One hundred years have passed since the discovery of “the internal reticular apparatus” by Camillo GOLGI. Investigations into the structure and function of the “Golgi apparatus” have raised more and more challenging issues for cell biologists. After long debate, many new findings have accumulated in the last 10 years as a result of the availability of elegant new genetic, biochemical and morphological tools. This, in turn, has raised many new questions to be solved. In addition, numerous new findings have led to some confusion on the understanding of the Golgi apparatus. This review article deals with several modern aspects of vesicular transport versus cisternal maturation. Disruption of the stacked structure in mitotic and drug-induced conditions is also discussed to demonstrate the importance of structural integrity in the Golgi apparatus.

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To understand the precise distribution of tripeptidyl peptidase I (TPP-I), a defect of which has been shown to induce late infantile neuronal ceroid lipofuscinosis, various tissues from rats and mice were analyzed using biochemical and immunohistochemical techniques. Western blot analyses showed that a protein band immunoreactive to anti-TPP-I appeared in tissue extracts of both animals at a molecular weight of approximately 47 kD. Protein levels of TPP-I differed among tissues; they were high in the rat brain, liver, stomach, kidney, thyroid and adrenal glands and in the mouse brain, stomach, kidney, and testis. The proteolytic activity of TPP-I was detectable; it differed in the tissues examined and did not always reflect the expression levels of the protein in the tissues. In particular, the TPP-I activity was low in the brains of both animals and high in the rat testis, although its protein levels were high in the former tissue and low in the latter. Double immunostaining showed the immunoreactivity for TPP-I to be well localized in granular structures of epithelial cells in renal tubules and the cerebral choroid plexus, both of which were also stained with lamp2, a lysosomal membrane protein marker, indicating that TPP-I is a lysosomal enzyme. The immunoreactivity was intense in F4/80-immunopositive macrophages/microglial cells located in various tissues including the thymus, spleen, liver, alimentary tract, and central nervous system. Although the immunoreactivity differed depending on the tissues and even within the same tissues between the species, it was detected in all tissues examined, especially in nerve cells, some types of endocrine cells, and oxyntic cells such as gastric parietal cells and bone osteoclasts. However, the immunoreactivity was faint and week in rat thyroid gland, although its protein level was high in the tissue. These lines of evidence suggest that TPP-I, a lysosomal serine proteinase, is widely distributed in rat and mouse tissues, although its expression levels vary among them.

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This study describes the unique distribution of Ruffini endings (RE) in the periodontal tissues of the guinea pig teeth with special references to their presence in the enamel-related aspects of the continuously growing incisors and molars. In guinea pig incisors, immunohistochemistry for PGP 9.5 and glia specific S-100 protein revealed a condensed distribution of well-developed RE in the bone-related part of the lingual periodontal ligament as has been reported in many other rodents. In most cases, some RE-like nerve elements characterized by dendritic ramification and rounded terminal Schwann cells were found to be located in the labial, enamel-related regions, where no periodontal ligament-like fiber arrangement was established. In the molar periodontal ligament, well-developed RE-like nerve elements were also distributed in the enamel-related part, but in intimate relation to thick periodontal fiber bundles inserted in the cementum pearls grown on the enamel surface. In some cases, few RE were located in the apical region of the alveolar socket, where no periodontal fiber bundles could be identified. Our data provide the first morphological evidence of the presence of RE-like nerve elements in the enamel-related, fibrous connective tissue of continuously erupting rodent incisors. These data indicate that RE in guinea pig periodontal tissues have variable spatial correlation to the surrounding fibers, implicating their diverse mechanoreceptive properties depending on the anatomical location.

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The distribution of endothelin B receptor (ETBR)-like immunoreactivity in the rat renal glomerulus was investigated using an affinity-purified antibody against a synthetic peptide corresponding to the amino acid residues 425-439 of the rat ETBR. Light microscopy showed ETBR-like immunoreactivity to be localized predominantly near the glomerular blood capillaries. By immunoelectron microscopy using the pre-embedding method, intense immunodeposits indicating ETBR were detected in podocytes, particularly in their foot processes, in contrast with the weak immunoreaction in endothelial cells of the glomerular blood capillaries and in the mesangial cells. In sections stained with the post-embedding method using immunogold particles, positive signals were also found on the plasma membrane of podocyte foot processes as well as the cytoplasm just beneath the cell membrane. These findings suggest that endothelin stimulates ETBR mainly on podocytes, thus resulting in a decrease of the glomerular blood flow and glomerular filtration rates.

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Kupffer cells constitute a major source of the heme-degrading enzyme, heme oxygenase (HO). This study examined the roles of Kupffer cells in the modulation of accelerated heme catabolism in ischemia-reperfused rat livers. Livers from rats treated with or without liposome-encapsulated dichloromethylene diphosphonate, a Kupffer cell-depleting reagent, underwent a 20-min ligation of the portal vein followed by reperfusion, The time course of the biliary output of bilirubin, the terminal heme-degrading product, and the expression of HO-1 mRNA and protein were monitored. HO-1 mRNA levels were elevated 3 to 12 h after ischemia/reperfusion in both control and Kupffer cell-depleted rats. Immunohistochemical analyses of control livers revealed that Kupffer cells expressed high levels of HO-1 while its expression in hepatocytes was low. In Kupffer cell-depleted livers, however, periportal hepatocytes displayed marked HO-1 expression. Under these conditions the two groups exhibited distinct profiles of biliary bilirubin excretion. In the controls, total bilirubin excretion increased 8-fold and peaked at 10 h after ischemia/reperfusion. In contrast, the Kupffer cell-depleting treatment resulted in a significant acceleration of the initial rise in bilirubin production, which peaked at 4 h. However, the total amount of bilirubin excreted within the initial 10 h after reperfusion was reduced by 50% as compared with that of the controls. In Kupffer cell-depleted rats, the levels of GOT and GPT as well as serum endotoxin concentrations were elevated after ischemia/reperfusion. These results suggest that Kupffer cells serve as an ischemia/reperfusion sensor that upregulates heme degradation and bilirubin excretion, and that Kupffer cells protect hepatocytes from gut-derived stressers - including endotoxin - following ischemia/reperfusion.

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The ultrastructure and distribution of hyalocytes were examined in guinea pig eyes by scanning electron microscopy (SEM) and transmission electron microscopy (TEM).
  Hyalocytes were distributed randomly on the vitreous surface of the retinal inner limiting membrane, where they were elongated in shape with a spherical perikaryon and a few stout processes. On the epithelial surface of the ciliary body, however, the cells were stellate with some short processes. The cells of both regions included typical dense hyalocyte granules in the cytoplasm.
  The surface morphology of hyalocytes indicates that the cells are wandering macrophages. The abundance of free cells in the ciliary body epithelium suggests that the area is a site for the emigration of hyalocytes or their precursors from the ciliary stroma.
  The homogeneous population of hyalocytes in the posterior part of the eyeball may be useful for experimental studies of the cell in vivo or their isolation for study in vitro.

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We investigated the effects of the leptin and ciliary neurotrophic factor (CNTF) on gap junction formation between folliculo-stellate cells in the anterior pituitary glands of male rats. Thirty-day-old Wistar-Imamichi strain male rats were castrated, and 30 days later they received intraperitoneal injections of either human recombinant leptin or recombinant rat CNTF. They were divided into seven groups according to the injected materials: PBS as a control, either 0.04, 0.2 or 1.0 mg/kg leptin or either 0.004, 0.02, or 0.1 mg/kg CNTF. Five rats from each group were killed 1, 2, 3, 4 and 5 days after the injections, and the pituitary gland was removed from each rat. Then the specimens were prepared for observation by transmission electron microscopy. We quantified the number of follicles and gap junctions and calculated the rate of occurrence of gap junctions as the ratio of the number of gap junctions existing between folliculo-stellate cells per intersected follicle profile in electron photomicrographs. The administration of 1.0 mg/kg leptin and 0.1 mg/kg CNTF to castrated male rats increased the number of gap junctions between folliculo-stellate cells. These observations indicate that the formation of gap junctions within the anterior pituitary gland of male rats is under the influence of leptin and CNTF.

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We examined the immunolocalization of isoforms of muscle proteins, myosin and troponin, in the cremaster muscle of the undescended testis (cryptorchidism). In cryptorchid rats induced by a nonsteroidal androgen antagonist, flutamide, the cremaster muscle contained embryonic myosin and embryonic/cardiac troponin T in both immunofluorescence microscopy and Western blotting using antibodies against myosin and troponin T specific for embryonic, cardiac and fast skeletal muscles. However, in muscles other than the cremaster muscle, i. e., the masseter, pectoral and abdominal muscles, embryonic isoforms of these proteins were undetectable by immunohistochemistry with these antibodies, even in the muscles from cryptorchid rats. Our results showing that high levels of embryonic isoforms of muscle proteins were specifically present in the cremaster muscle of cryptorchid rats induced by flutamide suggest that flutamide treatment of pregnant rats might affect genes controlling the development of the lumbar region of the fetus body resulting in the presence of embryonic protein isoforms in the cremaster muscles which are closely associated with undescended testes.

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