Structure and function of lymphatic postcapillaries (coordinating mechanism of the processes of interstitial transport and lymphatic resorption

Investigation performed on topography, structure and resorptive function of lymphatic microvessels in the serous membranes gives possibilities to work out a model on formation and transport of lymph. Lymphatic postcapillaries (LP) - thin-walled endothelial channels with valves make an important link of the lymphatic system. The LP structure is similar to that of capillaries. These microvessels are situated in the interstitial space area, that are characterized with their high content of plasma proteins and water. Interrelation in concentrations of protein in the LP lumen and in the tissue ensures the existence of the osmotic gradient pressure through the wall and contributes liquor resorption into the lumen of microvessels. Peritoneal lymphatic capillaries are situated in the areas of a higher hydrotation level of the interstitial space. They can control the rate of the liquor filtration from plasma into tissue and regulate the resorption level in the whole lymphatic network. The model provides a differentiated participation of the LP and capillaries in performing resorption of proteins and liquor from the interstitium. The resorption mechanisms are closely connected with processes of the lymph movement along the vessels.     

Electron microscopic study of 3H-leucine transport between the internal body environment and the enteral environment

Using electron microscopic autoradiography, a space-time characteristics of 3H-leucine transport, from the enteral to the inner environment, and from blood plasma to the enteral environment, has been given. Some gradient of intracellular leucin accumulation is determined, from the tip cells of a villus to epithelocytes of the basal membrane. Amino acid transport from blood plasma to the lumen of the intestine is accomplished mainly via the cytoplasm and intercellular space of epithelocytes of the villus basement. It is proposed that amino acid transport through membranes of microvilli and through basal-lateral membranes of cells being on various levels along the villus may start at different amino acid concentrations which may be, presumably, to different concentration and different conjugation of enzyme-transport complexes in these membranes.     

Endothelial cell barriers: Transport of molecules between blood and tissues

An endothelial cell monolayer separates interstitia from blood and lymph, and determines the bidirectional transfer of solutes and macromolecules across these biological spaces. We review advances in transport modalities across these endothelial barriers. Glucose is a major fuel for the brain and peripheral tissues, and insulin acts on both central and peripheral tissues to promote whole‐body metabolic signalling and anabolic activity. Blood‐brain barrier endothelial cells display stringent tight junctions and lack pinocytic activity. Delivery of blood glucose and insulin to the brain occurs through their respective carrier (Glucose transporter 1) and receptor (insulin receptor), enacting bona fide transcytosis. At supraphysiological concentrations, insulin is also likely transferred by fluid phase cellular uptake and paracellular transport, especially in peripheral microvascular endothelia. The lymphatic microvasculature also transports insulin but in this case from tissues to lymph and therefrom to blood. This serves to end the hormone's action and to absorb highly concentrated subcutaneously injected insulin in diabetic individuals. The former function may involve receptor‐mediated transcytosis into lymphatic endothelial cells, the latter fluid phase uptake and paracellular transport. Lymphatic capillaries also mediate carrier‐dependent transport of other nutrients and macromolecules. These findings challenge the notion that lymphatic capillaries only transport macromolecules through intercellular flaps.     

The intraorganic lymphatic bed of the mucous membrane of the tongue

The intraorganic lymphatic bed of the canine tongue mucous membrane is presented by the superficial and deep capillary networks and by the plexuses of the intraorganic lymphatic vessels. The capillaries of the superficial network in the thickness of the mucous membrane of the dorsal surface of the lingual tip, body and root form the lymphatic bed of the mechanical and gustatory papillas. The diameter of the capillaries in the both networks and in the intraorganic lymphatic vessels is greater in the mucous membrane of the ventral surface of the tip and of the lateral surfaces of the lingual body, and density of their arrangement is higher in the mucous membrane on the dorsal surface of the lingual tip, body and root. The capillaries of the superficial network in all areas of the lingual mucous membrane are thinner than those of the deep network, and the loops formed by the capillaries of the superficial network are less than the loops of the deep network.     

Electron microscopic characteristics of plant peroxidase transport pathways in the microcirculatory bed of the rabbit peritoneum

The dynamics of exogenic peroxidase transfer from blood into the roots of the rabbit mesenteric lymphatic system have been studied by means of electron microscopic methods in combination with the trasser technique. Light optic identification of the vascular segments and selection of samples for electron microscopic analysis make it possible to reveal certain differences in the pathways of protein transport via the walls of the blood capillaries and venules. The vesicular transport is the only means for peroxidase to be transferred via the walls of the mesenteric blood capillaries. The time for transendothelial transfer of the marker is more than 10 min. In the venules the vesicular transport of protein does not differ from that in the capillaries, however, the predominant leakage of peroxidase from blood into the interstitium is performed through open interendothelial contacts. The hemato-interstitial transport via the intercellular clefts takes less than 3 min. For transferring protein from the interstitium into the lumen of the lymphatic capillaries and postcapillaries, the vesicular mechanism is used, and to a less extent--the open intercellular contacts. A suggestion is made that the term "open contact" should be understood in functional meaning and this means should be considered as an intercellular pathway for transporting molecules of a definite size.     

Lymphatic vessels in lungfishes (Dipnoi)

 Lymphatic capillaries are distributed throughout the body of lepidosirenid and protopterid Dipnoi, except in the central nervous system. They form small, interconnected units which are individually evacuated into nearby blood capillaries by lymphatic micropumps. The number of lymphatic micropumps varies considerably in different parts of the body. In fin areas, 30–50 per mm³ tissue may be considered normal in Protopterus annectens, but up to 105 per mm³ have been counted in an anterior fin of Lepidosiren paradoxa. Lymphatic capillaries are formed by thin endothelial cells with fine processes into the surrounding interstitial space. Occasionally there is a faint, discontinuous basal lamina. Pericytes, however, are completely absent. Microfibrils establish contact between endothelial cells and surrounding connective tissue fibers. The lymphatic micropumps are essentially spherical, contractile organs of 35–55 μm in diameter. Their central lumen is lined by extensions of a single endothelial cell. Additional endothelial cells form inflow and outflow valves. The endothelial layer is surrounded by a single large, highly specialized muscle cell. This spherical muscle cell has many perforations, allowing the passage of thin outward processes of the endothelial cell which form part of the suspension apparatus of the lymphatic micropump. The muscle cell establishes a specialized end-to-end contact between opposing parts of its own cell membrane. This contact is very similar to an intercalated disc in vertebrate heart muscle. Each lymphatic micropump is suspended within a cell-free tissue area by microfibrils which radiate from the lymphatic micropump into the surrounding connective tissue. The microfibrils are occasionally reinforced by single collagen fibers. The cell-free area around each lymphatic micropump appears as a bright halo in both light and electron micrographs. No type of lymphatic vessel other than lymphatic capillaries could be detected in the Dipnoi studied. Lepidosireniform Dipnoi are the only Vertebrata besides the Tetrapoda in which lymphatic vessels and characteristic lymphatic pumps have been documented. In addition, these Dipnoi and all Tetrapoda share the same overall design of blood circulation, which is not divided into a primary and a secondary system of vessels, as it is in Actinopterygii, Chondrichthyes, and Agnatha. Since there are primary and secondary blood vessels in the gills of Latimeria chalumnae, while the existence of lymphatic vessels has not been confirmed, general angioarchitecture should be taken into account as an important character when phylogenetic relationships among extant Sarcopterygii are discussed. Accepted: 7 October 1997     

Occludin oligomeric assembly at tight junctions of the blood-brain barrier is disrupted by peripheral inflammatory hyperalgesia

Tight junctions (TJs) at the blood-brain barrier (BBB) dynamically alter paracellular diffusion of blood-borne substances from the peripheral circulation to the CNS in response to external stressors, such as pain, inflammation, and hypoxia. In this study, we investigated the effect of lambda-carrageenan-induced peripheral inflammatory pain (i.e., hyperalgesia) on the oligomeric assembly of the key TJ transmembrane protein, occludin. Oligomerization of integral membrane proteins is a critical step in TJ complex assembly that enables the generation of tightly packed, large multiprotein complexes capable of physically obliterating the interendothelial space to inhibit paracellular diffusion. Intact microvessels isolated from rat brains were fractionated by detergent-free density gradient centrifugation, and gradient fractions were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis/ Western blot. Injection of lambda-carrageenan into the rat hind paw produced after 3 h a marked change in the relative amounts of oligomeric, dimeric, and monomeric occludin isoforms associated with different plasma membrane lipid raft domains and intracellular compartments in endothelial cells at the BBB. Our findings suggest that increased BBB permeability (i.e., leak) associated with lambda-carrageenan-induced peripheral inflammatory pain is promoted by the disruption of disulfide-bonded occludin oligomeric assemblies, which renders them incapable of forming an impermeant physical barrier to paracellular transport.     

Intravital lectin perfusion analysis of vascular permeability in human micro- and macro- blood vessels

We previously applied intravital lectin perfusion in mouse models to elucidate mechanisms underlying vascular permeability. The present work transfers this technique to human models, analysing vascular permeability in macro- and microvessels. Human vascular endothelial surface carbohydrate biochemistry differs significantly from its murine counterpart, lacking alpha-galactosyl epitopes and expressing the L-fucose moiety in the glycocalyx; the poly-N-lactosamine glycan backbone is common to all mammals. We examined extensively lectin binding specificities in sections and in vivo, and then applied the poly-N-lactosamine-specific lectin LEA and the L-fucose-specific lectin UEA-I in human intravital perfusions. Transendothelial transport differed in macrovessels and microvessels. In microvessels of adult human fat tissue, rectal wall and rectal carcinomas, slow transendothelial transport by vesicles was followed by significant retention at the subendothelial basement membrane; paracellular passage was not observed. Passage time exceeded 1 h. Thus we found barrier mechanisms resembling those we described previously in murine tissues. In both adult and fetal macrovessels, the vena saphena magna and the umbilical vein, respectively, rapid passage across the endothelial lining was observed, the tracer localising completely in the subendothelial tissues within 15 min; vesicular transport was more rapid than in microvessels, and retention at the subendothelial basement membrane briefer.     

Formation of the external zone of the median eminence of rats in the perinatal period

The formation of interrelations of the axons of neurosecretory cells and of ependyme cells with the capillaries of primary portal plexus in rats from the 14th day of embryogenesis till the 9th day of postnatal life was studied using the light and electron microscope methods. During the whole period under study, the basal processes of the ependyme cells reach the primary portal plexus of the capillaries. The terminals of the basal processes are usually separated from the endothelium of the capillaries by two basal membranes and enclosed pericapillary space. After the birth, some basal process penetrate in the pericapillary space and terminate on the endothelium. The surface of contact of the ependyme cell processes with the external basal membrane increases with the age, this being accompanied by the increase of pinocytotic activity. The neurosecretory axons are found in the median eminence already on the 14th day of embryogenesis, but by the 20th day only they reach the external basal membrane and penetrate sometimes in the pericapillary space. After the birth, the number of axons reaching the external basal membrane and the surface of contact between them increase gradually with, apparently, a concomitant intensification of the transport of neurohormones in the portal circulatory system of the hypophysial-hypothalamic complex.     

A Model for Fluid Drainage by the Lymphatic System

This study investigates the fluid flow through tissues where lymphatic drainage occurs. Lymphatic drainage requires the use of two valve systems, primary and secondary. Primary valves are located in the initial lymphatics. Overlapping endothelial cells around the circumferential lining of lymphatic capillaries are presumed to act as a unidirectional valve system. Secondary valves are located in the lumen of the collecting lymphatics and act as another unidirectional valve system; these are well studied in contrast to primary valves. We propose a model for the drainage of fluid by the lymphatic system that includes the primary valve system. The analysis in this work incorporates the mechanics of the primary lymphatic valves as well as the fluid flow through the interstitium and that through the walls of the blood capillaries. The model predicts a piecewise linear relation between the drainage flux and the pressure difference between the blood and lymphatic capillaries. The model describes a permeable membrane around a blood capillary, an elastic primary lymphatic valve and the interstitium lying between the two.     

Degradation of the radula in the snails Biomphalaria glabrata say and Limnaea stagnalis L. (Gastropoda,Pulmonata)

Summary The radula of snails is formed at the posterior end of the radular gland or pocket, and degraded at the same rate at its anterior end. Degradation is due to different secretory activities of the inferior epithelium of the radular gland. Its secretions seem to degrade enzymatically the matrix of the radular membrane and basal plates of teeth, leaving only chitin containing microfibres and degradation products. The sclerotized parts of the teeth remain unchanged, but as they are now only loosely connected with the radular membrane. they are torn off easily during feeding movements. The rest of the degraded and frayed radular membrane and the subradular membrane are also lost by abrasion during feeding. The cells of the inferior epithelium are connected with each other by septate desmosomes and an elaborate system of deep lateral interdigitation which may provide tensile strength. Extrusion of degraded cells of the inferior epithelium into the subradular membrane takes place, although the thick basal lamina forms a continuous sheath which is closely adjoined to the basal parts of the inferior epithelium. Nerve fibres containing vesicles with electron dense neurosecretory material (deduced from the diameter of 200–250 nm) are attached to this sheath or penetrate into it; they may be involved in the regulation of production and degradation processes during radula replacement. Problems of the forward transport of radula and inferior epithelium are discussed.     

Development of the lymphatic bed of the human esophageal wall]

The lymphatic capillaries are firstly determined in fetuses at the age of 3-4 months in the oesophagus submucous lamina. In fetuses of 5-6 months of age transition of the lymphatic capillaries from the submucous lamina into the mucous membrane proper is noted. In fetuses of 6 months of age perivascular lymphatic capillaries and vessels appear. They form peculiar paths around arterioles, venules, arterial branches and venous tributaries. The lymphatic bed of the oesophageal wall is rather well developed in mature fetuses and newborns. In adult and old persons a partial reduction of the lymphatic bed in the oesophageal wall is observed.     

Microcirculatory disorders in the early period of autotoxic shock

Microcirculation in the rat mesenterium was studied in the early period of autotoxic shock, using light and electron microscopy. Progressive reduction in the number of functioning capillaries, blood flow slowing in microvessels, and increase in the number of leukocytes that adhere to the luminal surface of the endothelium have been noted. Cases of stasis, sludge syndrome, thrombosis of small venules and disturbances of vascular permeability for protein have been observed. Ultrastructural changes: the formation of transendothelial canals and destructive changes in the basal membrane testify to the disturbance of the barrier function in microvessels. The role of free radicals in the pathogenesis of autotoxic shock is discussed.     

Mechanistic interpretation of the influence of lipid phase transitions on transport functions.

In an attempt to understand the mechanism by which a structural change of membrane lipids affects transport functions, the temperature dependence of transport rate has been measured to below the low temperature end of the fluid in equilibrium ordered phase transition of the membrane lipids. The unsaturated fatty acid requiring Escherichia coli strain T105 was supplemented with either trans-delta9-octadecenoate or trans-delta9-hexadecenoate or supplemented with and subsequently starved for cis-delta9-octadecenoate. Fluid in equilibrium ordered phase transitions measured in whole cells using the fluorescence probe N-phenyl-1-naphthylamine were compared with the temperature dependence of beta-glucoside and beta-galactoside transport. In addition to the previously observed downward "break" in the Arrhenius plot of transport rate which occurred near the middle of the phase transition temperature range, a second upward "break" was observed which could be correlated with the low-temperature end of the phase transition. These experiments are interpreted in terms of a partitioning of transport proteins between ordered and fluid domains which is described by a lateral distribution coefficient, k. This distribution coefficient varies with the membrane lipid composition as well as with the transport system. Values for k suggest a 2-20-fold preference for the partitioning of transport proteins into the fluid parts of the membrane.     

Histotopography and composition of the microcirculatory bed of the sciatic nerve in the rat

By means of macro-microscopical preparation methods, horseradish peroxidase injection, semithin sections, electron microscopy histotopography and composition of microvessels of the sciatic nerve have been studied in 20 mature male white rats. The perineural membrane has been stated to have no its own vascular network. In composition of the sciatic nerve and its large branches muscular venules are revealed; they run longitudinally along the whole length of the endoneural space and are tributaries of the epineural veins. A suggestion is made that presence of the contractile apparatus in the venular wall can play an important role in formation of the postcapillary resistance, regulating the hydrostatic pressure value in the lumen of endoneural capillaries ad intensity of liquor filtration into the interstitial space.     

Ultrastructural regularities of new growth of secondary micro blood vessels in the prenatal period of human morphogenesis

By means of transmissive electron microscopy structural aspects of new formation of secondary blood microvessels in functionally different organs have been studied during fetal period of the human development. Growth buds appear on the basal surface of the endothelial lining of the blood microvessels predominantly in those areas of the vascular wall, where pericytes and adventitial cells are absent. The first stage in formation of the growth bud is separating of small compartments in the lumen of the maternal vessel. Then, connected by means of specialized contacts, endothelial cells of such a compartment move one by one towards periphery of the vessel and form the growth bud, protruding outside. In the area of the growth bud local destruction of the basal membrane is observed. Newly formed microvessels, anastomising, form vascular loops. Further processes in differentiation of separate segments of the newly formed capillary network into links of the hemomicrocirculatory bed are analogous to those, observed in the course of differentiation of the primary protocapillary network into the secondary intraorganic blood bed.     

Are close contacts between astrocytes and endothelial cells a prerequisite condition of a blood-brain barrier? The rat subfornical organ as an example*

The microvessels of the rat subfornical organ (SFO) are heterogeneous: those of the caudal part lack a blood-brain barrier (BBB) unlike those of the rostral part. The astroglial environment of these microvessels has been studied by combining an immunocytochemical technique employing an anti-GFAP (glial fibrillary acidic protein) antiserum with the morphological detection of a barrier to the protein-silver complex. All the SFO microvessels are surrounded by astrocytes characterized by a tumescent aspect; however, the relative proximity between the astrocytic feet and the endothelial cells varies considerably. The capillaries provided with a barrier (rostral SFO) are contiguous with the astrocytes from which they are only separated by a basement membrane. The capillaries devoid of BBB (caudal SFO) are surrounded by a pericapillary space that keeps the astrocytes at a short distance (capillaries with a very rich vesicular endothelium) or at a long distance (capillaries with a fenestrated endothelium). The astrocytes are absent in the choroid plexus where all microvessels are fenestrated and lack a barrier. These data suggest that the astrocytes release one or more signals which in their vicinity inhibit the expression of endothelial morphological characteristics (fenestrations, vesicles) responsible for the leakage of plasmatic proteins from the blood to the cerebral parenchyma of the circumventricular organs.     

Observations on the morphology of pancreatic secretory capillaries

Summary The fine morphology of, and the light microscopic distribution of ATPase in, pancreatic secretory capillaries suggest that they may be absorptive channels receiving material from the general blood circulation rather than secretory channels communicating with the duct system. This assumption is based upon many observations; 1. The pattern of microvilli of their wall differs from that of acinar lumen. 2. The material retained by them in ultrathin sections differs from that retained by acinar lumen. 3. They ramify, basally, to the level of Golgi region. 4. The lack of release of zymogen droplets in them. 5. Their points of origin from acinar lumen are controlled by zonulae occludentes. 6. Gulf-like extensions from the basal surface of acinar cell communicate with them. 7. Fenestrated blood capillaries run in their vicinity. 8. Blood capillaries and acinar cells share a joint basement lamina. 9. Light microscopically, ATPase activity is associated with their wall and not with the basal acinar cell surface.Publication No. 932 of the Division of Basic Health Sciences of Emory University; which was supported by the McCandless Research Funds and the School of Dentistry Fellowship Funds, of Emory University. The technical assistance of Miss Brenda Callahan, Mr. Peter Daly and Mr. Hugo Greiner is acknowledged. The use of the facilities of Dr. Wallace G. Campbell Jr. is appreciated.     

Structure of the skin of an air-breathing mudskipper, Periophthalmus magnuspinnatus

The epidermis of the mudskipper Periophthalmus magnuspinnatus consisted of three layers: the outermost layer, middle layer and stratum germinativum. Extensive vascular capillary networks were present near the superficial layer of epidermis and outermost layer. The diffusion distance between the vascular capillaries and the surface of epidermis was c . 1.5 ± 0.9μm. The middle layer consisted of small or voluminous cells swollen by epidermal cells. Due to the swollen cells, the thickness of the epidermis increased and the epidermis appeared web-like. The swollen cells contained tonofilaments, lucent contents and desmosomes. Fine blood capillaries were also discernible in this layer. Well-developed lymphatic spaces containing lymphocytes existed in the stratum germinativum. Numerous blood capillaries were present under the basement membrane. The dermis consisted of a stratum laxum and stratum compactum, and there was a definite area with acid mucopolysaccharides and a small scale in the stratum laxum. The skin had an epidermal pigment cell, dendritic melanophores (-cytes) containing melanin granules within their cytoplasm, and two kinds of dermal pigment cells, melanophores and colourless pigments containing reflecting platelets.     

Scanning electron microscope study of the liver microcirculation

During the experimental investigation performed in dogs and rats, by means of scanning electron microscopy of corrosive anatomical preparations, the spatial organization of all parts of the hepatic vascular bed (arterial, venous and lymphatic) has been studied, specific features of their components construction have been described. Within the limits of one hepatic lobule the number of vessels included in the portal vein system exceeds that of the arterial ones, originating from the proper hepatic artery system. In every part of the vascular bed the gradient of the form, orientation and pronouncement of the nuclei-containing zones in endotheliocytes and myocytes has been established. Various appliances participating in the blood and lymph stream regulation in different parts of the vascular bed have been revealed. As initial elements of the lymph bed, closed digital or loop-like capillaries should be regarded, they localize in the organ's connective tissue framework. Around the portal and hepatic veins and their branches, as well as around the biliary ducts, well developed plexuses of the lymphatic and blood capillaries and vessels localize, they are the main draining pathways of the organ. The degree of development and pronouncement of these plexuses depends on the lumen size in the formation they accompany.