RESUMO
The discovery of pits/caveolae in the plasmalemma advanced the study of macromolecule internalization. "Transcytosis" describes the transport of macromolecular cargo from one front of a polarized cell to the other within membrane-bounded carrier(s), via endocytosis, intracellular trafficking and exocytosis. Clathrin-mediated transcytosis is used extensively by epithelial cells, while caveolae-mediated transcytosis mostly occurs in endothelial cells. The internalization pathways were monitored by various markers, including radioisotopes, nanoparticles, enzymes, immunostains, and fluorophores. We describe an internalization pathway identified using a naturally-occurring biomarker, in vivo assembled ferritin, containing electron-dense iron cores. Iron, an essential trace metal for most living species and iron homeostasis, is crucial for cellular life. Ferritin is a ubiquitous and highly conserved archeoprotein whose main function is to store a reserve iron supply inside the cytoplasm in a non-toxic form. Ferritin is present in all organisms which have a metabolic requirement for iron and in even in organisms whose taxonomic rank is very low. The newborns of the blind mole, Spalax ehrenbergi, are born and live in a hypoxic environment and have significant iron overload in their liver and heart, but their iron metabolism has not been previously studied. These newborns, which are evolutionarily adapted to fluctuations in the environmental oxygen, have a unique ability to sequester transplacental iron and store it in ferritin without any signs of iron toxicity. Using the ferrihydrite cores of ferritin, we were able to monitor the ferritin internalization from portals of its entry into the cytosol of hepatocytes and cardiomyocytes and into the lysosomes.
Assuntos
Biomarcadores/metabolismo , Endocitose/fisiologia , Ferritinas/metabolismo , Substâncias Macromoleculares/metabolismo , Transdução de Sinais/fisiologia , Animais , Animais Recém-Nascidos , Compostos Férricos/química , Compostos Férricos/metabolismo , Ferritinas/química , Ferritinas/ultraestrutura , Hepatócitos/metabolismo , Hepatócitos/ultraestrutura , Hipóxia , Espaço Intracelular/metabolismo , Ferro/química , Fígado/citologia , Fígado/metabolismo , Fígado/ultraestrutura , Microscopia Eletrônica de Transmissão , Miocárdio/citologia , Miocárdio/metabolismo , Miocárdio/ultraestrutura , Miócitos Cardíacos/metabolismo , Miócitos Cardíacos/ultraestrutura , SpalaxRESUMO
The role of zinc, an essential element for normal brain function, in the pathology of Alzheimer's disease (AD) is poorly understood. On one hand, physiological and genetic evidence from transgenic mouse models supports its pathogenic role in promoting the deposition of the amyloid beta-protein (Abeta) in senile plaques. On the other hand, levels of extracellular ("free") zinc in the brain, as inferred by the levels of zinc in cerebrospinal fluid, were found to be too low for inducing Abeta aggregation. Remarkably, the release of transient high local concentrations of zinc during rapid synaptic events was reported. The role of such free zinc pulses in promoting Abeta aggregation has never been established. Using a range of time-resolved structural and spectroscopic techniques, we found that zinc, when introduced in millisecond pulses of micromolar concentrations, immediately interacts with Abeta 1-40 and promotes its aggregation. These interactions specifically stabilize non-fibrillar pathogenic related aggregate forms and prevent the formation of Abeta fibrils (more benign species) presumably by interfering with the self-assembly process of Abeta. These in vitro results strongly suggest a significant role for zinc pulses in Abeta pathology. We further propose that by interfering with Abeta self-assembly, which leads to insoluble, non-pathological fibrillar forms, zinc stabilizes transient, harmful amyloid forms. This report argues that zinc represents a class of molecular pathogens that effectively perturb the self-assembly of benign Abeta fibrils, and stabilize harmful non-fibrillar forms.
Assuntos
Doença de Alzheimer/metabolismo , Peptídeos beta-Amiloides/química , Zinco/química , Encéfalo/metabolismo , Humanos , Cinética , Microscopia Eletrônica de Transmissão , Modelos Químicos , Conformação Molecular , Peptídeos/química , Espalhamento de Radiação , Espectrofotometria/métodos , Fatores de Tempo , Difração de Raios XRESUMO
Macrophagic myofasciitis has been almost exclusively detected in adults only. We describe six children of Arab Moslem origin with this disorder. Three presented with hypotonia, developmental delay and seizures and were evaluated for a mitochondrial disorder. The other three children had hypotonia and predominantly motor delay. Five of the six families were consanguineous. A massive collection of macrophages was present in the fascia and adjacent epimysium in all biopsies. The macrophages were periodic-acid-Schiff positive and immunoreactive for CD68. One biopsy which was evaluated by electron microscopy and energy-dispersive X-ray microanalysis showed crystalline structures containing aluminum in macrophages. Two children with motor delay and hypotonia were treated with oral prednisone for 3 months with no clinical improvement. Genetic predisposition probably accounts for the variability in the prevalence of macrophagic myofasciitis in different populations. At least in childhood, there seems to be no connection between macrophagic myofasciitis as a pathological entity and the clinical symptoms and signs.
Assuntos
Consanguinidade , Macrófagos/patologia , Músculo Esquelético/patologia , Miosite de Corpos de Inclusão/etiologia , Miosite/patologia , Alumínio/metabolismo , Antígenos CD/metabolismo , Antígenos de Diferenciação Mielomonocítica/metabolismo , Biópsia/métodos , Pré-Escolar , Feminino , Humanos , Imuno-Histoquímica/métodos , Lactente , Macrófagos/química , Macrófagos/ultraestrutura , Masculino , Microscopia Eletrônica/métodos , Hipotonia Muscular/etiologia , Hipotonia Muscular/patologia , Debilidade Muscular/etiologia , Músculo Esquelético/química , Músculo Esquelético/fisiopatologia , Músculo Esquelético/ultraestrutura , Miosite/complicações , Miosite de Corpos de Inclusão/metabolismo , Miosite de Corpos de Inclusão/patologia , Vacinação/efeitos adversosRESUMO
We used a cell-free transcription/translation system to synthesize structural proteins of the T4 bacteriophage. We focused on two proteins that participate in the formation of the virus tail tube assembly. Synthesized separately, the proteins assembled into their in vivo forms, namely one polymerized into rigid hollow nanotubes approximately 20 nm thick and hundreds of nanometers long, the other assembled into 10 nm tube-capping hexameric rings. Co-synthesis of the two proteins, however, revealed a novel structure of a nanodoughnut with an outer diameter of approximately 50 nm and thickness of approximately 20 nm. Cell-free co-synthesis and assembly of T4 structural proteins can be extended in a combinatorial fashion. The addition of other structural genes offers control of native nanoassemblies and may reveal ones not observable by mixing purified components.
Assuntos
Nanoestruturas/química , Nanotubos de Peptídeos/química , Proteínas da Cauda Viral/biossíntese , Proteínas da Cauda Viral/química , Bacteriófago T4/metabolismo , Bacteriófago T4/ultraestrutura , Sistema Livre de Células , Microscopia Eletrônica , Nanoestruturas/ultraestrutura , Nanotecnologia , Nanotubos de Peptídeos/ultraestrutura , Proteínas da Cauda Viral/ultraestruturaRESUMO
The skeletons of adult echinoderms comprise large single crystals of calcite with smooth convoluted fenestrated morphologies, raising many questions about how they form. By using water etching, infrared spectroscopy, electron diffraction, and environmental scanning electron microscopy, we show that sea urchin spine regeneration proceeds via the initial deposition of amorphous calcium carbonate. Because most echinoderms produce the same type of skeletal material, they probably all use this same mechanism. Deposition of transient amorphous phases as a strategy for producing single crystals with complex morphology may have interesting implications for the development of sophisticated materials.