scholarly journals Rapid changes in synaptic vesicle cytochemistry after depolarization of cultured cholinergic sympathetic neurons.

1985 ◽  
Vol 101 (1) ◽  
pp. 217-226 ◽  
Author(s):  
M I Johnson ◽  
K Paik ◽  
D Higgins
Keyword(s):  
Synaptic Vesicles ◽  
Sympathetic Neurons ◽  
Uptake System ◽  
Vesicle Membrane ◽  
Vesicle Release ◽  
Cholinergic Function ◽  
Dense Cores ◽  
Amine Storage ◽  
Rapid Changes ◽  
Cervical Ganglia

Sympathetic neurons taken from rat superior cervical ganglia and grown in culture acquire cholinergic function under certain conditions. These cholinergic sympathetic neurons, however, retain a number of adrenergic properties, including the enzymes involved in the synthesis of norepinephrine (NE) and the storage of measurable amounts of NE. These neurons also retain a high affinity uptake system for NE; despite this, the majority of the synaptic vesicles remain clear even after incubation in catecholamines. The present study shows, however, that if these neurons are depolarized before incubation in catecholamine, the synaptic vesicles acquire dense cores indicative of amine storage. These manipulations are successful when cholinergic function is induced with either a medium that contains human placental serum and embryo extract or with heart-conditioned medium, and when the catecholamine is either NE or 5-hydroxydopamine. In some experiments, neurons are grown at low densities and shown to have cholinergic function by electrophysiological criteria. After incubation in NE, only 6% of the synaptic vesicles have dense cores. In contrast, similar neurons depolarized (80 mM K+) before incubation in catecholamine contain 82% dense-cored vesicles. These results are confirmed in network cultures where the percentage of dense-cored vesicles is increased 2.5 to 6.5 times by depolarizing the neurons before incubation with catecholamine. In both single neurons and in network cultures, the vesicle reloading is inhibited by reducing vesicle release during depolarization with an increased Mg++/Ca++ ratio or by blocking NE uptake either at the plasma membrane (desipramine) or at the vesicle membrane (reserpine). In addition, choline appears to play a competitive role because its presence during incubation in NE or after reloading results in decreased numbers of dense-cored vesicles. We conclude that the depolarization step preceding catecholamine incubation acts to empty the vesicles of acetylcholine, thus allowing them to reload with catecholamine. These data also suggest that the same vesicles may contain both neurotransmitters simultaneously.

scholarly journals Artificially imposed electrical potentials drive l-glutamate uptake into synaptic vesicles of bovine cerebral cortex

1990 ◽  
Vol 267 (1) ◽  
pp. 63-68 ◽  
Author(s):  
J Shioi ◽  
T Ueda
Keyword(s):  
Cerebral Cortex ◽  
Synaptic Vesicles ◽  
Glutamate Uptake ◽  
Electrical Potential ◽  
Potential Gradient ◽  
Protonmotive Force ◽  
Uptake System ◽  
Vesicle Membrane ◽  
Excitatory Neurotransmitter ◽  
Electrical Potentials

L-Glutamate is a major excitatory neurotransmitter in the central nervous system. MgATP-dependent glutamate uptake and H(+)-pumping ATPase activity were reported in highly purified synaptic vesicles [Naito & Ueda (1983) J. Biol. Chem. 258, 696-699; Shioi, Naito & Ueda (1989) Biochem. J. 258, 499-504], and it is hypothesized that an electrochemical H+ gradient across the vesicle membrane, the so-called protonmotive force, elicits the neurotransmitter uptake. An inside-positive diffusion potential across the vesicle membrane was established with valinomycin plus Rb+. This artificial electrical potential promoted the uptake of glutamate, but not aspartate, in the synaptic vesicles prepared from bovine cerebral cortex. The uptake was inhibited by the protonmotive-force dissipators carbonyl cyanide p-trifluoro-methoxyphenylhydrazone or nigericin, and was enhanced by concomitant imposition of a pH jump (alkalinization) in the external medium. Subcellular and subvesicular distributions showed the uptake system to be predominantly associated with small synaptic vesicles. The results support the hypothesis that glutamate uptake into synaptic vesicles is coupled with a H+ efflux down the electrochemical potential gradient, which is generated by H(+)-pumping ATPase.

scholarly journals AXONAL AGRANULAR RETICULUM AND SYNAPTIC VESICLES IN CULTURED EMBRYONIC CHICK SYMPATHETIC NEURONS

1973 ◽  
Vol 57 (1) ◽  
pp. 88-108 ◽  
Author(s):  
Saul Teichberg ◽  
Eric Holtzman
Keyword(s):  
Synaptic Vesicles ◽  
Sympathetic Neurons ◽  
Size Spectrum ◽  
Embryonic Chick ◽  
Membrane Systems ◽  
Axon Terminals ◽  
Golgi Membranes ◽  
Storage Vesicles ◽  
Adrenergic Systems ◽  
Catecholamine Storage

Cultured chick embryonic sympathetic neurons contain an extensive axonal network of sacs and tubules of agranular reticulum. The reticulum is also seen branching into networks in axon terminals and varicosities. The axonal reticulum and perikaryal endoplasmic reticulum resemble one another in their content of cytochemically demonstrable enzyme activities (G6Pase and IDPase) and in their characteristic membrane thicknesses (narrower than plasma membrane or some Golgi membranes). From the reticulum, both along the axon and at terminals, there appear to form dense-cored vesicles ranging in size from 400 to 1,000 Å in diameter. These vesicles behave pharmacologically and cytochemically like the classes of large and small catecholamine storage vesicles found in several adrenergic systems; for example, they can accumulate exogenous 5-hydroxydopamine. In addition, dense-cored vesicles at the larger (1,000 Å) end of the size spectrum appear to arise within perikaryal membrane systems associated with the Golgi apparatus; this is true also of very large (800–3,500 Å) dense-cored vesicles found in some perikarya.

VSM growth is stimulated in sympathetic neuron/VSM cocultures: role of TGF-β2 and endothelin

2000 ◽  
Vol 278 (2) ◽  
pp. H404-H411 ◽  
Author(s):  
Deborah H. Damon
Keyword(s):  
Transforming Growth Factor ◽  
Sympathetic Neurons ◽  
Sympathetic Nerves ◽  
Sympathetic Neuron ◽  
Vessel Growth ◽  
Coculture Model ◽  
Transforming Growth Factor Β2 ◽  
Cervical Ganglia ◽  
Stimulation Of

Sympathetic nerves are purported to stimulate blood vessel growth. The mechanism(s) underlying this stimulation has not been determined. With use of an in vitro coculture model, the present study tests the hypothesis that sympathetic neurons stimulate the growth of vascular smooth muscle (VSM) and evaluates potential mechanisms mediating this stimulation. Sympathetic neurons isolated from superior cervical ganglia (SCG) stimulated the growth of VSM. Growth of VSM in the presence of SCG (856 ± 81%) was significantly greater than that in the absence of SCG (626 ± 66%, P < 0.05). SCG did not stimulate VSM growth in transwell cocultures. An antibody that neutralized the activity of transforming growth factor-β2 (TGF-β2) inhibited SCG stimulation of VSM growth in coculture. SCG stimulation of VSM growth was also inhibited by an endothelin A receptor antagonist. These data suggest novel mechanisms for sympathetic modulation of vascular growth that may play a role in the physiological and/or pathological growth of the vasculature.

Active zone assembly and synaptic release

2006 ◽  
Vol 34 (5) ◽  
pp. 939-941 ◽  
Author(s):  
R.J. Kittel ◽  
S. Hallermann ◽  
S. Thomsen ◽  
C. Wichmann ◽  
S.J. Sigrist ◽  
...  
Keyword(s):  
Calcium Channels ◽  
Active Zone ◽  
Synaptic Vesicles ◽  
Spatial Arrangement ◽  
Transmission Characteristics ◽  
Vesicle Release ◽  
Synaptic Release ◽  
Voltage Gated Calcium Channels ◽  
Presynaptic Calcium ◽  
Chemical Synapses

Neurotransmitter release at chemical synapses occurs when synaptic vesicles fuse to the presynaptic membrane at a specialized site termed the active zone. The depolarization-induced fusion is highly dependent on calcium ions, and, correspondingly, the transmission characteristics of synapses are thought to be influenced by the spatial arrangement of voltage-gated calcium channels with respect to vesicle release sites. Here, we review the involvement of the Drosophila Bruchpilot (BRP) protein in active zone assembly, a process that is required for the clustering of presynaptic calcium channels to ensure efficient vesicle release.

scholarly journals Botulinum Neurotoxin a Blocks Synaptic Vesicle Exocytosis but Not Endocytosis at the Nerve Terminal

1999 ◽  
Vol 147 (6) ◽  
pp. 1249-1260 ◽  
Author(s):  
Elaine A. Neale ◽  
Linda M. Bowers ◽  
Min Jia ◽  
Karen E. Bateman ◽  
Lura C. Williamson
Keyword(s):  
Synaptic Vesicle ◽  
Nerve Terminal ◽  
Neurotransmitter Release ◽  
Synaptic Vesicles ◽  
Botulinum Neurotoxin ◽  
Vesicle Membrane ◽  
Botulinum Neurotoxin A ◽  
Synaptic Vesicle Exocytosis ◽  
Vesicle Exocytosis ◽  
Botulinum Neurotoxins

The supply of synaptic vesicles in the nerve terminal is maintained by a temporally linked balance of exo- and endocytosis. Tetanus and botulinum neurotoxins block neurotransmitter release by the enzymatic cleavage of proteins identified as critical for synaptic vesicle exocytosis. We show here that botulinum neurotoxin A is unique in that the toxin-induced block in exocytosis does not arrest vesicle membrane endocytosis. In the murine spinal cord, cell cultures exposed to botulinum neurotoxin A, neither K+-evoked neurotransmitter release nor synaptic currents can be detected, twice the ordinary number of synaptic vesicles are docked at the synaptic active zone, and its protein substrate is cleaved, which is similar to observations with tetanus and other botulinal neurotoxins. In marked contrast, K+ depolarization, in the presence of Ca2+, triggers the endocytosis of the vesicle membrane in botulinum neurotoxin A–blocked cultures as evidenced by FM1-43 staining of synaptic terminals and uptake of HRP into synaptic vesicles. These experiments are the first demonstration that botulinum neurotoxin A uncouples vesicle exo- from endocytosis, and provide evidence that Ca2+ is required for synaptic vesicle membrane retrieval.

Chromogranin A, dopamine β-hydroxylase and secretion from the adrenal medulla

1971 ◽  
Vol 261 (839) ◽  
pp. 279-289 ◽  
Keyword(s):  
Adrenal Medulla ◽  
Synaptic Vesicles ◽  
Adenine Nucleotides ◽  
Sympathetic Nerves ◽  
Neural Stimulation ◽  
Secretory Process ◽  
Vesicle Membrane ◽  
The Past ◽  
Storage Vesicles ◽  
Experimental Approaches

Studies of the biosynthesis, storage and secretion of catecholamines by the adrenal medulla have served as models for similar studies of the adrenergic neuron. For example, the synthesis of noradrenaline and the intracellular distribution of the biosynthetic enzymes was first described in the adrenal medulla and subsequently shown to be the same in sympathetic nerves (Blaschko 1939; Kirshner 1957, 1959; Levin, Levenberg & Kaufman i960; Potter & Axelrod 1963; Nagatsu, Levitt & Udenfriend 1964; Stjarne & Lishajko 1966; Oka et al. 1967; Musacchio 1968; Laduron & Belpaire 1968). The storage vesicles of the adrenal medulla have counterparts in the synaptic vesicles (Blaschko & Welch 1953; Hillarp, Lagerstedt & Nilson 1953; von Euler & Hillarp 1956; Schumann 1958) and the incorporation of catecholamines into the storage vesicles, and the storage complex itself, seems to be similar in both tissues, (Kirshner 1962; Carlsson, Hillarp & Waldeck 1963; von Euler & Lishajko 1963; von Euler, Lishajko & Stjarne 1963; Stjarne 1964). Recently it has been demonstrated that proteins specifically localized in the storage vesicles of the adrenal medulla are also present in the storage vesicles of sympathetic nerve endings (Hopwood 1967, 1968; Geffen, Livett & Rush 1969; Banks, Helle & Major 1969; de Potter, de Schaepdryver, Moerman & Smith 1969). There are obvious differences between the two types of vesicles (Stjarne 1964; Potter 1967), but the similarities are such as to suggest that the vesicles from both tissues serve the same physiological functions—to synthesize and store adrenaline or noradrenaline and to release these compounds in response to neural stimulation. Secretion from the adrenal medulla appears to be a good model for release of neurotransmitters at synapses in the sense that it provides and suggests experimental approaches to the problem (Geffen et al. 1969; de Potter et al. 1969). In general, the secretion of substances which are synthesized in cells and stored in subcellular organelles have many features in common (Douglas 1968; Stormorken 1969) and release of neurotransmitters at synapses may be another example of this generalized biological process. During the past few years, evidence has been presented from several laboratories that secretion from the adrenal medulla occurs by exocytosis. The simultaneous release of catecholamines, adenine nucleotides, chromogranins and soluble dopamine β-hydroxylase contained within the storage vesicles and the retention of dopamine-β- hydroxylase firmly bound to the vesicle membrane have provided critical information on this secretory process.

scholarly journals Identification of a transmembrane glycoprotein specific for secretory vesicles of neural and endocrine cells.

1985 ◽  
Vol 100 (4) ◽  
pp. 1284-1294 ◽  
Author(s):  
K Buckley ◽  
R B Kelly
Keyword(s):  
Synaptic Vesicles ◽  
Endocrine Cells ◽  
Electric Organ ◽  
Cell Types ◽  
Cross Reactivity ◽  
Secretory Vesicles ◽  
Vesicle Membrane ◽  
Electron Microscopic ◽  
Transmembrane Glycoprotein ◽  
Immunoperoxidase Labeling

Several types of cells store proteins in secretory vesicles from which they are released by an appropriate stimulus. It might be expected that the secretory vesicles in different cell types use similar molecular machinery. Here we describe a transmembrane glycoprotein (Mr approximately 100,000) that is present in secretory vesicles in all neurons and endocrine cells studied, in species from elasmobranch fish to mammals, and in neural and endocrine cell lines. It was detected by cross-reactivity with monoclonal antibodies raised to highly purified cholinergic synaptic vesicles from the electric organ of fish. By immunoprecipitation of intact synaptic vesicles and electron microscopic immunoperoxidase labeling, we have shown that the antigenic determinant is on the cytoplasmic face of the synaptic vesicles. However, the electrophoretic mobility of the antigen synthesized in the presence of tunicamycin is reduced to Mr approximately 62,000, which suggests that the antigen is glycosylated and must therefore span the vesicle membrane.

scholarly journals Pineal Gland Transplants into the Cerebral Hemisphere of Newborn Rats: A Study of the Blood Brain Barrier and Innervation

1991 ◽  
Vol 2 (2) ◽  
pp. 113-124 ◽  
Author(s):  
J. A. McNulty ◽  
L. M. Fox ◽  
P. L. Shaw ◽  
V. E. Alones ◽  
B. S. Klausen ◽  
...  
Keyword(s):  
Sympathetic Neurons ◽  
Peripheral Zone ◽  
Volume Density ◽  
Newborn Rats ◽  
Neural Input ◽  
Neural Transplants ◽  
Cervical Ganglia ◽  
To Receive ◽  
Host Influence

Pineal glands from neonatal (0-1 day) Long-Evans black-hooded rats were transplanted into the cerebral hemispheres of litter mates for periods of 1 to 5.5 months. Grafts exhibited differentiated pinealocytes that were intensely immunoreactive for serotonin. Transplant vasculature was permeable to endogenous IgG, comprised fenestrated endothelia with wide pericapillary spaces typical ofin situglands, and had a volume density intermediate to that of surrounding cortex andin situpineals. Along the periphery, transplant capillaries tended to have continuous endothelia similar to those of host cortex. This peripheral zone was impermeable to endogenous IgG and appeared to increase in size in older grafts. The presence of noradrenergic-like fibers within the perivascular compartment suggested that transplants were innervated by peripheral sympathetic neurons from the superior cervical ganglia. In animals which had been superior cervical ganglionectomized, noradrenergic-like fibers were absent or degenerating. Neural regulation of transplant metabolic activity was suggested by the increased frequency of pinealocyte synaptic ribbons in denervated grafts. These findings are consistent with the hypothesis that factors from both graft and host influence vasculature physiology and differentiation in neural transplants. Furthermore, grafts appeared to receive appropriate neural input from the peripheral sympathetic system.

scholarly journals Viral proteins E1B19K and p35 protect sympathetic neurons from cell death induced by NGF deprivation.

1995 ◽  
Vol 128 (1) ◽  
pp. 201-208 ◽  
Author(s):  
I Martinou ◽  
P A Fernandez ◽  
M Missotten ◽  
E White ◽  
B Allet ◽  
...  
Keyword(s):  
Cell Death ◽  
Molecular Mechanisms ◽  
Sympathetic Neurons ◽  
Neuronal Cell Death ◽  
Neuronal Cell ◽  
Viral Proteins ◽  
Lymphoid Cells ◽  
Expression Vectors ◽  
Cervical Ganglia ◽  
Adenoviral Proteins

To study molecular mechanisms underlying neuronal cell death, we have used sympathetic neurons from superior cervical ganglia which undergo programmed cell death when deprived of nerve growth factor. These neurons have been microinjected with expression vectors containing cDNAs encoding selected proteins to test their regulatory influence over cell death. Using this procedure, we have shown previously that sympathetic neurons can be protected from NGF deprivation by the protooncogene Bcl-2. We now report that the E1B19K protein from adenovirus and the p35 protein from baculovirus also rescue neurons. Other adenoviral proteins, E1A and E1B55K, have no effect on neuronal survival. E1B55K, known to block apoptosis mediated by p53 in proliferative cells, failed to rescue sympathetic neurons suggesting that p53 is not involved in neuronal death induced by NGF deprivation. E1B19K and p35 were also coinjected with Bcl-Xs which blocks Bcl-2 function in lymphoid cells. Although Bcl-Xs blocked the ability of Bcl-2 to rescue neurons, it had no effect on survival that was dependent upon expression of E1B19K or p35.

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