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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

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

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

In vitro development of core cells of the inner cell mass of the mouse blastocyst: effects of conditioned medium.

Blastocysts submitted to two rounds of immunosurgery give rise to cores of presumptive ectoderm cells, many of which do not survive for more than 48 hours when cultured individually. Precoating of the culture plates with conditioned medium (CM) from PYS-2 cells increases the incidence with which cores regenerate an outer layer. This procedure also improves the survival frequency of the cores, but only for a limited period of time. The small number of cores which survive for two weeks or more, either in uncoated or CM-coated plates, give rise to any array of cell types, including giant cells resembling trophoblast.

Animals

Surface differentiation antigens of human mammary epithelial cells carried on the human milk fat globule.

Rabbit antibodies against components of the human milk fat globule bind specifically to normal human breast epithelial cells and cell lines derived from breast carcinomas, as well as to the outer surface of the human milk fat globule. Variation in indirect immunofluorescence staining in both intensity per cell and percentage of cells stained is observed for the different brest cell lines. Cells derived from other epithelial and other ectodermal tissues, fetal fibroblasts, cells of the blood buffy coat, and even fibroblasts of the breast itself do not bind the antibodies. This suggests that these antibodies are detecting cell-type-specific antigens. These normal breast epithelial cell antigens are on the cell surface and their expression is stable in long-term cultured cell lines, even after much chromosomal variation in a given line. By affinity chromatography, three distinct antigenic components can be isolated from the milk fat globule, one of which contains carbohydrate. These differentiation antigens of the human breast epithelial cell are not only useful as specific cell-type markers, but also can provide a tool to study the role of the cell surface in normal and neoplastic mammary development.

Animals

Differential expression of two cadherins in Xenopus laevis.

Using a cadherin fraction from Xenopus tissue culture cells as an immunogen, two monoclonal antibodies were obtained that allowed the characterization of two distinct cadherins in the Xenopus embryo. The two cadherins differ in molecular weight, in their time of appearance during development and in their spatial pattern of expression. One of the antigens was identified as E-cadherin. It appears in the embryonic ectoderm during gastrulation when epidermal differentiation commences and it disappears from the neural plate area upon neural induction. The second antigen could not be allocated to any of the known cadherin subtypes and was termed U-cadherin. It is present in the egg and becomes deposited in newly formed inner cell membranes during cleavage, the outer apical membranes of the embryo remaining devoid of the cadherin throughout development. U-cadherin is found on membranes of all cells up to the late neurula stages. A conspicuous polarized expression of the antigen on the membranes of individual inner cells suggests its participation in the segregation of cell layers and organ anlagen. These findings are discussed in the context of current hypotheses on the role of cadherins in establishing the spatial structure of the embryo.

Animals

[Changes in the expression of prekeratins with molecular weight of 55 and 40 kD in monolayer epithelium during skin morphogenesis in rats].

It has been shown by indirect immunofluorescence using monoclonal antibodies against adult rat simple epithelial prekeratins with the molecular weight of 55 kD (PK55) and 40 kD (PK40) that PK55 was expressed in the covering ectoderm until the 11th day of the antenatal period. PK55 expression markedly increased in ectodermal cells lining the heart region, with PK40 appearing in the same cells on day 11. Beginning from the 16th day gradual loss of both prekeratins started with the parallel formation of squamous differentiated epidermis. These proteins were retained in the outer layers of peridermal cells and in some cells of the basal layer (probably Merkel cells). PK55 was re-expressed on the 18th day in cells migrating into dermal layer during hair folliculi formation. PK55 disappeared again in mature folliculi. Thus, a correlation between distinct morphogenetic events and PK55 and PK40 expression has been found.

Animals

Ornithine aminotransferase distribution in ocular tissues and retinas of cat and mouse.

Ornithine aminotransferase (OAT), a mitochondrial matrix enzyme, is genetically deficient in patients with gyrate atrophy of the choroid and retina. Histologically defined micro-samples (10 ng-6.8 micrograms dry weight) were dissected out from freeze-dried tissue sections of eyeballs of cat and mouse, and the OAT activities were determined by a newly developed microassay method in the ocular tissues and retinal layers. Very high specific activities of OAT, expressed on a dry weight basis, were found in the feline ocular tissues of ectodermal origin, that is, neuroretina, retinal pigment epithelium, ciliary processes and epithelium of iris. In cat and mouse retinas, high OAT activities were distributed in the inner retina with an activity peak in the inner plexiform and ganglion cell layers. Very low activity was present in the outer nuclear layer. The inner segments of photoreceptor cells, which are very rich in mitochondria, contained the highest OAT activity. In contrast, the outer segments of photoreceptor cells contained the low activity resulting from contamination by small pieces of inner segments.

Animals

Histochemical localization of transforming growth factor-beta 1 in developing rat molars using antibodies to different epitopes.

In this study transforming growth factor-beta 1 (TGF-beta 1) has been immunolocalized in developing rat molars using two well characterized polyclonal antibodies, Anti-CC and Anti-LC, that recognize extracellular and intracellular TGF-beta 1, respectively. With immunohistochemical methods and the ABC-peroxidase system of detection, the growth factor was immunolocalized within the ectodermally derived enamel organ and the neural crest-derived dental papilla at the early and advanced bell stages of development. With Anti-CC, widespread and abundant extracellular TGF-beta 1 was found associated with the stellate reticulum and within central and apical regions of dental papilla mesenchyme. In contrast, Anti-LC localized TGF-beta 1 intensely within the cells of the outer dental epithelium. Moderate immunostaining for TGF-beta 1 with Anti-LC was also evident within the apical cytoplasm of inner dental epithelial cells and odontoblasts. These findings support the hypothesis that TGF-beta 1 may play a paracrine role in tooth development by regulating the epithelial-mesenchymal interactions that influence growth and cytodifferentiation events.

Animals

Insulin-like growth factor II may play a local role in the regulation of ocular size.

The ultimate size and shape of the eye has a profound influence on its refraction and function. However, the role of growth factors in normal ocular development is poorly understood. Insulin-like growth factors IGF-I and -II have major effects on cell growth and differentiation in tissue culture. Recently their importance for in vivo development has been studied; IGF-II is predominant prenatally, with a probable local role in the differentiation of some mesodermally derived tissues. Ocular development and size is partially dictated by the condensation of the outer collagenous scleral coat (the 'white') of the eye from orbital mesoderm. We investigated IGF-II expression and IGF-II receptor distribution during normal ocular development in the mouse fetus using in situ hybridization and immunohistochemistry. IGF-II mRNA was expressed by the loose mesenchymal orbital tissue as it differentiated to form the sclera, but not in the compact mature sclera or cornea, or in the ectodermally derived retina or skin. IGF-II gene expression was seen in the orbit at E14, reached a peak just before parturition and then declined to background levels after birth. Similarly, type 2 IGF receptors were shown with immunohistochemistry to be present on developing scleral cells and to be modulated in parallel with IGF-II mRNA expression. We suggest the IGF-II expression by differentiating cells that compact to form the collagenous ocular coat plays a local role in determining the ultimate shape and size of the developing eye.

Animals

In vitro development of inner cell masses isolated immunosurgically from mouse blastocysts. I. Inner cell masses from 3.5-day p.c. blastocysts incubated for 24 h before immunosurgery.

This paper describes the in vitro development of inner cell masses isolated immunosurgically from mouse blastocysts which had been collected on 3.5 days p.c. and then incubated for 24 h. The inner cell masses continue to grow in culture and develop through a series of stages with increasing complexity of internal organization. By day 1 all of the cultured ICMs have an outer layer of endoderm, and by day 3 some of them have two distinct kinds of inside cells; a columnar epithelial layer and a thin hemisphere of elongated cells. Later, mesodermal cells appear to delaminate from a limited region of the columnar layer, close to where it forms a junction with the thinner cells. By day 5, about 25% of the cultured ICMs have a striking resemblance to normal 7.5-day p.c. C3H embryos, with embryonic ectoderm, extra-embryonic ectoderm and chorion, embryonic and extra-embryonic mesoderm, and visceral endoderm. When mechanically disrupted and grown as attached clumps of cells in a tissue dish, these embryo-like structures give rise to trophoblast-like giant cells. These results suggest that the inner cell mass of 4.5-day p.c. blastocysts contains cells which can give rise to trophoblast derivates in culture.

Animals

Ontogeny of basic fibroblast growth factor binding sites in mouse ocular tissues.

Basic fibroblast growth factor (bFGF) binding to ocular tissues has been studied by autoradiographical and biochemical approaches directly performed on sections during mouse embryonic and postnatal development. Frozen sections of embryos (9 to 18 days), newborns, and adults (1 day to 6 months) were incubated with iodinated bFGF. One specific FGF binding site (KD = 2.5 nM) is colocalized with heparan sulfate proteoglycans of the basement membranes and is heparitinase sensitive. It first appears at Day 9 around the neural tube, the optic vesicles, and below the head ectoderm and by Day 14 of embryonic development is found in all basement membranes of the eye. At Day 16, very intensely labeled patches appear, corresponding to mast cells which have been characterized by metachromatic staining of their heparin-rich granulations with toluidine blue. In addition to the latter binding, we have also observed a general diffuse distribution of silver grains on all tissues and preferentially in the ecto- and neuroectodermic tissues. From Days 17-18, there is heterogeneous labeling inside the retina, localized in the pigmented epithelium and in three different layers colocalized with the inner and outer plexiform layers and with the inner segments of the photoreceptors. This binding is heparitinase resistant but N-glycanase sensitive and may represent a second specific binding site corresponding to cellular FGF receptors (KD = 280 pM). Both types of binding patterns observed suggest a significant role for bFGF in eye development and physiology.

Animals

Epidermoid cysts of the cranial bones.

Ectodermally derived tissue in ectopic sites is seen in the head and neck. Extracranial dermoid and epidermoid tumors are relatively rare. While most otolaryngologists are familiar with the cholesteatoma of the otic area, not very many are exposed to this lesion in other sites. Those of the cranial bones are even rarer still. These tumors may expand (1) laterally in the cranial bones, (2) externally to present as masses in the scalp or facial region, or (3) internally to involve the intracranial contents. A patient with a frontal bone epidermoid, whose tumor had expanded in all three directions to erode both inner and outer cortex of cranium and laterally to rupture into the frontal sinus, is discussed. The literature is reviewed and the management of these rare masses is discussed.

Adult

Electron microscopic studies on primary mesenchyme cell ingression and gastrulation in relation to vegetal pole cell behavior in sea urchin embryos.

Early morphogenetic events of primary mesenchyme cell (PMC) ingression and gastrulation were examined by scanning and transmission electron microscopy, with special attention directed to changes in the shape of vegetal pole cells, the length of their microvilli, and interactions between microvilli and the hyaline layer (HL). Eight cells (vegetal pole cells) with elongated microvilli remained in the vegetal pole region while surrounding cells ingressed into the blastocoel to form the primary mesenchyme. These vegetal pole cells indented with the surrounding cells at the stage of gastrulation. The outer surface area with elongated microvilli of vegetal pole cells expanded at the stage of PMC ingression, but was considerably reduced at gastrulation. Microvilli on vegetal pole cells continued to adhere to the HL up to the stage of PMC ingression, but ceased to do so at the time of gastrulation. Thus, the area with separated HL, which is restricted to the region of the PMC released at the stage of PMC ingression, spreads almost entirely throughout the area of the indenting vegetal plate at gastrulation. The apical lamina, apparently consisting of fibrous material intertwinning the stalks of the microvilli, filled the space between the HL and ectodermal cells. The cells surrounding those of the vegetal pole and indenting with those at the stage of gastrulation appeared to behave in the same way as ingressing PMCs in both cell-shape and loss of adhesion of microvilli to HL. The role of vegetal pole cells in early morphogenetic events is discussed.

Animals

Transforming growth factor alpha and its receptor in neural retina.

Transforming growth factor alpha (TGF-alpha) stimulates mitosis of many ectodermal cells but has not previously been studied for its role in neural tissues such as retina. We examined bovine retina for the presence of TGF-alpha mRNA, TGF-alpha protein and for the presence and location of the TGF-alpha/EGF receptor. Biochemical studies demonstrated a high level (770 fmol/mg protein) of specific, high affinity (Kd = 2 nM) TGF-alpha/EGF receptors in membrane homogenates of neural retina, but undetectable binding to homogenates of retinal pigment epithelium. Light microscopic autoradiograms of sections of neural retinal tissue incubated with 125I-EGF indicated that specific TGF-alpha/EGF receptors were present on one or more cell types of the retina with the exception of the outer segments of the photoreceptor cells. In addition, retinal cells appear to synthesize TGF-alpha since both mRNA for TGF-alpha and TGF-alpha protein (4.2 ng/mg protein) were detected in retinal extracts using cDNA hybridization and TGF-alpha RIA techniques. The role(s) of TGF-alpha and its receptor in retina is unknown, but it is possible that they interact via an autocrine/paracrine mechanism to influence retinal regeneration, proliferative retinopathies or neural transmission.

Animals

Cellular morphology and architecture during early morphogenesis of the ascidian egg: an SEM study.

1. Cellular morphology and architecture during early morphogenesis of the ascidian embryo were examined by SEM. 2. The outer surface of the embryo was essentially smooth. The blastocoel could be seen in the dissected blastula. On the cell surface bordering the blastocoel, numerous pseudopodia extended from cells onto adjacent cells. These pseudopodia were suggested to contribute to cell-to-cell adhesion. 3. Before the initiation of gastrulation, a layer of the cells of the animal (ventral) hemisphere and that of the cells of the vegetal (dorsal) hemisphere adhered closely together. The blastocoel could no longer be observed. 4. The gastrulation began during the seventh cleavage. The gastrocoel was formed by a folding of the two layers of the cells. Examinations of the dissected gastrulae suggested two cooperative forces for the gastrulation: first, the epibolic or enfolding movement of the ventral ectoderm cells and secondly, the change in shape of the constituent cells. 5. The neural tube formation progressed in a similar fashion to that of vertebrates.

Animals

Expression of a high molecular weight cell surface glycoprotein (LETS protein) by preimplantation mouse embryos and teratocarcinoma stem cells.

The expression of a high molecular weight cell surface glycoprotein (LETS, fibronectin) by preimplantation mouse embryos as well as cultured teratocarcinoma stem cells was detected by using indirect immunofluorescent staining. When each stage of preimplantation embryonic development was tested for the presence of LETS protein, none was observed on two-cell, four-cell, or eight-cell embryos, or on the morula or the outer cell layer (trophectoderm) of the early or late blastocyst. However, when the inner cell mass was isolated by immunosurgery, positive staining was observed. The intensity of the staining was significantly greater on the inner cell mass isolated from the expanded (day 4) blastocyst than on that from the early (day 3) blastocyst. Certain established cell lines of teratocarcinoma stem cells (embryonal carcinoma cells) also express cell surface LETS protein. "Nullipotent" (Nulli-SCC-1) as well as pluripotent (PSA 1) embryonal carcinoma cell lines have deposits of LETS protein concentrated in areas of cell-cell contact. In addition, a teratocarcinoma-derived endodermal cell line (PYS) was found to be capable of depositing LETS onto the substratum in a fibrillar network.Taken together, our results indicate that LETS protein is synthesized at a specific stage of preimplantation mouse embryonic development. In particular, they suggest that LETS protein is a product of the embryonic ectoderm, and that some types of embryonic endoderm are also capable of synthesizing this protein.

Animals