In spite of a claim by KOBAYASHI (1990) that they do not correspond to the cells originally depicted by CAJAL, a particular category of fibroblast-like cells have been identified in the gut by electron microscopy (FAUSSONE-PELLEGRINI, 1977;THUNEBERG, 1980) and by immunohistochemistry for Kit protein (MAEDA et al., 1992) under the term of the “interstitial cells of Cajal (ICC)”. Generating electrical slow waves, the ICC are intercalated between the intramural neurons and the effector smooth muscular cells, to form a gastroenteric pacemaker system.
  ICC at the level of the myenteric plexus (IC-MY) are multipolar cells forming a reticular network. The network of IC-MY which is believed to be the origin of electrical slow waves is morphologically independent from but associated with the myenteric plexus. On the other hand, intramuscular ICC (IC-IM) usually have spindle-shaped contours arranged in parallel with the bulk smooth muscle cells. Associated with nerve bundles and blood vessels, the IC-IM possess receptors for neurotransmitters and such circulating hormones as cholecystokinin, suggesting their roles in neuro-muscular and hormone-muscular transmissions. In addition, gap junctions connect the IC-MY and IC-IM, thereby realizing the electrically synchronized integrity of ICC as a pacemaker system in the gut. The smooth muscle cells are also coupled with ICC via gap junctions, and the functional unit thus formed enables rhythmically synchronized contractions and relaxations.
  It has recently been found that a lack of Kit-expressing cells may induce hyper-contractility of the tunica muscularis in vivo, whereas a decrease in Kit expression within the muscle wall causes dysmotility-like symptoms in vivo. The pacemaker system in the gut thus seems to play a critical role in the maintenance of both moderate and normal motility of the digestive tract. A loss of Kit positive cells has been detected in several diseases with an impaired motor activity, including diabetic gastroenteropathy. Pathogenesis of these diseases is thought to be accounted for by impaired slow waves and neuromuscular transmissions; a pacemaker disorder may possibly induce a dysmotility-like symptom called ‘gastroenteric arrhythmia’.
  A knowledge of the structure and function of the ICC and the pacemaker system provides a basis for clarifying the normal mechanism and the pathophysiology of motility in the digestive tract.

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The diazepam binding inhibitor (DBI) was originally isolated from the brain as an intrinsic ligand of the benzodiazepine binding site on the type-A γ-aminobutyric acid receptor (GABA_A receptor). Its wide-spread distribution in non-neural tissues outside the brain suggests that DBI has various functions other than GABA-mediated neurotransmission. Since DBI is identical with the acyl-CoA binding protein, which has the ability to bind long chain acyl-CoA esters, the major function of DBI may possibly be related to lipid metabolism. This idea was supported by our previous study showing the consistent coexpression of DBI and fatty acid binding proteins (FABPs) in epithelia throughout the gastrointestinal tract. The present histochemical study focused on the distribution of DBI in neural tissues, and revealed a definite existence of DBI in non-neuronal supporting cells in both the central and peripheral nervous systems. In the brain, intense immunoreactivity for DBI was detected in the cerebellar Bergmann glia, olfactory ensheathing glia, subgranular layer of the dentate gyrus, and retinal Müller cells. In the peripheral nervous system, satellite cells in sensory/autonomic ganglia, Schwann cells, and sustentacular cells in the adrenal medulla were immunoreactive to a DBI antibody. Moreover, the colocalization of DBI and brain-type FABP (B-FABP) was observed in most of the non-neuronal supporting cells mentioned above, indicating that DBI and B-FABP are cooperatively involved in the energy metabolism of astrocytes and related cells, which are thought to support neuronal development and functions.

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The mutual interactions of small proteoglycans with collagen fibrils in the extracellular matrix remain to be completely understood. The present research investigated the extracellular matrix of the rat tail tendon by atomic force microscopy (AFM) as well as by scanning electron microscopy (SEM). Observations showed simply dehydrated specimens made of large heterogeneous fibrils, tightly packed in mutual contact with no visible interfibrillar spaces. Proteoglycans usually extended onto neighboring fibrils, forming an intricate interfibrillar weaving highly sensitive to chondroitinase digestion. Pre-treatment with cupromeronic blue only affected the proteoglycans side chains, which appeared better preserved but somewhat thickened. Observation of hydrated specimens by AFM confirmed the close packing of collagen fibrils and the abundance of collagen-bound proteoglycans. Interfibrillar bridges were only occasionally observed in this tissue, whose fibrils are instead tightly bound together by proteoglycans in a structure quite consistent with its functional requirements. The molecular machinery responsible for these interactions is the subject of ongoing research.

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The neurotoxic effects of inorganic lead are known to include peripheral neuropathy in adults and encephalopathy in children. The purpose of this study was to determine the effect of inorganic lead (PbCl₂) administration on norepinephrinergic neurons of the locus ceruleus in neonatal rats by immunocytochemical and electron microscopic analyses.
  Lead chloride solutions, 0.05%, 0.1% and 0.2% in concentrations, were prepared in distilled water and administered orally via drinking water. After 4, 8, or 12 weeks of continuous administration, the rats were sacrificed and brains were immunostained with the tyrosine hydroxylase antibody. The number of immunoreactive cell bodies in the locus ceruleus was estimated. Densitometric analysis of immunoreactive profiles visualized by electron microscopy was performed using an image analyzer.
  The numbers of immunoreactive neurons in the locus ceruleus were increased statistically by lead administration. The intensity of the immunoreaction, both under the light and electron microscopes was also increased.
  Degenerative changes, including intra-axonal vacuole formation and widening of the extracellular spaces, were found by electron microscopy in and around the tyrosine hydroxylase immunoreactive axons.
  Increased tyrosine hydroxylase immunoreactivity may correlate with the hyper-reactivity of lead intoxicated children. Degenerative changes may account for the reported deficits in intellectual attainment and achievement in lead intoxicated children.

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Structural changes in the developing rat lung were studied by a combined use of light microscopy including immunohistochemistry for α-smooth muscle actin (α-SMA) and scanning electron microscopy (SEM) using the KOH-collagenase digestion method. In the embryonic stage (E11-E13), the lung bud appeared as an outgrowth from the ventral wall of the foregut which grew caudally into the splanchnic mesoderm to form a pair of bronchial buds at the end. At E13, the airway smooth muscle cells first appeared around the bifurcation of the trachea. These smooth muscle cells were restricted to the dorsal surface of the tracheal epithelium, suggesting a difference in character between the dorsal and ventral sides of the mesenchymal cells in this region. During the pseudoglandular stage (E13-E18.5), the bronchial buds repeatedly gave off branches in the mesenchymal tissue. The smooth muscle cells in the bronchioles were spindle-shaped and arranged completely circularly around the epithelial tube, except that the terminal bud of bronchioles lacked the smooth muscles. The neck of the terminal bud was constantly surrounded by flat and irregularly-shaped immature smooth muscle cells, representing an early event in the smooth muscle cell differentiation from mesenchymal cells. In the canalicular to saccular stages (E18.5 to birth), the terminals of bronchioles became saccular, thus forming prospective alveolar acini. At birth, the alveolar wall became thinner than before birth, and the individual smooth muscle cells in bronchioles were elongated like a tape.As to the blood vessel differentiation, various sized sinusoidal spaces indicating the primitive blood vessels were already present in the mesenchymal tissue at E11.5. The endothelial cells of these sinusoidal spaces were irregularly shaped and sometimes extended their processes into the lumen. The network of tubular vessels appeared from E14.5. These vessels had tapering ends as well as transluminal trabeculae, suggesting that capillary growth proceedsby both the sprouting and partitioning (i.e., intussusception) of vessels in the pseudoglandular stage.

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The hepatic portal vein has been known to make a spontaneous peristaltic movement in some mammals, including the mouse and rat. To investigate the fine structure of the portal vein in relation to its physiological characteristics, we observed the mouse portal vein by using various histological techniques including conventional light microscopy, videomicroscopy, transmission and scanning electron microscopy, and real-time confocal laser scanning microscopy.
  The mouse hepatic portal vein was provided with a spiral fold which was produced by the inner layer, i.e. the endothelium and smooth muscles of the wall protruding into the lumen. Longitudinal smooth muscle cells spanned the interval of the fold, like a spirally arranged palisade around the vessel wall.
  The longitudinal muscle fibers ended at the spiral fold, being partly connected with a network of irregularly shaped smooth muscle cells. This network, hitherto unknown, was recognized to be restricted to the fold in distribution and characterized by numerous gap junctions connecting the muscle cells.
  Real-time confocal laser scanning microscopy using a Ca²⁺ sensitive fluorescent dye revealed that a transient and periodic increase in Ca²⁺ concentration occurred in the longitudinal smooth muscle cells and was transmitted spirally from the intestinal to the hepatic side. These findings indicate that, during the peristaltic movement, the contraction of smooth muscle cells is transmitted along the longitudinal smooth muscles of the portal vein wall toward the liver, presumably controlled by the network of the irregularly-shaped smooth muscle cells in the fold of the portal vein.
Light microscopic observation in some specimens indicated an occurrence of cardiac muscle cells outside the smooth muscle layer. Restricted to the site of the porta hepatis in distribution, their involvement in the peristaltic contraction of the portal vein seemed unlikely.

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The expression and localization of the aquaporin-1 (AQP1) water channel were examined in the glomeruli of the human kidney. A ribonuclease protection assay showed the expression of AQP1 mRNA in human glomeruli but not in rat glomeruli. Western blot analysis revealed 28 kDa and 35 kDa bands corresponding to unglycosylated and glycosylated AQP1 proteins in human glomeruli. Immunoreactive AQP1 was demonstrated almost exclusively in the mesangium in the human glomeruli by immunohistochemistry. The endothelium of glomerular capillaries was only partly immunostained while podocytes and Bowman’s capsule epithelia were not immunolabeled. Immunoelectron microscopy localized the immunoreactive AQP1 on the plasma membrane of mesangial cells in human glomeruli. The immouno-gold labeling was dense on the projections of mesangial cells protruding to the glomerular capillary lumen or to endothelial cells, but was sparse on other parts of the mesangial cell surface. No immunoreactivity for AQP1 was demonstrated in rat glomeruli. This study showed the distinct localization of AQP1 in the mesangial cells of human glomeruli, suggesting its role in water movement through these cells.

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Taste-mGluR4, cloned from taste tissues, is a truncated variant of brain-expressed mGluR4a (brain-mGluR4), and is known to be a candidate for the receptor involved in the umami taste sense. Although the expression patterns of taste- and brain-mGluR4 mRNAs have been demonstrated, no mention has so far been made of the expression of these two mGluR4 proteins in taste tissues. The present study examined the expression of taste-mGluR4 and brain-mGluR4 proteins in rat taste tissues by using a specific antibody for mGluR4a which shared a C-terminus of both taste- and brain-mGluR4, for immunoblot analysis and immunohistochemistry.
  Immunoblot analysis showed that both brain-mGluR4 and taste-mGluR4 were expressed in the taste tissues. Taste-mGluR4 was not detected in the cerebellum. The immunoreactive band for brain-mGluR4 protein was much stronger than that for taste-mGluR4 protein. In the cryosections of fungiform, foliate and circumvallate papillae, the antibody against taste-mGluR4 exhibited intense labeling of the taste pores and taste hairs in all the taste buds of gustatory papillae examined; the immunoreaction to the antibody against brain-mGluR4 was more intense at the same sites of the taste buds. The portions of the taste bud cells below the taste pore and surrounding keratinocytes did not show any immunoreactivities.
  The results of the present study strongly suggest that, in addition to taste-mGluR4, brain-mGluR4 may function even more importantly than the former as a receptor for glutamate, i.e. the umami taste sensation.

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The laboratory shrew, Suncus murinus, which lacks such gut associated lymph organs as the appendix and Peyer’s plates, was recently demonstrated (KUBO and ISOMURA, 1996) to possess a pair of anal tonsils at the end of its rectum. The present paper deals with the development of this lymphoid organ as observed by light and electron microscopy. The anal tonsil was characterized by the initial postnatal development. On neonatal Day 1, a pair of epithelial crypts formed at the dorsal boundary between the anus and the ostium urogenitoanale. On Day 2 after birth, Iymphocytes began to accumulate in the subepithelial mesenchymal tissue under the crypt. From Day 3 on, the lymphocytes increased to form a lymph nodule, from which, on Day 5, some lymphocytes began to penetrate into the crypt epithelium. The crypt and the nodule were fused together between Days 6 and 8. A germinal center-like structure was observed on Day 20 after birth. Around Day 40, the invading cells comprised cellular units consisting of large and small lymphocytes and plasma cells. High endothelial venules were observed in the parafollicular area at this time. These findings indicate that the anal tonsil originates from an accumulation of lymphocytes in the mesenchymal tissue close to a particular epithelium of the crypt, presumably in response to antigens in foods; the tonsilar structure is then gradually completed by fusion of the lymphoid and epithelial elements. This paper further reports on an electron microscope finding on Day 8 where the anal tonsillar crypt epithelium was seen to contain some basal-granulated cells of the open type.

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