Ba2+ does not support synaptic vesicle retrieval in rat cerebrocortical synaptosomes.
AuthorsCousin MA, Robinson PJ
JournalNeurosci Lett
PubMed ID9754790
To investigate whether any specific requirement for extracellular Ca2+ exists in the control synaptic vesicle retrieval, we examined the ability of the divalent cation Ba2+ to substitute for Ca2+ in both vesicle exocytosis and endocytosis. Ba2+ stimulated glutamate release from rat cerebrocortical synaptosomes. Ba2+-evoked release was inhibited by bafilomycin A1, ... More
Frequency dependence of synaptic vesicle exocytosis in aortic baroreceptor neurons and the role of group III mGluRs.
AuthorsPamidimukkala J, Hay M
JournalBrain Res
PubMed ID15051525
Synaptic transmission between baroreceptor afferents and the nucleus tractus solitarius (NTS) is essential for reflex regulation of blood pressure. High frequency stimulation of the afferents in vivo leads to a decrease in synaptic strength and is generally attributed to reduction in presynaptic neurotransmitter release. It has been hypothesized that during ... More
Long-term staining of live Merkel cells with FM dyes.
AuthorsFukuda J, Ishimine H, Masaki Y
JournalCell Tissue Res
PubMed ID12658440
'Live Merkel cells in the skin and hair follicles are known to incorporate a fluorescence dye, quinacrine, which has been utilized to identify and dissect the cells for experiments. Quinacrine fluorescence of the cells is, however, quickly lost and quinacrine-stained Merkel cells soon become difficult to identify in tissue culture. ... More
Two mechanisms of synaptic vesicle recycling in rat brain nerve terminals.
AuthorsCousin MA, Robinson PJ
JournalJ Neurochem
PubMed ID10987846
'KCl and 4-aminopyridine (4-AP) evoke glutamate release from rat brain cortical nerve terminals by voltage clamping or by Na(+) channel-generated repetitive action potentials, respectively. Stimulation by 4-AP but not KCl is largely mediated by protein kinase C (PKC). To determine whether KCl and 4-AP utilise the same mechanism to release ... More
Calcium triggers calcineurin-dependent synaptic vesicle recycling in mammalian nerve terminals.
AuthorsMarks B, McMahon HT
JournalCurr Biol
PubMed ID9651678
'BACKGROUND: Following exocytosis at the synapse, synaptic vesicle components are recovered by endocytosis. Morphological analysis has suggested that this occurs by a clathrin-mediated pathway, and the GTPase dynamin is thought to be involved in ''pinching off'' endocytosing vesicles. The finding that the calcium-dependent phosphatase calcineurin can dephosphorylate dynamin and two ... More
The timing of synaptic vesicle endocytosis.
AuthorsRyan TA, Smith SJ, Reuter H
JournalProc Natl Acad Sci U S A
PubMed ID8643616
'Alternative models to describe the endocytosis phase of synaptic vesicle recycling are associated with time scales of vesicle recovery ranging from milliseconds to tens of seconds. There have been suggestions that one of the major models, envisioned as a slow process that occurs only after complete fusion of the vesicle ... More
Evidence for early endosome-like fusion of recently endocytosed synaptic vesicles.
AuthorsRizzoli SO, Bethani I, Zwilling D, Wenzel D, Siddiqui TJ, Brandhorst D, Jahn R
JournalTraffic
PubMed ID17004320
'Early endosomes are well-established acceptor compartments of endocytic vesicles in many cell types. Little evidence of their existence or function has been obtained in synapses, and it is generally believed that synaptic vesicles recycle without passing through an endosomal intermediate. We show here that the early endosomal SNARE proteins are ... More
Monitoring secretory membrane with FM1-43 fluorescence.
AuthorsCochilla AJ, Angleson JK, Betz WJ
JournalAnnu Rev Neurosci
PubMed ID10202529
'FM1-43 and similar styryl dyes have proven useful as probes for membrane trafficking because they reversibly stain membranes, are impermeable to membranes, and are more fluorescent when bound to membranes than when in solution. Because these dyes stain membranes in an activity-dependent manner, they are ideal for studies of neurotransmitter ... More
Rapid reuse of readily releasable pool vesicles at hippocampal synapses.
AuthorsPyle JL, Kavalali ET, Piedras-Rentería ES, Tsien RW
JournalNeuron
PubMed ID11086996
'Functional presynaptic vesicles have been subdivided into readily releasable (RRP) and reserve (RP) pools. We studied recycling properties of RRP vesicles through differential retention of FM1-43 and FM2-10 and by varying the time window for FM dye uptake. Both approaches indicated that vesicles residing in the RRP underwent rapid endocytosis ... More
Syndapin I is the phosphorylation-regulated dynamin I partner in synaptic vesicle endocytosis.
AuthorsAnggono V, Smillie KJ, Graham ME, Valova VA, Cousin MA, Robinson PJ
JournalNat Neurosci
PubMed ID16648848
'Dynamin I is dephosphorylated at Ser-774 and Ser-778 during synaptic vesicle endocytosis (SVE) in nerve terminals. Phosphorylation was proposed to regulate the assembly of an endocytic protein complex with amphiphysin or endophilin. Instead, we found it recruits syndapin I for SVE and does not control amphiphysin or endophilin binding in ... More
Using FM1-43 to study neuropeptide granule dynamics and exocytosis.
In the study of neuropeptide secretion and membrane trafficking, the fluorescent dye FM1-43 provides the ability to label selectively those structures that are undergoing exocytosis and endocytosis in living cells in real time. This review describes the unique properties of the FM dyes that make them ideal for studying neuropeptide ... More
Imaging synaptic activity in intact brain and slices with FM1-43 in C. elegans, lamprey, and rat.
AuthorsKay AR, Alfonso A, Alford S, Cline HT, Holgado AM, Sakmann B, Snitsarev VA, Stricker TP, Takahashi M, Wu LG
JournalNeuron
PubMed ID10624945
The fluorescent probe FM1-43 has been used extensively for imaging vesicle recycling; however, high nonspecific adsorption resulting in elevated background levels has precluded its use in certain tissues, notably brain slices. We have found that a sulfobutylated derivative of beta-cyclodextrin (ADVASEP-7) has a higher affinity for FM1-43 than the plasma ... More
"Kiss and run" exocytosis at hippocampal synapses.
AuthorsStevens CF, Williams JH
JournalProc Natl Acad Sci U S A
PubMed ID11050187
We have combined electrophysiology and imaging to measure the release of neurotransmitter and fluorescent dye at synapses of cultured hippocampal neurons. These experiments have revealed a "kiss and run" mode of exocytosis in which synaptic vesicles release glutamate normally but do not permit dye to enter or escape from the ... More
Two endocytic recycling routes selectively fill two vesicle pools in frog motor nerve terminals.
AuthorsRichards DA, Guatimosim C, Betz WJ
JournalNeuron
PubMed ID11055437
We have identified and characterized two vesicle recycling pathways in frog motor nerve terminals. We exploited the differential staining properties of FM dyes of varying hydrophobicity to label selectively two different vesicle pools, using optical imaging and electron microscopy of photoconverted dyes. During a 1 min tetanus, a rapidly recycling ... More
Imaging exocytosis and endocytosis.
AuthorsBetz WJ, Mao F, Smith CB
JournalCurr Opin Neurobiol
PubMed ID8794083
From the secretion of neurotransmitters via synaptic vesicles to the expulsion of cellular waste via contractile vacuoles, exocytosis and its sequel, endocytosis, are being explored with a variety of new optical tools. Fluorescent markers, especially styryl dyes such as FM1-43 (which reversibly labels endosomal membranes), have been used to follow ... More
SNARE function analyzed in synaptobrevin/VAMP knockout mice.
AuthorsSchoch S, Deák F, Königstorfer A, Mozhayeva M, Sara Y, Südhof TC, Kavalali ET
JournalScience
PubMed ID11691998
SNAREs (soluble NSF-attachment protein receptors) are generally acknowledged as central components of membrane fusion reactions, but their precise function has remained enigmatic. Competing hypotheses suggest roles for SNAREs in mediating the specificity of fusion, catalyzing fusion, or actually executing fusion. We generated knockout mice lacking synaptobrevin/VAMP 2, the vesicular SNARE ... More
Optical measurements of activity-dependent membrane recycling in motor nerve terminals of mammalian skeletal muscle.
AuthorsRibchester RR, Mao F, Betz WJ
JournalProc Biol Sci
PubMed ID8153137
Motor nerve terminals in a variety of rat and mouse skeletal muscles were stained in an activity-dependent fashion using the styryl dyes FM1-43 or FM2-10. Low-light video microscopy and digital image processing techniques were used to evaluate destaining of the preparations during application of depolarizing stimuli. Best results were obtained ... More
Kinetics and regulation of fast endocytosis at hippocampal synapses.
AuthorsKlingauf J, Kavalali ET, Tsien RW
JournalNature
PubMed ID9707119
Presynaptic nerve terminals often contain as few as a hundred vesicles and so must recycle them soon after exocytosis to preserve synaptic transmission and presynaptic morphology during repetitive firing. The kinetics and mechanisms of vesicular endocytosis and repriming have therefore been studied. Vesicles in hippocampal nerve terminals can become available ... More
Characterization of rapid membrane internalization and recycling.
AuthorsHao M, Maxfield FR
JournalJ Biol Chem
PubMed ID10809763
Lipids and other membrane constituents recycle between the plasma membrane and intracellular endocytic compartments. In CHO cells, approximately half of the internalized C(6)-NBD-SM, a fluorescent lipid analogue widely used as a membrane maker, recycles via the endocytic recycling compartment with a t(12) of approximately 12 min (Mayor, S., Presley, J. ... More
Two modes of exocytosis from synaptosomes are differentially regulated by protein phosphatase types 2A and 2B.
AuthorsBaldwin ML, Rostas JA, Sim AT
JournalJ Neurochem
PubMed ID12753078
The inhibitors okadaic acid (OA), fostriecin (FOS) and cyclosporin A (CsA), were used to investigate the roles of protein phosphatases in regulating exocytosis in rat brain synaptosomes by measuring glutamate release and the release of the styryl dye FM 2-10. Depolarization was induced by 30 mM KCl, or 0.3 mM ... More
Properties of the demarcation membrane system in living rat megakaryocytes.
The demarcation membrane system (DMS) is the precursor of platelet cell membranes yet little is known of its properties in living megakaryocytes. Using confocal microscopy, we now demonstrate that demarcation membranes in freshly isolated rat marrow megakaryocytes are rapidly stained by styryl membrane indicators such as di-8-ANEPPS and FM 2-10, ... More
Synaptic vesicle recycling in cultured cerebellar granule cells: role of vesicular acidification and refilling.
AuthorsCousin MA, Nicholls DG
JournalJ Neurochem
PubMed ID9349537
The role of the transvesicular protonmotive force in synaptic vesicle recycling was investigated in cultured cerebellar granule cells. The vesicular V-ATPase was inhibited by 1 microM bafilomycin A1; as an alternative, the pH component of the gradient was selectively collapsed by equilibration of the cells with 10 mM methylamine and ... More
Fluorescent styryl dyes FM1-43 and FM2-10 are muscarinic receptor antagonists: intravital visualization of receptor occupancy.
AuthorsMazzone SB, Mori N, Burman M, Palovich M, Belmonte KE, Canning BJ
JournalJ Physiol
PubMed ID16728454
The fluorescent styryl dyes FM1-43 and FM2-10 have been used to visualize the endocytic and exocytic processes involved in neurotransmission in a variety of central and peripheral nerve preparations. Their utility is limited to some extent by a poorly understood vesicular-independent labelling of cells and tissues. We show here that ... More
Effects of 2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one on synaptic vesicle cycling at the frog neuromuscular junction.
AuthorsRizzoli SO, Betz WJ
JournalJ Neurosci
PubMed ID12486161
Inositol phospholipids are thought to play an important regulatory role in synaptic membrane traffic. We investigated the effects of perturbing 3-phosphoinositide metabolism on neurotransmission at the frog neuromuscular junction. We used the reversible phosphoinositide-3 kinase (PI3K) inhibitor 2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one [LY294002 (LY)] and we examined its effects by intracellular recording, fluorescence imaging ... More
Perturbation of synaptic vesicle delivery during neurotransmitter release triggered independently of calcium influx.
AuthorsCongar P, Trudeau LE
JournalJ Physiol
PubMed ID12154178
Although much evidence suggests that calcium (Ca(2+)) usually triggers synaptic vesicle exocytosis and neurotransmitter release, the role of Ca(2+) in vesicle endocytosis and in the delivery of fusion-competent vesicles (i.e. mobilisation and/or priming) in nerve terminals remains unclear. To address this issue, we have studied synaptic vesicle dynamics in cultured ... More
In situ identification and visualization of neurons that mediate enteric and enteropancreatic reflexes.
AuthorsKirchgessner AL, Liu MT, Gershon MD
JournalJ Comp Neurol
PubMed ID8835732
To identify neurons participating in enteric and enteropancreatic reflexes, we validated the use of the activity-dependent markers FM1-43 and FM2-10 as "on-line" probes for the visualization of activated guinea pig enteric and pancreatic neurons. FM1-43 or FM2-10 labeling of neuronal perikarya and processes was induced by KCl (70 mM), veratridine ... More
Dynamic Growth and Shrinkage of the Salmonella-Containing Vacuole Determines the Intracellular Pathogen Niche.