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Biomedical subjects

D Ribatti

Publications and source records attributed to D Ribatti.

At least 181 records · Page 10Linked to original sources

Effects of the chronic hypoxia on the developing blood-brain barrier in the chick embryo optic tectum.

The blood-brain barrier to horseradish peroxidase has been investigated microscopically and ultrastructurally in the optic tectum of hatching chicken embryos which were kept under hypoxic conditions from the 3rd incubation day. The results indicated that the weakness sites of the blood-brain barrier correspond to the tight interendothelial junctions, whose breakdown allows extravasation of the marker into limited areas of the tectum wall.

Animals↗

[Use of horseradish peroxidase as a marker in studies of the formation of the blood-brain barrier (optic tectum of the normal and experimentally-treated chick embryo)].

The development of the blood brain barrier (BBB) and the vessel permeability to horseradish peroxidase (HRP) have been analyzed in the optic tectum of chick embryos developed under normal and hypoxic conditions, normal chickens, and chickens born from fertilized eggs incubated under hypoxia but kept in the open air after hatching. The development of chick embryos under a situation of chronic hypoxia was obtained by covering, a half of the shell of fertilized eggs with a thick layer of melted paraffin to obtain a reduction of the exchanges normally occurring between embryonic blood vessels and open air. In the tecta developed in normal conditions the BBB to HRP begins to form on the 14th i.d. and it is complete on the 17th i.d. The O2 deprivation, producing remarkable alterations of the neural substratum, does not affect the development of the BBB to HRP, since in chicks of 17 i.d., grown up under hypoxic conditions, the tectal microvessels are not permeable to the tracer, being it mainly confined within the vessel lumina. Nevertheless in specimens kept under hypoxia until hatching, areas of perivascular spread of the marker have been observed corresponding to the vessel wall tracts presumably damaged by the experimental conditions along which the BBB to HRP is not complete.

Animals↗

Blood vessel ultrastructure in the chick optic tectum developing under chronic hypoxia conditions.

The maturation process of blood vessels has been ultrastructurally investigated in the optic tectum of chick embryos kept in a condition of aerogenic hypoxia and of chickens born from fertilized eggs incubated under hypoxia but kept in the open air after hatching. By comparing the fine structure of the intratectal vessels of chick embryos exposed to hypoxia to that of embryos developed under normal conditions, the conclusion has been drawn that O2 deprivation does not prevent the temporal sequence of appearance and/or differentiation of the various vascular wall components (endothelium, endothelial basement lamina, pericytes, perivascular glia), but it produces, at least in a part of the latter, modifications, the type and degree of which apparently depend upon hypoxia duration.

Animals↗

Vasculogenesis in the chick embryo optic tectum.

The initial formation and further development of the intraneural blood vessel network in the tectum opticum of the chick from the 4th to the 14th incubation day have been analyzed and some quantitative data morphometrically recorded. Vessels have been filled by intracardial injection of India ink in vivo. As inferred from our previous investigations on the vasculogenesis of several districts of the central and peripheral nervous system in the chick embryo, also in the developing optic tectum growth and distribution pattern of the vessels seem to unfold step by step under the local influence of earlier occurring morpho-histogenetic processes of the corresponding neural substratum.

Animals↗

Ultrastructural basis of the vessel wall differentiation in the chick embryo optic tectum.

The mode and the temporal sequence of the growth and differentiation processes of the intraneural blood vessels have been ultrastructurally analyzed in optic tecta of 4, 8, 12, 16 and 19 incubation day chick embryos and of chicks 15 days after hatching. The findings suggest that the morphological maturation of the various components of the vascular wall and perivascular spaces, leading to the formation of the blood-brain barrier, unfold according to a spatio-temporal gradient, radially extending from the surface of the tectum rudiment.

Animals↗

[Effects of chronic hypoxia on microcirculation of the central nervous system during embryonic development].

In the attempt to know the factors influencing the vasculogenesis and to verify whether the vessel formation and growth are influenced, during different ontogenetic periods, by oxygen deficiency, the intraneural vascular network has been morphometrically analyzed in chicken embryo optic tectum under conditions of aerogenic hypoxia. Chicken eggs, incubated under routine conditions, have been half-painted with melted wax at the 2nd incubation day (i.d.). Fragments of optic tectum, isolated from living embryos at the 8th, 14th and 17th i.d., have been fixed and embedded according to the usual E/M procedures. A parallel series of normally developed embryos of 8, 14 and 17 incubation days has been likewise prepared. On semithin sections from the hypoxic specimens and normal control embryos the area occupied by vessels (Av), the number of vessels (Nv) and the diameter of the radially directed ones (Dv) have been evaluated. The preliminary results indicate that hypoxia evokes, from the 8th to the 17th i.d., an increment of the Av parameter, due first to microvessel neoformation and enlargement from the 8th to the 14th i.d., then from the 14th to the 17th i.d. only to a growth of new capillary branches. The response of the vascular network to the hypoxic condition is more marked from the 8th to the 14th i.d., t.i. when, in relation to the differentiation and the stratification of the neurons, the metabolic requirements of the developing tectum are presumably increased. O2 deficiency causes severe developmental disorders of the cyto- and mieloarchitecture of the tectum: the vascular response can apparently prevent actual damages only when the hypoxic condition lasts for a relatively short time.

Animals↗

[Structural changes of fenestrated capillaries during development of various endocrine glands in the chicken].

The ultrastructure of the blood capillaries has been analyzed in the adenohypophysis, the pineal body, the thyroid and the suprarenal gland of chick embryos of 7, 14 and 19 days of incubation and, respectively, of the adult fowl. During the organogenesis of these endocrine glands the vascular primordia, precociously appearing among the epithelial components, undergo morphological maturative changes leading to the formation of capillaries built of a fenestrated endothelium and an underlying thin layer of pericytes included in a splitting of the endothelial basement lamina. The capillary differentiation processes, almost identical in the mentioned glands, follow a different chronological sequence, occurring precociously in thyroid and suprarenal and later on in adenohypophysis and pineal body. The relationships between the structural modifications of the capillary wall and the morphological and functional differentiation of the endocrine cells during the organogenesis of the glands have been briefly discussed.

Adrenal Glands↗

[Ultrastructural aspects of vasculogenesis in the optic tectum of the chick embryo].

In chick embryos of 4,8,12 and 19 incubation days the ultrastructure of the intrinsic vessels of the optic lobes has been analyzed. During the early periods of the optic tectum vasculogenesis morphological differences have been observed between the vessels which, arising from the external vascular plexus, radially run through the mantle layer, and the vessels of the deep vascular plexus, which precociously arises by branching and anastomosing of the radial trunks immediately beneath the neuroepithelium. Such differences concerning the endothelium thickness, the characteristics of its luminal and abluminal outline, the time of appearance of the endothelial basement lamina and the glial layer, and the width of the perivascular spaces have been related to the growing patterns of the radial vessels which, at least during the first developmental days, are unbranched trunks and precociously differentiate elongating as the mantle layer become more and more thick, and, respectively, of the vessels of the deep plexus, the morphological maturation of which is belated owing to the continuous sprouting of new vascular sprouts.

Animals↗

Ultrastructural study of the developing blood capillaries in the pineal gland of the domestic fowl.

The ultrastructure of the pineal gland capillaries in chick embryos of 7, 14 and 19 days of incubation and in three month chicks has been analyzed. The results show that vascular primordia are detectable among the pineal developing vesicles on the 7th incubation day. Significant maturative changes in the capillary wall (flattening of the endothelial cells, appearance of pores, narrowing of the perivascular spaces and formation of the endothelial basement lamina) take place during the last days of the embryonic life and are concluded only after the hatching. Relationships between the vasculogenetic and the morpho-histogenetic processes in the pineal body, and some differences in the maturative changes of the vascular wall of the pineal and other chick embryo endocrine glands, have been briefly discussed.

Animals↗

Preliminary investigation on the submicroscopical structure of blood capillaries of the pineal body in the embryonic and adult domestic fowl.

The morphological changes of the blood network capillaries in the pineal gland of the domestic fowl during embryonic development and in the first post-hatching year have been analyzed. Our preliminary results indicate that the first intrinsic vessels, showing an immature aspect, appear among the epithelial vesicles on 7th-8th incubation day (i.d.), when the pinealoblasts are actively proliferating; then, they drastically increase in number (10th i.d.), but they begin to undergo significant maturative changes only on the 19th i.d., in parallel with the unfolding of the pinealocytes differentiation. Finally they reach the definitive set-up within the first post-hatching year, while the pineal is acquiring its full morphological and functional maturity.

Aging↗

[Perivascular spaces in organogenesis of the cephalic lobe of the hypophyseal pars distalis of the chick].

The modifications of the extent and ultrastructure of the perivascular spaces during the development of blood vessel networks in the pars distalis of the adenohypophysis have been analyzed in normally developing chick embryos, from the 7th incubation day to hatching. The quantitative evaluation of the changes undergone by the perivascular spaces, carried out on semithin sections, showed that their extent gradually decreases and that this reduction is paralleled by an increment of the extent of the capillary bed. The ultrastructural study of the hypophyseal sinusoids allowed a precise determination of the size and the minute morphology of these spaces, which attain their definitive characters only during the late incubation stages, when the endothelial basement lamina forms.

Animals↗

[The vascular network and the perivascular spaces of the pars distalis of the hypophysis in developing chick embryos].

A quantitative analysis of the modifications undergone by the vascular networks and the perivascular spaces in the rudiment of the adenohypophysis has been carried out in normally developing chick embryos from the 7th incubation day to hatching. The present results point to the existence, during the developmental period mentioned, of a relationship between the extent of the capillary bed and, respectively, of the perivascular spaces, the gradual increment of the former being paralleled by a decrease of the latter. The rate of these events appears to be higher between the 14th and the 16th incubation days, namely when the vessels begin to acquire some of the morphological features typical of sinusoids, their thin walls getting gradually into closer contact with the secretory epithelial cells.

Animals↗

The role of fibroblast growth factor-2 in the vascularization of the chick embryo chorioallantoic membrane.

The CAM is an extraembryonic membrane which serves as a gas exchange surface and its respiratory function is provided by an extensive capillary network. The development of the vascular system of the CAM is a complex, highly regulated process that depends on genetic and epigenetic factors expressed by endothelial and non-endothelial cells. In spite of the evidence that several growth factors are angiogenic in the CAM assay, poorly investigated is their role in the development of the CAM's vascular system. This article reviews our studies concerning the role of exogenous and endogenous fibroblast growth factor-2 (FGF-2) in the CAM vascularization. The findings in all these studies support the importance of FGF-2 as an autocrine paracrine stimulator of angiogenesis and its key role in the development of the vascular system in the avian embryo.

Allantois↗

Angiogenesis spectrum in the stroma of B-cell non-Hodgkin's lymphomas. An immunohistochemical and ultrastructural study.

Samples of lymph nodes from 88 patients with B-cell non-Hodgkin's lymphoma (B-NHL) grouped by the Working Formulation (WF) and from 15 patients with benign lymphadenopathies were investigated immunohistochemically and ultrastructurally for changes in angiogenesis and stromal distribution of two subendothelial basement membrane (BM) components, namely laminin and type IV collagen. The microvessel number was usually low in lymphadenopathies, and increased significantly in low-grade B-NHL. Intermediate-grade tumors displayed a further significant increase that was mainly due to their diffuse subtypes rather than to the follicular subtype. High-grade B-NHL showed the highest counts. By contrast with the lymphadenopathies studied, the stroma of B-NHL reacted intensely with both BM components, whose linear co-expression was significantly associated with low-grade and follicular intermediate-grade B-NHL, while expression of laminin alone in a granular pattern was detected in diffuse intermediate-grade and high-grade tumors. Ultrastructural analysis revealed immature vessels more frequently in diffuse intermediate-grade, and in high-grade B-NHL. These in situ data suggest that angiogenesis occurring in B-NHL increases along their progression path, and emphasize the importance of angiogenesis as an epigenetic phenomenon of B-NHL progression.

Analysis of Variance↗

Cell-mediated delivery of fibroblast growth factor-2 and vascular endothelial growth factor onto the chick chorioallantoic membrane: endothelial fenestration and angiogenesis.

Fibroblast growth factor-2 (FGF2) and vascular endothelial growth factor (VEGF) exert their angiogenic activity by interacting with endothelial cells in a distinct manner. In this study, we investigated the morphological features of endothelial cells of the chick embryo chorioallantoic membrane (CAM) microvasculature after stimulation with FGF2 or VEGF. In order to provide a continuous delivery of the growth factor, we utilized a recently developed gelatin sponge/CAM assay in which a limited number of FGF2- or VEGF-transfected cells were adsorbed onto gelatin sponges and applied on the top of the CAM on day 8 of development. Their angiogenic activity was compared to that exerted by a single bolus of the corresponding growth factor. All the angiogenic stimuli induced a comparable vasoproliferative response, as demonstrated by the appearance of similar numbers of immature blood vessels within the sponge on day 12. No angiogenic response was observed in CAMs implanted with the corresponding parental cell lines or vehicle. Electron microscopy demonstrated that VEGF-overexpressing cells modified the phenotype of the endothelium of the blood vessels at the boundary between the implant and the surrounding CAM mesenchyme. The endothelial lining of 30% of these vessels showed segmental attenuations, was frequently interrupted and became fenestrated, mimicking what is observed in tumor vasculature. In contrast, the vessels consisted of continuous endothelium sealed by tight junctions in all the other experimental conditions. These results indicate that FGF2 and VEGF interact with endothelial cells of the CAM in a distinct manner. Both growth factors induce a potent angiogenic response, but only VEGF delivered in a continuous manner by its transfectants can modify the phenotype of the otherwise quiescent endothelium of CAM blood microvessels. The gelatin sponge/CAM assay may constitute a new model to study the mechanisms leading to endothelial fenestration in tumor growth.

Adsorption↗