Differential expression of Tenomodulin and Chondromodulin-1 at the insertion site of the tendon reflects a phenotypic transition of the resident cells.

The insertion site of the tendon to the skeletal element is hypovascular and is one of the most common sites of dysfunction in the musculoskeletal system. However, the resident cells have been poorly defined due to a lack of a specific marker for tenocytes. We previously reported that Tenomodulin (Tnmd) and Chondromodulin-1 (Chm1) are homologous angiogenesis inhibitors and predominantly expressed in the avascular region of tendons and cartilage, respectively. In this study, we analyzed the expression of Tnmd, Chm1, alpha 1 chain of the type I collagen (Col1a1) and alpha 1 chain of the type II collagen (Col2a1) at the insertion site of the Achilles, patellar, or rotator cuff tendons of 1-week-old rabbits by in situ hybridization analysis. Tnmd was co-expressed with Col1a1 in tenocytes of these tendons, while Chm1 and Col2a1 were detected in chondrocytes of the hyaline cartilage. Interestingly, the cell population between Tnmd/Col1a1 positive tenocytes and Chm1/Col2a1 positive chondrocytes expressed Col1a1 but none of the other markers (Tnmd, Chm1, and Col2a1). Red blood cells were exclusively present at the interface between the tendon substance and cartilage in the insertion site of the Achilles tendon. Lack of Tnmd and Chm1 in this newly characterized cell population may allow the transitional zone between the poorly vascularized tendon and cartilage to establish the unique vascular pattern for blood supply.

Surgical anatomy and efficient modification of procedures for selective extradural anterior clinoidectomy.

OBJECTIVE: The surgical anatomy of structures surrounding the anterior clinoid process (ACP) was examined in 39 cadavers to enable safe performance of selective extradural anterior clinoidectomy. METHODS: The lateral portion of the optic canal (OC) at the end of the orbit was defined as point A, and the lateral margin of the dural insertion into the superior orbital fissure (SOF) as B. A vertical line from A to the lateral margin of the ACP was assumed to define a crossing, defined as C. Distances between A and B, C and B, and C and A were measured. The length and width of OC and the optic strut were also measured. RESULTS: The mean distances between A and B, C and B, and C and A were 11.8, 8.9 and 6.3 mm, respectively. The mean length and width of OC were 8.9 and 5.7 mm, and those of the optic strut 5.4 and 2.4 mm, respectively. CONCLUSION: We propose a modified, efficient procedure for selective extradural anterior clinoidectomy, as follows. Drilling is started from the point approximately 9 mm posterior to the lateral margin of the dural insertion into the SOP, and pointed medially in the direction with a right angle to the lateral margin of the ACP. After drilling about 6 mm to reach the lateral border of the OC, unroofing of the OC is carried out to remove the ACP en bloc by fracturing of the optic strut. Using the present procedure, the distance of drilling of the lesser wing of the sphenoid bone is minimized.

A micro-anatomical model of the distribution of myocardial endomysial collagen.

Myocardial connective tissue probably provides passive support for regulating heart tensile strength and stiffness and ultimately for controlling heart mechanics through its endomysial part. However, endomysial collagen micro-arrangement is still a matter of debate. In order to define the fine distribution of left ventricle endomysial collagen, we applied the NaOH-scanning electron microscopy (SEM) maceration method (one of the techniques of choice for studying collagen micro-arrangement) to rabbit heart. Gomori-reticulum staining was used for correlated light microscopy (LM) observations. The SEM-NaOH method allowed isolation of collagen by removing other extracellular matrix components and cells and preserved collagen structure and position. Endomysial collagen appeared arranged in laminae that delimited the lacunae that were left empty by macerated myocytes and small vessels (mostly capillaries). These laminae were formed by reticular fibers, as confirmed by LM observations of Gomorireticulum-stained samples, and were organized in irregularly meshed networks made of thin (single) and thick (composed) filaments. In longitudinal views, collagen laminae extended the entire length of lacunae. In transversal views, the cut surface of the laminae appeared to be made of collagen bundles. These observations provide an updated microanatomical view of endomysial collagen distribution, which integrates previous studies. This model is based on the evidence that collagen laminae enveloped the surface of small vessels and myocytes. Thus, a type of myocyte-myocyte or capillary-myocyte "laminar connection" anchored to the entire cell length here is emphasized, rather than a type of "strut connection" anchored to defined loci, as usually described. This structure explains better how endomysium may provide the necessary support for heart compliance and protection against overstretch.

Phylogeny and ontogeny of the lymphatic stomata connecting the pleural and peritoneal cavities with the lymphatic system--a review.

This paper reviews the phylogeny and ontogeny of "lymphatic stomata" through which fluids and cells in the pleural and peritoneal cavities enter the lymphatic system. In amphibians, the pleuroperitoneal cavity is connected through numerous pores with the wide subvertebral lymphatic sinus corresponding to the thoracic duct in mammals. In reptiles, direct connections of the pleural and peritoneal cavities with the lymphatic system have been reported. In mammals, the pleural and peritoneal cavities are directly connected with lymphatics through lymphatic stomata. How do lymphatic stomata develop in mammals? In the rat, distinct lymphatics are noted in the subpleural space of the diaphragm periphery in 16 days old embryo. With age, the supleural lymphatics increase and form a polygonal network. They show a tubular appearance and possess many valves. Some of them become endowed with smooth muscle cells. In 19 days old embryos, some lymphatics appear in the subperitoneal space of the diaphragm. They extend centripetally and form many lateral projections that later elongate and connect with those from adjacent lymphatics, thus forming a lattice-like network or "lymphatic lacunae". During early postnatal days, the lymphatic lacunae project many bulges that subsequently come into contact with the pores among mesothelial cells lining the diaphragmatic peritoneum, thus forming lymphatic stomata. They increase until postnatal week 10. The lymphatic stomata in the costal pleura also develop during early postnatal days.

Light and electron microscopic study of cholinergic and noradrenergic elements in the basolateral nucleus of the rat amygdala: evidence for interactions between the two systems.

Pharmacological studies have suggested that the cholinergic (ACh) and noradrenergic (NA) systems in the amygdala (AM) play an important role in learning and memory storage and that the two systems interact to modulate memory storage. To obtain anatomical evidence for the interaction, the organization of the ACh and NA fibers in rat AM was investigated by immunocytochemistry for choline acetyltransferase (ChAT) and dopamine-beta-hydroxylase (DBH) in conjunction with light, confocal laser scanning, and electron microscopy (LM, CLSM, and TEM, respectively). LM showed that the ChAT immunoreactivity was densest in the basolateral nucleus (BL), whereas the DBH immunoreactivity was densest in the posterior BL. CLSM demonstrated that the ChAT-immunoreactive profiles in the BL were frequently located in juxtaposition to the DBH-immunoreactive axons. The TEM observations were as follows: The majority of the synapses formed by ChAT-immunoreactive terminals were symmetric, but DBH-immunoreactive axons formed both asymmetric and symmetric synapses. The ChAT-immunoreactive terminals usually established the symmetric synaptic contacts with the DBH-immunoreactive terminals and varicosities. The DBH-immunoreactive terminals formed the asymmetric synapses with the ChAT-immunoreactive dendrites of the intrinsic neurons within the AM. The results provide anatomical substrates for mnemonic functions of the ACh and NA systems and for the interactions between the two systems in the AM.

Postnatal development of lymphatic vessels and their smooth muscle cells in the rat diaphragm: a confocal microscopic study.

This paper reports on how lymphatic vessels and their smooth muscle cells develop in the diaphragm of postnatal rats. Lymphatic endothelial cells in the diaphragm were labeled by an intraperitoneal injection of DiI-labeled acetylated low-density lipoprotein (DiIac-LDL). During postnatal week 1, DiI-ac-LDL was detected in many free cells in addition to distinct endothelial cells that formed lymphatic vessels. Occasionally, saccular lymphatics isolated from previously formed lymphatics were recognized; these were referred to as lymphatic islands. The DiI-ac-LDL-labeled free and lymphatic endothelial cells showed immunoreactivity for CD 34 and Flt-4, but most of them did not express either OX 62 or ED 1 immunoreactivity, with only some showing ED 1 immunoreactivity. This suggests that most of the DiI-ac-LDL-labeled elements were lymphatic endothelial cells, and that some were macrophages. After postnatal week 1, the DiI-ac-LDL positive cells were restricted to lymphatic vessels. Until postnatal week 6, lymphatic vessels increased as the diaphragm enlarged. Towards the end of postnatal week 2, free cells expressing alpha-smooth muscle actin (alpha-SMA) immunoreactivity increased in the diaphragm, and some of these were in contact with lymphatics. A coarse plexus of smooth muscle cells surrounding the lymphatic vessels first appeared at postnatal week 2, and this plexus became denser with age. Our findings indicate that lymphatic vessels are formed not only by sprouting from previously formed lymphatic vessels but also by migrating endothelial cells, and that smooth muscle cells may be differentiated from mesenchymal cells to form a plexus surrounding the lymphatic vessels.

Lectin binding patterns in rat nasal-associated lymphoid tissue (NALT) and the influence of various types of lectin on particle uptake in NALT.

We investigated the binding of four types of lectin to follicle-associated epithelium overlying the nasal-associated lymphoid tissue (NALT) of rats in order to identify M-cell specific surface markers and to determine the influence of lectin administration to NALT on the uptake of a particulate antigen. The NALT tissues were incubated with a panel of four types of lectin conjugated to horseradish peroxidase (HRP). Ulex europaeus-1 (UEA-1) and Dolichos biflorus (DBA) lectin stained the surface of M-cells and goblet cells. Uniform staining by Triticum vulgaris (WGA) was detected in the M-cells, ciliated cells and goblet cells. In contrast, staining of Griffonia simplicifolia I isolectin-B4 (GSI-B4) was almost exclusively M-cell specific. The administration of M-cell specific lectin (GS I-B4) to NALT suppressed the uptake of baker's yeast particles administered later, whereas the non-specific one (UEA-1) had no influence on the uptake. These results indicate that GS I-B4 is a useful marker for the identification of rat NALT M-cells and that such a specific expression of surface glycoconjugates by M-cells may permit the targeting of vaccines and drugs to the antigen sampling sites of the nose. It also appears possible to block the uptake of pathogens by an administration of M-cell specific lectin to NALT.

A corrosion casting/scanning electron microscope method that simultaneously demonstrates clear outlines of endothelial cells and three-dimensional vascular organization.

This paper describes a method that can definitively demonstrate endothelial cell boundaries on corrosion casts of arteries, veins, and capillaries. After perfusion with silver nitrate, a casting medium was injected into the entire vascular bed. The injected tissues were either exposed to light or immersed in the photographic developer to develop the silver halide, and corroded in a 5% NaOH solution at 60 degrees C overnight. Observations of the casts containing water in a low vacuum scanning electron microscope equipped with a cooling stage clearly showed endothelial cell boundaries on casts of every type of vessel as well as their three-dimensional architecture. The low vacuum scanning electron microscope images of wet casts were almost identical in quality to the back-scattered electron images of dried casts without any coating. Secondary electron images of the dried casts with metal coating clearly showed endothelial cell outlines and nuclear imprints. The secondary electron images at high magnification indicated that silver granules were precipitated in the grooves along endothelial cell boundaries on the casts. Since this method can demonstrate endothelial cell boundaries of every type of vessel in addition to their three-dimensional architecture, it will be a powerful tool for examining endothelial cell morphology and microvascular organization in pathological as well as normal tissues.

Correlation of glutamate-induced apoptosis with caspase activities in cultured rat cerebral cortical neurons.

In cultured rat cortical neurons lactate dehydrogenase (LDH) activity in the medium, a cell-death marker, increased gradually after exposure to glutamate (100 microM to 1 mM) for 60 min and reached a plateau at 24 to 30 h. Neuronal death was mainly apoptotic as suggested by typical electron microscopic findings, fluorescent double staining with membrane-permeating and nonpermeating chromatin dyes, nick end labeling, and assessment of DNA fragmentation by agarose gel electrophoresis. After 1 mM glutamate exposure, a rise of interleukin-1beta converting enzyme (ICE)-like protease activity in neurons was parallel to cysteine protease p32 (CPP32)-like protease activity and declined before CPP32-like protease activity reached the peak (at 6 h). LDH activity in the medium of glutamate-exposed neurons was decreased by specific ICE and/or CPP32 inhibitors, acetyl-L-tyrosyl-L-valyl-L-alanyl-L-aspart-1-al (Ac-YVAD-CHO) and acetyl-L-aspartyl-L-glutamyl-L-valyl-L-aspart-1-al (Ac-DEVD-CHO), respectively, in a dose-dependent manner. Fluorescent double staining of nuclei also demonstrated that at 100 microM each inhibitor prevented neuronal apoptosis and that this effect was additive. Among agonists corresponding to various glutamate receptor subtypes, N-methyl-D-aspartate (NMDA) and kainate induced apoptosis in cortical neuronal cultures while alpha-amino-3-hydroxy-5-methylisoxazole-4-propinate (AMPA) did not. The metabotropic glutamate receptor agonist, 1-aminocyclopentane-1S, 3R-dicarboxylate (ACPD) prevented apoptosis. Interestingly, apoptosis at 24 h after agonist or antagonist exposure correlated closely with caspase activity 6 h after exposure.

Inhibitory effect of nilvadipine on disruption of blood-aqueous barrier induced by prostaglandin E2 application in pigmented rabbits: A morphologic study.

The purpose of the present study is to investigate the morphologic sites of breakdown in eyes pretreated with nilvadipine (a calcium channel blocker) that has been shown to inhibit the acute rise of aqueous flare induced by prostaglandin E2 (PGE2). Nilvadipine (100 microg/kg body weight) was injected intravenously in pigmented rabbits. Thirty minutes later, vehicle or PGE2 (10, 50 or 250 microg/ml) was applied on the cornea by use of a glass cylinder. Forty-five minutes later, the animals received horseradish peroxidase (HRP) intravenously and the eyes were enucleated. Distribution of HRP in the anterior segments was observed by electron microscopy. Without nilvadipine pretreatment, HRP was seen in the intercellular space of nonpigmented cells of the eyes treated with 50 microg/ml PGE2 and in the iris stroma of the eyes treated with 250 microg/ml PGE2. With nilvadipine pretreatment, HRP was not observed in these sites. Our results indicate that nilvadipine suppresses disruption of the different sites of the blood-aqueous barrier.

Development of diaphragmatic lymphatics: the process of their direct connection to the peritoneal cavity.

The development of the lymphatic system in the rat diaphragm was studied from embryonic day 16 to 25 weeks after birth by histochemistry for 5'-nucleotidase, scanning electron microscopy of KOH-treated or intact tissues, and transmission electron microscopy of thin sections. On embryonic day 16, distinct lymphatics were noted in the subpleural space of the diaphragm periphery. The endothelial cells at this stage contained an abundance of rough endoplasmic reticulum, a developed Golgi apparatus and mitochondria, and fewer pinocytotic vesicles than those in adults. The subpleural lymphatics subsequently increased and formed a polygonal network. They possessed many valves, and by postnatal week 6, some thick collecting lymphatics became endowed with smooth muscle cells. On embryonic day 19, some lymphatics appeared in the subperitoneal space. They extended centripetally and had many lateral projections that subsequently became elongated and connected with those from adjacent lymphatics, thus forming a lattice-like network. During the early postnatal days, the subperitoneal lymphatics projected many bulges that subsequently became elongated, and came into contact with the pores among the mesothelial cells, thus forming lymphatic stomata connecting the lymphatic lacunae to the peritoneal cavity. The lymphatic stomata increased until postnatal week 10. The results show that lymphatics appear as early as embryonic day 16 in the subpleural space of the diaphragm periphery, and develop with age by sprouting to form networks in both the subpleural and the subperitoneal spaces, and that the direct connection of the lymphatic lacunae to the peritoneal cavity is formed after birth.

Lymphatics of the rat mammary gland during virgin, pregnant, lactating and post-weaning periods.

The distribution and ultrastructure of the lymphatics of the rat mammary gland in virgin, pregnant, lactating and post-weaning periods were examined by enzyme-histochemistry for 5'-nucleotidase (5'-Nase) and transmission electron microscopy. Enzyme-histochemistry for 5'-Nase stained lymphatics in dark brown. In the lactating period, lymphatics abounded in the interlobular connective tissues, but in other periods they were few. The interlobular lymphatics drained into collecting lymphatics running along the mammary ducts. Gaps between lymphatic endothelial cells were significantly wider in lactating period than in other periods, while both number and area of vesicles in the lymphatic endothelial cells were significantly larger in the virgin period than in other periods. In the pregnant and lactating period, the lymphatics contained many lymphocytes and lipid droplets. The results show that during lactating period, the interlobular lymphatics increase and that gaps between lymphatic endothelial cells serve as a major route through which tissue fluids and particulate matters enter the lymphatics, while vesicles seem to be main trans-endothelial transport route during the virgin period. The results will provide basic information for our next investigation on lymphangiogenesis in association with breast cancer.

Organization of the blood and lymphatic microvasculature of the gallbladder in the guinea pig: a scanning electron microscopic study.

The organization of the blood and lymphatic microvessels of the gallbladder in the guinea pig is demonstrated by scanning electron microscopy (SEM) of vascular corrosion casts, and SEM of KOH-macerated tissues. In the lamina propria of the gallbladder, there is a dense network of subepithelial capillaries. The network is supplied by the arterioles that come off the arterial plexus located deep in the lamina propria. The network gathers into the postcapillary venules continuous with the collecting venular plexus located immediately below the subepithelial capillary network. The precapillary arterioles are sparsely surrounded by a single layer of circularly oriented extensions of smooth muscle cells. The terminal arterioles are endowed with circularly oriented fusiform smooth muscle cells. The nervous plexus is also noticed along the terminal arterioles. The capillaries are embraced by flat prolongations of pericytes. The postcapillary venules are sparsely surrounded by stellate pericytes and the collecting venules are sparsely surrounded by elongated or branched spindle-shaped, primitive smooth muscle cells which extend their long process in various directions along the vascular wall. The lymphatics are mostly located in the subserosal layer. The tips of the initial lymphatics are closed by endothelial cells, although there are frequently some gaps between them. The thin flaps of the lymphatic endothelial cells overlap or interdigitate with each other. The luminar surfaces of the lymphatics show oval nuclear protrusions, while the abluminal surfaces showed numerous microfolds except for the oval and flat nuclear portions. The lymphatics possess neither smooth muscle cells nor pericytes.

Comparative analysis of insulo-acinar portal system in rats, guinea pigs, and dogs.

The insulo-acinar portal system in the rat, guinea pig, and dog was comparatively analyzed using corrosion casting method in scanning electron microscopy and confocal laser scanning microscopy. In all animals examined, there were three types of arterioles according to their destination: 1) the arteriole which supplied the capillary glomerulus of the islet, 2) the arterioles which directly branched out into capillaries around the acini, and 3) the arterioles which supplied the duct system. In the rat, the afferent vessel usually ended in the cortical layer of the islet and its main branches ran along this layer before giving secondary capillary branches into the deeper regions, while in the dog and guinea pig, the region where the afferent arterioles branched out into secondary capillary branches varied among individual islets. There were three types of efferent vessels of the islet: 1) the insulo-acinar portal vessels that radiated from the islet to join the capillary network in the exocrine pancreas, 2) the emissary venules of the islet, leading directly into the systemic circulation, and 3) the insulo-ductal portal vessels which drained into the peri-ductal capillary network. In the rat and guinea pig, the intralobular islets possessed both the insulo-acinar portal vessels and the emissary venules, while the interlobular islets possessed emissary venules with occasionally occurring insulo-acinar portal vessels. In the dog, most of the islets were located within the lobule and possessed preferentially the insulo-acinar portal vessels. In this animal, the lobule was supplied by several microvascular units, in the center of which was located the capillary glomerulus of the islet. The peri-insular zone of the unit was mainly supplied by the insulo-acinar portal vessels, while the periphery, the tele-insular zone, was directly supplied by arterioles as well. The venules originated at the periphery of the unit. The islet in the dog had virtually no emissary venules. Confocal laser scanning microscopy of the rat islets showed that B cells occupied the core of all islets. The microvascular architecture within the rat islet appeared to be organized as to drain blood from the A and D cell area to the B cell area of the islet.

Sites of disruption of the blood-aqueous barrier after application of prostaglandin E2 in pigmented rabbits.

We examined the disruption of the blood-aqueous barrier following prostaglandin (PG)-E2 application in rabbits. Vehicle or PGE2 in 10, 50 or 250 microg/ml concentration was applied to the cornea of pigmented rabbit with the use of a glass cylinder. After PGE2 administration, horseradish peroxidase (HRP) was injected intravenously. Then the eyes were enucleated, and distribution of HRP in the anterior segments was examined by electron microscopy. In control eyes, diffusion of HRP was blocked by vascular endothelial cells in the iris and by nonpigmented epithelial cells in the iridial and ciliary processes. In the iridial and ciliary processes of the eyes treated with 10 microg/ml PGE2, no HRP reaction product was seen in intercellular spaces of the nonpigmented epithelial cells, but it was found in pinocytotic vesicles. In the eyes treated with 50 microg/ml PGE2 HRP reaction product was found in intercellular spaces of the nonpigmented cells in the iridial processes. In the eyes treated with 250 microg/ml PGE2 HRP reaction product was further distributed in the iris stroma. The present study demonstrated that the sites of breakdown of the blood-aqueous barrier depended upon the doses of exogenous PGE2.

Distribution and ultrastructure of the stomata connecting the pleural cavity with lymphatics in the rat costal pleura.

We investigated the detailed distribution and ultrastructure of the stomata connecting the pleural cavity and the lymphatics in the rat costal pleura by scanning electron, transmission electron and light microscopy. The mesothelial cells lining the costal pleura appeared as both flattened and thick cell bodies. The thick cells possessed more rough endoplasmic reticula, Golgi complexes, mitochondria, and free ribosomes than the flattened cells. The thick cells were distributed in the intercostal regions each cephalic to the junction of the costal cartilage and bone, and in the band-like regions along the cephalic and caudal sides of each rib in the lateral and dorsal thoracic walls. In the regions lined with thick cells, there were stomata [12.9 +/- 10.3 microns2 (mean +/- SD) in area] consisting of prolongations of thick mesothelial cells and funnel-like projections of lymphatic endothelial cells that came up along the rims of the pores (5.9 +/- 3.2 microns2 in average area) in the submesothelial collagen fiber network. At the stomata, the basal lamina of the mesothelium was continuous with that of the endothelium. The mesothelial cells forming the stomata were mostly in close contact with the endothelial cells, but some gaps also existed between them. Valve-like endothelial flaps were frequently observed wherever endothelial cells constituting the stomata merged into the submesothelial lymphatics. Also present were lymphatic bulges that were either in close contact with the base of the thick mesothelial cells or exposed through the mesothelial pores. The lymphatic network was especially well developed in the submesothelial layer at and around the thick-cell regions. The initial lymphatics drained into the intercostal collecting lymphatics, which in turn led into either the parasternal or paravertebral lymphatic trunk. Our results suggest that the stomata play a major role in absorbing fluids and particulates in the pleural cavity. The thick mesothelial cells appear to secrete chemotactic substances to the endothelial cells. Understanding the heterogeneous distribution of the stomata could prove to be important clinically in inflammatory diseases and tumors in the chest.

Obliteration of the lymphatic trunks draining diaphragmatic lymph causes peritoneal fluid to enter the pleural cavity.

Pathways of peritoneal fluids to the pleural cavity in the rat were investigated by light microscopy and transmission electron microscopy (TEM). Intraperitoneally injected India ink was demonstrated to enter the subperitoneal lymphatics through lymphatic stomata, and to drain through the subpleural collecting lymphatics, into the parasternal, paravertebral and mediastinal lymphatic trunks as well as the thoracic duct. Five to 10 min after the intraperitoneal injection of India ink, the parasternal lymphatic trunk was ligated at the third intercostal space. Thirty minutes, 1 h, or 2 h after the ligation of either the right or the left trunk, India ink was macroscopically recognized only around the ligated trunk. When the right and left trunks were simultaneously ligated, India ink leaked around both trunks. Five hours after the ligation of both trunks, a massive amount of ink was located in the interstitium of the anterior thoracic wall. TEM revealed carbon particles passing through gaps of the lymphatic endothelial cells into the interstitial space, and partly reaching the mesothelial surface lining the anterior thoracic wall. Results show that obstruction or narrowing of the lymphatic trunks draining the diaphragmatic lymph causes a hydrothorax, indicating that this is at least one mechanism causing this during continuous ambulatory peritoneal dialysis and diseases with ascites.

Distribution of lymphatics in human palatine tonsils: a study by enzyme-histochemistry and scanning electron microscopy of lymphatic corrosion casts.

The distribution of lymphatics in human palatine tonsils was studied by enzyme-histochemistry for 5'-nucleotidase (5'-Nase) and scanning electron microscopy (SEM) of lymphatic corrosion casts. The palatine tonsils were found to possess lymphatics in the parafollicular area (i.e., interfollicular, interfolliculo-septal, and folliculo-septal area), in the connective tissue septa, and in the capsules, but not in the subepithelial area between the follicles and the follicle-associated epithelia or within the follicles. The tubular lymphatics originated some 200-300 microns below the epithelium and formed a three-dimensional network in the parafollicular area. Some lymphatics around the lower part of the follicle were flat, wide, and irregular in shape, and thus appeared to be lymphatic sinuses, referred to as perifollicular lymphatic sinuses. The lymphatics in the parafollicular area drained into the septal lymphatics, which ran rather straight in the connective tissue septa. The septal lymphatics finally gathered into the broader capsular lymphatics. Most of the septal and capsular lymphatics were endowed with valves. Our results indicate that lymphocytes and fluid from the follicles and the subepithelial region enter the perifollicular lymphatic sinuses and/or the interfollicular lymphatics, pass through the interfolliculo- and folliculo-septal lymphatics, and finally enter the septal and capsular lymphatics to leave the tonsil.