Submicroscopic analysis of the microcirculatory system of the pancreas in several vertebrates]

The metabolic link of the microcirculatory system of the exocrinous part of the pancreas studied electron microscopically in the frog, chicken and rat has a general plan of the structure. It consists of capillaries, pericapillary gap and intercellular clefts of glandular cells connected with it. But in the frog and chicken the adventitional layer was found to be absent from the blood capillary wall, the luminal surface of endothelial cells was increased. The width of the basal layer and intercellular clefts in the rat was less than in other objects. The existence of cytoplasmic spiculae of exocrinous pancreocytes in the pancreas of different vertebrates allows to consider them as an element of the exocrinous part microcirculatory system.     

Analysis of the pathways of substance transport into the acinar cells]

In the exchange link of the microcirculation system of the exocrine part of the pancreas of Rana temporaria the substances moved from the blood capillary into the pericapillary space, then into the intercellular clefts and into the acinar cells by active transport. This is confirmed by the electron microscope studies of the ATP-ase activity localization in the exchange link: there are numerous lead phosphate granules in the endothelium of blood capillaries, on the fibrillae structures of the pericapillary space interstitium, on the lateral plasmic membrane of the exocrine pancreacytes, and on the cytoplasmic plates forming pinocytotic vacuoles.     

Permeability of muscle capillaries to microperoxidase

In this study we attempted to identify a morphologic counterpart of the small pore of muscle capillaries. The existence of such a pore has been postulated by physiologists to explain the permeability of muscle capillaries to small macromolecules. We injected mice intravenously with microperoxidase (MP) and fixed specimens of diaphragm at intervals of 0-250 s after the injection to localize the tracer by electron microscopy. The small size of MP (1,900 mol wt and 20 A molecular diameter [MD]) ensures its ready passage through the small pore since the latter is thought to be either a cylindrical channel 90 A in diameter or a slit 55 A wide. MP appears in the pericapillary interstitium within 30 s of initiation of its intravenous injection. The patterns of localization of MP observed within clefts between adjacent capillary endothelial cells indicate that some endothelial junctions are permeable to this tracer. Although small vesicles transfer MP across the endothelium, we do not believe that the vesicles transfer substantial amounts of MP into the pericapillary interstitium. We did not obtain evidence that MP crosses the endothelium of capillaries through channels formed either by a single vesicle or by a series of linked vesicles opening simultaneously at both surfaces of the endothelial cell. From our observations we conclude that some endothelial junctions of capillaries are permeable to MP, and that these permeable junctions are a plausible morphologic counterpart of the small pore.     

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.     

Change in the myocardial capillary endothelium in the early periods after x-ray irradiation (an electron microscopic study)

According to the data of quantitative electron microscopic analysis in rats 1-48 h after total x-ray irradiation (450 rad) the number of microvesicles in endothelial capillaries decreases. Increasing size of the microvesicles and appearance of vacuoles does not compensate decreased total transport volume of these structures. After irradiation, in most capillaries the endothelial layer does not change its thickness; neither lesions in intracellular contacts nor formation of holes are observed. Both width of pericapillary zones and total volume of interstitial space in the myocardium do not change during the time of investigation.     

Ultrastructure of the renal corpuscle of fresh water fishes

By means of light (semithin slices) and electron microscopes structural organization of the corpuscle and glomerulus of the capsule has been studied in freshwater fishes--carp (Cyprinus carpio) and crucian carp (Carassius carassius). The diameter of their glomeruli are in the carp from 80.8 +/- 1.4 mcm up to 59.8 +/- 2.4 mcm and in the crucian carp--from 86.4 +/- 1.8 mcm up to 69.25 +/- 0.2 mcm. Between these dimensions there are significant differences. Components of the external wall of the capsule in the fishes studied have similar structure. In epitheliocytes, situating on the basement membrane, electron opaque inclusions have been revealed. They resemble, by their ultrastructure, granules of peripolar cells, described in mammals and in man. In places, where epitheliocytes have contacts, thin folds are formed, they turn into the capsule lumen. Touching the cell surface, they can form border vacuoles. The main peculiarity of endotheliocytes in the glomerular capillaries is presence of diaphragmated and open fenestrae (their size is from 30 to 50 nm) in the peripheral zone of their cytoplasm. Unlike the higher vertebrates, the fishes have a well developed pericapillary mesangium. It contains mesangial matrix and rare processes of mesangiocytes.     

Extravascular circulation in the pituitary of Mugil cephalus (Teleostei)

Summary Extravascular circulation in the pituitary of Mugil cephalus was investigated by injecting live fish with horseradish peroxidase and studying the distribution of the enzyme in the gland. The principal components of the extravascular circulatory system are the pericapillary spaces, and, arising from them, the interlobular and circumhypophyseal spaces. Extensions of these spaces penetrate the glandular parenchyma of the pars distalis, where they merge with pericellular spaces. In the neurohypophysis, pericapillary spaces are connected to the periaxonal spaces.Capillaries penetrating from the proximal neurohypophysis into the pars distalis are accompanied by neurosecretory axons. These axons form a mass of tissue which is limited near the capillaries by the pericapillary spaces and near the adenohypophysis by the interlobular spaces. Toward the interior of the adenohypophysis the amount of nervous tissue accompanying the capillaries progressively diminishes, thus reducing the distance between pericapillary and interlobular spaces. Within the pars distalis, the neurosecretory axons accompanying the capillaries are sparse, and the secretory and stellate cells are mostly located directly adjacent to the pericapillary spaces. In the neuro-intermediate lobe, interlobular spaces outline the neuro-adenohypophyseal boundary.The relationship between extravascular spaces and hormone-secreting cells varies in the different regions of the adenohypophysis depending upon the type of neurosecretory innervation in the respective region. In the regions of prolactin and gonadotropin cells, where neurosecretory axons are in direct contact with the secretory cells, the hormone-secreting and stellate cells are adjacent to the pericapillary spaces. In the regions of ACTH and STH cells, secretory and stellate cells are found adjacent to the interlobular spaces, which are interposed between the cells and the neurosecretory axons.Abbreviations AH adenohypophysis - CH circumhypophyseal - DNH distal neurohypophysis - HRP horseradish peroxidase - NH neurohypophysis - NS neurosecretory - PD pars distalis - PI pars intermedia - PPD proximal pars distalis - RNH rostral neurohypophysis - RPD rostral pars distalis This research was supported by a grant from the National Council for Research and Development, Israel, and the GKSS Geesthacht-Tesperhude, Federal Republic of Germany     

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.     

Electron microscopic study of monoaminergic fibers in the posterior lobe of the rat hypophysis]

Monoaminergic fibers and their swellings containing dense core vesicles, 700-1000 A in diameter, were found by electron microscopic studies in the posterior pituitary of rat. Most of them are situated far from capillaries and are surrounded by neurosecretory fibers and pituicytes. There are usually no specialized structures at the sites of contact between them. Single monoaminergic fibers come in contact with the capillaries and even penetrate the pericapillary space. Large swelling of these fibers which sometimes occur contain different polymorphous inclusions, degenerating mitochondria and numerous small tubules along with the dense core vesicles.     

Fine structure of the paracellular junctions of terminal villous capillaries in the perfused human placeta

Summary Selected lobules of human term placentae were extracorporeally perfused for a recovery period of 20 min, fixed by perfusion and mordanted with ferrocyanide prior to processing for transmission electron microscopy. The lateral membranes of the endothelial cells of the terminal villous capillaries were found to be separated by paracellular clefts of mean width 15.6 nm. At tight junctional regions (1–4 sites per cleft) the two membranes approached each other more closely and frequently appeared to fuse. However, tilting of the sections in the electron microscope stage showed that the membranes were separated by a gap of mean width 4.1 nm in at least 94% of tight junctional profiles. When individual tight junctions were studied by a combination of serial sectioning and goniometric tilting, they were seen to widen abruptly within a distance of three to seven consecutive thin sections, indicating they were not continuous throughout the axial length of the capillaries. The wide regions of the clefts usually showed linkers, strands of glycocalyx-like material spanning the gap. Linkers may contribute to cell adhesion and possibly form part of a filter within the tortuous paracellular pathway provided by the discontinuous network of tight junctional strands. Human term placental capillaries appear to resemble closely other continuous non-brain capillaries.     

Ultrastructural characterization of exocytotic release sites in different layers of the median eminence of the rat

Summary The release of neuronal secretory products by exocytosis in different layers of the median eminence of the rat was investigated ultrastructurally after perfusion with Ringer solution containing tannic acid. Exocytotic images were observed in all layers studied. Neurohaemal release sites were found in the pars externa of the palisade layer, where they occurred not only against the basal lamina of the pericapillary space, but also opposite, adjacent to neuronal and glial elements. In the lateral portion of the pars externa of the palisade layer most release sites were separated from the pericapillary space or the pial surface by ependymal or glial processes. In the pars interna of the palisade layer, and in the reticular, fibre and subependymal layers, release was observed in different types of axonal processes without morphological synaptic specializations. We suggest that products released in the pars externa of the palisade layer are destined to reach the capillaries of the primary portal plexus. Although the non-vascular release sites may serve a similar hormonal function, they may alternatively represent the morphological correlate of axoaxonal contacts or of paracrine, non-synaptic release sites.     

Ultrastructural study of neurovascular contacts in the median eminence of the rat

Summary In the median eminence of the rat axons of the supraoptic-paraventricular-hypophyseal tract with elementary neurosecretory granules (150–200 m) traverse the internal zone. Terminals containing dense core vesicles 60–120 m in diameter end on the portal capillaries of the median eminence. A unique organisation of the primary portal capillaries is shown. Endothelial cells have many fenestrae. The pericapillary space has numerous extensions all of which represent a special zone around the endothelial tube. The fine structural organisation and function of the neurovascular contacts in the median eminence are discussed.     

Morphology of the endocrine portion of the teleost pancreas]

Endocrine component of the crucian (Carassius carassius), carp (Cyprinus carpio), tench (Tinca tinca) and silurus (Silurus glanis) pancreas is structurally organized in the form of pancreatic islets. Gorbusha (Oncorhynchus gorbuscha) has, besides the islets, some Brockman's bodies. Endocrine component of the pancreas of Teleostei possesses A-, B-, D and acinar-islet cells "B". All types of cells are shoot-shaped and all have contacts with the capillaries. Extrusion of the hormones from the endocrine cells is carried out via emiocytosis, and in gorbusha at the time of migration--by microapocrine means. Secretory granules were observed to get into the capillaries and make hormonal storage necessary for fish migration. It was demonstrated that endocrine component of the pancreas in Teleostei is highly rich in innervation, neuronal fibers containing small granular vesicles.     

Junctions between pericytes and the endothelium in rat myocardial capillaries: A morphometric and immunogold study

Summary Pericytes are cells of mesodermal origin which are closely associated with the microvasculature. Despite numerous studies little is known about their function. We have studied the relationship between pericytes and the endothelium in rat myocardial capillaries employing ultrastructural and immunogold techniques. 14% of the subendothelial cell membrane is covered by comparatively small pericytic cell processes. About half of these processes are completely embedded in baseement membrane material, whereas the remaining half forms closer contacts with the endothelium. These contacts are devoid of anti-laminin immunogold label, a marker for basement membranes. A small fraction of these contacts has been identified as tight junctions resembling those seen between endothelial cells in capillaries of the same tissue. The remaining majority of junctions reveals a cleft of approximately 18 nm between the apposed membranes in which a succession of cleft-spanning structures can often bedetected. It was also found that pericytic processes are preferentially located close to interendothelial junctions. We suggest that the high frequency of intimate junctions between pericytes and the endothelium and the preferential localisation near paracellular clefts may have functional significance.     

Subcellular compartmentalization of the luteal cell in the ovary of the dog

The compartmentalization of the parenchyma of the corpus luteum in the dog was studied by both 100 and 1000 KV electron microscopy. The organells within the luteal cell are oriented with a high degree of consistency towards the pericapillary space. Characteristically, the avascular pole and the lateral margins of the cell posses predominantly stacked and whorled cisternae of agranular ER. In the central medial portions of the cell, pleomorphic mitochondria with tubulo-vesicular cristae and anastomosing tubules of agranular ER predominate. However, the distribution of organelles in this compartment is graded. Mitochondria predominate in the central medial areas while tubular ER is more dominant peripherally. Microfilaments are ubiquitous in this compartment and run a longitudinal course between and around the subcellular components towards the pericapillary space. The Golgi apparatus is large and prominent and is positioned over the pole of the nucleus that faces the basal lamina. Coated vesicles are abundant in the Golgi regions and along the lateral surface of the cell. Three distinct regional specializations of the cell surface exist. The basal surface contains long pleomorphic cytoplasmic folds that fill the pericapillary space, are interconnected by small gap junctions and contain abundant multivesicular bodies. The lateral cell surface is covered with microvilli and is organized into tortuous intercellular channels and canaliculi. These are interrupted at intervals by cytoplasmic protrusions that extend from one cell well into the cytoplasm of the next. Large, well-developed gap junctions line the margins of the cells furthest removed from the pericapillary space. Finally, the individual cells exhibit heterogeneity with respect to the amount one subcellular organelle or compartment is expressed relative to another.These observations are discussed in relation to the subcellular compartmentalization of progesterone synthesis and release.     

Light and electronmicroscopic study of area M, the chemosensitive area of the ventral medulla oblongata

An electron and light microscopic study was performed to elucidate the structure of area M, an area on the ventral surface of the medulla oblongata which participates in the central respiratory control of the cat. We found no particular morphological barriers to fluid transport from the cerebrospinal fluid space towards the capillaries of this area. The astroglial contacts proved to be mostly gap junctions; furthermore, no tight junctions could be found here. Multipolar ganglion cells could be localized at a depth of 400-800 micron. These cells form numerous synaptic contacts with thin, unmyelinated axons running superficially, parallel to the pial surface in area M. Our findings support the contention that the ventral chemosensitive areas of the medulla oblongata are influenced by the chemical constitution of the cerebrospinal fluid.     

Structure of spinal cord capillaries in hypokinesia

Changes in spatial interrelations of the spinal cord capillaries and motoneurons and capillary ultrastructure were studied under hypokinesia. Spatial interrelations between the capillaries and neurons were not demonstrated to change under hypokinesia. They were estimated by the following parameters: area of neuronal profile field, number of capillaries, their length, distance from the nerve cell body, capillary bed area and index of capillary-neuronal interrelations. Quantitative investigation revealed capillary stenosis: their diameter was one and a half times less under hypokinesia. Morphologically, capillary stenosis was accompanied by the basal membrane thickening and endothelial cytoplasm vacuolization. There was a direct relation between endothelial villi and the places of the endothelial cells contacts, dilatation of the contact interstices and solidifying of their borders. Changes in the capillaries were followed by reactions in the pericapillary structures, such as: fibrillae were formed, mitochondrii accumulated in the perivascular glial projections, the membrane next to capillary astrocyte projections underwent desmosome-like condensation. Mitochondrial accumulations were also observed in the nerve cell projections and in their cytoplasm sites contacting with the blood vessels.     

The structure and vascularization of the pars tuberalis of the hypophysis of Rana temporaria

Summary The pars tuberalis of the hypophysis of Rana temporaria presents the general structural and the cytological characteristics of an endocrine gland. It is composed of elongated cells with long, branching processes ending on the external basement membrane of the pericapillary space. The pars tuberalis cells produce secretory granules which are accumulated in the pericapillary endings of the processes.Corresponding to its separate localization, the pars tuberalis of Rana temporaria has a separate vascularization of which the efferent capillaries anastomose with the capillary plexus of the median eminence. The general direction of the blood flow of the pars tuberalis is towards the capillaries of the median eminence. Also, the secretory products of the pars tuberalis pass into the blood stream of the hypophysial portal system.Several characteristics of the pars tuberalis show that its function must be different from that of the pars distalis of the hypophysis. Moreover, in contrast with the pars distalis, the activity of the pars tuberalis is not regulated by neurohumoral factors.The results show that a role of the pars tuberalis in the regulation of the activity of the pars distalis of the hypophysis is not excluded.     

The vascularization of the organon vasculosum hypothalami ofRana temporaria

Summary InRana temporaria, the blood capillaries of the organon vasculosum hypothalami are the bilateral origin of the hypothalamic branch of the encephalo-posthypophysial portal vein. In the organon vasculosum hypothalami, pericapillary accumulations of monoamines occur. The peculiar Vascularization of the organon vaseulosum and the pericapillary accumulation of monoamines suggest a possible functional relation between the organon vasculosum and the neuro-intermediate lobe of the hypophysis via the encephalo-posthypophysial portal system.     

Development of the vasculature of the pars distalis in a tumor-susceptible strain of rat (Fischer 344) differs from vascular development in a non-tumor-susceptible strain (Lewis)

The development of the vasculature of the pars distalis of two strains of rat, Fischer 344 (F344) and Lewis (LEW), was followed in 16-day (16d) and 20-day (20d) fetuses, and in 1-day (1d), 5d, 20d, 50d, and 6-month-old females. No differences in the two strains were apparent in 16d fetuses; and the capillaries that were present were immature, i.e., tall, non-fenestrated endothelial cells, and were surrounded by poorly delineated pericapillary spaces. Immature capillaries also were predominant in 20d fetuses of both strains. Agranular folliculo-stellate cells were identifiable, projecting endfeet to the parenchymal basal lamina in 20d F344 fetuses, but not in LEW fetuses. Postnatally, the capillaries of LEW rats became progressively more thin-walled and fenestrated, and were surrounded by a pericapillary space that was well delimited by basal laminae at 20d. In 50d and 6-month LEW rats, capillaries were intact and surrounded by well-defined pericapillary spaces. By comparison in F344 rats, the capillaries remained more immature even in 50d rats and older. In addition, in F344 rats focal disruptions in endothelial cells and disruptions in parenchymal and capillary basal laminae were present in all postnatal stages, and a dramatic accumulation of plasma was evident within the pericapillary spaces at 20d. Endfeet processes of folliculo-stellate cells were abundant at the parenchymal basal lamina of 1d and 5d F344 neonates, but only rarely were identified in LEW neonates. Some activation of folliculo-stellate cells, i.e., increased numbers of lysosomes and dilated endoplasmic reticulum, was present in 50d F344 rats. Connective-tissue cells within the pericapillary space also were numerous and activated in F344 rats. Discrete gaps in the parenchymal basal lamina were evident subjacent to the folliculo-stellate cell endfeet in F344 rats but not in LEW rats. The vascular bed of F344 rats differs in its development from that of LEW rats. Characteristic of the F344 strain is a persistence of more immature capillaries, an inherent vascular fragility, and an activated state of folliculo-stellate cells.