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Characteristics of necrosis Type Definition Pathophysiology Microscopic appearance Example Coagulative necrosis A type of necrosis caused by tissue ischemia that occurs in most tissues except the brain.
Vessel wall damage caused by immune complex deposition e. Rheumatoid arthritis Peptic ulcer disease Immune vasculitis e. Overview of calcification Metastatic calcification Dystrophic calcification Description Diffuse calcification of normal tissue Localized calcification e. Increased pH levels promote calcium deposition. Abnormal necrotic tissues or degenerated inflammatory sites Etiology Secondary to h ypercalcemia , for example: Primary hyperparathyroidism Hypervitaminosis D e. Cardiovascular conditions: arteriosclerosis , heart valve disease , thrombi , calcification of atherosclerotic plaques Fat necrosis e.
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Lipodystrophy Leukodystrophy Osteodystrophy Chondrodystrophy Myotonic dystrophy Duchenne muscular dystrophy Becker muscular dystrophy. Increased tissue size via enlargement of cells due to an increase in organelles and structural proteins.
Physiological hypertrophy Increased muscle mass through sport Uterus enlargement due to hormonal changes Pathological hypertrophy : hypertrophic cardiomyopathy due to arterial hypertension.
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These results suggest that the induced apoptotic spermatogenic cells are phagocytosed by Sertoli cells and used to produce ATP in vivo. Each cell type predominantly utilizes one of the two energy substrates for ATP production.
In the presence of 2-MP 0. Given that the lipids formed in Sertoli cells after engulfment of apoptotic spermatogenic cells are mainly consisted of LCFAs, we conclude that apoptotic germ cells are an energy source of Sertoli cells.
Expression of key enzyme genes involved in metabolism in Sertoli cells. To determine whether primary cultured Sertoli cells can retain their differentiated functions, we examined the phagocytotic ability of Sertoli cells through ingesting latex beads, and expression of two functional markers of Sertoli cells, androgen binding protein ABP and androgen receptor AR , during in vitro culture.
By contrast, treatment with cytochalasin B resulted in a marked decrease in the phagocytosis of latex beads by the Sertoli cells. These data suggest that the Sertoli cells retain their major differentiated functions during culture. Characterizations of Sertoli cells. A Phagocytosis of latex beads LB by Sertoli cells. Sertoli cells engulfed latex beads arrows were identified under a fluorescence microscope upper panel , and the ratio of the Sertoli cells having internalized latex beads lower panel.
Inhibition of phagocytosis by cytochalasin B CB was performed parallely. Phagocytic clearance of apoptotic spermatogenic cells and residual bodies by Sertoli cells is necessary for efficient production of sperm Maeda et al. In this study, we demonstrate that phagocytosis of apoptotic spermatogenic cells results in the formation of lipids, which are further metabolized to produce ATP in Sertoli cells.
This energy source could be physiologically important for Sertoli cells to support spermatogenesis. Sertoli cells provide essential physical and tropic support for the development of spermatogenic cells Griswold We found that Sertoli cells produce the high ATP level, which corresponds to the functions of these cells.
This result is also in agreement with previous observations that an active energy metabolism occurs within Sertoli cells Floridi et al. The meaning of the death of such a large proportion of spermatogenic cells is unclear. One explanation is that the number of spermatogenic cells that Sertoli cells can support for maturation is limited, and the elimination of apoptotic cells provides space in the seminiferous epithelium for the development of healthy spermatogenic cells.
It is possible that the engulfment of apoptotic spermatogenic cells enables Sertoli cells to support spermatogenesis. How the functions of Sertoli cells can be conferred by the clearance of dying germ cells remains to be clarified. The results in the present study showed that apoptotic spermatogenic cells can be used to produce ATP by Sertoli cells. Notably, the phenomenon that the dying cells are used as an energy source is specific to Sertoli cells, but not to other phagocytes such as macrophages.
This energy source could be particularly important for Sertoli cells considering that they are seldom reached by blood circulation in vivo owing to the basement membrane, blood—testis barrier and lacking blood vessels in the seminiferous epithelium. Moreover, this energy resource may confer the meaning of the most spermatogenic cells to undergo apoptosis during spermatogenesis. Lipids and glucoses are common energy substrates for the production of ATP within cells.
Under physiological conditions, most cells predominantly use glucoses to produce ATP, and lipids function as the storage of energy. The result is in agreement with a recent report that the lipid droplets in Sertoli cells mainly consist of LCFAs Huyghe et al. The lipid exchange between Sertoli cells and the periphery are currently not well understood. Sertoli cells are largely separated from the peripheral circulation due to the barriers of basement membrane, tight junction between Sertoli cells and lacking of blood capillaries in seminiferous epithelium.
The simplest way for Sertoli cells to have sufficient lipids for supporting spermatogenesis would be to recycle the lipid contents in the residual bodies and apoptotic spermatogenic cells. Massive apoptosis of germ cells also occurs in ovary.
It is presumed that only less than 0. Death of the majority of germ cells is thus an event common to the gametogenesis in both males and females. The significance of gametocytes dying by apoptosis could be an important issue that is worthwhile to investigate further. The study of this issue may open a novel avenue to understand the meaning of germ cell death.
The animals were maintained in a temperature- and humidity-controlled room on 12h light: 12h darkness cycle, and had free access to food and water. All the measures taken for the mice were in accordance with guidelines for the Care and Use of Laboratory Animals approved by the Chinese Council on Animal Care.
Three-week-old mice were used for isolation of testicular cells based on a procedure described previously Wang et al. Briefly, decapsulated testes were incubated in D-Hanks' solution with 0. The tubules were then incubated in D-Hanks' solution with 0. Twenty-four hours later, the Sertoli cells were collected and subjected to experiments. The epididymal epithelial cell culture was prepared as previously described Moore et al.
Epididymal tissue was obtained from 8-week old mice and washed in PBS to remove blood. The caput epididymidis were minced into small fragments followed by three rinses in PBS to remove spermatozoa. Three days latter, the epididymal epithelial cells formed monolayer and subjected to experiments. The peritoneal macrophages were isolated based on a previous approach Chong et al. The phagocytosis assay was performed based on a previous protocol Wang et al.
Briefly, Sertoli cells or macrophages were co-cultured with apoptotic germ cells, living germ cells, FITC-labeled inactivated bacteria E.
It is known that Sertoli cells and macrophages are phagocytic cells, and most interstitial cells and epididymal cells are non-phagocytic cells. To distinguish the phagocytic from non-phagocytic Sertoli cells, lipid droplet formation was determined by ORO staining that can be a criterion to evaluate phagocytosis of apoptotic germ cells by phagocytes Wang et al. Fluorescent-labeled bacteria and latex beads engulfed by the phagocytes were examined under a fluorescence microscope IX, Olympus.
The cellular ATP was measured by a modified luciferin-luciferase assay Yang et al. The fragments of the tubules were subjected to ATP measurement. The lipid droplets were visualized by ORO staining Wang et al. The lipid droplets in Sertoli cells were analyzed under a microscope IX71, Olympus.
The area ratio of lipid droplets to cell nucleus was used to evaluate phagocytotic ability of Sertoli cells. In total, Sertoli cells from 5 repeat wells were analyzed for each occasion, and the mean values were presented in results.
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