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C Bélisle

Publications and source records attributed to C Bélisle.

12 recordsLinked to original sources

A modified silver impregnation technique for the observation of thick sections of lymph nodes perfused with colloidal carbon.

For many years, a variant of the silver impregnation technique of Bielchowsky has been used to study the lymph node because it clearly outlines the various structures which are usually hard to contrast with standard staining methods. Like other variants of silver impregnation, this method blackens the cell nuclei as well as the reticular fibers; however, it inhibits the impregnation of the nuclear chromatin immediately adjacent to fibers. Hence, this variant selectively darkens the lymphoid cell populations of the nodal structures which contain a loose fiber network. To study the blood vascular network of the lymph node based on perfusion of colloidal carbon, a staining procedure was needed which would contrast nodal structures on thick sections, while allowing the carbon-filled small blood vessels to be distinguished from the impregnated coarse reticular fibers. In an attempt to adapt this variant of Bielchowsky's technique, 10, 20, 40 and 60 microns thick sections from rat nodes, fixed in a solution of Bouin-Hollande for 72 hr, were silver impregnated with serial dilutions (1:2 to 1:128) of the ammoniacal silver solution. Forty-micrometer thick sections impregnated with a 1:16 dilution of the original silver solution at 37 C and for 30 min provided the best results for the conditions.

Animals↗

Blood vascular network of the rat lymph node: tridimensional studies by light and scanning electron microscopy.

Many aspects of the blood vascular network of the lymph node are unknown, and others need confirmation. We have studied the blood vasculature of rat peripheral lymph nodes by means of carbon perfusion and vascular cast corrosion techniques. At the hilus of the node, an artery gives off arterioles running in medullary cords towards the cortex. Some reach the peripheral cortex directly, branching there into slender cortical vessels. Other arterioles enter the periphery of the deep cortex units, and then head towards the peripheral cortex. Upon reaching it, they curve part way above the center of the deep cortex units and provide slender branches to the overlying peripheral cortex. Dense plexuses of capillaries arise from arterioles in the medullary cords, in the periphery of the deep cortex units, and in the outermost stratum of the extrafollicular zone of the peripheral cortex. In the cortex, the draining high endothelial venules are restricted to the extrafollicular zone and to the periphery of the deep cortex units. At the cortico-medullary junction, these peculiar venules transform into regular medullary venules which form the hilar veins. In contrast, the folliculo-nodules and center of the deep cortex units are little vascularized by a loose capillary network, while no vessels occur in the subsinus layer. These features of the node vascular network are of interest in relation to the node architecture.

Animals↗

The narrowing of high endothelial venules of the rat lymph node.

The lymph node contains blood vessels of a special type, termed "high endothelial venules" (HEVs), which are involved in the process of lymphocyte recirculation. In standard tissue sections, many HEVs exhibit a nearly closed or closed lumen containing small lymphocytes but few, if any, erythrocytes. The question arose as to whether the appearance of HEVs in tissue sections is influenced by the routine method of animal sacrifice and/or of tissue processing. Therefore, the present work investigated the effects on HEVs of sacrificing rats as well as of excising and fixing their nodes with various procedures. It was observed that procedures involving animal bleeding or blood loss from nodes increase the percentage of HEVs exhibiting a nearly closed or closed lumen. The results further revealed that the endothelial thickness and other morphological features of HEVs are modified by this artifactual narrowing of HEVs. The possible significance of the phenomenon is discussed.

Animals↗

Tridimensional study of the deep cortex of the rat lymph node. VI. The deep cortex units of the germ-free rat.

The deep cortex of the normal rat lymph node consists of semirounded lymphocytic structures, termed deep cortex "units," each being centered on the opening of an afferent lymphatic. The aim of the present work was to investigate the morphologic features of the units in germ-free animals, in an attempt to evaluate the influence on natural exogenous antigenic stimulation on the development of the units. For this, the lymph nodes from various anatomic locations from 8-week-old Sprague-Dawley germ-free rats were analysed tridimensionally. The observations revealed that, in comparison with the lymph nodes of normal rats, the units of the cervical and mesenteric lymph nodes of the germ-free animals were underdeveloped, while those of the brachial, inguinal, and popliteal lymph nodes were unchanged. Moreover, the germ-free state modified the units of the mesenteric lymph nodes in a manner not encountered in the remaining lymph nodes. Other morphologic features of the peripheral cortex of the lymph nodes of germ-free rats also differed from those of normal ones. The significance of these differences is discussed with respect to immune responses and the process of lymphocyte recirculation. They are of interest because they support previous proposals regarding some aspects of the functioning of the normal lymph node, accounting for the features of the structures and overall architecture of the organ.

Animals↗

Tridimensional study of the deep cortex of the rat lymph node. I: Topography of the deep cortex.

Diagrams of the lymph node currently represent its deep cortex (paracortex) as a layer of rather uniform thickness underlying the whole peripheral cortex. However, this concept has not been supported by actual observations; previous investigators have observed, instead, related structures whose appearance varied greatly from nodule-like to ill-defined components. Clearly, the present knowledge of the histology of the deep cortex is inadequate and confusing. Therefore, we undertook a tridimensional study of the region in different nodes of rats. The present work, bearing on the topography of the region, revealed that the deep cortex of the rat node is formed of one to several basic "units." Each unit is a semi-rounded structure, varying from semispheric to semi-ovoid in shape and contiguous to a portion of peripheral cortex. The work further showed that two to several units can fuse to form a "complex." The data indicated that the number, the size and the shape of the units and/or of the complexes of a node differ to some extent according to its anatomical location. These differences probably reflect corresponding variations in the nature and importance of the antigenic stimulation in the different sites of the organism. Finally, the study demonstrated the necessity of tridimensional examination of a node to obtain adequate information on its overall architecture and, particularly, on its deep cortex topography.

Animals↗

Tridimensional study of the deep cortex of the rat lymph node. II: Relation of deep cortex units to afferent lymphatic vessels.

Recently, we reported that the deep cortex of the rat lymph node is formed of semi-rounded structures, the "deep cortex units," contiguous to the peripheral cortex and bulging into the medulla. It was suggested that a unit represents an accumulation of lymphocytes centered on the opening of an afferent lymphatic vessel. To verify the proposal, we carried out a tridimensional analysis of serially sectioned rat nodes, fixed by perfusion and trimmed in such a way as to preserve their lymphatics. The tridimensional analysis revealed that a constant topographical relationship exists between the units and the openings of the afferent lymphatics. The results demonstrated that the topographical organization of the deep cortex of a rat node correlates with the distribution pattern of the opening(s) of its afferent lymphatic(s). The overall observations suggested the following explanation for the shape and topography of the units: factor(s) present in the lymph would spread in a radial manner from the opening(s) of an afferent lymphatic through the underlying cortex. The factor(s) would induce morphological modifications in the stimulated semi-rounded area which, in turn, would provoke a local accumulation of circulating lymphocytes.

Animals↗

Tridimensional study of the deep cortex of the rat lymph node. III. Morphology of the deep cortex units.

Recently we reported that the deep cortex of the rat lymph node is made up of semi-rounded "units," some of which are partially fused into "complexes." We further found that each unit is centered on the opening(s) of an afferent lymphatic vessel, the topographical organization of the deep cortex of a node correlating with the distribution pattern of the opening(s) of its afferent lymphatic(s). The present study aims to clarify the morphology of the deep cortex unit, particularly with regard to its reticular framework, its lymphatic sinuses, as well as its network of postcapillary venules. For that purpose, we analyzed rat nodes from various locations by way of tridimensional reconstruction. The observations revealed that each unit is formed of a "center" and a "periphery," distinguishable from one another on the basis of their morphological features. The center is nearly devoid of reticular fibers, whereas the periphery exhibits a dense framework of fibers. Moreover, the periphery is the site of concentration of most postcapillary venules of a unit and contains lymphatic sinuses which, peculiarly, are often loaded with small lymphocytes. While both regions are populated mainly by small lymphocytes, the periphery usually contains a lower concentration of these cells than the center. The overall findings support the view that the center is a site of cellular retention and proliferation, whereas the periphery is a site of rapid lymphocyte migration in and out of the unit.

Animals↗

Tridimensional study of the deep cortex of the rat lymph node. IV. Differential labelling of the deep cortex units with 3H-uridine.

Using tridimensional reconstruction, it was recently found that the deep cortex of rat lymph nodes comprises one to several basic "units." Each unit is a semi-rounded structure contiguous to the peripheral cortex and bulging into the medulla of a node. Other investigators reported that transfused lymphocytes, heavily labelled in vitro by 3H-uridine, became concentrated in an ill-defined region of nodes, referred to as the mid and deep cortex. This suggested to us that the in vivo labelling of nodes with 3H-uridine might allow to further characterize the units on a physiological basis. Therefore, rats were injected intravenously with a dose of 1--20 muCi of 3H-uridine/gm body weight and sacrificed 1 hour to 3 days later. The radioautographs of their nodes were exposed up to 535 days. The observations revealed that a large dose of 3H-uridine combined with a long exposure of the radioautographs yielded a differential labelling of the cell population of the units, characterized by a much more intense reaction than that of the surrounding structures. This demonstrated that the physiology of the lymphocyte population of the deep cortex units differs from the morphologically similar lymphocyte population of the extrafollicular zone of the peripheral cortex. The possible reason(s) for the differential labelling of the units is discussed.

Animals↗

Tridimensional study of the deep cortex of the rat lymph node. V: Postnatal development of the deep cortex units.

Recently, the deep cortex of the adult rat lymph node was shown to be made up of semirounded lymphocytic "units." Each unit is contiguous to the peripheral cortex, bulges into the medulla, and is centered on the opening(s) of an afferent lymphatic vessel of a node. Furthermore, each unit comprises a "center" and a "periphery," bearing distinct morphological features. The present study investigated the postnatal development of the units in rats of various ages. One minute after birth, no lymphocytic structures were detected in the nodes. One day after birth, tiny rounded lymphocytic areas were detected in the developing cortex. These areas were topographically related to the openings of afferent lymphatic vessels. One week after birth, small semirounded lymphocytic areas with some morphological features of the adult deep cortex units were observed. Two weeks after birth, typical units were present in the nodes. The observations indicated that the rounded lymphocytic areas observed in nodes of rats aged 1 week or less were actually developing deep cortex units. The overall findings further provided information on the morphological processes involved during the postnatal development of the deep cortex units. Key works: lymph node, deep cortex, development of deep cortex, rat.

Aging↗

Topography of the deep cortex of the lymph nodes of various mammalian species.

In preceeding studies, we clarified the histology of the deep cortex of the rat lymph node. It was shown that the deep cortex is made up of basic elements termed "deep cortex units,' some of which are fused to one another into "deep cortex complexes.' Each unit is a semirounded lymphocytic structure, centered on the opening of an afferent lymphatic, contiguous to the peripheral cortex, and bulging into the medulla of a node. Moreover, each unit comprises a "center' and a "periphery,' bearing distinct morphological features. The present work was undertaken to verify whether the histology of the deep cortex of nodes form various species of mammals, currently used or experimental immunology, is comparable to that of the rate. The observations yielded a positive answer to the question.

Adolescent↗