Microscopy of vascular architecture and arteriovenous communications in the spleen of two odontocetes

The red pulp of the spleens of the short-finned pilot whale (Globicephala macrorhynchus) and the Pacific bottle-nosed dolphin (Tursiops truncatus gilli) (Odontoceti) were examined by light and electron microscopy and found to comprise two venous layers, an inner and an outer. The inner layer is homologous to the intermediate zone (IZ) of primitive-type mammalian spleens and contains sinusoids consisting of endothelial cells and a thin layer of extracellular deposits. Its vascular structure is unclear. The venous vessels of this layer eventually communicate with veins of the perivenous outer layer. The perivenous layer contains veins of various sizes, interstitial elements, and trabeculae. It is filled with blood cells, particularly plasma cells, but no myeloid cells. The perivenous layer (PVL) is homologous to the red pulp of common mammalian spleens but shows signs of involution. The white pulp gives origin to arterial terminals that end in the red pulp, where they communicate directly with the sinusoidal veins producing a closed circulation. The arterial terminals do not show ellipsoids. The presence of an IZ with a closed circulation and the involution of the red pulp makes the spleen of Odeontoceti another example of a mammalian spleen of the primitive type that has been altered by the evolutionary process. Vascular remodelling of the spleen of Odontoceti seems to follow the pattern noted in the spleens of nonmammalian vertebrates. © 1994 Wiley-Liss, Inc.     

Studies of a primitive mammalian spleen, the opossum (Didelphis virginiana)

Light microscopic sections of the adult opossum (Didelphis virginiana) spleen were observed to lack venous sinuses; this primitive mammalian spleen may be classified as non-sinusal in nature. In the spleen of the opossum, the capillary segments of the penicillar arteries lacked ellipsoid sheaths characteristic of certain mammalian spleens. Separating the lymphoid nodules from the surrounding red pulp was a distinct band of vascular tissue, the marginal zone. Arising from the central artery within the lymphoid nodule, vessels of capillary dimension were observed to terminate within the marginal zone and the area between lymphoid nodule and marginal zone. In addition to the vascular channels established by the terminal arterial vessels within the red pulp, the system of vessels within the marginal zone has been implicated as an important intermediate vascular channel within the spleen.     

Macrophages and Reticulum Cells in the Spleen of the Dogfish,Scyliorhinus canicula

The presence and ultrastructural features of reticulum cells and macrophages were studied in the spleen of the dogfish Scyliorhinus canicula. Three morphologically distinguishable regions of the spleen were identified: the white pulp, the red pulp and the ellipsoids. In all three, the splenic parenchyma was a meshwork supported by reticulum cells and fibres. Reticulum cells in both the white and the red pulp are irregular elements, the processes of which are joined by cell junctions and embrace developing reticular fibres. The ellipsoids of the dogfish spleen are terminal branches of the splenic arteries of the white pulp, with a sheath consisting of reticulum cells, reticular fibres, ground substance, macrophages and occasional lymphocytes. Isolated melanomacrophages also occur in the ellipsoid walls as well as in the red pulp. In both the white and the red pulp phagocytic reticulum cells, and macrophages appear frequently forming cell associations with surrounding blood cells, mainly lymphocytes. The functional significance of the ellipsoids and the cell-cell clusters of the white and the red pulp is discussed in relation to the immune capacities demonstrated in elasmobranchs.     

Phagocytosis in the spleen of the sunfish Lepomis spp.

A histological investigation of the filtering function of the spleen of the sunfish Lepomis spp. was conducted by light, scanning, and transmission electron microscopy. The parenchyma of the organ is predominantly red pulp, a system of splenic cords and sinuses. The white pulp consists of loose lymphoid tissue which forms a cuff around the pulp arteries. Filtering of particulate matter from the blood occurs in the red pulp by phagocytes of the pulp cords and ellipsoids (periarterial macrophage sheaths). The ellipsoids are pale-staining cuffs of macrophages and reticular cells in a framework of reticular fibres surrounding the arterial capillaries. Destruction of effete blood cells (especially erythrocytes) is confined to the pigment nodules; particulate matter is not taken up by the nodules. These yellow-brown bodies are dispersed throughout the red pulp and are bounded by a reticular capsule. They contain masses of phagocytes and have the appearance of a morula. They are associated with blood vessels and are surrounded by sinusoids. Prussian Blue stain shows the presence of haemosiderin within their phagocytes. The phagocytes of the pigment nodules are filled with inclusions such as residual bodies, siderosomes, and fragments of erythrocytes. The early filtering of particulate matter by the phagocytes of the pulp cords and ellipsoids may allow for a more efficient phagocytosis of erythrocytes by the pigment nodules, followed by storage and reutilization of iron-containing compounds uncontaminated by other phagocytosed material.     

The spleen of the cervidae (Gray, 1821). Quantitative-morphological studies on the spleens of stags (Cervus elaphus, L.1785) and of deers (Capreolus capreolus, L. 1758)]

The macroscopical and microscopical structure of 17 spleens of Cervus elaphus and 9 spleens of Capreolus capreolus is described. The spleens of both species exhibit structural characteristics which resemble those of the reticular "non-sinusoidal" type. These include: spleen arterial ramifications of the "magistral type" (SCHABADASCH), bilayered capsule, well developed smooth muscle cells containing trabecular networks, numerous muscle cells in the red pulp, poorly developed white pulp, splenic vein of large diameter, lack of veins associated with trabeculae, a thick tunica media in trabecular arteries and in arterial vessels of the hilus region, poorly developed SCHWEIGGER-SEIDEL sheaths, and splenic nerve trunks of considerable diameter. These structural features are comparable to those in other ruminant species having storage type spleens. However, there are differences in certain quantitative parameters between the spleen of Cervus elaphus and that of Capreolus capreolus, i.e., spleen weight versus body weight, relative volume of trabecular networks and capsular tissue, and relative amount of smooth muscle cells in trabecular tissue. On the basis of these quantitative parameters the spleen of Cervus elaphus and that of Capreolus capreolus can be classified into the system of spleen types as described by v. Herrath(1953) and should thus be ordered between the extreme storage type spleen and the extreme metabolic type spleen. The quantitative data observed in the spleen of Cervus elaphus are similar to those seen in the horse, whereas the data of the spleen of Capreolus capreolus can be compared to that of the sheep and the cow.     

Luminal morphology of small arterial vessels in the contracted spleen,studied by scanning electron microscopy of microcorrosion casts

Unique luminal configurations exhibited by small arterial vessels in contracted spleens of dog and cat were studied by means of vascular corrosion casts examined by scanning electron microscopy. Concertina-like pleating was seen in casts of trabecular arteries/arterioles, whereas within lymphatic nodules arteriolar casts lacked pleating and were smooth and uniformly cylindrical (as were all small arterial vessels in distended spleens). Morphological details of arterial vessels observed in histological sections indicated that pleating is not due to contraction of specially arranged vascular smooth muscle but to overall shortening of trabecular arterial vessels, caused by contraction of longitudinal smooth muscle in trabeculae. Another phenomenon observed in casts from contracted spleens was an almost complete "pinching-off" of many arteriolar lumens; histological evidence indicated that this is due to contraction of vascular smooth muscle, which selectively diverts flow away from certain regions of the organ. Also noted was a markedly convoluted, tortuous configuration of arterioles (penicilli) in the red pulp of contracted spleens.     

The architecture of the spleen of the yellow-bellied toad, Bombina variegata

The spleen of the yellow-bellied toad, Bombina variegata , consists of distinct white and red pulps. The well-developed white pulp is formed by a large central lymphocytic region around the numerous blood vessels and by its smaller peripheral ramifications, both surrounded by the more or less developed connective tissue boundary layer. Large peripheral sinuses of the white pulp, filled mostly with lymphocytes, are usually present at the inner side of this boundary. At the outer side of the boundary layer, the lymphocytic marginal zone is often observed. This zone merges into the erythrocyte-rich red pulp formed by cellular cords and small venous sinusoids.
The structure of the spleen of Bombina variegata differs considerably from the spleens of other anuran species studied so far. The highly developed white pulp and its distinct separation from the red pulp may be connected with the important role of the spleen as the main secondary lymphoid organ of B. variegata. The splenic compartmentalization makes the yellow-bellied toads a useful model for experimental immunobiological studies.     

Structure of the spleen of the sea bass (Dicentrarchus labrax): A light and electron microscopic study

The spleen of sea bass (Dicentrarchus labrax) is composed mainly of red pulp, whereas the white pulp is poorly developed. The red pulp consists of clear reticular cells intermingled with blood cells, sinusoids, and melanomacrophage centers (MMCs). The MMCs are enclosed by an interrupted connective tissue capsule and show some areas in continuity with the adjacent pulp. The MMCs are formed by the association of free macrophages that have phagocytosed some blood cells. Sparse white pulp is diffuse, forming a cuff around the pulp arteries and MMCs, or occurring in small groups between the splenic cords. A longitudinal artery and vein, lying side by side, extend the length of the spleen. Frequently the capillaries are surrounded by a sheath of macrophages or ellipsoids. These macrophages may contain erythrocytes in varying degrees of degradation. Lymphopoiesis and plasmapoiesis occur in the sparse lymphold areas. Abundant plasma cell groups may indicate the presence of antibody production.     

Spleen of the snake (Elaphe climacophora) and intrasplenic vascular architecture

The spleen of the blue-green snake (Elaphe climacophora) lies at the head of the pancreas and is separated from it by a fibrous capsule. A hilus is not clearly distinguished. Arteries and veins enter or leave the spleen through the capsule, but no collateral relationship is observed between these vessels. Histologically, the spleen consists of the capsule-septal tissue, lymphoid tissue (the white pulp), and a fibrous zone (the perilymphoid fibrous zone: PLFZ) around the lymphoid tissue that includes many small veins. The PLFZ probably represents the involuted red pulp. The border between the white pulp and PLFZ is unclear in routine histological sections due to diffuse infiltration of lymphocytes into the latter region, but the border could be distinguished clearly in silver-impregnated specimens. Arteries enter the spleen, run within the septa, and divide into terminal arteries in the lymphoid tissue that form end branches. There are no ellipsoids around the terminal arteries. The end branches communicate with veins of the PLFZ through transitional vessels within the lymphoid tissue (closed circulation). The veins of the PLFZ anastomose with drainage veins in the capsule. The snake spleen retains the basic histological characteristics of a spleen and is morphologically distinguishable from a lymph node. It may represent an extreme example of a spleen modified by the remodelling of the intrasplenic vasculatures during evolution. © 1995 Wiley-Liss, Inc.     

The blood circulation inside the spleen. I. The arterial blood vessels general distribution in the rat spleen

As an attempt to investigate the different pathways followed by the blood into the spleen and to analyse their functional significance, a technique was used mainly based on the intraarterial perfusion of a Prussian blue "solution" added of some chemical mediators and vasoactive substances. Such technique provides results which may be analysed taking into account the effect of the anaesthetic used, that may influence the findings. From the anaesthetic used, the sulfuric ether and the barbital sodium produce vasoconstriction of the white pulp blood vessels, whereas the chlorpromazine-promethazine doesn't have this effect, and so the Prussian blue appears inside these vessels. The vasodilator drugs, such as succinonitrile and papaverine hydrochloride, show a general vasodilator effect on the splenic arterial system. Teh arterial vessels of the white and the red pulp, including those placed at the subcapsular areas, become enlarged; into the white pulp, either the central or the peripheral blood vessel plexus of the lymphatic follicle becomes evident. The latter readily constitutes the perifollicular and the pericolumnar plexus. The blood vessels of this plexus become permeable to the Prussian blue "solution" by the heparin sodium effect, and so the dye particles enter the marginal zone and the splenic sinuses. In addition, from the white pulp arteries arise 2 types of anastomotic arterioles which appear enlarged after succinonitrile treatment: The short anastomotic arterioles that crosses the marginal zone entering the red pulp near the white pulp; the long anastomotic arterioles which enter the red pulp and after a long course end up into or around a collector sinus. The addition of histamine dihydrochloride to the perfusion solutions shows a slight vasodilator effect mainly on the subcapsular penicillar arterioles, including the helicine arterioles. The adrenergic stimulation of the splenic blood vessels induces a generalized arterial constriction, except of the anastomotic arterioles, that becomes open; in such way, the blood pathway follows the course of the anastomotic arterioles and the collector arterioles also become constricted. The adrenergic vasoconstrictor effect is inhibited by the phenoxy-benzamine hydrochloride. The addition of acetylcholine chloride, in the dosage, used, induces a generalized arterial vessel constriction, mainly of the perifollicular plexus. This effect is inhibited by atropine sulfate which, on the other hand, produces evident enlargement of the perifollicular and pericolumnar arterial plexus.     

Anatomical pathways from the white to the red pulp in the human spleen

In the human spleen we failed to find marginal zone bridging channels which in rats and mice are said to serve as return routes for lymphocytes from the white into the red pulp. In human spleens, using anticartilaginous antisera which distinctly visualized extracellular structures, in some parts we found the periarterial lymphocyte sheath to be closely attached to the red pulp, so that lymphocytes and other material could pass from the white pulp directly into the red pulp and vice versa. The strips of reticular fibres that seemed to bridge the marginal zone between the follicles and the red pulp proved to be components of reticular structures around the arteries, passing from the periarterial lymphocyte sheath into the follicles or from the follicles through the marginal zone into the red pulp.     

Ultrastructural analysis of normal and immunized spleen of the snapping turtle,Chelydra serpentine

In the ultrastructural comparison of normal, unimmunized spleens with immunized spleens at key intervals after antigenic stimulation with keyhole limpet hemocyanin (KLH), we noted cellular and cytological features which reflect the cellular kinetics of the primary immune response, particularly with respect to plasma cell production. Although lymphoblasts and mature plasma cells are present in the white and red pulp, respectively, intermediate stages of the plasma cell line are rarely found in normal spleen. Following antigenic challenge, we found a marked increase in lymphoblasts in the white pulp, most of them containing short segments of rough endoplasmic reticulum suggesting initial differentiation toward plasma cells. Following an apparent migration of cells from the white to the red pulp, we found plasma cells in various stages of maturation in the red pulp cords and sinuses. The ultrastructural features of these cells reflect 'the differentiation of lympho blasts into mature plasma cells. Both immature and mature plasma cells usually possess dilated cisternae of rough endoplasmic reticulum, suggesting that they are capable of producing and storing a secretory product, presumably antibody. We also noted a large number of immature macrophages and monocytes in immunized spleens. These cellular events and their cytological characteristics are compared to those described in other vertebrate classes.     

The argentophil reticular cells of the mammalian spleen. II. The argentophil reticular cell arrangement inside the red pulp and its relationship with the endothelial lining cells of the splenic sinuses

The red pulp's argentophil reticular cell network of the spleen is composed by 3 types of fixed cells: 1. the primitive reticular cell, slightly argentophil; 2. the small reticular cell; 3. the larger reticular cell, strongly argentophil and phagocytic. This latter shows the classical morphological characteristics attributed to the reticular cells of the spleen. The large argentophil reticular cell may become free, constituting a 4th cell type, the free macrophage. A 5th reticular cell type is the dendritic cell found into the lymphatic follicles of the white pulp. The argentophil reticular cells of the red pulp assemble together to form the reticular cells' network, that occurs inside the red pulp cords. The primitive and the small reticular cell form the fundamental network on which the large cells are apposed. The reticular cells of this network maitain relationship with the arterial terminal vessels of the red pulp, being responsible by the ellipsoid structure. In those arteriolar segments without ellipsoid and in those mammalian species devoid of ellipsoid, the white pulp reticular cells, that surround the blood vessel as a part of the lymphoid periarteriolar sheath, mix with the red pulp's reticular cells and both can hardly be discriminated. The ellipsoids are formed by large argentophil cells arranged in concentrical layers around its lumen that sometimes appear devoid of endothelial lining cells. The red pulp's argentophil reticular cells, either the small or the large ones, contributed to the structure of the splenic sinuses' wall; its thin processes surround the sinus wall outside the endothelial lining cell as fibrillar structures that cross the back side of the lining cells. Two or more argentophil reticular cells send fibrillar processes to a single sinus. The perisinusal reticular cells may send a process between adjacent endothelial lining, cells that insinuate and attain the sinus lumen; this process becomes thick and eventually, the reticular cell enter the sinus lumen as a free macrophage. The argentophil reticular cells of the red pulp make connection between the capsule or the trabeculae and the reticular cell network. The endothelial lining cells of the splenic sinuses are not argentophil.     

The marginal zone and marginal sinus in the spleen of the gerbil. A light and electron microscopy study

The spleen of the adult gerbil (M. unguiculatus) is characterized by the absence of venous sinuses and ellipsoid sheaths. The follicle (white pulp) is separated from the surrounding red pulp by a distinct marginal zone. The cell types in the marginal zone are common to both the follicle and red pulp. Separating the marginal zone from the follicle is a vascular channel of capillary dimension, the marginal sinus. A number of terminal segments of the arterial vessels within the follicle were observed to form a direct connection with the marginal sinus. Ultrastructurally, discontinuities were evident within the walls of the marginal sinus that would permit passage of both cellular and plasma components from the marginal sinus to either marginal zone or the follicle.     

The spleen ofmustelus schmitti (chondrichthyes, triakidae): A light and electron microscopic study

The spleen ofMustelus schmitti is described, being an elongated organ running dorsally along the stomach and surrounded by a thin capsule without muscular tissue. The classical division between red and white pulps was evident, with a marginal sinus surrounding the latter. Numerous ellipsoids were located in the red pulp, inside nodular-like structures, or in the marginal sinus. Two types of reticular cells were apparent as well as macrophages and melanomacrophages. Hemopoiesis was present through immature and mature cells of the erythroid and thrombocytoid lineages, but no evidence of granulopoiesis was found. Comparison amongMustelus species and between chondrichthyan and mammalian spleens are made.     

Microcirculatory pathways in normal human spleen, demonstrated by scanning electron microscopy of corrosion casts

Confusion regarding microcirculatory pathways in normal human spleen has arisen due to extrapolation from pathological material and from other mammalian spleens, not to mention difficulties in tracing intricate three-dimensional routes from the study of thin sections or cut surfaces of tissue. We examined microcirculatory pathways in normal human spleens freshly obtained from organ transplant donors. A modified corrosion casting procedure was used to obtain an open view of vessels and their connections. Our results demonstrate: 1) "arteriolar-capillary bundles" within lymphatic nodules and extensive branching of arterioles in the marginal zone (MZ); 2) the marginal sinus around lymphatic nodules; 3) the peri-marginal cavernous sinus (PMCS) outside the MZ or immediately adjacent to the nodule itself; the PMCS receives flow via ellipsoid sheaths and MZ, or directly from arterial capillaries, and drains into venous sinuses; 4) fast pathways for flow into venous sinuses via ellipsoid sheaths; 5) arterial capillary terminations in the reticular meshwork of the red pulp or MZ ("open" circulation); direct connections to venous sinuses also occur ("closed" circulation), although rarely; and 6) numerous open-ended venous sinuses in the MZ, allowing a large proportion of the splenic inflow to bypass the red cell filtration sites in the reticular meshwork and at venous sinus walls.     

Light microscopic study of the alligator (Alligator mississippiensis) spleen with special reference to vascular architecture

The spleen of the American alligator (Alligator mississippiensis) was studied histologically. The alligator spleen is a bean-shaped organ covered by a thick capsule. The concave side of the spleen faces the pancreas. Many venous vessels are present in the capsule. The stem segment of a large intestinal artery, the lieno-intestinal artery, enters the organ from its upper pole, runs within the organ at the axial center (axial artery) and leaves it from the lower pole. Many peripheral branches originate from the axial artery towards the capsule, but the artery has no associated collateral veins. The capsule/trabecula and white and red pulp may be distinguished. The capsular veins appear to be continuous with venous vessels that sheathe the axial artery and its peripheral branches. Surrounding the axial artery are trabeculae containing leiomyocytes and nerves. The white pulp consists of lymphoid tissue and occurs around terminal arterioles and sheathed capillaries. The materials examined do not show germinal centers. The large red pulp is composed of venous vessels and splenic cords rich in reticular fibers. Two venous routes, hilar and capsular, are present. The structural characteristics of the alligator spleen are similar to spleens of other reptiles; however, its vascular architecture is primitive, suggesting that the alligator spleen may be a portal spleen. J Morphol 233:43–52, 1997. © 1997 Wiley-Liss, Inc.     

The intermediate circulation in the nonsinusal spleen of the cat, studied by scanning electron microscopy of microcorrosion casts

Scanning electron microscopy of microcorrosion casts was used to visualize circulatory pathways of the intermediate circulation in nonsinusal spleen of cat. The marginal sinus (MS) around lymphatic nodules is a distinct vascular space which fills preferentially before the filling of the marginal zone (MZ) and surrounding red pulp occurs. The MS, which has a plentiful vascular supply, does not usually enclose the nodule completely. From the MS, flow occurs radially outwards into the MZ. Corrosion casts and histological sections both showed that a diversity of forms of the MZ exists: The thickness of MZ and the arrangement of its reticulum vary among nodules and between different areas of the same nodule, from a complete absence to a region of up to 50 microns in width. No direct arteriovenous connections were found (in contrast to dog spleen: Schmidt et al., '83b). Aside from capillary endings in the MS and MZ, all arterial capillaries terminate in the reticular spaces of the red pulp, i.e., the circulation appears to be entirely "open." From each capillary termination a great variety of flow pathways through the reticular meshwork to the pulp venules is available; some of these routes are quite long but others may involve distances as short as 15-25 microns. Evidence of flow into ellipsoid sheaths was abundant in casts from dilated spleens, but scarce in contracted spleens. In contrast to the extensive system of interconnected venous sinuses in dog spleen, the pulp venules found in cat spleen are nonanastomosing, shorter, and much smaller in caliber, and all receive flow freely from the reticular meshwork via open ends and fenestrations in their walls.     

Microvascularization of the spleen in larval and adult Xenopus laevis: Histomorphology and scanning electron microscopy of vascular corrosion casts

Microvascular anatomy and histomorphology of larval and adult spleens of the Clawed Toad, Xenopus laevis were studied by light microscopy of paraplast embedded serial tissue sections and scanning electron microscopy (SEM) of vascular corrosion casts (VCCs). Histology showed i) that white and red pulp are present at the onset of metamorphic climax (stage 57) and ii) that splenic vessels penetrated deeply into the splenic parenchyma at the height of metamorphic climax (stage 64). Scanning electron microscopy of VCCs demonstrated gross arterial supply and venous drainage, splenic microvascular patterns as well as the structure of the interstitial (extravasal) spaces representing the “open circulation routes.” These spaces identified themselves as interconnected resin masses of two distinct forms, namely “broccoli‐shaped” forms and highly interconnected small resin structures. Arterial and venous trees were clearly identified, as were transitions from capillaries to interstitial spaces and from interstitial spaces to pulp venules. Venous sinuses were not diagnosed (nonsinusal spleen). The splenic circulation in Xenopus laevis is “open.” It is hypothesized that red blood cells circulate via splenic artery, central arteries, penicillar arteries, and red pulp capillaries primarily via “broccoli‐shaped” interstitial spaces, pulp venules and veins into subcapsular veins to splenic veins while lymphocytes circulate also via the interstitial spaces represented by the highly interconnected small resin structures in vascular corrosion casts. In physiological terms, the former most likely represent the fast route for blood circulation, while the latter represent the slow route. J. Morphol. 277:1559–1569, 2016. © 2016 Wiley Periodicals, Inc.