The three-dimensional structure of the Golgi complex in human and mouse cultured fibroblasts was investigated ultracytochemically by demonstrating the acid phosphatase (ACPase) activity in the Golgi complex and observing ultrathin or thick (0.2-3μm) sections with a conventional (80 or 100KV) or high voltage (1, 000KV) electron microscope. The stereoscopic observation of stereo-pair pictures taken by tilting the thick sections with a goniometer has helped the understanding of the three-dimensional structure of the Golgi complex. The ACPase activity is positive in three different cross-sectional views of the Golgi complex: 1) the cisternal part, which is composed of three to several cisternae arranged in parallel; 2) the polygonal array of thin (75nm) tubules, which may be the face view of a Golgi cisterna; and 3) the reticular part, which is located at the inner (mature) face of the Golgi complex and is composed of flat portions and thick (100-160nm) tubular portions. The reticular part of the Golgi complex may correspond to the GERL in other cells (16, 19, 21). The continuity of the reticular part of the Golgi complex (possibly GERL) throughout the cell was clearly demonstrated as a result of the use of thick sections cut parallel to the substratum, which enabled us to observe the whole Golgi complex system in a cell.

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Granulomatous lesions which were induced with living BCG, heat-killed BCG and mycobacterial fraction (glycolipid) in rabbit lungs were studied by electron microscopy. The transformation of macrophages into epithelioid cells and giant cells observed in granulomatous lesion was named “the epithelioid cell system”. According to lysosomal function, this system can be divided into three stages: (1) heterophagic stage, (2) intermediate stage and (3) autophagic stage. At every stage dense cuplike structures in the plasma membrane and the Golgi-associated tubular system in cytoplasm were observed. By biochemical assay of granulomatous lesion lysosomal cathepsin B revealed a correlative increment of specific activity on time course after pulmonary inoculation. Cathepsin B and lysozyme showed a high level of specific activity. However, specific activity of lysozyme in granulomatous lesion induced with living BCG was higher than that in granulomatous lesion induced with mycobacterial glycolipid.

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The morphogenesis of ACPase activity was observed in the following experimental materials:
1) The parenchymal cells and macrophages of the adrenal cortex of cholesterol-treated rabbits.
2) The same cells of the adrenal cortex of aged rats.
3) The macrophages of the mesenteric lymph nodes of rats having received a large amount of macromolecular dextran.
In these experiments, (1) the increase or occurrence of activity as a function of biological significance was difficult to detect in cells displaying a degenerative process, notwithstanding their apparent histochemical stainability as “ceroid pigment”, and (2) in the macrophages, under the present experimental conditions, the increase or occurrence of activity could be observed in more or less extensive areas of the cytoplasm, especially on the membrane system sharply demarcating these areas; the so-called residual bodies were seen to be produced there in groups concomitant with the appearance of their histochemical stainability as ceroid pigment.
This membrane system, judging by the evidence given by the plane ultrastructural pictures, seems to be intimately related to the smooth or rough ER system.

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For the purpose of clarifying intralysosomal accumulation of intermediary catabolites in the reticuloendothelial cells of sphingolipidoses, 23 cases of Gaucher's disease, 7 of Fabry's disease, 2 of Sandhoff's disease, 4 of generalized gangliosidosis, 2 of Farber's disease and 11 of Niemann-Pick disease were studied histopathologically, histochemically or electron microscopically. In Gaucher cells characteristic of Gaucher's disease, the storage inclusions contained tubules of about 200 to 600Å in diameter. In Fabry's disease, pleomorphic osmiophilic lamellated inclusions, which were histochemically of heterogenous nature, were stored, while Sandhoff's disease was characterized by accumulation of miniature but somewhat pleomorphic membranous cytoplasmic bodies in foam cells. As for generalized gangliosidosis, vacuolar inclusions similar to those seen in genetic mucopolysaccharidoses accumulated in the infantile type, but Gaucher-like cells predominated in the juvenile type, which contained fibrillar inclusions. In Farber's disease, curvilinear tubular structures were characteristic in the inclusions of foam cells. Myelin-like figures were found in the storage inclusions of Niemann-Pick cells. The variegated inclusions stored in the reticuloendothelial storage cells of these diseases were mostly surrounded by a delimiting membrane and enzyme cytochemically showed localization of acid phosphatase, indicating that they are transformed secondary lysosomes. Possible sources of the inclusions are discussed in most of the diseases.

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An enzymic digestive method using either purified mould lipase or snake venom phospholipase A₂ was applied to ultracytochemical observations on both newly developed lipid droplets in the parenchymal cells of Ethionine-evoked triglyceride fatty liver and characteristic cytoplasmic bodies in the hepatocytes of Coralgil-induced phospholipid fatty liver. The reaction products in the author's method indicated that PL might be synthesized in ER of the hepatic cells, that the synthesis of liver TG might be restricted to the smooth ER and TG-rich particles originating in the same smooth compartment of ER where TG would be packed into the droplets by a coat of polar lipids, PL, and that the inclusions induced by Coralgil might be digested in the participation of lysosomal enzymes of the hepatic cells and they are easily acted upon by phospholipase. This clarifies the nature of PL-rich bodies.

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A comparative study was made on lipid inclusions of human and rabbits. Atherosclerotic lesions were produced in rabbits fed 0.67% cholesterol for seven months. The loci of the inclusions produced in tissue, three-dimensional configuration, crystallographic characteristics, physical properties and chemical compositions were compared, and found to be similar and even identical between man and rabbit. The lipid inclusions were thus presumed to be formed in arterial tissues by a cellular metabolism common to man and rabbit, and shaped by a physico-chemical effect independent of difference in species. Hence, rabbits are found usable as a model for the study of lipid inclusions in experimentally induced atherosclerosis.

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