Ultrastructural study of microsporidian invasion into cells

Abstract

Nosema michaelis spores were primed to discharge extracellularly or into culture medium 199 containing ascites leukemia EL4 cells, mouse macrophages, neuroblastoma C1300 adapted to ascites, human red blood cells, and blue crab epithelial cells and hemocytes. Before discharge, the polar tube has a single membrane enveloping a glycoprotein-like matrix. After extrusion, there are two membrane envelopes surrounded by a glycoprotein sheath. This sheath is silver methenamine negative, trypsinsensitive, has low solubility to 1% SDS treatment, and binds ferritin-conjugated concanavalin A. Before spore discharge, the sporoplasm is dispersed, uncompartmentalized by any membrane and exterior to the extrusion apparatus. After passage through the extrusion tube, the sporoplasm ends up in vesicle formed at the end of extruded tube. The sporoplasm nucleus maintains its integrity during extrusion; both the cytoplasm and nuclear components of the discharged sporoplasm are compartmentalized within membranes. After 15-20 min within the host cytoplasm, the sporoplasm shows obvious cytoplasmic reorganization and endoplasmic reticulum synthesis. Within the host cell, the sporoplasm is surrounded by two envelopes, the outer of which appears continuous with the polar tube sheath. Two envelopes surround the parasite when it is injected into extracellular medium, into human red blood cells, or into ascites leukemia EL4 cells. One envelope is lost when the sporoplasm is injected into normal host cells such as blue crab hemocytes and epithelial cells. A fibrous corona surrounds the sporoplasm in ascites leukemia EL4 cells, neuroblastoma C1300 or mouse macrophages. This corona is believed to be host reaction material since it is absent when parasites are injected into blue crab gut epithelial cells and hemocytes, into extracellular medium, or into human red blood cells. © 1972 Springer-Verlag.