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The nuclear envelope; its structure and relation to cytoplasmic membranes.

An electron microscope study of thin sections of interphase cells has revealed the following:- Circular pores are formed in the double nuclear envelope by continuities between the inner and outer membranes which permit contact between the nucleoplasm and the cytoplasm unmediated by a well defined membrane. The pores, seen in sections normal to the nuclear envelope, are profiles of the ring-shaped structures described by others and seen in tangential section. The inner and outer nuclear membranes are continuous with one another and enclose the perinuclear space. The pores contain a diffuse, faintly particulate material. A survey of cells of the rat derived from the embryonic ectoderm, mesoderm, and endoderm, and of a protozoan and an alga has revealed pores in all tissues examined, without exception. It is concluded that pores in the nuclear envelope are a fundamental feature of all resting cells. In certain cells, the outer nuclear membrane is continuous with membranes of the endoplasmic reticulum, hence the perinuclear space is continuous with cavities enclosed by those membranes. There are indications that this is true for all resting cells, at least in a transitory way. On the basis of these observations, the hypothesis is made that two pathways of exchange exist between the nucleus and the cytoplasm; by way of the perinuclear space and cavities of the endoplasmic reticulum and by way of the pores in the nuclear envelope.

Animals↗

Radial intercalation of ciliated cells during Xenopus skin development.

Cells with motile cilia cover the skin of Xenopus tadpoles in a characteristic spacing pattern. This pattern arises during early development when cells within the inner layer of ectoderm are selected out by Notch to form ciliated cell precursors (CCPs) that then radially intercalate into the outer epithelial cell layer to form ciliated cells. When Notch is inhibited and CCPs are overproduced, radial intercalation becomes limiting and the spacing of ciliated cells is maintained. To determine why this is the case, we used confocal microscopy to image intercalating cells labeled using transplantation and a transgenic approach that labels CCPs with green fluorescent protein (GFP). Our results indicate that inner cells intercalate by first wedging between the basal surface of the outer epithelium but only insert apically at the vertices where multiple outer cells make contact. When overproduced, more CCPs are able to wedge basally, but apical insertion becomes limiting. We propose that limitations imposed by the outer layer, along with restrictions on the apical insertion of CCPs, determine their pattern of radial intercalation.

Animals↗

[Ultrastructural aspects of the migration of somite cells to the posterior limb buds of mice].

Migrating cells originating selectively in the ventral lateral edge of the somites adjacent to the mouse hindlimb bud have been studied in transverse sections by transmission and scanning electron microscopy. Collected during the 10th and 11th gestational days, the embryos have been classified according to the number of metameres. As soon as the 28 somite stage, discrete cytological modifications occur in a limited caudal area of the ventro-lateral somitic edge. Loosing the typical epithelial arrangement characteristic of the dermatome cells, these ventral cells show large areas of close contact between their plasma membrane and a superficial microfilamentous material accumulates in the contact areas. At the 33 somite stage, the same groups of cells elongate and form long cellular trails invading the proximal area of the limb bud mesoderm. The migrating cells become polarized along the migrating axis and they retain large and smooth intercellular contacts with each other. Very selective ultrastructural features of the migrating somitic cells can be interpreted in relation to their cinetic activity or to their early myogenic differentiation. In addition to their mutual superficial relationships, the migrating cells are characterized by the presence of numerous oriented microtubules, of a high number of active mitochondria, of an abundant granular endoplasmic reticulum and of an hypertrophied Golgi apparatus regularly located near the nucleus in the "trailing" edge of the cells. Several dense granules with a diameter of 8 nm are present in the mitochondrial matrix. The extensive Golgi apparatus is associated to numerous thick walled vesicles, which increase from 60 to 150 nm in diameter as they become closer and closer to the plasma membrane. These vesicles are absent in the mesodermal cells of somatopleural origin; their presence in the migrating somitic cells is probably related to an early myogenic differentiation. The observation made near the distal end of the somitic cellular trails suggests that the more distal somitic cells rapidly loose their ultrastructural particularities as soon as they are dispersed in the limb bud mesoderm; this aspect of the processus, however, requires the study of later developmental stages. Other observations made in the same material bring some precisions to the ecto-mesodermal relationships which are established in the apical area of the limb bud. Scanning electron microscopic observations of thick sections reveal that the outer mesodermal cells of this area send numerous filopodia which make contact with the basement membrane underlying the apical ectodermal ridge.

Animals↗

Cell proliferation during the early stages of human eye development.

The distribution as well as the ultrastructural and biochemical characteristics of proliferating cells in the human eye were investigated in five conceptuses of 5-9 postovulatory weeks, using morphological techniques and Ki-67 immunostaining. The Ki-67 nuclear protein was used as a proliferation marker because of its expression in all phases of the cell cycle except the resting phase (G0). The labelling indices of Ki-67-positive cells were analysed by means of the Kruskal-Wallis ANOVA test and the Wilcoxon matched-pairs test. In the 5th week, mitotic cells were the most numerous between the two layers of the optic cup, the optic cup and stalk, and between the lens pit and the surface ectoderm. During the 6th week, cells were observed in the lens epithelium covering the whole cavity of the lens vesicle as well as in the neuroblast zone and the pigmented epithelium of the retina. At later stages (7th-9th weeks), Ki-67-positive cells were restricted to the anterior lens epithelium, the outer neuroblast zone, and the pigmented retina. Throughout all stages examined, mitotic figures were found lying exclusively adjacent to the intraretinal space. Early in the lens pit, they were confined to the free epithelial surface, and later were facing the cavity of the lens vesicle. The proliferative activity was the most intensive in the 6th week, whereas it decreased significantly in the later stages. Additionally, when proliferative activities were compared, the peripheral retina appeared to be less mature than the central before the 9th week. In the earliest analysed stage, cell proliferation might be associated with the sculpturing of the optic cup and stalk, the cornea, and the lens. In the 6th week, the most intensive proliferation seems to be involved not only in the further morphogenesis of the optic cup and the lens vesicle but also in the retinal neurogenesis. At later stages, the decreased proliferation might participate in the neurogenesis of the outer neuroblast zone and the secondary lens fibre formation.

Biomarkers↗

Expression of anterior Hox genes during larval development of the gastropod Haliotis asinina.

We report the spatial expression patterns of five anterior Hox genes during larval development of the gastropod mollusc Haliotis asinina, an unsegmented spiralian lophotrochozoan. Molecular alignments and phylogenetic analysis indicate that these genes are homologues of Drosophila HOM-C genes labial, proboscipedia, zen, Deformed, and Sex combs reduced; the abalone genes are named Has-Hox1, -Hox2, -Hox3, -Hox4, and -Hox5. Has-Hox transcripts are first detected in the free-swimming trochophore larval stage and restricted to the posttrochal ectoderm. Has-Hox2, -Hox3, and -Hox4 are expressed in bilaterally symmetrical and overlapping patterns in presumptive neuroectodermal cells on the ventral side of the trochophore. Has-Hox1 expression is restricted to a ring of cells on the dorsoposterior surface, corresponding to the outer mantle edge where new larval shell is being synthesized. There appears to be little change in the expression domains of these Has-Hox genes in pre- and posttorsional veliger larvae, with expression maintained in ectodermal and neuroectodermal tissues. Has-Hox2, -Hox3, -Hox4, and-Hox5 appear to be expressed in a colinear manner in the ganglia and connectives in the twisted nervous system. This pattern is not evident in older larvae. Has-Hox1 and-Hox4 are expressed in the margin of the mantle in the posttorsional veliger, suggesting that Hox genes play a role in gastropod shell formation.

Amino Acid Sequence↗

The role of fibroblast growth factor in early Xenopus development.

In early amphibian development, the mesoderm is formed around the equator of the blastula in response to an inductive signal from the endoderm. A screen of candidate substances showed that a small group of heparin-binding growth factors (HBGFs) were active as mesoderm-inducing agents in vitro. The factors aFGF, bFGF, kFGF and ECDGF all show similar potency and can produce inductions at concentrations above about 100 pM. The product of the murine int-2 gene is also active, but with a lower specific activity. Above the induction threshold there is a progressive increase of muscle formation with dose. Single blastula ectoderm cells can be induced and will differentiate in a defined medium to form mesodermal tissues. All inner blastula cells are competent to respond to the factors but outer cells, bearing oocyte-derived membrane, are not. Inducing activity can be extracted from Xenopus blastulae and binds to heparin like the previously described HBGFs. Antibody neutralization and Western blotting experiments identify this activity as bFGF. The amounts present are small but would be sufficient to evoke inductions in vivo. It is not yet known whether the bFGF is localized to the endoderm, although it is known that inducing activity secreted by endodermal cells can be neutralized by heparin. The competence of ectoderm to respond to HBGFs rises from about the 128-cell stage and falls again by the onset of gastrulation. This change is paralleled by a rise and fall of binding of 125I-aFGF. Chemical cross-linking reveals that this binding is attributable to a receptor of relative molecular mass about 130 x 10(3).(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Association of SPARC (osteonectin, BM-40) with extracellular and intracellular components of the ciliated surface ectoderm of Xenopus embryos.

SPARC (Secreted Protein, Acidic, Rich in Cysteine) was detected by immunohistochemistry in the sensorial layer of the bilayered embryonic epidermis of Xenopus laevis during neurulation, when a subset of the sensorial cells are selected to differentiate into ciliated cell precursors. After the ciliated cells had intercalated into the outer layer and had undergone ciliogenesis, intense SPARC immunostaining was associated with the cilia and remained associated with the cilia throughout their persistence on the epidermis. Circumferential SPARC immunostaining was also detected at the interface between surface epithelial cells. Animal cap explants indicated that the embryonic activation of SPARC expression in the dorsal ectoderm does not require signaling from factors secreted by the underlying mesoderm. Immunoelectron microscopy revealed that SPARC is intimately associated with the 9 + 2 microtubule arrays of cilia. Our data indicate that SPARC plays a role in the development and function of the surface ciliated epidermis of Xenopus embryos. We propose that the counter-adhesive activity of SPARC facilitates the intercalation of ciliary cell precursors to the surface epithelial layer, where its Ca(2+)-binding abilities promote cell-cell adhesion. Based on its association with ciliary microtubule arrays, we also propose that intracellular SPARC may play a role in regulating ciliary beat frequency and polarity.

Animals↗

The Notch signalling pathway in hair growth.

The Notch signalling pathway is an important mediator of cell fate selection whose involvement in epidermal appendage formation is now becoming recognised. Hair follicle development and hair formation involve the co-ordinated differentiation of several different cell types in which Notch appears to have a role. We report intricate expression patterns for the Notch-1 receptor and three ligands, Delta-1, Jagged-1 and Jagged-2 in the hair follicle. Notch-1 is expressed in ectodermal-derived cells of the follicle, in the inner cells of the embryonic placode and the follicle bulb, and in the suprabasal cells of the mature outer root sheath. Delta-1 is only expressed during embryonic follicle development and is exclusive to the mesenchymal cells of the pre-papilla located beneath the follicle placode. Expression of Jagged-1 or Jagged-2 overlaps Notch-1 expression at all stages. In mature follicles, Jagged-1 and Jagged-2 are expressed in complementary patterns in the follicle bulb and outer root sheath, Jagged-1 in suprabasal cells and Jagged-2 predominantly in basal cells. In the follicle bulb, Jagged-2 is localised to the inner (basal) bulb cells next to the dermal papilla which do not express Notch-1, whereas Jagged-1 expression in the upper follicle bulb overlaps Notch-1 expression and correlates with bulb cell differentiation into hair shaft cortical and cuticle keratinocytes.

Animals↗

Human TIMP-3 is expressed during fetal development, hair growth cycle, and cancer progression.

We studied the expression and regulation of TIMP-3, a recently cloned member of the tissue inhibitor of the metalloproteinase family, during human fetal development and in various human tissues, with emphasis on epithelial structures. Expression of TIMP-3 mRNA was detected by in situ hybridization in developing bone, kidney, and various mesenchymal structures. At 16 weeks of gestation, ectoderm-derived cells of hair germs expressed TIMP-3 mRNA, and beginning from the twentieth week consistent expression was detected in epithelial outer root sheath cells of growing hair follicles. In normal adult human skin, expression of TIMP-3 mRNA was limited to hair follicles, starting at the early anagen (growing) phase and vanishing at the catagen (regressing) phase. TIMP-3 mRNA was not detected in benign hair follicle-derived tumors but was present in tumor cells of infiltrative basal cell carcinomas and in surrounding stromal cells in squamous cell carcinomas. Human primary keratinocytes in culture expressed TIMP-3 mRNAs, the levels of which were upregulated by transforming growth factor-beta (TGF-beta), whereas interleukin-1beta (IL-1beta) and tumor necrosis factor-alpha (TNF-alpha) had no effect. Our results suggest a role for TIMP-3 in connective tissue remodeling during fetal development, hair growth cycle, and cancer progression.

Adult↗

Cornea-lens transdifferentiation in the anuran, Xenopus tropicalis.

Previously, the only anuran amphibian known to regenerate the lens of the eye was Xenopus laevis. This occurs during larval stages through transdifferentiation of the outer cornea epithelium under control of factors presumably secreted by the neural retina. This study demonstrates that a distantly related species, X. tropicalis, is also able to regenerate lenses through this process. A transgenic line of X. tropicalis was used to examine the process of cornea-lens transdifferentiation in which green fluorescent protein (GFP) is expressed in differentiated lens cells under the control of the Xenopus gamma1-crystallin promoter element. Unlike X. laevis, the process of cornea-lens transdifferentiation typically occurs at a very low frequency in X. tropicalis due to the rapid rate at which the inner cornea endothelium heals to recover the pupillary opening. The inner cornea endothelium serves as a key physical barrier that normally prevents retinal signals from reaching the outer cornea epithelium. If this barrier is circumvented by implanting outer cornea epithelium of transgenic tadpoles directly into the vitreous chamber of non-transgenic X. tropicalis larval eyes, a higher percentage of cases formed lenses expressing GFP. Lenses were also formed if these tissues were implanted into X. laevis larval eyes, suggesting the same or similar inducing factors are present in both species. When pericorneal ectoderm and posteriolateral flank ectoderm were implanted into the vitreous chamber, only in rare cases did pericorneal ectoderm form lens cells. Thus, unlike the case in X. laevis, competence to respond to the inducing factors is tightly restricted to the cornea epithelium in X. tropicalis. As controls, all these tissues were implanted into the space located between the inner and outer corneas. None of these implants, including outer cornea epithelium, exhibited GFP expression. Thus, the essential inductive factors are normally contained within the vitreous chamber. One explanation why this type of lens regeneration is not seen in some other anurans could be due to the rapid rate at which the inner cornea endothelium heals to recover the pupillary opening once the original lens is removed. These findings are discussed in terms of the evolution of this developmental process within the anurans.

Animals↗

Transplantation studies to investigate mesoderm-ectoderm adhesive cell interactions during gastrulation.

Experiments involving transplantation of the roof of the blastocoel in Rana pipiens embryos reveal that the inner surface of the roof of the blastocoel must be coated with a fibrous extracellular matrix (F-ECM) to serve as a substratum for mesodermal cell migration. When the roof of the blastocoel is inverted the original outer surface, now projecting toward the blastocoel, does not become coated with F-ECM and does not support mesodermal cell migration. When the roof of the blastocoel is removed from a normal embryo and transplanted into an interspecific arrested hybrid embryo known to be deficient in F-ECM synthesis, the grafted ectodermal fragment does not become coated with F-ECM and does not support normal mesodermal cell migration. When a hybrid graft is placed in a normal embryo, the grafted ectodermal fragment becomes coated with F-ECM and supports mesodermal cell migration. In normal control embryos migrating mesodermal cells are polarized due to formation of lamellipodia on their leading but not their trailing edges. These cells are arranged in overlapping layers. The leading cells form lamellipodia on the roof of the blastocoel and trailing cells form lamellipodia on one another.

Animals↗

Abnormalities of ERG in congenital aniridia.

Congenital aniridia is generally associated with nystagmus, corneal pannus, cataract, ectopia lentis, glaucoma, macular hypoplasia, optic nerve hypoplasia and compromised visual function. Many theories have been proposed, including a failure in the development of the neural ectoderm and/or an aberrant development of mesoderm. We observed the ERG from 19 patients with congenital aniridia. Fourteen patients had abnormal ERG, including the reduced a wave trough under dark adapted red stimuli with dark adaptation; abnormal a or/and b waves in dark or/and light adapted ERG or abnormal oscillatory potentials. The ERG abnormalities reflected the disturbances either in the outer retina such as photoreceptors or in the structures of inner retina. The preserved visual function and several ocular abnormalities show that the aniridia may be a consequence of several pathogenetic mechanisms. This fact supported the opinion that congenital aniridia would be a concurrent failure of both ectoderm and mesoderm in embryonate development.

Adolescent↗

Cell allocation and lineage in the early mouse embryo.

In the early mammalian embryo, initially asymmetric cell contacts appear to induce blastomere polarization, elements of which can persists through cytokinesis. This leads to the generation of inner and outer populations of blastomeres which may subsequently diverge as a result of residing in distinct microenvironments. Similar processes may account for the generation of primitive endoderm versus primitive ectoderm, and that of trophectoderm versus inner cell mass. However, if this is the case, the response of cells to positional cues must change as a function of either their previous positional history or the number of cycles they have completed. Once these primary tissues have been established, specific interactions between them lead to further cellular diversification.

Animals↗

Spatiotemporal distribution of SPARC/osteonectin in developing and mature chicken retina.

Expression of SPARC (Secreted Protein, Acidic, Rich in Cysteine), a counteradhesive, calcium-binding extracellular matrix (ECM) glycoprotein, is associated with several morphogenetic events during early development. In this study, changes in the spatiotemporal distribution of SPARC transcripts and the protein during chicken retinal development were documented by in situ hybridization and indirect immunofluorescence microscopy. SPARC transcripts were first detected within the proliferating neural ectoderm at embryonic day 4. 5 (E4.5), followed short thereafter (E5) by appearance of SPARC. SPARC was enriched within the inner plexiform layer (IPL) by E10 and within the outer plexiform layer (OPL) by E14, several days after these layers became morphologically distinct. Significant levels of SPARC transcripts were first observed within the ganglion cell layer (GCL) at E17 prior to accumulation of SPARC within the nerve fiber layer, seen first at E20. SPARC protein was first detected within the developing retinal pigment epithelium (RPE) at E10 and increased significantly at RPE cells ceased to proliferate and continued differentiating. Of special note was the restriction of SPARC to the basal-half of the RPE cells. SPARC transcripts were similarly distributed in the adult retina, but at lower levels than in the period just prior to hatching. In the adult retina SPARC was retained in the nerve fiber layer and present in the inner nuclear layer (INL) and outer nuclear layer (ONL), but lost from the IPL and OPL. These changes in expression pattern with time indicate that SPARC is developmentally regulated and therefore may have important function(s) in both morphological development of the retina and functioning of the mature eye.

Amino Acid Sequence↗

Prenatal diagnosis of ventriculo-coronary communications in a second-trimester fetus using transvaginal and transabdominal color Doppler sonography.

We report on the prenatal diagnosis of ventriculo-coronary communication associated with pulmonary atresia and an intact interventricular septum. The diagnosis was made by transvaginal color Doppler sonography at 17 weeks' gestation and confirmed by transabdominal sonography at 19 weeks. Color Doppler demonstrated the communication between the distal hypoplastic right ventricle and the right coronary artery, with the course of this vessel situated along the outer heart wall. Spectral Doppler assessment showed bidirectional arterial flow in this vessel. Extracardiac skeletal anomalies (cleft hands and feet) were also detected and classified as an ectrodactyly-ectodermal dysplasia clefting (EEC) syndrome. This led the parents to opt for termination of pregnancy. All findings were confirmed on autopsy. Using stereomicroscopy the malformations of the coronary system and main connections to the lumen of the right ventricle were additionally demonstrated. To our knowledge this is the first report on the identification of coronary artery malformations in a midtrimester fetus using transvaginal color Doppler. The early appearance enabled us to investigate the pathophysiological sequence of the combined defect. Further investigation might elucidate whether ventriculo-coronary communications arise secondary to pulmonary atresia and intact inter-ventricular septum, due to increased ventricular pressure, or are the primary defect leading to intrauterine pulmonary atresia or, indeed, whether both defects appear simultaneously.

Adult↗

Xenopus mesoderm induction: evidence for early size control and partial autonomy for pattern development by onset of gastrulation.

Experiments are described that examine the state of organisation of the presumptive mesoderm and ectoderm of the Xenopus embryo at stages up to the onset of gastrulation. It is shown that a process during blastula stages, establishing the normal proportions in which this cell population is partitioned to found the two outer 'germ layers', has a positive regulative property. An operation has been performed to excise the yolky endodermal core, at the beginning of gastrulation, leaving only the presumptive territories of mesoderm, neural tissue, epidermis and supra-blastoporal endoderm. This reveals that by this time a stable capacity exists within the induced tissue to express the craniocaudal sequence of the normal pattern, including the proper numbers of somite segments. The mediolateral organisation of such body patterns is however abnormal. The relevance of the observations to understanding mechanisms of axial pattern control is discussed.

Animals↗

Distribution and structure of dental enamel in incisors of Tabby mice.

OBJECTIVE: In Tabby mice, the Ta (EDA) gene is mutated. The resulting syndrome is homologous to hypohidrotic ectodermal dysplasia in humans. The Tabby phenotype is characterized by developmental defects of ectodermally derived structures. The teeth show aberrations in number, size and morphology. Dental enamel is a product of specialized epithelial cells, the ameloblasts. It was the aim of the present study to investigate the dental enamel phenotype in Tabby incisors, with emphasis on its distribution and structure. DESIGN: The incisors from five female Tabby and three female wild-type mice were sectioned and ground transversely, etched for 45s with 0.1% nitric acid, sputter-coated with gold-palladium, and observed in SEM. RESULTS: All measured dimensions were more variable in Tabby mice, as was the outline of the enamel-dentin junction. Maxillary incisors were wider in Tabby mice, while mandibular incisors were wider in wild-type mice. No significant difference in enamel thickness was observed. The enamel on the mesial aspect tended to extend further lingually in Tabby incisors in both jaws. On the lateral aspect, this tendency was only significant in mandibular incisors. The enamel-dentin junction often lacked the mesial concavity. Instances of hypoplastic enamel were observed. The complex mouse enamel structure was generally well preserved in Tabby mice, only few instances of aberrant structure were observed. CONCLUSIONS: It is suggested that the reciprocal expression pattern of Ta and Edar (the Ta ligand receptor gene) in outer and inner enamel epithelium, respectively, may influence the position of the enamel-cementum junction.

Animals↗

Multiple fibrofolliculomas (Birt-Hogg-Dubé) associated with a large connective tissue nevus.

Multiple follicular tumors of the type recently described by Birt and colleagues as fibrofolliculomas were observed in the dorsal skin of a white man in association with a large connective tissue nevus. No signs of systemic disease or phakomatosis were present. This type of lesions represents benign neoplasia of the fibrous root sheath of the hair follicle in association with proliferations of the infundibular portion of the outer epithelial root sheath. Its existence supports the view that the mesodermal portions of the pilar apparatus can become neoplastic, and also provides another example of the close interactions existing between the ectodermal and mesodermal components of the hair follicle.

Hair↗