Current knowledge on the Ruffini endings, primary mechanoreceptors in the periodontal ligament is reviewed with special reference to their cytochemical features and regeneration process. Morphologically, they are characterized by extensive ramifications of expanded axonal terminals and an association with specialized Schwann cells, called lamellar or terminal Schwann cells, which are categorized, based on their histochemical properties, as non-myelin-forming Schwann cells. Following nerve injury, the periodontal Ruffini endings of the rat incisor ligament can regenerate more rapidly than Ruffini endings in other tissues. During regeneration, terminal Schwann cells associated with the periodontal Ruffini endings migrate into regions where they are never found under normal conditions. Also during regeneration, alterations in the expression level of various bioactive substances occur in both axonal and Schwann cell elements in the periodontal Ruffini endings. Neuropeptide Y, which is not detected in intact periodontal Ruffini endings, is transiently expressed in their regenerating axons. Growth-associated protein-43 (GAP-43) is expressed transiently in both axonal and Schwann cell elements during regeneration, while this protein is localized in the Schwann sheath of periodontal Ruffini endings under normal conditions. The expression of calbindin D28k and calretinin, both belonging to the buffering type of calcium-binding proteins, was delayed in periodontal Ruffini endings, compared to their morphological regeneration. As the importance of axon-Schwann cell interaetions has been proposed, further investigations are needed to elucidate their molecular mechanism-particularly the contribution of growth factors-during the regeneration as well as development of the periodontal Ruffini endings.

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The cytoarchitecture of smooth muscle cells and pericytes in the rat cardiac vessels was studied by scanning electron microscopy after the removal of connective tissue matrices using a modified KOH-collagenase digestion method. The initial stem of the coronary arteries had groups of smooth muscle cells which ran in various directions on the outermost layer of the media. Although smooth muscle cells in coronary arteries of more than 100 μm in the outer diameter were arranged in a rough circle around the vessel axis, oblique and/or longitudinal muscle bundles were often present in the medio-adventitial border of the vessels. The presence of irregularly oriented muscular bundles is probably connected with resistance against the stretching force induced by the beating of the heart. As the vessel size decreased toward the periphery, almost all of the smooth muscle cells became spindle-shaped with several tiny processes and ran circularly or helicaly to the vessel axis. In the precapillary arterioles (6-12 μm), smooth muscle cells acquired various cytoplasmic processes which helicaly surrounded endothelial cells. Unmyelinated nerves were often associated with arterioles. Blood capillaries were morphologically divided into three segments: arterial capillaries which had pericytes with wide and circularly oriented processes, true capillaries whose pericytes extended long and thin primary processes bilaterally along the vessel axis, and venous capillaries surrounded irregularly and loosely by wide pericytic processes. The stellate pericytes in the postcapillary venules (10-30 μm) gradually changed into flat tape-like smooth muscle cells, which ran circularly in the collecting venules and veins (30-200 μm). The large collecting veins were finally overwhelmed by superficial thin layer of the myocardium, their own smooth muscle cells being very sparse. This suggests that large veins have poor ability to contract by themselves but are influenced by the surrounding myocardial cells.

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The present study was designed to analyze the subfibrillar structure of corneal and scleral collagen fibrils by scanning electron microscopy (SEM) and atomic force microscopy (AFM). Isolated collagen fibrils of the bovine cornea and sclera were fixed with 1% OsO₄, stained with phosphotungstic acid and uranyl acetate, dehydrated in ethanol, critical point-dried, metal-coated, and observed in an in-lens type field emission SEM. Some isolated collagen fibrils were fixed with 1% OsO₄, dehydrated, critical point-dried and observed without metal-coating in an AFM. Isolated collagen fibrils treated with acetic acid were also examined by SEM and AFM. SEM and AFM images revealed that corneal and scleral collagen fibrils had periodic transverse grooves and ridges on their surface; the periodicity (i. e., D-periodicity) was about 63 nm in the cornea and about 67 nm in the sclera. Both corneal and scleral collagen fibrils contained subfibrils running helicoidally in a rightward direction to the longitudinal axis of the fibril; the inclination angle was about 15° in the corneal fibrils and 5° in the scleral fibrils. These findings indicate that the different D-periodicity between corneal and scleral fibrils depends on the different inclinations of the subfibrils in each fibril. The present study thus showed that corneal collagen fibrils differ from scleral collagen fibrils not only in diameter but also in substructure.

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The development and cytoarchitectures of liver hematopoiesis in the mouse from 10 to 19 days of gestation were examined by light and electron microscopy. In fetal liver hematopoiesis, four stages were identified: Stage I, the onset of hematopoiesis at 10 days; Stage II, expansion of the volume of the hematopoietic compartment at 11 and 12 days; Stage III, the peak in the volume of the hematopoietic compartment at 13 and 14 days; and Stage IV, the involution of hematopoiesis after 15 days. During Stages I-II, hematopoietic stem cells appeared to move from the sinusoidal lumina into primitive hepatic cell cords through the sinusoidal endothelium to give rise to colonies among hepatoblasts. At Stage III, the hematopoietic colonies formed ellipsoidal foci as a structural unit of hematopoiesis. These foci were 35-70 × 20-40 μm in size, and erythroblastic islands could be observed in the center of each. Each island contained central macrophages surrounded by a ring of erythroblasts. The macrophages underwent mitosis, showing close contact with the erythroblasts, after which the hematopoietic foci appeared as cords. At Stage IV, these cord-shaped hematopoietic foci became disrupted, and round solitary foci including macrophages appeared within the hepatic cell cords on meandering sinusoids. In fetal liver hematopoiesis, macrophages could be one of the major cell components comprising the hematopoietic microenvironment, especially at Stages II and III.

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To understand the roles of thymic stromal cells in T-lymphocyte development, we semiquantitatively analysed rat thymi recovering from irradiation (6 Gy), using a transmission electron microscope. The most striking findings were that the percentage of subcapsular epithelial cells significantly increased in the cortex on day 3 after irradiation compared with the control; the percentage of intermediate epithelial cells significantly increased in the cortex on days 3 and 5 after irradiation and in the medulla on days 5 and 7 compared with the control; the interdigitating cells disappeared from the medulla by day 7 after irradiation and reappeared on day 9. The present data thus reveal that during recovery after irradiation (6 Gy), marked changes occur in the relative proportions of different epithelial cell subtypes in the cortex and medulla of the rat thymus. In addition, the percentages of macrophages and interdigitating cells also changed during the recovery. These changes, which may be associated with the abrupt proliferation of thymocytes after irradiation, should shed light on the significance of stromal cells in the T cell development.

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While the intestine has abundant intraepithelial lymphocytes (IELs) including extrathymically differentiated T-cell populations and natural killer (NK) cells, the stomach contains only a few IELs. To elucidate whether the gastric epithelium is capable of inducing predominant lymphocyte lodging and subsequent differentiation within, we counted the number of IELS and lamina propria lymphocytes (LPLs) and calculated the percentage of IELS to total lymphocytes for each alpha-beta (αβ) T cell, gamma-dalta (γδ) T cell, CD4⁺ cell, CD8⁺ cell and NK cell in normal and regenerating gastric mucosa as well as the intestinal mucosa of the rat. In the normal rat pylorus, a few αβT cells but no γδT cells were found in the epithelium and lamina propria. In regenerating gastric mucosa, all subsets of LPLs increased in number to a degree comparable to those in intestinal mucosa, whereas every IEL subset, though slightly increased, was much smaller in number than in the intestinal mucosa, consequently giving lower percentages of IELs. Electron microscopic observations revealed that all IELs in regenerating gastric mucosa were agranular, while 55% of intestinal IELs were large granular lymphocytes positively stained for an NK-cell, αβ-cell or γδT-cell marker. The present results indicate that, unlike the intestinal epithelium, the gastric epithelium does not induce the preferential localization of T cells/NK cells and T-cell differentiation into granular lymphocytes in the epithelium even under conditions of prominent LPL infiltration.

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To investigate the effects of tissue fluid on the growth of enamel crystals, upper and lower incisors extracted from 3-week-old Wistar rats were removed of the enamel organ, implanted subcutaneously in the dorsal portion of animals from the same litter, and harvested at 72 h or 1 week after implantation. The grafts were chemically fixed with surrounding tissues and prepared for light and electron microscopy, X-ray microanalysis, or for the immunohistochemistry of amelogenin. Mineralization of implanted enamel layers was examined by contact X-ray microradiography.
  The immunoreactivities for 25 kD amelogenin in immature enamel decreased sequentially, starting from the surface to the deeper layers; by 1 week after implantation, no positive reactivities remained in the entire enamel layers at the stages of matrix formation and early maturation. In accordance with the loss of enamel proteins, immature enamel gained mineral density until it attained higher radio opacity than that of the adjacent dentin by 1 week. In contrast, the radio opacity of the full thickness of the enamel at early maturation remained low except for a surperficial thin layer.
  Electron microscopy revealed no sign of growth of original enamel crystals, but showed heavy precipitation of electron-dense fine granules of calcium phosphate in all layers of the secretory enamel and the superficial layer of enamel at early maturation, which showed high radio opacity. The Ca/P ratio and electron diffraction patterns of the granular materials precipitated between intrinsic enamel crystals indicated the property of hydroxy apatite or octacalcium phos-phate though a characteristic ribbon-like profile of enamel crystals was lacking.
  These data indicate that the enamel organ blocks exogenous mineral precipitates in growing enamel during the stage of matrix formation and plays an essential regulatory role for fine enamel crystallites to grow into large hexagonal crystals.

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Some species of soft-shelled turtle have been known to use a conspicuous mass of villiform processes of the pharyngeal mucosa as an aquatic respiratory organ when staying underwater for prolonged periods, such as hibernation. Using hibernating turtles, Trionyx sinensis japonicus, the present study employed scanning electron microscopy to demonstrate for the first time the detailed morphology and distribution of these villiform processes. Two types of processes, complex and simple, could be identifited.
  Light microscope observation of the transverse sections of the villi demonstrated a rich vascularization in the connective tissue of the villi, comprising arterioles and venules running in the core and capillaries in the periphery. Most of the capillaries were invaginated into the multilayered cuboidal epithelium. Near the tip of the villi they became swollen, forming sinusoidal capillaries.
  Transmission electron microscopy clarified the fine structure of the blood-water barrier, which consisted of a non-fenestrated endothelium and an attenuated epithelium that sandwiched a connective tissue with a discontinuous subendothelial and a continuous subepithelial basement lamina. The epithelium consisted of secretory cells, mitochondria-rich cells, and basal cells.
  The mitochondria-rich cells contained a cytoplasmic area filled with tubulovesicular elements. Based on their ultrastructural resemblance with the chloride cells in the fish and tadpole, these cells are suggested to be involved in the uptake of Na⁺ and Cl^- from fresh water for keeping ionic balance in the blood.

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Following our observations of the fine structure of the pharyngeal villiform processes of the hibernating soft-shelled turtle, Trionyx sinensis japonicus (YOKOSUKA et al., 2000), this paper deals with a scanning electron microscope study of the resin casts of blood vessels supplying those processes.
  Each villiform process contained arterioles and venules which ran in the axial portion of the process; capillaries formed a network at the periphery of the connective tissue core of the villus. In the distal portions of the villus, the capillaries increased markedly in their caliber to form sinusoidal capillaries. Such a vascular architecture supports the view that the villi-form processes serve in the aquatic respiration of the soft-shelled turtle.
  The casts indicated an occurrence of sphincters in the vascular bed of the villi.

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