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F Harrisson

Publications and source records attributed to F Harrisson.

At least 37 records · Page 2Linked to original sources

Immunohistochemical localization of epidermal growth factor in the ovary of the adult Japanese quail.

The present study focuses on the immunohistochemical localization of epidermal growth factor in the ovary of the adult Japanese quail. Immunoreactivity was predominantly found in the smooth muscle cells of blood vessels and of chordae, in granulosa cells of pre-lampbrush follicles, in interstitial cells, in the Balbiani complex of pre-lampbrush oocytes, and in ganglia. In developing follicles, immunoreactivity was also detected in some granulosa and thecal cells, in the zona radiata, and especially in cell clusters localized in the thecal periphery. The number of immunostained cells in the granulosa decreased during folliculogenesis, and increased after ovulation. In the ooplasm of oocytes, immunoreactivity was shifted from the Balbiani complex to the zona radiata during development. These observations support the hypothesis that epidermal growth factors acts primarily on less differentiated follicles. It is also suggested that epidermal growth factor can modulate ovarian contractility. Finally, in one ovary, we detected immunostained bodies in the ooplasm of small developing oocytes.

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Persistence after hypothalamectomy of a tamoxifen-like effect in the ovary of quail embryos exposed cyclically in ovo to low incubation temperatures.

Three days old quail embryos were hypothalamectomized in ovo by the decapitation technique according to Callebaut (1993). Half of these embryos was exposed to day cyclically repeated prolonged low subnormal incubation temperatures (25 degrees C). The other half of these decapitated embryos was normally continuously incubated. As is the case for not decapitated embryos the ovaries of the first group presented a tamoxifen like effect: cortical rim atrophy with proportional hypertrophy of the medulla and a pronounced decrease in the number of oogonia and oocytes. By contrast the ovaries of the second group of decapitated embryos presented a normal aspect without sterilization of the ovigerous sex cords. It was concluded that: 1. the hypophysis, the hypothalamus or other prosencephalic tissues play no intermediary role in the establishment of a tamoxifen like effect in the ovary; 2. the early embryonic development of the quail ovary during the considered period takes place without the intervention of the hypothalamus.

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Behaviour of dissociated hypoblast cells on the basal lamina and on extracellular fibrils in the gastrulating chicken embryo.

The spreading behaviour of dissociated hypoblast cells on and besides a band of aligned fibrils associated with the basal lamina of the epiblast was investigated by the use of scanning electron microscopy. A horse-shaped band of aligned fibrils, first demonstrated by Wakely and England (1979), is present during the gastrulation stages of chicken embryos on the ventral side of the epiblast at the cranial and lateral borders of the area pellucida. The basal lamina of the area pellucida situated inside the fibrillar band enables the spreading and probably the locomotion of dissociated cells, which appeared as polarized cells. Numerous cells were also found on the fibrillar band, and these cells lacked distinct lamellae and a polarized shape. Extensions of the cells contacted the extracellular fibrils and, at these sites of contact, the pattern of the fibrils was frequently deformed. From these observations and from previous results emerged the concept that spreading and locomotion of dissociated hypoblast cells, as well as single mesoblast cells and healing hypoblast epithelium, are inhibited by the band of extracellular fibrils, which acts as a physical barrier. The cell biological basis of the mechanism by which extracellular fibrils associated with the basal lamina arrest the migration of hypoblast and mesoblast cells, but guide the migration of primordial germ cells, is discussed.

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Microinjection of antifibronectin antibodies in the chicken blastoderm: inhibition of mesoblast cell migration but not of cell ingression at the primitive streak.

The involvement of fibronectin in adhesion and migration of individual mesoblast cells during chicken gastrulation was examined after microinjection of functional and nonfunctional antifibronectin antibodies in the blastoderm during the period of rapid migration of mesoblast cells. The injection of affinity-purified polyclonal antihuman fibronectin antibody (total IgG or Fab fragment) or of monoclonal antichicken cellular fibronectin caused a thickening of the primitive streak, which was composed of loosely connected cells. This effect was most evident at the level of Hensen's node, and very few mesoblast cells were observed migrating in the space between upper layer and deep layer. The obvious explanation of this effect was that the de-epithelialization of upper layer cells persisted in the presence of antibodies, but ingressed cells failed to emigrate from the primitive streak. Immunostaining of microinjected antibodies showed binding to the basement membrane, to the cell surface of mesoblast cells that had migrated before microinjection occurred, and to the cell surface of deep layer cells. Cells that ingressed and detached in the course of reincubation of the embryo possessed little immunolabelling along their cell surface. The results suggest that the failure of ingressed cells to emigrate from the primitive streak and to form mesoblast was due (1) to alterations in adhesion between newly ingressed primitive streak cells, which had the ability to detach but possessed relatively little fibronectin along their cell surfaces and a small number of cell protrusions, and (2) probably to a lack of adhesion of detached cells to the basement membrane, which was blocked by the presence of antifibronectin antibodies. We conclude that the presence of fibronectin in the basement membrane is required for emigration of ingressed cells and migration of mesoblast cells to occur. Once migration has commenced, fibronectin is also deposited along the cell surface of migrating cells, a factor that may increase their mutual adhesion.

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In vitro study of the contractility of the wall of the preovulatory follicle in the Japanese quail.

In the present study, we examined in vitro the potassium-induced contractions of isolated parts of the wall of the quail preovulatory follicle through use of an electromagnetic force-length transducer system. We studied longitudinal and transverse strips of the whole wall, of the theca, and of the outer layer (tunica albuginea and surface epithelium). It was observed that force development and shortening of strips of the whole wall and those of the theca are similar. Statistical analysis of maximal isometric force, peak shortening length, and percentage of reduction of follicular volume showed that the orientation of strips is without importance. Results from different orientations were pooled, and the different layers were compared statistically. The general result of this analysis was that the contractile activity of the outer layer was significantly lower than the performance of either the whole wall or the theca. We concluded that contraction of the follicular wall is mainly dependent on the contractile activity of the theca externa. This contractile system is comparable to that of mammals. We suggest that the main function of the theca externa during ovulation is to maintain a constant intrafollicular pressure.

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Cellular origin of the basement membrane in embryonic chicken/quail chimeras.

The cellular origin of the components of the basal lamina of an epithelium, the chicken epiblast, has been investigated in a model system resulting from the transplantation of metabolically labeled quail hypoblast (an epithelium without basal lamina), associated or not with individual mesoblast cells, into an unlabeled chicken blastoderm deprived of its own hypoblast. The ability to discriminate chicken from quail cells after nuclear staining, combined with autoradiographic labeling of the basal lamina components, made it possible to determine the origin of the cells and labeled compounds in tissue sections of the chimeras. The transplantation of 3H-glucosamine or 3H-fucose-labeled quail hypoblast into a chicken host embryo, and subsequent culture of the chimeric embryo for 5 h, led to the transfer of labeled macromolecules from the quail graft to the chicken basal lamina. Pre-treatment of sections with several glycosaminoglycan-degrading enzymes with different substrate specificities suggested that the 3H-glucosamine-labeled compounds that are deposited in the basal lamina are glycoproteins and/or heparan sulfate proteoglycan. However, in view of our current knowledge of the cellular origin of the latter compound, this molecule probably originates from the epithelium itself. Transfer of 3H-proline and 3H-hydroxyproline-containing molecules (mainly collagens) from the graft to the host basal lamina was not observed. Chasing the labeled compounds with unlabeled precursor during culture of the chimeras did not influence the final autoradiographic pattern. It is concluded that the basal lamina of the epiblast has a dual epithelial origin, resulting from the interaction of epiblast-derived materials and non-collagenous glycoproteins synthesized by the hypoblast. Evidence supporting the case of a non-epithelial, mesoblastic origin of non-collagenous glycoproteins was not found. An extensive review of literature on the epithelial vs non-epithelial origin of basement membrane components, mainly in mammalian species, is also provided.

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Smooth muscle cells in the walls of ovarian follicles in the Japanese quail.

The walls of pre-ovulatory follicles of the Japanese quail were examined at the ultrastructural level for the presence of cells displaying the typical morphological features of smooth muscle cells. These characteristics were found in the cells of the chordae, the tunica albuginea, and the theca externa. Small, elongated cells, containing microfilaments, were observed in the theca of prelampbrush follicles localized in the ovarian cortex. These thecal cells were considered as the putative precursors of the thecal smooth muscle cells of the pre-ovulatory follicle. The difference between the smooth muscle cells of the pre-ovulatory follicle and those in the wall of the most recent post-ovulatory follicle is the contracted state of the latter, which is most evident in the cells of the theca externa. It can be concluded that the cells of the theca externa are smooth muscle cells which are mainly contracted during the ovulatory process. A comparison was made with other vertebrate species.

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Immunohistochemical localisation of monoclonal antibody R 24-recognized ganglioside Glac2 in early chick embryos.

The spatio-temporal cellular expression and biosynthesis of ganglioside Glac2 was investigated in early chick embryogenesis. For demonstration of embryonic Glac2-biosynthesis, chick embryos of stage 0 and of stages 4-5 were incubated in vitro in the presence of radioactive sugar precursors. It was found that chick embryos synthesize Glac2 as early as at the blastula stage as well as at the gastrula stage, both within the area pellucida and the area opaca. In contrast to the biosynthetical findings immunohistochemical staining of the chick embryos at various stages by aid of the mouse monoclonal antibody (mAb) R 24, specific for the immunoepitope NeuAc alpha, 8NeuAc alpha, 3Gal beta less than, as present on the ganglioside Glac2, revealed a spatio-temporal cellular pattern of expression of this ganglioside in early chick embryos. Immunohistochemical staining of the chick embryo at stage 0 shows that all cells of the embryo, the extraembryonic epiblast and the yolk endoderm included, are mAb R 24-positive. At the intermediate streak stage (stage 3), the cranial part of the deep layer, the so-called endophyll, is strongly mAb R 24-positive, whereas at the end of gastrulation (stage 5), mAb R 24-recognized epitopes appear to be restricted to a narrow band of deep-layer cells in the endophyllic crescent and to the yolk endoderm of the area opaca. At this stage, no labelling by the antibody is observed in cell layers of the future embryo. The beginning of neurulation (stage 7) is characterized by the expression of the mAb R 24-recognized epitope in the notochord, whilst the deep layer in the cranial part of the neural fold still expresses this epitope. No ecto- or mesodermal structures are stained by the antibody at this developmental stage. During further development (stage 12 and 13), mAb R 24-reactivity is restricted to the cranial part of the embryo with a preferential staining of cells of endodermal origin. At these stages, the notochord expresses mAb R 24 binding sites only in its cranial region. The spatial and temporal correlation between the presence of mAb R 24-recognized epitopes and the morphogenetic positioning of tissues may be indicative for a possible role of the ganglioside Glac2 in corresponding cellular interactions.

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Immunolocalization of smooth muscle-like cells in the quail ovary.

We localized alpha-smooth muscle actin (alpha-SMA) in the quail ovary, using the peroxidase-anti-peroxidase technique. Special attention was paid to the influence of fixation on the immunoreactivity of the antigen. The immunostaining of alpha-SMA largely depended on the nature of the fixative. The antigen could most successfully be localized in ovaries fixed in Carnoy's fluid. We also localized alpha-SMA and desmin in semi-thin glycol methacrylate sections of the pre-ovulatory follicle, using the immunogold-silver staining method. The sections were pretreated with Lugol's iodine or sodium metaperiodate to enhance the immunoreactivity. alpha-SMA was demonstrated in the cells of the chordae, the tunica albuginea, and the theca externa of each follicle. These structures were inter-connected, forming an ovarian suspensory apparatus. The thecal cells of prelampbrush follicles also expressed alpha-SMA. In the wall of the pre-ovulatory follicle, desmin was found in the cells of the chordae and the tunica albuginea, and in a few cells of the theca externa. In the theca interna, desmin, and sometimes alpha-SMA, was observed in cells adjacent to the endothelium of sinusoids, which are probably pericytes. Our results support the hypothesis that in birds the ovarian follicles possess a thecal contractile system, that is presumably involved in the ovulatory process.

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The arrest of cell migration in the chicken blastoderm: experimental evidence for the involvement of a band of extracellular fibrils associated with the basal lamina.

This article overviews our current knowledge of the occurrence and distribution of oriented extracellular fibrils associated with the basal lamina, and their presumptive role in contact guidance of cells in early embryos. To investigate the role of the band of extracellular fibrils situated at the basal side of the epiblast at the cranial edge of the area pellucida of the chicken blastoderm, we determined the precise location and morphology of the fibrils using TEM and SEM, described the relationship between migrating mesoblast cells and the fibrillar band, and, finally, tested experimentally the behavior of homologous and heterologous tissues in the vicinity of the fibrillar band. The descriptive analysis demonstrated that a horseshoe-shaped, 170 microns-wide band of fibrils occurs at the cranial and lateral edges of the area pellucida and area opaca, the highest density being found in the area pellucida. Migrating mesoblast cells presented a surface morphology that was different from the morphology of cells reaching the fibrils at the lateral edge of the area pellucida. Mesoblast cells never crossed the fibrils, an observation that may explain why during gastrulation, mesoblast cells invade the area opaca only in the caudal region, where no fibrillar band is present. The experimental analysis, which involved transplantation and healing experiments, demonstrated that the arrest of cell migration, that occurred in all cases in the vicinity of the fibrillar band, was correlated with changes in surface morphology suggesting a decreased cell adhesion to the fibrils. From these observations emerged the view that the horseshoe-shaped fibrillar band functions as a barrier inhibiting migration of individual mesoblast cells and expansion of tissue sheets, rather than as an extracellular substrate mediating the oriented guidance of cells. In addition to its inhibitory role in cell migration, the extracellular band may also be regarded as a factor that stabilizes the polarity of the early embryo by determining the cranial and lateral limits between embryonic and extraembryonic tissues.

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Features of polyingression and primitive streak ingression through the basal lamina in the chicken blastoderm.

The de-epithelialization of cells of the upper layer during the phenomena of polyingression and primitive streak ingression was studied by analyzing, from the time of laying to the end of gastrulation, the ultrastructure of the basal lamina underlying the upper layer. The electron density of the basal lamina and associated extracellular materials was enhanced by addition of tannic acid to the fixative. Special attention was also paid to the spatial and temporal distribution of blebs at the basal surface of the upper layer, and to the contribution of the de-epithelialized cells to the formation of the deep layer. The results indicate that a nascent basal lamina is already present at the time of laying, especially beneath regions of the area pellucida where polyingression is not apparent. From the onset of incubation, the basal lamina rapidly develops, and it is interrupted by a large number of blebs. However, during the first 6-8 h of incubation, i.e., stages 1-2 of Vakaet (Arch. Biol. (Liège) 81:387-426, 1970), a downward movement of de-epithelialized cells that insert into the deep layer and form the endophyll persists cranially. This phenomenon of polyingression, which starts during the intrauterine period, probably extends from caudal to cranial and comes to an end by stage 3. During these first three stages, the number of blebs progressively decreases, especially in the cranial part of the area pellucida, and a thicker, continuous basal lamina associated with numerous interstitial bodies is laid down. The caudal part of the upper layer is still actively blebbing at that time. Due to the convergence of this area toward the axis of the blastoderm, which leads to ingression at and elongation of the primitive streak up to and including stage 6, the number of blebs at the basal surface of the upper layer progressively decreases. From stage 7 on, blebs are virtually absent; shortening of the primitive streak and formation of the head process begin. At the level of the head process, primitive streak ingression has ceased and a novel basal lamina is progressively deposited beneath the upper layer. By stage 9, a thick, smooth basal lamina physically separates the upper layer from the head mesenchyme. Summarizing, at the time of gastrulation, the presence of blebs that perforate the basal lamina is correlated with the de-epithelialization of cells. Before incubation, however, de-epithelialization of upper-layer cells occurs before the assembly of the basal lamina.(ABSTRACT TRUNCATED AT 400 WORDS)

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Localization of smooth-muscle markers in the ovaries of some ectothermic vertebrates.

In the present study, we have localized desmin and alpha-smooth-muscle actin in the ovaries of the zebrafish, the axolotl, and the red-eared turtle, using the unlabelled antibody peroxidase-antiperoxidase technique. In the axolotl ovary both smooth-muscle markers were demonstrated in cord-like structures, extended along ovarian blood vessels, and in some inner ovarian epithelium cells. In the ovaries of the teleost, smooth-muscle-like cells are detected in a suspensory apparatus formed by venous cords, the tunica albuginea, and the coat around the ovarian artery. Also, in the turtle ovary, smooth-muscle-like cells were found in a suspensory apparatus formed by chordae, the tunica albuginea, and the theca externa of the ovarian follicles. At the present time, the prevailing hypothesis is that, in addition to a role in the mechanical support, the smooth-muscle-like cells in the ovaries of these vertebrates seem to be important with respect to ovarian contractile processes.

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Primary cilia associated with striated rootlets in granulated and folliculo-stellate cells of the avian adenohypophysis.

During observation of the ultrastructure of adenohypophyses of normal and experimentally-manipulated quails, primary cilia were found in secretory cells as well as in non-granulated, folliculo-stellate cells of both cephalic and caudal lobes of the gland. These solitary cilia shared morphological characteristics with those observed in other cell types and species, i.e. they arose from a basal body, which basically had a centriolar structure and 9 doublets of microtubules and no central tubules in their axoneme. A 8 + 1 arrangement of microtubules was exceptionally observed. A 9 + 2 pattern, which is commonly described in motile cilia, was never found. The cilia extended in extracellular spaces between secretory cells, but not in the follicular cavities nor in the blood vessels. In addition to the basal body, a single centriole was frequently present in its vicinity. The basal body was often associated with a basal foot or satellite from which microtubules radiated, and with ladderlike structures corresponding to the classical description of striated rootlets. The presumptive roles of primary cilia in general, and of their morphological features in particular, are discussed in view of our results and compared to the several observations reported in mammalian adenohypophyses. As the evidence gained in favour of a given function of primary cilia has, so far, always been circumstantial, extreme caution in interpretation must be exercise.

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Immunohistochemical localization of desmin in the quail ovary. Demonstration of a suspensory apparatus.

We have localized desmin in the quail ovary, by the unlabelled antibody peroxidase-antiperoxidase technique, using two monoclonal and one polyclonal antisera. Special attention has been paid to the influence of fixation and of proteolytic pretreatment of sections. It appeared that the immunostaining of desmin largely depends on the nature of the fixative. Carnoy fluid, Bouin's fixative, and a paraformaldehyde-acetic acid fixative preserved the histological structure very efficiently. However, trypsin pretreatment proved to be necessary to unmask the antigenic sites in the ovaries fixed in Bouin's fixative and the paraformaldehyde-acetic acid fixative. Desmin immunoreactivity was detected in the tunica albuginea and the chordae, a number of which surrounding the blood vessels, from the hilus to the thecal surface of the follicles. Small branches of chordae connected them with the tunica albuginea, forming a suspensory apparatus. Desmin was also localized in the smooth-muscle cells of the blood vessels. In the theca, immunoreactivity was detected in the wall of arterioles, of venules, and of capillaries. Further experimental and immunohistochemical research have to be performed to establish if the suspensory apparatus is a myoid tissue.

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The chicken blastoderm: in vitro methods of investigation of developmental processes.

A brief description of the structure and a critical survey of the methods of explantation, cultivation, and experimental manipulation of chicken blastoderms are presented. Special attention has been paid to those methods that have enabled major developments in cell and developmental biology of gastrulation. The data suggest that the chicken blastoderm is an appropriate system for the study of developmental processes that are often analogous to medical problems.

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The extracellular matrix and cell surface, mediators of cell interactions in chicken gastrulation.

This article reviews the factors that are involved in cell-cell and cell-matrix interactions during chicken gastrulation. The chemical nature of the extracellular matrix, the structure, composition, cellular origin and remodeling of the basement membrane, and the nature of the cell surfaces are successively analyzed in relation to a variety of cell biological processes, such as cell-to-cell and cell-to-substrate adhesion, specific binding to biological macromolecules and to receptors of the cell surface, promotion of cell migration and positioning, transmembrane triggering of intracellular events, and modulation of cell shape. These processes are the cellular basis of morphogenesis in the chicken blastoderm.

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Morphological and immunocytochemical studies of fibronectin-coated, plasma membrane-limited vesicles in the early chicken embryo.

Vesticles with a mean outside diameter of 32.8 nm have been observed in the early chicken embryo after fixation with a mixture of glutaraldehyde and tannic acid. Densitometric tracing has revealed that the vesicles are limited by a unit membrane. The presence of complex carbohydrates is suggested by the increased electron density of the vesticles after addition of tannic acid to the fixative. Immunocytochemical staining with a monoclonal antibody directed against chicken cellular fibronectin demonstrated the presence of this glycoprotein along the surface of the vesicles. These results suggest a cellular origin of the vesicles, since their surface shares morphological and biochemical similarities with the cell surfaces of the embryonic tissue layers. Recycling of plasma-membrane vesicles may occur, as vesicles were found in the vicinity of coated vesicles. We postulate that extracellular materials of the cell surface, which may affect cell and tissue interactions, are shed in the environment together with plasma-membrane vesicles. The difficulties encountered in observing the vesicles stems from the facts that an adequate visualization method is necessary and that they are few in number. The latter reason suggests their transient nature. The vesicles probably rapidly disintegrate in the extracellular milieu or are recycled by the cell surface.

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