scholarly journals Single-particle fluorescence tracking combined with TrackMate assay reveals highly heterogeneous and discontinuous lysosomal transport in freely orientated axons

2022 ◽  
Author(s):  
Yongyang Liu ◽  
Yaxin Lu ◽  
Zhiyong Tang ◽  
Yuheng Cao ◽  
Dehua Huang ◽  
...  
Keyword(s):  
Axonal Transport ◽  
Single Particle ◽  
Neuronal Development ◽  
Synapse Formation ◽  
Transport Model ◽  
Gene Transfection ◽  
Axon Growth ◽  
Physical Factors ◽  
Size Of Particles ◽  
Number Of Bends

Axonal transport plays a significant role in the establishment of neuronal polarity, axon growth, and synapse formation during neuronal development. The axon of a naturally growing neuron is a highly complex and multifurcated structure with a large number of bends and branches. Nowadays, the study of dynamic axonal transport in morphologically complex neurons is greatly limited by the technological barrier. Here, a sparse gene transfection strategy was developed to locate fluorescent mCherry in the lysosome of primary neurons, thus enabling us to track the lysosome-based axonal transport with a single-particle resolution. Thereby, several axonal transport models were observed, including forward or backward transport model, stop-and-go model, repeated back-and-forth transport model, and cross-branch transport model. Then, the accurate single-particle velocity quantification by TrackMate revealed a highly heterogeneous and discontinuous transportation process of lysosome-based axonal transport in freely orientated axons. And, multiple physical factors, such as the axonal structure and the size of particles, were disclosed to affect the velocity of particle transporting in freely orientated axons. The combined single-particle fluorescence tracking and TrackMate assay can be served as a facile tool for evaluating axonal transport in neuronal development and axonal transport-related diseases.

scholarly journals MicroExonator enables systematic discovery and quantification of microexons across mouse embryonic development

2020 ◽  
Author(s):  
Guillermo E. Parada ◽  
Roberto Munita ◽  
Ilias Georgakopoulos-Soares ◽  
Hugo Fernandez ◽  
Emmanouil Metzakopian ◽  
...  
Keyword(s):  
Neuronal Development ◽  
Synapse Formation ◽  
De Novo ◽  
Axon Growth ◽  
Developmental Time ◽  
Rna Seq ◽  
Mouse Tissues ◽  
Comprehensive Survey ◽  
Mouse Transcript ◽  
Mouse Visual Cortex

AbstractMicroexons, exons that are ≤30 nucleotides, were shown to play key roles in neuronal development, but are difficult to detect and quantify using standard RNA-Seq alignment tools. Here, we present MicroExonator, a novel pipeline for reproducible de novo discovery and quantification of microexons. We processed 289 RNA-seq datasets from eighteen mouse tissues corresponding to nine embryonic and postnatal stages, providing the most comprehensive survey of microexons available for mouse. We detected 2,984 microexons, 332 of which are differentially spliced throughout mouse embryonic brain development, including 29 that are not present in mouse transcript annotation databases. Unsupervised clustering of microexons alone segregates brain tissues by developmental time and further analysis suggest a key function for microexon inclusion in axon growth and synapse formation. Finally, we analysed single-cell RNA-seq data from the mouse visual cortex and we report differential inclusion between neuronal subpopulations, suggesting that some microexons could be cell-type specific.

Hikaru genki protein is secreted into synaptic clefts from an early stage of synapse formation in Drosophila

Development ◽  
1996 ◽  
Vol 122 (2) ◽  
pp. 589-597 ◽  
Author(s):  
M. Hoshino ◽  
E. Suzuki ◽  
Y. Nabeshima ◽  
C. Hama
Keyword(s):  
Critical Period ◽  
Synapse Formation ◽  
Neural Circuits ◽  
Motor Behavior ◽  
Early Stage ◽  
Axon Growth ◽  
Number Of Factors ◽  
Protein Functions ◽  
Abnormal Motor ◽  
Immunohistochemical Analyses

The development of neural circuits is regulated by a large number of factors that are localized at distinct neural sites. We report here the localization of one of these factors, hikaru genki (hig) protein, at synaptic clefts in the pupal and adult nervous systems of Drosophila. In hig mutants, unusually frequent bursting activity of the muscles and abnormal motor behavior during the adult stage suggest the misfunction of neuromuscular circuitry. Our immunohistochemical analyses revealed that hig protein, produced by neurons, is secreted from the presynaptic terminals into the spaces between the presynaptic and postsynaptic terminals. In addition, we have found that the localization of this protein in the synaptic spaces temporally correlates with its functional requirement during a critical period that occurs in the middle stage of pupal formation, a period when a number of dendrite and axon growth cones meet to form synapses. These findings indicate that hig protein functions in the formation of functional neural circuits from the early stages of synapse formation.

scholarly journals βII-spectrin promotes mouse brain connectivity through stabilizing axonal plasma membranes and enabling axonal organelle transport

2019 ◽  
Vol 116 (31) ◽  
pp. 15686-15695 ◽  
Author(s):  
Damaris N. Lorenzo ◽  
Alexandra Badea ◽  
Ruobo Zhou ◽  
Peter J. Mohler ◽  
Xiaowei Zhuang ◽  
...  
Keyword(s):  
Axonal Transport ◽  
Plasma Membranes ◽  
Brain Connectivity ◽  
Axon Growth ◽  
Membrane Skeleton ◽  
Lipid Binding ◽  
Organelle Transport ◽  
Double Knockout ◽  
Micrometer Scale

βII-spectrin is the generally expressed member of the β-spectrin family of elongated polypeptides that form micrometer-scale networks associated with plasma membranes. We addressed in vivo functions of βII-spectrin in neurons by knockout of βII-spectrin in mouse neural progenitors. βII-spectrin deficiency caused severe defects in long-range axonal connectivity and axonal degeneration. βII-spectrin–null neurons exhibited reduced axon growth, loss of actin–spectrin-based periodic membrane skeleton, and impaired bidirectional axonal transport of synaptic cargo. We found that βII-spectrin associates with KIF3A, KIF5B, KIF1A, and dynactin, implicating spectrin in the coupling of motors and synaptic cargo. βII-spectrin required phosphoinositide lipid binding to promote axonal transport and restore axon growth. Knockout of ankyrin-B (AnkB), a βII-spectrin partner, primarily impaired retrograde organelle transport, while double knockout of βII-spectrin and AnkB nearly eliminated transport. Thus, βII-spectrin promotes both axon growth and axon stability through establishing the actin–spectrin-based membrane-associated periodic skeleton as well as enabling axonal transport of synaptic cargo.

Emerging roles for HOX proteins in synaptogenesis

2018 ◽  
Vol 62 (11-12) ◽  
pp. 807-818 ◽  
Author(s):  
Françoise Gofflot ◽  
Benoit Lizen
Keyword(s):  
Cell Fate ◽  
Synapse Formation ◽  
Neural Circuit ◽  
Target Selection ◽  
Axon Growth ◽  
Current Data ◽  
Cell Fate Specification ◽  
Hox Proteins ◽  
Circuit Formation ◽  
Vertebrate Central Nervous System

Neural circuit formation requires the intricate orchestration of multiple developmental events including cell fate specification, cell migration, axon guidance, dendritic growth, synaptic target selection, and synaptogenesis. The HOX proteins are well-known transcriptional regulators that control embryonic development. Investigations into their action in the vertebrate central nervous system have demonstrated pivotal roles in specifying neural subpopulations, but also in several successive steps required for the assembly of neuronal circuitry, such as neuron migration, axon growth and pathfinding and synaptic target selection. Several lines of evidence suggest that the HOX transcription factors could also regulate synaptogenesis processes even after the process of axonal and dendritic guidance has concluded. Here we will review the current data on HOX proteins in neural circuit formation in order to evaluate their potential roles in establishing neuronal connectivity with specific emphasis on synapse formation and maturation.

scholarly journals Correct Laminar Positioning in the Neocortex Influences Proper Dendritic and Synaptic Development

2018 ◽  
Vol 28 (8) ◽  
pp. 2976-2990 ◽  
Author(s):  
Fanny Sandrine Martineau ◽  
Surajit Sahu ◽  
Vanessa Plantier ◽  
Emmanuelle Buhler ◽  
Fabienne Schaller ◽  
...  
Keyword(s):  
Neuronal Migration ◽  
Cortical Neurons ◽  
Neuronal Development ◽  
Synapse Formation ◽  
Functional Organization ◽  
Functional Integration ◽  
Proper Function ◽  
Cortical Circuits ◽  
Gabaergic Synapses ◽  
Dendrite Morphogenesis

Abstract The neocortex is a 6-layered laminated structure with a precise anatomical and functional organization ensuring proper function. Laminar positioning of cortical neurons, as determined by termination of neuronal migration, is a key determinant of their ability to assemble into functional circuits. However, the exact contribution of laminar placement to dendrite morphogenesis and synapse formation remains unclear. Here we manipulated the laminar position of cortical neurons by knocking down doublecortin (Dcx), a crucial effector of migration, and show that misplaced neurons fail to properly form dendrites, spines, and functional glutamatergic and GABAergic synapses. We further show that knocking down Dcx in properly positioned neurons induces similar but milder defects, suggesting that the laminar misplacement is the primary cause of altered neuronal development. Thus, the specific laminar environment of their fated layers is crucial for the maturation of cortical neurons, and influences their functional integration into developing cortical circuits.

scholarly journals Sensitivity of oxygen dynamics in the water column of the Baltic Sea to external forcing

Ocean Science ◽  
2010 ◽  
Vol 6 (2) ◽  
pp. 461-474 ◽  
Author(s):  
S. Miladinova ◽  
A. Stips
Keyword(s):  
Baltic Sea ◽  
Transport Model ◽  
Meteorological Data ◽  
Biogeochemical Model ◽  
Physical Factors ◽  
Observation Data ◽  
Intermediate Water ◽  
The Baltic Sea ◽  
The Baltic ◽  
Oxygen Dynamics

Abstract. A 1-D biogeochemical/physical model of marine systems has been applied to study the oxygen cycle in four stations of different sub-basins of the Baltic Sea, namely, in the Gotland Deep, Bornholm, Arkona and Fladen. The model consists of the biogeochemical model of Neumann et al. (2002) coupled with the 1-D General Ocean Turbulence Model (GOTM). The model has been forced with meteorological data from the ECMWF reanalysis project for the period 1998–2003, producing a six year hindcast which is validated with datasets from the Baltic Environmental Database (BED) for the same period. The vertical profiles of temperature and salinity are relaxed towards both profiles provided by 3-D simulations of General Estuarine Transport Model (GETM) and observed profiles from BED. Modifications in the parameterisation of the air-sea oxygen fluxes have led to a significant improvement of the model results in the surface and intermediate water layers. The largest mismatch with observations is found in simulating the oxygen dynamics in the Baltic Sea bottom waters. The model results demonstrate the good capability of the model to predict the time-evolution of the physical and biogeochemical variables at all different stations. Comparative analysis of the modelled oxygen concentrations with respect to observation data is performed to distinguish the relative importance of several factors on the seasonal, interannual and long-term variations of oxygen. It is found that natural physical factors, like the magnitude of the vertical turbulent mixing, wind speed and the variation of temperature and salinity fields are the major factors controlling the oxygen dynamics in the Baltic Sea. The influence of limiting nutrients is less pronounced, at least under the nutrient flux parameterisation assumed in the model.

Slow axonal transport: the subunit transport model

1997 ◽  
Vol 7 (10) ◽  
pp. 384-388 ◽  
Author(s):  
Nobutaka Hirokawa ◽  
Sumio TeradaTakeshi Funakoshi ◽  
Sen Takeda
Keyword(s):  
Axonal Transport ◽  
Transport Model ◽  
Slow Axonal Transport

scholarly journals Stages in axon formation: observations of growth of Aplysia axons in culture using video-enhanced contrast-differential interference contrast microscopy.

1986 ◽  
Vol 103 (5) ◽  
pp. 1921-1931 ◽  
Author(s):  
D J Goldberg ◽  
D W Burmeister
Keyword(s):  
Axonal Transport ◽  
Growth Cone ◽  
Axon Growth ◽  
Leading Edge ◽  
Main Body ◽  
Differential Interference Contrast ◽  
Differential Interference Contrast Microscopy ◽  
Fast Axonal Transport ◽  
Contrast Microscopy ◽  
Interference Contrast Microscopy

The regenerative growth in culture of the axons of two giant identified neurons from the central nervous system of Aplysia californica was observed using video-enhanced contrast-differential interference contrast microscopy. This technique allowed the visualization in living cells of the membranous organelles of the growth cone. Elongation of axonal branches always occurred through the same sequence of events: A flat organelle-free veil protruded from the front of the growth cone, gradually filled with vesicles that entered by fast axonal transport and Brownian motion from the main body of the growth cone, became more voluminous and engorged with organelles (vesicles, mitochondria, and one or two large, irregular, refractile bodies), and, finally, assumed the cylindrical shape of the axon branch with the organelles predominantly moving by bidirectional fast axonal transport. The veil is thus the nascent axon. Because veils appear to be initially free of membranous organelles, addition of membrane to the plasmalemma by exocytosis is likely to occur in the main body of the growth cone rather than at the leading edge. Veils almost always formed with filopodial borders, protruding between either fully extended or growing filopodia. Therefore, one function of the filopodia is to direct elongation by demarcating the pathway along which axolemma flows. Models of axon growth in which the body of the growth cone is pulled forward, or in which advance of the leading edge is achieved by filopodial shortening or contraction against an adhesion to the substrate, are inconsistent with our observations. We suggest that, during the elongation phase of growth, filopodia may act as structural supports.

scholarly journals Superfluous Role of Mammalian Septins 3 and 5 in Neuronal Development and Synaptic Transmission

2008 ◽  
Vol 28 (23) ◽  
pp. 7012-7029 ◽  
Author(s):  
Christopher W. Tsang ◽  
Michael Fedchyshyn ◽  
John Harrison ◽  
Hong Xie ◽  
Jing Xue ◽  
...  
Keyword(s):  
Synaptic Transmission ◽  
Synaptic Vesicle ◽  
Hippocampal Neurons ◽  
Neuronal Development ◽  
Synapse Formation ◽  
Vesicle Traffic ◽  
Axon Development ◽  
Central Synapses ◽  
Synaptic Vesicle Fusion

ABSTRACT The septin family of GTPases, first identified for their roles in cell division, are also expressed in postmitotic tissues. SEPT3 (G-septin) and SEPT5 (CDCrel-1) are highly expressed in neurons, enriched in presynaptic terminals, and associated with synaptic vesicles. These characteristics suggest that SEPT3 or SEPT5 might be important for synapse formation, maturation, or synaptic vesicle traffic. Since Sept5 −/− mice do not show any overt neurological phenotypes, we generated Sept3 −/− and Sept3 −/− Sept5 −/− mice and found that SEPT3 and SEPT5 are not essential for development, fertility, or viability. Changes in the expression of septins were noted in the absence of SEPT3, SEPT5, and both septins. SEPT5 association with other septins in brain tissue was unaffected by the removal of SEPT3. No abnormalities were observed in the gross morphology and synapses of the hippocampus. Similarly, axon development and synapse formation were unaffected in vitro. In cultured hippocampal neurons, the size of the recycling synaptic vesicle pool was unaltered in the absence of SEPT3. Furthermore, synaptic transmission at two different central synapses was not significantly affected in Sept3 −/− Sept5 −/− mice. These results indicate that SEPT3 and SEPT5 are dispensable for neuronal development as well as for synaptic vesicle fusion and recycling.

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