人心房肌细胞特殊颗粒的电镜酶细胞化学和免疫电镜研究

本研究应用电镜CMP酶细胞化学和心钠素免疫电镜胶体金,分别标记人心房肌细胞中初级溶酶体和心钠素,进行鉴别定位观察和定量分析。心钠素颗粒数量居多,占78.7～79.3％,大小均一,直径0.20±0.05μm(x±s),分布于整个细胞质内,多成簇,尤以肌膜下,核极区多见。而溶酶体大小不一,直径0.30±0.19μm(x±s),显著大于心钠素颗粒(P<0.001),一般在核极区成簇分布,也见肌原纤维之间和肌膜下单个散在分布。     

溶酶体保护蛋白/组织蛋白酶A基因敲除小鼠听力和耳形态学初步研究

目的研究溶酶体保护蛋白/组织蛋白酶A(protective protein/cathepsin A,PPCA)基因敲除小鼠听功能和耳形态学改变,探讨半乳糖唾液酸沉积症听力损害的病理生理机制。方法应用听性脑干反应(ABR)测试和颞骨连续切片,观察1月和2月龄的PPCA基因敲除纯合子(PPCA-/-)小鼠ABR反应阈和光镜下外耳、中耳及内耳形态改变,并以野生型(PPCA / )小鼠作对照。结果1月龄PPCA-/-小鼠ABR反应阈和耳形态无明显改变;2月龄时,短声和短音8、163、2 kHz反应阈较PPCA / 提高40~45 dB SPL,中耳黏膜增厚、听骨细胞囊泡化、变形和关节腔融合,血管纹增厚、螺旋神经节细胞、螺旋缘纤维细胞、前庭膜、基底膜及沿前庭阶外淋巴隙的间皮细胞囊泡化,但Corti器毛细胞及支持细胞正常。结论溶酶体保护蛋白/组织蛋白酶A缺乏可导致听力损害、中耳及耳蜗形态改变、中耳炎、听骨改变以及耳蜗螺旋神经节、血管纹、螺旋缘、前庭膜和基底膜等细胞的溶酶体储积,可能分别是传导性聋和感觉神经性聋的形成机制。     

Les formes psychiatriques de la maladie de Niemann-Pick de type C

Niemann-Pick type C disease diagnosed in adult neurology departments may be infantile or juvenile forms with prolongated life spans or forms starting at adolescence or adulthood. Psychomotor retardation is practically constant. In some cases, a psychosis may be the only manifestation for several years. The diagnosis is often made when extrapyramidal symptoms appear as well as cerebellar ataxia and ophtalmoplegia in the vertical gaze. The treatment with psychotropic drugs raises the question of a superimposition of a drug induced lipidosis. Hepatosplenomegaly is often discrete contrary to classical infantile cases. Foam cells as well as sea blue histiocytes are features of the disease. The filipin test evidences the lysosomal storage of cholesterol.     

“Reducing body”-like inclusions in skeletal muscle in childhood-onset acid maltase deficiency

Summary Unusual inclusions with some of the features of reducing bodies were encountered in the skeletal muscle biopsy of a 2.5-year-old boy with childhoodonset acid maltase deficiency. The biopsy revealed a vacuolar myopathy with lysosomal storage of glycogen and eosinophilic refractile inclusions in myofibers, which appeared dark blue with the menadione-nitroblue tetrazolium reaction. The significance of the association of inclusions with reducing properties in the setting of acid maltase deficiency is discussed.     

溶酶体在实验性心肌肥大过程中的作用

本实验用大鼠主动脉结扎的心肌肥大模型,结合电镜酶细胞化学技术,观察心肌肥大发展过程中溶酶体的变化。结果表明:心肌肥大发展时,溶酶体参与心肌细胞改建;到心肌肥大后期,出现大量自噬现象,造成心肌不可逆损伤。     

Ultrastructure of cell death in bulbar cushions of chick embryo heart

Summary The ultrastructure of the physiological cell death was studied in distal ventral bulbar cushions of 15 chick embryo hearts on the 4th and 5th day of incubation. Microperfusion fixation was performed. The ultracytochemistry of a lysosomal hydrolytic enzyme acid phosphatase was also investigated in another 15 embryonic hearts.In the course of the cell degeneration an increase in cellulr autophagy was observed without previous cytoplasmic or nuclear changes or phagocyte ingestion. A cytoplasmic diffusion of acid phosphatase outside of lysosomes was observed.Besides the cell death with the marked participation of the lysosomal system, another kind of dying cells was found, characterized by their nuclear pycnosis and cytoplasmic condensation. Starting from the 5th day of incubation the dying and dead cells were found phagocytized by some of their neighbouring viable mesenchymal cells. A formation of ribosomal crystals was not observed.The formation and fate of cytolysomes as well as the fate of phagocytes are discussed. The presence of pre-necrotic cells with important autophagy and of necrotic cells with nuclear changes was related to the possibility of a dual cause of the cell death. In the case of pre-necrotic cells the epigenetic factors like the biomechanic action of hemodynamics were considered, while the necrotic cells seem to be programmed to death by their genome.Finally the uniformity of cell death ultrastructure in different organs and species was noticed.     

Lysosomal augmentation during aminoglycoside uptake in cochlear hair cells

Aminoglycoside antibiotics, such as kanamycin, have ototoxic side effects, which often result in degeneration of cochlear and vestibular hair cells in the inner ear. Cytotoxic effects of aminoglycosides, however, do not appear immediately after cellular uptake of aminoglycosides. In order to understand the mechanisms responsible for the delayed emergence of aminoglycoside ototoxicity, changes in lysosomal activities in cochlear hair cells were evaluated during a repeated administration of kanamycin by two methods. Electron microscopic localization of acid phosphatase (AcPase) revealed that AcPase started to accumulate in vesicles 27 h after the start of kanamycin administration. In addition, the number and size of AcPase-filled vesicles increased with repeated kanamycin doses. Confocal microscopic localization of the LysoTracker probe, a vital lysosomal marker, showed an increase in the size of lysosomes in hair cells that were treated with kanamycin. The temporal changes in the augmentation of lysosomes paralleled those in intracellular kanamycin levels. These results suggest that the intralysosomal compartments can accumulate extensive amounts of aminoglycosides, which might lead to lysosomal swelling and subsequent rupture.     

Adaptive immunity at the crossroads of autophagy and metabolism

The function of lymphocytes is dependent on their plasticity, particularly their adaptation to energy availability and environmental stress, and their protein synthesis machinery. Lymphocytes are constantly under metabolic stress, and macroautophagy/autophagy is the primary metabolic pathway that helps cells overcome stressors. The intrinsic role of autophagy in regulating the metabolism of adaptive immune cells has recently gained increasing attention. In this review, we summarize and discuss the versatile roles of autophagy in regulating cellular metabolism and the implications of autophagy for immune cell function and fate, especially for T and B lymphocytes.