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Ultrastructural Characteristics of DHA-Induced Pyroptosis.

ABSTRACT: Microglial cells are resident macrophages of the central nervous system (CNS) that respond to bioactive lipids such as docosahexaenoic acid (DHA). Low micromolar concentrations of DHA typically promote anti-inflammatory functions of microglia, but higher concentrations result in a form of pro-inflammatory programmed cell death known as pyroptosis. This study used scanning electron microscopy (SEM) and transmission electron microscopy (TEM) to investigate the morphological characteristics of pyroptosis in BV-2 microglial cells following exposure to 200 µM DHA. Vehicle-treated cells are characterized by extended processes, spine-like projections or 0.4 to 5.2 µm in length, and numerous extracellular vesicles (EVs) tethered to the surface of the plasma membrane. In contrast to vehicle-treated cells, gross abnormalities are observed after treating cells with 200 µM DHA for 4 h. These include the appearance of numerous pits or pores of varying sizes across the cell surface, structural collapse and flattening of the cell shape. Moreover, EVs and spines were lost following DHA treatment, possibly due to release from the cell surface. The membrane pores appear after DHA treatment initially measured?~?30 nm, consistent with the previously reported gasdermin D (GSDMD) pore complexes. Complete collapse of cytoplasmic organization and loss of nuclear envelope integrity were also observed in DHA-treated cells. These processes are morphologically distinct from the changes that occur during cisplatin-induced apoptosis, such as the appearance of apoptotic bodies and tightly packed organelles, and the maintenance of EVs and nuclear envelope integrity. Cumulatively, this study provides a systematic description of the ultrastructural characteristics of DHA-induced pyroptosis, including distinguishing features that differentiate this process from apoptosis.

SUBMITTER: Herr DR

PROVIDER: S-EPMC7230060 | biostudies-literature | 2020 Jun

REPOSITORIES: biostudies-literature

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Ultrastructural Characteristics of DHA-Induced Pyroptosis.

Herr Deron R DR Yam Ting Yu Amelia TYA Tan Wan Shun Daniel WSD Koh Sally Shuxian SS Wong Wai Shiu Fred WSF Ong Wei-Yi WY Chayaburakul Kanokporn K

Neuromolecular medicine 20200104 2

Microglial cells are resident macrophages of the central nervous system (CNS) that respond to bioactive lipids such as docosahexaenoic acid (DHA). Low micromolar concentrations of DHA typically promote anti-inflammatory functions of microglia, but higher concentrations result in a form of pro-inflammatory programmed cell death known as pyroptosis. This study used scanning electron microscopy (SEM) and transmission electron microscopy (TEM) to investigate the morphological characteristics of pyro ...[more]

PMID: 31902115

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Project description:The giant panda (Ailuropoda melanoleuca) is a vulnerable species and a charismatic member of zoological collections worldwide. Despite its importance as a representative species for global wildlife conservation efforts, no studies to date have described normal cell morphology or cytoplasmic constituents by traditional techniques such as cytochemical staining and evaluation of ultrastructural features. The objective of this study was to accurately identify and characterize the leukocytes and platelets of clinically healthy giant pandas using routine Wright-Giemsa stain, eight cytochemical stains, immunocytochemistry (CD3), and transmission electron microscopy (TEM) to further the collective understanding of normal cellular morphological features, cytochemical reactivity, and cytoplasmic contents found in health. Voluntary venipuncture was performed on four healthy individual animals (two adults and two juveniles), as part of routine preventive health evaluation. Blood was collected for routine and cytochemical stains, and into 2.5% glutaraldehyde for TEM. On Wright-Giemsa-stained blood films, leukocytes were differentiated into granulocytes (neutrophils, eosinophils, basophils) and mononuclear cells (lymphocytes, monocytes). Cytochemical staining revealed similar leukocyte and platelet staining patterns to those reported in other mammals, with some notable differences. By TEM, leukocytes with nuclear and cytoplasmic features of mononuclear cells were readily differentiated from granulocytes, and platelets had similar ultrastructural features to those reported in other mammals. Neutrophils were the predominant cell type followed by lymphocytes, while basophils were rare. Rare large or reactive lymphocytes, rare reactive monocytes, and rare large platelets were noted in apparently healthy giant pandas of this study. A unique mononuclear cell, with a moderately indented nucleus and shared cytochemical and ultrastructural characteristics of lymphocytes and monocytes, was discovered in this species. The combined cytochemical, immunocytochemical (CD3), and ultrastructural features of these unique cells more closely resemble those of monocytes, but the definitive cell lineage remains unknown at this time. This study provides novel information on giant panda leukocyte morphology and cellular constituents in health, shows the importance of manual blood film review, has important implications for hemogram interpretation in future clinical cases and research, and provides a baseline for future characterization and understanding of hemogram changes in response to disease.

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Ultrastructural Characteristics of DHA-Induced Pyroptosis.

Publications

Ultrastructural Characteristics of DHA-Induced Pyroptosis.

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