scholarly journals Dynamin I phosphorylation and the control of synaptic vesicle endocytosis.

2005 ◽  
Vol 72 ◽  
pp. 87-97 ◽  
Author(s):  
Karen J. Smillie ◽  
Michael A. Cousin
Keyword(s):  
Synaptic Vesicle ◽  
Nerve Terminal ◽  
Current Knowledge ◽  
Direct Proof ◽  
Nerve Terminals ◽  
Cyclin Dependent Kinase ◽  
Calcium Dependent ◽  
Dependent Protein ◽  
Dynamin I ◽  
Cyclin Dependent Kinase 5

The GTPase dynamin I is essential for synaptic vesicle endocytosis in nerve terminals. It is a nerve terminal phosphoprotein that is dephosphorylated on nerve terminal stimulation by the calcium-dependent protein phosphatase calcineurin and then rephosphorylated by cyclin-dependent kinase 5 on termination of the stimulus. Because of its unusual phosphorylation profile, the phosphorylation status of dynamin I was assumed to be inexorably linked to synaptic vesicle endocytosis; however, direct proof of this link has been elusive until very recently. This review will describe current knowledge regarding dynamin I phosphorylation in nerve terminals and how this regulates its biological function with respect to synaptic vesicle endocytosis.

Enhanced activation of Ca2+/Calmodulin-dependent protein kinase II upon downregulation of cyclin-dependent kinase 5-p35

2006 ◽  
Vol 84 (4) ◽  
pp. 747-754 ◽  
Author(s):  
Tomohisa Hosokawa ◽  
Taro Saito ◽  
Akiko Asada ◽  
Toshio Ohshima ◽  
Makoto Itakura ◽  
...  
Keyword(s):  
Protein Kinase ◽  
Cyclin Dependent Kinase ◽  
Dependent Protein Kinase ◽  
Protein Kinase Ii ◽  
Dependent Protein ◽  
Cyclin Dependent Kinase 5

scholarly journals Use of phosphosynapsin I-specific antibodies for image analysis of signal transduction in single nerve terminals

2000 ◽  
Vol 113 (20) ◽  
pp. 3573-3582 ◽  
Author(s):  
A. Menegon ◽  
D.D. Dunlap ◽  
F. Castano ◽  
F. Benfenati ◽  
A.J. Czernik ◽  
...  
Keyword(s):  
Signal Transduction ◽  
Protein Kinase ◽  
Cyclic Amp ◽  
Nerve Terminal ◽  
Polyclonal Antibodies ◽  
Second Messengers ◽  
Nerve Terminals ◽  
Dependent Protein Kinase ◽  
Spatially Resolved ◽  
Dependent Protein

We have developed a semi-quantitative method for indirectly revealing variations in the concentration of second messengers (Ca(2+), cyclic AMP) in single presynaptic boutons by detecting the phosphorylation of the synapsins, excellent nerve terminal substrates for cyclic AMP- and Ca(2+)/calmodulin-dependent protein kinases. For this purpose, we employed polyclonal, antipeptide antibodies recognising exclusively synapsin I phosphorylated by Ca(2+)/calmodulin-dependent protein kinase II (at site 3) or synapsins I/II phosphorylated by either cAMP-dependent protein kinase or Ca(2+)/calmodulin-dependent protein kinase I (at site 1). Cerebellar granular neurones in culture were double-labelled with a monoclonal antibody to synapsins I/II and either of the polyclonal antibodies. Digitised images were analysed to determine the relative phosphorylation stoichiometry at each individual nerve terminal. We have found that: (i) under basal conditions, phosphorylation of site 3 was undetectable, whereas site 1 exhibited some degree of constitutive phosphorylation; (ii) depolarisation in the presence of extracellular Ca(2+) was followed by a selective and widespread increase in site 3 phosphorylation, although the relative phosphorylation stoichiometry varied among individual terminals; and (iii) phosphorylation of site 1 was increased by stimulation of cyclic AMP-dependent protein kinase but not by depolarisation and often occurred in specific nerve terminal sub-populations aligned along axon branches. In addition to shedding light on the regulation of synapsin phosphorylation in living nerve terminals, this approach permits the spatially-resolved analysis of the activation of signal transduction pathways in the presynaptic compartment, which is usually too small to be studied with other currently available techniques.

scholarly journals Distinct functions of a cGMP-dependent protein kinase in nerve terminal growth and synaptic vesicle cycling

2019 ◽  
Vol 132 (7) ◽  
pp. jcs227165 ◽  
Author(s):  
Jeffrey S. Dason ◽  
Aaron M. Allen ◽  
Oscar E. Vasquez ◽  
Marla B. Sokolowski
Keyword(s):  
Protein Kinase ◽  
Synaptic Vesicle ◽  
Nerve Terminal ◽  
Dependent Protein Kinase ◽  
Vesicle Cycling ◽  
Cgmp Dependent Protein Kinase ◽  
Dependent Protein

scholarly journals Synapsin I (protein I), a nerve terminal-specific phosphoprotein. III. Its association with synaptic vesicles studied in a highly purified synaptic vesicle preparation.

1983 ◽  
Vol 96 (5) ◽  
pp. 1374-1388 ◽  
Author(s):  
W B Huttner ◽  
W Schiebler ◽  
P Greengard ◽  
P De Camilli
Keyword(s):  
Ionic Strength ◽  
Synaptic Vesicle ◽  
Synaptic Vesicles ◽  
Immunological Methods ◽  
Synapsin I ◽  
Tail Region ◽  
Electron Microscopic ◽  
Experimental Conditions ◽  
Calcium Dependent ◽  
Dependent Protein

Synapsin I (protein I) is a neuron-specific phosphoprotein, which is a substrate for cAMP-dependent and Ca/calmodulin-dependent protein kinases. In two accompanying studies (De Camilli, P., R. Cameron, and P. Greengard, and De Camilli, P., S. M. Harris, Jr., W. B. Huttner, and P. Greengard, 1983, J. Cell Biol. 96:1337-1354 and 1355-1373) we have shown, by immunocytochemical techniques at the light microscopic and electron microscopic levels, that synapsin I is present in the majority of, and possibly in all, nerve terminals, where it is primarily associated with synaptic vesicles. In the present study we have prepared a highly purified synaptic vesicle fraction from rat brain by a procedure that involves permeation chromatography on controlled-pore glass as a final purification step. Using immunological methods, synapsin I concentrations were determined in various subcellular fractions obtained in the course of vesicle purification. Synapsin I was found to copurify with synaptic vesicles and to represent approximately 6% of the total protein in the highly purified synaptic vesicle fraction. The copurification of synapsin I with synaptic vesicles was dependent on the use of low ionic strength media throughout the purification. Synapsin I was released into the soluble phase by increased ionic strength at neutral pH, but not by nonionic detergents. The highly purified synaptic vesicle fraction contained a calcium-dependent protein kinase that phosphorylated endogenous synapsin I in its collagenase-sensitive tail region. The phosphorylation of this region appeared to facilitate the dissociation of synapsin I from synaptic vesicles under the experimental conditions used.

scholarly journals Cdk5 and the mystery of synaptic vesicle endocytosis

2003 ◽  
Vol 163 (4) ◽  
pp. 697-699 ◽  
Author(s):  
Chan Nguyen ◽  
James A. Bibb
Keyword(s):  
Protein Phosphorylation ◽  
Synaptic Vesicle ◽  
Careful Examination ◽  
Cyclin Dependent Kinase ◽  
Synaptic Vesicle Recycling ◽  
Vesicle Recycling ◽  
Important Regulator ◽  
Cyclin Dependent Kinase 5

Regulation of endocytosis by protein phosphorylation and dephosphorylation is critical to synaptic vesicle recycling. Two groups have now identified the neuronal kinase Cdk5 (cyclin-dependent kinase 5) as an important regulator of this process. Robinson and coworkers recently demonstrated that Cdk5 is necessary for synaptic vesicle endocytosis (SVE) (Tan et al., 2003), whereas a new report in this issue claims that Cdk5 negatively regulates SVE (Tomizawa et al., 2003). Careful examination of the data reveals a model that helps resolve the apparently contradictory nature of these reports.

scholarly journals Syndapin I, a Synaptic Dynamin-binding Protein that Associates with the Neural Wiskott-Aldrich Syndrome Protein

1999 ◽  
Vol 10 (2) ◽  
pp. 501-513 ◽  
Author(s):  
Britta Qualmann ◽  
Jack Roos ◽  
Paul J. DiGregorio ◽  
Regis B. Kelly
Keyword(s):  
Synaptic Vesicle ◽  
Nerve Terminal ◽  
Specific Interaction ◽  
Sh3 Domain ◽  
High Molecular Weight Complex ◽  
Wiskott Aldrich Syndrome ◽  
C Terminus ◽  
Dynamin I ◽  
Syndrome Protein

The GTPase dynamin has been clearly implicated in clathrin-mediated endocytosis of synaptic vesicle membranes at the presynaptic nerve terminal. Here we describe a novel 52-kDa protein in rat brain that binds the proline-rich C terminus of dynamin. Syndapin I (synaptic, dynamin-associated protein I) is highly enriched in brain where it exists in a high molecular weight complex. Syndapin I can be involved in multiple protein–protein interactions via a src homology 3 (SH3) domain at the C terminus and two predicted coiled-coil stretches. Coprecipitation studies and blot overlay analyses revealed that syndapin I binds the brain-specific proteins dynamin I, synaptojanin, and synapsin I via an SH3 domain-specific interaction. Coimmunoprecipitation of dynamin I with antibodies recognizing syndapin I and colocalization of syndapin I with dynamin I at vesicular structures in primary neurons indicate that syndapin I associates with dynamin I in vivo and may play a role in synaptic vesicle endocytosis. Furthermore, syndapin I associates with the neural Wiskott-Aldrich syndrome protein, an actin-depolymerizing protein that regulates cytoskeletal rearrangement. These characteristics of syndapin I suggest a molecular link between cytoskeletal dynamics and synaptic vesicle recycling in the nerve terminal.

scholarly journals Clathrin-coated vesicles in nervous tissue are involved primarily in synaptic vesicle recycling.

1992 ◽  
Vol 118 (6) ◽  
pp. 1379-1388 ◽  
Author(s):  
P R Maycox ◽  
E Link ◽  
A Reetz ◽  
S A Morris ◽  
R Jahn
Keyword(s):  
Synaptic Vesicle ◽  
Nerve Terminal ◽  
Synaptic Vesicles ◽  
Nervous Tissue ◽  
Coated Vesicles ◽  
Nerve Terminals ◽  
Coat Proteins ◽  
Synaptic Vesicle Recycling ◽  
Vesicle Recycling ◽  
Vesicle Proteins

The recycling of synaptic vesicles in nerve terminals is thought to involve clathrin-coated vesicles. However, the properties of nerve terminal coated vesicles have not been characterized. Starting from a preparation of purified nerve terminals obtained from rat brain, we isolated clathrin-coated vesicles by a series of differential and density gradient centrifugation steps. The enrichment of coated vesicles during fractionation was monitored by EM. The final fraction consisted of greater than 90% of coated vesicles, with only negligible contamination by synaptic vesicles. Control experiments revealed that the contribution by coated vesicles derived from the axo-dendritic region or from nonneuronal cells is minimal. The membrane composition of nerve terminal-derived coated vesicles was very similar to that of synaptic vesicles, containing the membrane proteins synaptophysin, synaptotagmin, p29, synaptobrevin and the 116-kD subunit of the vacuolar proton pump, in similar stoichiometric ratios. The small GTP-binding protein rab3A was absent, probably reflecting its dissociation from synaptic vesicles during endocytosis. Immunogold EM revealed that virtually all coated vesicles carried synaptic vesicle proteins, demonstrating that the contribution by coated vesicles derived from other membrane traffic pathways is negligible. Coated vesicles isolated from the whole brain exhibited a similar composition, most of them carrying synaptic vesicle proteins. This indicates that in nervous tissue, coated vesicles function predominantly in the synaptic vesicle pathway. Nerve terminal-derived coated vesicles contained AP-2 adaptor complexes, which is in agreement with their plasmalemmal origin. Furthermore, the neuron-specific coat proteins AP 180 and auxilin, as well as the alpha a1 and alpha c1-adaptins, were enriched in this fraction, suggesting a function for these coat proteins in synaptic vesicle recycling.

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