scholarly journals The supramolecular landscape of growing human axons

2020 ◽  
Author(s):  
Patrick C. Hoffmann ◽  
Stefano L. Giandomenico ◽  
Iva Ganeva ◽  
Michael R. Wozny ◽  
Magdalena Sutcliffe ◽  
...  
Keyword(s):  
Endoplasmic Reticulum ◽  
Light Microscopy ◽  
Brain Development ◽  
Electron Tomography ◽  
Molecular Architecture ◽  
High Demand ◽  
Unique Biology ◽  
Structural Details ◽  
Intricate Shape ◽  
Native Context

AbstractDuring brain development, human axons must extend over great distances in a relatively short amount of time. How the subcellular architecture of the growing axon sustains the requirements for such rapid build-up of cellular constituents has remained elusive. Human axons have been particularly inaccessible to imaging at molecular resolution in a near-native context. Here we apply cryo-correlative light microscopy and electron tomography to growing axonal tracts from human cerebral organoids. Our data reveal a wealth of structural details on the arrangement of macromolecules, cytoskeletal components, and organelles in elongating axon shafts. In particular, the intricate shape of the endoplasmic reticulum is consistent with its role in fulfilling the high demand for lipid biosynthesis to support growth. Furthermore, the scarcity of ribosomes within the growing shaft suggests limited translational competence during expansion of this compartment. These data provide an unprecedented resource and reveal a molecular architecture that helps explain the unique biology of growing human axons.

Some Unresolved Problems concerning the Cochlear Nerve

1976 ◽  
Vol 85 (6_suppl2) ◽  
pp. 1-28 ◽  
Author(s):  
R. Lorente de Nó
Keyword(s):  
Light Microscopy ◽  
Hair Cells ◽  
Organ Of Corti ◽  
Bushy Cell ◽  
Afferent Fibers ◽  
Cells Of Origin ◽  
Structural Details ◽  
Synaptic Junctions ◽  
Ventral Nucleus ◽  
Spiral Fibers

Three types of afferent fibers innervate the hair cells of the organ of Corti: 1) specific radial fibers which establish contacts with a very few neighboring internal hair cells; 2) spiral fibers, each one of which establishes contact with a number of external hair cells distributed throughout long segments of the cochlea; and 3) unspecific radial fibers which are collaterals arising radially at irregular intervals from fibers of the ganglionic spiral bundles and which establish contact with internal hair cells. The existence of spiral ganglionic bundles of fibers oriented apicalward has long been described, and the fact that a number of ganglionic spiral fibers give off radial collaterals to innervate internal hair cells was illustrated by Cajal and by Lorente de Nó. However, those structural details are not mentioned in the modern literature. In the ventral nucleus there are neurons with efferent axons which join the trapezoid body and cells with short axons ramified within the ventral nucleus itself. Two types of cells with efferent axons are illustrated and described, the spherical or bushy cell and the basket cell; and it is shown that branches of division of the two types of efferent axons form association tracts which end in the tuberculum acusticum. Also, the fact is illustrated that fibers having their cells of origin in that tuberculum form association paths which end in the ventral nucleus by means of extensive ramifications which form numerous synaptic endings. The dendritic and fibrillar plexuses in the ventral nucleus are described, an analysis is made of the relationships between the two plexuses and of the synaptic junctions that mediate transmission of nerve impulses. The synaptic junctions belong to a considerable number of types and in all illustrations the important fact repeatedly appears that one and the same fiber may form synaptic endings of widely different sizes and shapes located either on the same neuron or, more frequently, on different neurons. The intimate structure of each type of synaptic ending cannot be revealed by light microscopy, but only light microscopy can reveal to which kind of fiber the synaptic endings do belong. The presentation is concluded with a brief and preliminary discussion of physiological corollaries of certain general features of the anatomy of the acoustic system.

scholarly journals Ultrastructural studies of the hemocytes of Panstrongylus megistus (Hemiptera: Reduvidae)

1989 ◽  
Vol 84 (2) ◽  
pp. 171-188 ◽  
Author(s):  
Margherita A. Barracco ◽  
Clarice T. Loch
Keyword(s):  
Endoplasmic Reticulum ◽  
Light Microscopy ◽  
Rough Endoplasmic Reticulum ◽  
Lipid Droplets ◽  
Panstrongylus Megistus ◽  
Ultrastructural Studies ◽  
Hemocyte Count

Ultrastructural analyses revealed the presence of six hemocyte types in the hemolymph of Panstrogylus megistus, partially confirming our previous results obtained through light microscopy. Prohemocytes: small, round hemocytes with a thin cytoplasm layer, espcieally rich in free ribosomes and poor in membranous systems. Plasmatocytes: polymorphic cells, whose cytoplasm contains many lysosomes and a well developed rough endoplasmic reticulum (RER).They are extremely phagocytic. Sometimes, they show a large vacuolation. Granulocytes: granular hemocytes whose granules show different degrees of electrondensity. Most of them, have an internal structuration. Coagulocytes: oval or elongated hemocytes, which show pronounced perinuclear cisternae as normally observed in coagulocytes. The cytoplasm is usually electrondense, poor in membranous systems and contains many labile granules. Oenocytoids: large and very stable hemocytes, whose homogeneous cytoplasme is rich in loose ribosomes and poor in membranous systems. Adipohemocytes: large cells, containing several characteristic lipid droplets. The cytoplasm is also rich in glycogen, RER and large mitochondria. The total and differential hemocyte count (THC and DHC) were also calculated for this reduviid. THC increases from 2,900 hemocytes/cubic millimeter of hemolymph in the 4th intar to 4,350 in the 5th and then, decreases to 1,950 in the adults. Plasmatocytes and coagulocytes are the predominant hemocyte types.

scholarly journals Immuno- and Correlative Light Microscopy-Electron Tomography Methods for 3D Protein Localization in Yeast

Traffic ◽  
2014 ◽  
Vol 15 (10) ◽  
pp. 1164-1178 ◽  
Author(s):  
Muriel Mari ◽  
Willie J.C. Geerts ◽  
Fulvio Reggiori
Keyword(s):  
Light Microscopy ◽  
Electron Tomography ◽  
Protein Localization

scholarly journals MIA3/TANGO1 enables efficient secretion by constraining COPII vesicle budding

2021 ◽  
Author(s):  
Janine McCaughey ◽  
Judith M. Mantell ◽  
Chris R. Neal ◽  
Kate Heesom ◽  
David J. Stephens
Keyword(s):  
Endoplasmic Reticulum ◽  
Light Microscopy ◽  
Secretory Pathway ◽  
Genome Engineering ◽  
High Volume ◽  
Vesicle Budding ◽  
Early Secretory Pathway ◽  
Copii Vesicle ◽  
Intermediate Compartment ◽  
Golgi Organization

AbstractComplex machinery is required to drive secretory cargo export from the endoplasmic reticulum. In vertebrates, this includes transport and Golgi organization protein 1 (TANGO1), encoded by the Mia3 gene. Here, using genome engineering of human cells light microscopy, secretion assays, and proteomics, we show loss of Mia3/TANGO1 results in formation of numerous vesicles and a loss of early secretory pathway integrity. This restricts secretion not only of large proteins like procollagens but of all types of secretory cargo. Our data shows that Mia3/TANGO1 constrains the propensity of COPII to form vesicles promoting instead the formation of the ER-Golgi intermediate compartment. Thus, Mia3/TANGO1 facilities the secretion of complex and high volume cargoes from vertebrate cells.

scholarly journals Zika virus replication in glioblastoma cells: electron microscopic tomography shows 3D arrangement of endoplasmic reticulum, replication organelles, and viral ribonucleoproteins

2021 ◽  
Author(s):  
Johannes Wieland ◽  
Stefan Frey ◽  
Ulrich Rupp ◽  
Sandra Essbauer ◽  
Rüdiger Groß ◽  
...  
Keyword(s):  
Endoplasmic Reticulum ◽  
Cell Line ◽  
Zika Virus ◽  
Structural Changes ◽  
Electron Tomography ◽  
Glioblastoma Cells ◽  
Electron Microscopic ◽  
N Protein ◽  
Ribonucleoprotein Complexes ◽  
Infected Cells

AbstractStructural changes of two patient-derived glioblastoma cell lines after Zika virus infection were investigated using scanning transmission electron tomography on high-pressure-frozen, freeze-substituted samples. In Zika-virus-infected cells, Golgi structures were barely visible under an electron microscope, and viral factories appeared. The cytosol outside of the viral factories resembled the cytosol of uninfected cells. The viral factories contained largely deranged endoplasmic reticulum (ER), filled with many so-called replication organelles consisting of a luminal vesicle surrounded by the ER membrane. Viral capsids were observed in the vicinity of the replication organelles (cell line #12537 GB) or in ER cisternae at large distance from the replication organelles (cell line #15747 GB). Near the replication organelles, we observed many about 100-nm-long filaments that may represent viral ribonucleoprotein complexes (RNPs), which consist of the RNA genome and N protein oligomers. In addition, we compared Zika-virus-infected cells with cells infected with a phlebovirus (sandfly fever Turkey virus). Zika virions are formed in the ER, whereas phlebovirus virions are assembled in the Golgi apparatus. Our findings will help to understand the replication cycle in the virus factories and the building of the replication organelles in glioblastoma cells.

Light Microscopy and Cryo-Electron Tomography of motile Malaria Parasites

2007 ◽  
Vol 13 (S03) ◽  
Author(s):  
F Frischknecht ◽  
M Kudryashev ◽  
S Lepper ◽  
S Münter ◽  
S Hegge ◽  
...  
Keyword(s):  
Light Microscopy ◽  
Electron Tomography ◽  
Malaria Parasites ◽  
Cryo Electron Tomography

scholarly journals The Origin, Dynamic Morphology, and PI4P-Independent Formation of Encephalomyocarditis Virus Replication Organelles

mBio ◽  
2018 ◽  
Vol 9 (2) ◽  
pp. e00420-18 ◽  
Author(s):  
C. E. Melia ◽  
H. M. van der Schaar ◽  
A. W. M. de Jong ◽  
H. R. Lyoo ◽  
E. J. Snijder ◽  
...  
Keyword(s):  
Endoplasmic Reticulum ◽  
Rna Synthesis ◽  
Electron Tomography ◽  
Membrane Vesicles ◽  
Encephalomyocarditis Virus ◽  
Escape Mutant ◽  
Membrane Structures ◽  
Double Membrane ◽  
Single Membrane ◽  
Phosphatidylinositol 4

ABSTRACTPicornaviruses induce dramatic rearrangements of endomembranes in the cells that they infect to produce dedicated platforms for viral replication. These structures, termed replication organelles (ROs), have been well characterized for theEnterovirusgenus of thePicornaviridae. However, it is unknown whether the diverse RO morphologies associated with enterovirus infection are conserved among other picornaviruses. Here, we use serial electron tomography at different stages of infection to assess the three-dimensional architecture of ROs induced by encephalomyocarditis virus (EMCV), a member of theCardiovirusgenus of the family of picornaviruses that is distantly related. Ultrastructural analyses revealed connections between early single-membrane EMCV ROs and the endoplasmic reticulum (ER), establishing the ER as a likely donor organelle for their formation. These early single-membrane ROs appear to transform into double-membrane vesicles (DMVs) as infection progresses. Both single- and double-membrane structures were found to support viral RNA synthesis, and progeny viruses accumulated in close proximity, suggesting a spatial association between RNA synthesis and virus assembly. Further, we explored the role of phosphatidylinositol 4-phosphate (PI4P), a critical host factor for both enterovirus and cardiovirus replication that has been recently found to expedite enterovirus RO formation rather than being strictly required. By exploiting an EMCV escape mutant, we found that low-PI4P conditions could also be overcome for the formation of cardiovirus ROs. Collectively, our data show that despite differences in the membrane source, there are striking similarities in the biogenesis, morphology, and transformation of cardiovirus and enterovirus ROs, which may well extend to other picornaviruses.IMPORTANCELike all positive-sense RNA viruses, picornaviruses induce the rearrangement of host cell membranes to form unique structures, or replication organelles (ROs), that support viral RNA synthesis. Here, we investigate the architecture and biogenesis of cardiovirus ROs and compare them with those induced by enteroviruses, members of the well-characterized picornavirus genusEnterovirus. The origins and dynamic morphologies of cardiovirus ROs are revealed using electron tomography, which points to the endoplasmic reticulum as the donor organelle usurped to produce single-membrane tubules and vesicles that transform into double-membrane vesicles. We show that PI4P, a critical lipid for cardiovirus and enterovirus replication, is not strictly required for the formation of cardiovirus ROs, as functional ROs with typical morphologies are formed under phosphatidylinositol 4-kinase type III alpha (PI4KA) inhibition in cells infected with an escape mutant. Our data show that the transformation from single-membrane structures to double-membrane vesicles is a conserved feature of cardiovirus and enterovirus infections that likely extends to other picornavirus genera.

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