The lymphatic system plays important roles in maintaining tissue fluid homeostasis, immune surveillance of the body, and the taking up dietary fat and fat-soluble vitamins A, D, E and K. The lymphatic system is involved in many pathological conditions, including lymphedema, inflammatory diseases, and tumor dissemination. A clear understanding of the organization of the lymphatic vessels in normal conditions would be critically important to develop new treatments for diseases involving the lymphatic vascular system. Therefore, the present paper reviews the organization of the lymphatic vascular system of a variety of organs, including the thyroid gland, lung and pleura, small intestine, cecum and colon in the rat, the diaphragm in the rat, monkey, and human, Peyer's patches and the appendix in the rabbit, and human tonsils. Methods employed include scanning electron microscopy of lymphatic corrosion casts and tissues with or without treatment of alkali maceration technique, transmission electron microscopy of intact tissues, confocal microscopy in conjunction with immunohistochemistry to some lymphatic-specific markers (i.e., LYVE-1 and VEGFR-3), and light microscopy in conjunction with enzyme-histochemistry to 5'-nucleotidase. Some developmental aspects of the lymphatic vessels and lymphedema are also discussed.

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In the present study, we found that leucocyte function-associated antigen-1 (LFA-1), and integrin (heterodimer complex of CD11a and CD18), which are abundant in immunological synapse, were expressed in developing hippocampal neurons. The expression of LFA-1 in hippocampal neurons was in the period of synaptogenesis, and synaptogenesis was inhibited by the blocking antibodies of anti-CD11a or anti-CD18 in vitro. Since it is known that LFA-1 has an important role in the immunological response, especially in immunological synapse, LFA-1 is considered to have an important role in neuronal synapse and is highly involved in synaptogenesis in the early developmental stage in vitro. In vivo, we also confirmed that CD18 was expressed in hippocampus in the early developmental stage. Telencephalin, which is a candidate for postsynaptic elements to contact LFA-1, was precisely opposed to CD18-positive structures in presynaptic elements, and telencephalin was considered to be involved in synaptogenesis. The present study showed that 17β-estradiol of steroid hormones, which are well known to have various effects on hippocampal neurons, has a significant influence on the presynaptic expression of CD18 in synaptogenesis and inhibited synaptogenesis in the early developmental stage in vitro. These results suggest that LFA-1 plays some mechanisms in synaptic contacts and synaptogenesis of hippocampal neurons.

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The present study was carried out to determine the effect of decorin in the process of collagen assembly. Collagen fibrils were obtained in vitro by aggregation from commercialized acid-soluble type I collagen with the addition of different concentrations of decorin (0-25 μg/ml). All specimens were observed by scanning electron microscopy (SEM) and transmission electron microscopy (TEM). The distribution of collagen fibril diameters was also analyzed by TEM. In samples without or with low concentrations of decorin, highly porous collagen fiber networks were formed. On the other hand, dense networks were observed in samples treated with high concentrations of decorin. The influence of decorin secreted by cells on collagen fibrils was observed by SEM, and the fiber network elasticity was measured using a rheometer. SEM images showed that collagen fiber networks without fibroblasts were much looser than those cultured with normal fibroblasts. The networks cultured with the fibroblasts were composed of straight fibers with large diameters. On the other hand, collagen fiber networks cultured with siRNA-decorin-transfected (siDT) fibroblasts had loose, meandering fibers with small diameters. The elasticity of collagen fiber networks cultured with untransfected fibroblasts showed no significant difference over the 7-day incubation period. However, significantly lower elastic values were obtained for collagen fiber networks treated with siDT cells on days 3 and 7. In addition, after treatment with 5.0 or 25 μg/ml decorin, the l collagen fiber networks cultured with siDT cells exhibited an altered structure that showed a dense structure similar to that of the fiber networks cultured with untransfected fibroblasts. In conclusion, this in vitro study showed that decorin is a regulatory and architecturally small leucine-rich repeat proteoglycan in the process of collagen fibril assembly.

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The skeletal muscle capillary supply (capillarity) dynamically changes in response to muscle conditions such as growth, atrophy, and hypertrophy. The capillary number-to-fiber ratio is reported to correlate closely with the muscle fiber cross sectional area. However, little information is available regarding the capillarity of neonatal and very young skeletal muscles. In this study, the vascular endothelium was reliably stained with an anti-PECAM-1 antibody, and relationships between the capillarity and muscle fiber parameters were analyzed. For assessment of the capillarity, we used the capillary length-to-fiber ratio, due to the presence of transversely running vessels. In young and adult rats, the capillary length-to-fiber ratio was proportional to both the muscle fiber cross sectional area and muscle fiber radius. However, when these data were analyzed together with data from neonatal and very young rats, the capillary length-to-fiber ratio correlated more closely with the muscle fiber radius than the muscle fiber cross sectional area in the tibialis anterior muscle. The capillary number-to-fiber ratio demonstrated results very similar to the capillary length-to-fiber ratio. During muscle atrophy after denervation, the number of capillaries was decreased in a non-apoptotic manner as revealed by electron microscopy, maintaining the close relationship between the parameters described above. In conclusion, capillarity was closely correlated with the muscle fiber radius (which represents the perimeter) during growth and atrophy. This indicates that the capillarity is linked to the muscle fiber surface area (which is determined by perimeter and section thickness), in agreement with the essential role of the cell membrane in the transport of materials by simple diffusion or active transport.

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The present study employed immunohistochemistry for single-stranded DNA (ssDNA) to detect apoptotic cells in taste buds of the rat circumvallate papilla. Double-labeling of ssDNA and markers for each cell type - phospholipase C β2 (PLCβ2) and α-gustducin for type II cells, neural cell adhesion molecule (NCAM) for type III cells, and Jacalin for type IV cells - was also performed to reveal which types of cells die by apoptosis. We detected approximately 16.8% and 14.0% of ssDNA-immunoreactive nuclei among PLCβ2-immunoreactive and α-gustducinimmunoreactive cells, respectively, but rarely found ssDNA-immunoreactive cells among NCAM-immunoreactive or Jacalin-labeled cells, indicating that type II cells die by apoptosis. We also applied double labeling of ssDNA and human blood group antigen H (AbH) - which mostly labels type I cells as well as other cell types - and found that approximately 78% of ssDNA-immunoreactive cells were labeled with AbH, indicating that apoptosis also occurs in type I cells. The present results revealed that apoptosis occurs in both type I cells (dark cells) and type II cells (light cells), suggesting that there are two major cell lineages (dark cell and light cell lineages) for the differentiation of taste bud cells. In summury, type IV cells differentiate into dark and light cells and type III cells differentiate to type II cells within the light cell line.

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