellular matrix components during the morphogenesis of human gastric mucosa.

BACKGROUND: Components of extracellular matrix play a crucial role in morphogenesis and epithelial cell differentiation. In this study, we examined the spatiotemporal expression and distribution of several major extracellular macromolecules of developing and adult human gastric mucosa, with particular emphasis on the pit-gland axis. METHODS: Indirect immunofluorescence was performed on cryosections of developing and adult gastric tissues by using specific antibodies. RESULTS: From 8 weeks of gestation onward, including adults, heparan sulfate proteoglycan, type IV collagen, and laminin alpha 1, beta 1, and gamma 1 chains were systematically and uniformly located at the basement membrane of the stratified epithelium (8-11 weeks) and of surface and pit-gland epithelia. Between 8 and 20 weeks of gestation, fibronectin and tenascin were colocalized throughout the entire mesenchyme. In adult mucosa, fibronectin was distributed at all levels of surface and glandular epithelium, whereas tenascin expression was restricted to surface and pit epithelial cells. The expression of the alpha 2 laminin chain was first detected at 12 weeks at the base of forming gastric glands. In the adult, the alpha 2 as opposed to the alpha 1 chain of laminin was confined to the basement membrane of the glandular and lower part of gastric pit epithelia. CONCLUSIONS: This analysis of the composition of the extracellular matrix in human gastric mucosa suggests an important role for some of its components in morphogenesis and maintenance of gastric glands.

Collagen↗
PubMed1999

Expression of green fluorescent protein in the ureteric bud of transgenic mice: a new tool for the analysis of ureteric bud morphogenesis.

The growth and branching of the ureteric bud is a complex process that is ultimately responsible for the organization of the collecting duct system as well as the number of nephrons in the metanephric kidney. While the genes involved in the regulation of this process have begun to be elucidated, our understanding of the cellular and molecular basis of ureteric bud morphogenesis remains rudimentary. Furthermore, the timing and sequence of branching and elongation that gives rise to the collecting system of the kidney can only be inferred from retrospective staining or microdissection of fixed preparations. To aid in the investigation of these issues, we developed strains of transgenic mice in which a green fluorescent protein (GFP) is expressed in the ureteric bud under the control of the Hoxb7 promoter. In these mice, GFP is expressed in every branch of the ureteric bud throughout renal development, and in its derivative epithelia in the adult kidney. As GFP fluorescence can be easily visualized in living tissue, this allows the dynamic pattern of ureteric bud growth and branching to be followed over several days when the kidneys are cultured in vitro. Using confocal microscopy, branching of the ureteric bud in all three dimensions can be analyzed. These mice represent an extremely powerful tool to characterize the normal patterns of ureteric bud morphogenesis and to investigate the response of the bud to growth factors, matrix elements, and other agents that regulate its growth and branching.

Animals↗
PubMed2001

Role of the actin bundling protein fascin in growth cone morphogenesis: localization in filopodia and lamellipodia.

Growth cones at the distal tips of growing nerve axons contain bundles of actin filaments distributed throughout the lamellipodium and that project into filopodia. The regulation of actin bundling by specific actin binding proteins is likely to play an important role in many growth cone behaviors. Although the actin binding protein, fascin, has been localized in growth cones, little information is available on its functional significance. We used the large growth cones of the snail Helisoma to determine whether fascin was involved in temporal changes in actin filaments during growth cone morphogenesis. Fascin localized to radially oriented actin bundles in lamellipodia (ribs) and filopodia. Using a fascin antibody and a GFP fascin construct, we found that fascin incorporated into actin bundles from the beginning of growth cone formation at the cut end of axons. Fascin associated with most of the actin bundle except the proximal 6--12% adjacent to the central domain, which is the region associated with actin disassembly. Later, during growth cone morphogenesis when actin ribs shortened, the proximal fascin-free zone of bundles increased, but fascin was retained in the distal, filopodial portion of bundles. Treatment with tumor promoter 12-O-tetradecanoylphorbol-13-acetate (TPA), which phosphorylates fascin and decreases its affinity for actin, resulted in loss of all actin bundles from growth cones. Our findings suggest that fascin may be particularly important for the linear structure and dynamics of filopodia and for lamellipodial rib dynamics by regulating filament organization in bundles.

Actins↗
PubMed2001

Microtubule integrity is essential for apical polarization and epithelial morphogenesis in the thyroid.

In this study, we examined the contribution of microtubules to epithelial morphogenesis in primary thyroid cell cultures. Thyroid follicles consist of a single layer of polarized epithelial cells surrounding a closed compartment, the follicular lumen. Freshly isolated porcine thyroid cells aggregate and reorganize to form follicles when grown in primary cultures. Follicular reorganization is principally a morphogenetic process that entails the assembly of biochemically distinct apical and basolateral membrane domains, delimited by tight junctions. The establishment of cell surface polarity during folliculogenesis coincided with the polarized redistribution of microtubules, predominantly in the developing apical poles of cells. Disruption of microtubule integrity using either colchicine or nocodazole caused loss of defined apical membrane domains, tight junctions and follicular lumina. Apical membrane and tight junction markers became randomly distributed at the outer surfaces of aggregates. In contrast, the basolateral surface markers, E-cadherin and Na(+),K(+)-ATPase, remained correctly localized at sites of cell-cell contact and at the free surfaces of cell aggregates. These findings demonstrate that microtubules play a necessary role in thyroid epithelial morphogenesis. Specifically, microtubules are essential to preserve the correct localization of apical membrane components within enclosed cellular aggregates, a situation that is also likely to pertain where lumina must be formed from solid aggregates of epithelial precursors.

Actins↗
PubMed2000

Intracellular signaling pathways that regulate dendritic spine morphogenesis.

Rac is a member of the Rho family of small GTPases and acts as a molecular switch. When GTP-bound, Rac binds specific effectors to induce downstream signaling events, including actin cytoskeletal rearrangements (Hall, Science 1998;279:509-514). Herein we review the recent evidence suggesting that Rac is involved in the morphogenesis of dendritic spines (Luo et al., Nature 1996;379:837-840; Nakayama et al., J Neurosci 2000; 20:5329-5338). In addition, we discuss how Rac activity is regulated by guanine nucleotide exchange factors, which may be further regulated by extracellular factors. Thus, the Rac signal transduction pathway may provide links between extracellular ligands or synaptic activity and the regulation of the actin cytoskeleton in spine morphogenesis.

Actins↗
PubMed1990

New insights on the mechanism of testis differentiation from the morphogenesis of experimentally induced testes in genetically female chick embryos.

Embryonic testes grafted in the extraembryonic coelom of 3-day-old genetically female chick embryos may induce total and definitive reversal of gonadal sex differentiation. In this experimental condition, the left gonad becomes a testis instead of an ovary. This makes it possible to compare testicular and ovarian morphogenesis in animals having the same genetic sex and to discount what is due to differences in the genetic determination between male and female. The morphogenesis of such testes is marked by a disappearance of the cortical germinal epithelium. The medullary sex cords keep a narrow lumen instead of becoming large lacunae. The germ cells remain few in the sex cords and do not become meiotic. Furthermore, interstitial cell development is known to be very slow. As a consequence the gross size of the gonad is much smaller than that of an ovary. All these morphogenetic phenomena are unlike those observed during normal ovarian differentiation and evidence an inhibiting influence of the grafted testes. Since inhibition and masculinization are concomitant, inhibition appears to be the mechanism responsible for gonadal sex reversal. The extraembryonic situation of the grafted testes and their relation with the embryo only via the blood stream demonstrates the role of a secreted substance or substances still to be exactly identified. Previous data suggest that this could be the anti-Müllerian-hormone (AMH). Furthermore, previous and present results show that testis differentiation can be actively induced in a bird. This does not agree with the hypothesis that the gonads of the homogametic sex, i.e., the testes in birds, do not need any inducer in order to differentiate.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1994

Temporally and spatially restricted expression of apolipoprotein J in the developing heart defines discrete stages of valve morphogenesis.

During cardiac valve morphogenesis, a series of interactions between the mesodermal-derived myocardium and the overlying endothelium lead to condensed leaflet structure formation. At the atrioventricular (AV) canal, endocardial cells are transformed by specialized underlying myocardial cells into endocardial cushions, and then remodeled into mitral and tricuspid valves. Aortic and pulmonary valves develop by a similar mechanism in the primitive outflow tract. Few genes exhibit restricted spatiotemporal expression in these critical embryonic structures, thus limiting the clues to the sequence of molecular events necessary for valvulogenesis. Apolipoprotein J (ApoJ), a secreted glycoprotein expressed in a variety of cell types at tissue interfaces, exhibits a highly restricted and dynamic expression pattern in the developing heart. ApoJ transcripts were detected in mice at day 9.0 of gestation in the wall of the developing truncus arteriosus. By day 10, intense signal occurred in a thin layer of myocardial cells adjacent to developing endocardial cushions of both atrioventricular canal and truncus arteriosus. No apoJ mRNA was present in the overlying endocardial cushions until day 13.5 when prevalvular condensation begins. Intense expression occurred in the stromal connective tissue throughout leaflet formation. The highly restricted spatiotemporal expression pattern of apoJ in the developing heart implicates its role in the morphogenesis of the AV canal and outflow tract into cardiac valves.

Animals↗
PubMed1982

The morphogenetic triangle: a new conceptual tool for application to problems in dental morphogenesis.

A hypothetical mathematical model of human dental crown height attainment during morphogenesis was developed. The model is based on well-known histologic stages of tooth development from initiation to crown completion, with special emphasis on the dynamics and life-cycle of the cells of the inner enamel epithelium. From three temporal events (P, initiation of proliferation; D, initiation of differentiation; C, crown completion), and crown height (H) measured at the dentino-enamel junction, a morphogenetic triangle PDZ can be constructed where Z = coordinates of H and C at the time of crown completion. The slope of lines PZ and DZ represent average proliferation rate (Rp) and average differentiation rate (Rd) of the inner enamel epithelium, respectively, during crown formation. From data available in the literature, and based on morphogenetic triangle principles, Rp and Rd were calculated for the deciduous and succedaneous dentitions. A strong positive correlation between Rp and Rd was observed throughout the 20-tooth series (r = 0.996); however, Rp and Rd showed moderately strong negative associations with H, P, D, C, and the intervals between PD, PC, and DC. The smaller deciduous tooth in each deciduous:permanent pair was associated with faster Rp and Rd, and shorter intervals between initiation of proliferation, initiation of differentiation, and crown completion. The data generated in this analysis of a hypothetical dentition, while not definitive, provide new insights and a theoretical basis for a broader understanding of the basic mechanisms involved in the attainment of certain tooth dimensions during morphogenesis.

Dental Enamel↗