Mutation of the atrophin2 gene in the zebrafish disrupts signaling by fibroblast growth factor during development of the inner ear.
AuthorsAsai Y, Chan DK, Starr CJ, Kappler JA, Kollmar R, Hudspeth AJ
JournalProc Natl Acad Sci U S A
PubMed ID16754885
'The development of the vertebrate inner ear depends on the precise expression of fibroblast growth factors. In a mutagenesis screen for zebrafish with abnormalities of inner-ear development and behavior, we isolated a mutant line, ru622, whose phenotypic characteristics resembled those of null mutants for the gene encoding fibroblast growth factor ... More
Regulation of human organic cation transporter hOCT2 by PKA, PI3K, and calmodulin-dependent kinases.
AuthorsCetinkaya I, Ciarimboli G, Yalçinkaya G, Mehrens T, Velic A, Hirsch JR, Gorboulev V, Koepsell H, Schlatter E
JournalAm J Physiol Renal Physiol
PubMed ID12388397
'Properties and regulation of the human organic cation (OC) transporter type 2 (hOCT2) expressed in HEK-293 cells were extensively characterized using the fluorescent OC 4-[4-(dimethylamino)styryl]-N-methylpyridinium (ASP(+)). ASP(+) uptake was electrogenic and inhibited by TPA(+) (EC(50) = 2.7 microM), tetraethylammonium (EC(50) = 35 microM), cimetidine (EC(50) = 36 microM), or quinine ... More
Differential physiology and morphology of motor axons to ventral longitudinal muscles in larval Drosophila.
AuthorsKurdyak P, Atwood HL, Stewart BA, Wu CF
JournalJ Comp Neurol
PubMed ID7884051
'Morphological and physiological characteristics of the two major motor axons supplying the commonly studied ventral longitudinal muscle fibers (6 and 7) of third-instar Drosophila melanogaster larvae were investigated. The innervating terminals of the two motor axons differ in the size of their synapse-bearing varicosities. The terminal with the larger varicosities ... More
Synaptic physiology and mitochondrial function in crayfish tonic and phasic motor neurons.
AuthorsNguyen PV, Marin L, Atwood HL
JournalJ Neurophysiol
PubMed ID9242280
'Phasic and tonic motor neurons of crustaceans differ strikingly in their junctional synaptic physiology. Tonic neurons generally produce small excitatory postsynaptic potentials (EPSPs) that facilitate strongly as stimulation frequency is increased, and normally show no synaptic depression. In contrast, phasic neurons produce relatively large EPSPs with weak frequency facilitation and ... More
Intracellular delivery mediated by an ethosomal carrier.
AuthorsTouitou E, Godin B, Dayan N, Weiss C, Piliponsky A, Levi-Schaffer F
JournalBiomaterials
PubMed ID11575480
'The goal of this work was to investigate the efficiency of transcellular delivery into Swiss albino mice 3T3 fibroblasts of molecules with various physico-chemical characteristics from ethosomes, phospholipid vesicular carriers containing ethanol. The probes chosen were: 4-(4-diethylamino) styryl-N-methylpyridinium iodide (D-289), rhodamine red dihexadecanoylglycerophosphoethanolamine (RR) and fluorescent phosphatidylcholine (PC*). The penetration ... More
In vivo observations of pre- and postsynaptic changes during the transition from multiple to single innervation at developing neuromuscular junctions.
AuthorsBalice-Gordon RJ, Lichtman JW
JournalJ Neurosci
PubMed ID8426240
'Synaptic rearrangements in developing muscle were studied by visualizing individual neuromuscular junctions in the sternomastoid muscle of living neonatal mice as they underwent the transition from multiple to single innervation. Vital staining of ACh receptors (AChRs) with rhodamine-conjugated alpha-bungarotoxin showed that while junctions were still multiply innervated (usually by two ... More
The remodeling of synaptic extracellular matrix and its dynamic relationship with nerve terminals at living frog neuromuscular junctions.
AuthorsChen LL, Folsom DB, Ko CP
JournalJ Neurosci
PubMed ID1715392
'The question of whether the synaptic extracellular matrix undergoes remodeling and how this remodeling is related to nerve terminal plasticity was examined in living neuromuscular junctions of adult frogs. Sartorius muscles were double stained with a fluorescent nerve terminal dye 4-(4-diethylamino-styryl)-N-methylpyridinium iodide (4-Di-2-Asp) and rhodamine-tagged peanut agglutinin (PNA) which recognizes ... More
Binding and transport in norepinephrine transporters. Real-time, spatially resolved analysis in single cells using a fluorescent substrate.
AuthorsSchwartz JW, Blakely RD, DeFelice LJ
JournalJ Biol Chem
PubMed ID12499385
'Monoamine transporters, the molecular targets for drugs of abuse and antidepressants, clear norepinephrine, dopamine, or serotonin from the synaptic cleft. Neurotransmitters, amphetamines, and neurotoxins bind before being transported, whereas cocaine and antidepressants bind to block transport. Although binding is crucial to transport, few assays separate binding from transport, nor do ... More
The concentration jump method. Kinetics of vital staining of mitochondria in HeLa cells with lipophilic cationic fluorescent dyes.
AuthorsIrion G, Ochsenfeld L, Naujok A, Zimmermann HW
JournalHistochemistry
PubMed ID7682211
'Lipophilic cationic fluorescent dyes (D) specifically stain the mitochondria of living cells. A perfusion chamber for cell cultures is described, which can be used to determine the kinetics of vital staining of the mitochondria of single selected cells in situ. In these experiments styrylpyridinium dyes and cultures of HeLa cells ... More
Regenerating muscle fibers induce directional sprouting from nearby nerve terminals: studies in living mice.
Authorsvan Mier P, Lichtman JW
JournalJ Neurosci
PubMed ID8083762
'The principal aim of this work was to better understand how regenerating muscle fibers become innervated in adult animals. To induce muscle regeneration, individual identified muscle fibers in a mouse were damaged with a laser focused through a microscope. The muscle fiber that degenerated and the muscle fiber that was ... More
Regular and photodamage-enhanced remodelling in vitally stained frog and mouse neuromuscular junctions.
AuthorsLangenfeld-Oster B, Dorlöchter M, Wernig A
JournalJ Neurocytol
PubMed ID7691996
'Repeated in vivo observations of vitally stained neuromuscular junctions allow direct monitoring of ongoing structural changes, although, normally occurring changes (remodelling) and those inflicted by the illumination itself (photodamage) need to be dissociated. In frog cutaneus pectoris muscles, stained in vivo with 4Di-2ASP twice within four to five weeks, growth ... More
Validation of a fluorescence-based high-throughput assay for the measurement of neurotransmitter transporter uptake activity.
'Pre-synaptic dopamine, norepinephrine and serotonin transporters (DAT, NET and SERT) terminate synaptic catecholamine transmission through reuptake of released neurotransmitter. Common approaches for studying these transporters involve radiolabeled substrates or inhibitors which, however, have several limitations. In this study we have used a novel neurotransmitter transporter uptake assay kit. The assay ... More
Functional live cell imaging of the pulmonary neuroepithelial body microenvironment.
AuthorsDe Proost I, Pintelon I, Brouns I, Kroese AB, Riccardi D, Kemp PJ, Timmermans JP, Adriaensen D,
JournalAm J Respir Cell Mol Biol
PubMed ID18367726
Pulmonary neuroepithelial bodies (NEBs) are densely innervated groups of neuroendocrine cells invariably accompanied by Clara-like cells. Together with NEBs, Clara-like cells form the so-called
Interlayer neurones in the rat superior colliculus: a tracer study using Dil/Di-ASP.
AuthorsHilbig H, Schierwagen A
JournalNeuroreport
PubMed ID8003679
Five different populations of interlayer neurones (ILNs) can be described after DiI/Di-ASP tracing in rat superior colliculus (SC). All of these labelled neurones preferentially lay in the rostro-medial part of the SC. Most of them are located in the stratum opticum and in the stratum griseum superficiale. Our results indicate ... More
A simple and rapid method for staining the enteric ganglia: application for Hirschsprung's disease.
AuthorsHanani M, Udassin R, Ariel I, Freund HR
JournalJ Pediatr Surg
PubMed ID8229573
A simple and reliable method is described for staining the nerve plexuses of the human gastrointestinal tract in whole-mounts. The staining does not require fixation and consists of a single step of 6 to 10 minutes incubation in a solution of the fluorescent dye 4-(4-diethylaminostyryl)-methylpyridinium iodide. Aganglionic bowel specimens from ... More
Fluorescent probes that stain living nerve terminals.
AuthorsMagrassi L, Purves D, Lichtman JW
JournalJ Neurosci
PubMed ID3572476
We have evaluated the efficacy of 18 cationic mitochondrial dyes that, as a class, show some ability to stain living nerve terminals. Several of these agents provide excellent staining of neuromuscular junctions in a wide range of species. More detailed studies of the most effective of these dyes--4-(4-diethylaminostyryl)-N-methylpyridinium iodide (4-Di-2-ASP)--indicate ... More
Visualization of neuromuscular junctions over periods of several months in living mice.
AuthorsLichtman JW, Magrassi L, Purves D
JournalJ Neurosci
PubMed ID3572477
Identified neuromuscular junctions were followed in the sternomastoid muscle of living mice for several months by repeated staining with the fluorescent dye 4-(4-diethylaminostyryl)-N-methylpyridinium iodide (4-Di-2-ASP; Magrassi et al., 1987). Overall terminal growth occurred at many endplates; however, the branching pattern of presynaptic arbors was largely unchanged, even after several months. ... More
Recordings from human myenteric neurons using voltage-sensitive dyes.
AuthorsVignali S, Peter N, Ceyhan G, Demir IE, Zeller F, Senseman D, Michel K, Schemann M,
JournalJ Neurosci Methods
PubMed ID20691728
Voltage-sensitive dye (VSD) imaging became a powerful tool to detect neural activity in the enteric nervous system, including its routine use in submucous neurons in freshly dissected human tissue. However, VSD imaging of human myenteric neurons remained a challenge because of limited visibility of the ganglia and dye accessibility. We ... More
Vital imaging of glomeruli in the mouse olfactory bulb.
AuthorsLaMantia AS, Pomeroy SL, Purves D
JournalJ Neurosci
PubMed ID1545246
We have monitored the pattern of identified glomeruli in the olfactory bulbs of newborn, juvenile, and adult mice over intervals of several hours to several weeks. Our purpose was to assess the development and stability of these complex units in the mammalian brain. Glomeruli can be observed by vital fluorescent ... More
Assessment of nerve ultrastructure by fibre-optic confocal microscopy.
AuthorsCushway TR, Lanzetta M, Cox G, Trickett R, Owen ER
JournalMicrosurgery
PubMed ID9393664
Fibre-optic technology combined with confocality produces a microscope capable of optical thin sectioning. In this original study, tibial nerves have been stained in a rat model with a vital dye, 4-(4-diethylaminostyryl)-N-methylpyridinium iodide, and analysed by fibre-optic confocal microscopy to produce detailed images of nerve ultrastructure. Schwann cells, nodes of Ranvier ... More
Confocal microscopy of the lizard motor nerve terminals.
AuthorsMelamed N, Rahamimoff R
JournalJ Basic Clin Physiol Pharmacol
PubMed ID1723895
Confocal imaging was performed on the ceratomandibularis nerve muscle preparation of the lizard Anolis carolinensis, using 4-Di-2-ASP as a fluorescent probe. The imaging system consisted of a Sarastro Phoibos 1000 (Molecular Dynamics) scanning system and a Zeiss Universal microscope. The data were analyzed using the VANIS set of programs on ... More
Visualization of enteric and gallbladder ganglia with a vital fluorescent dye.
AuthorsHanani M
JournalJ Auton Nerv Syst
PubMed ID1619213
A method is described here for the vital staining of the nerve plexuses of the gastrointestinal tract with the fluorescent dye 4-(4-diethylaminostyryl)-methylpyridinium iodide (4-Di-2-ASP). This dye stained nerve fibers in the myenteric and submucosal plexuses and in the intramural ganglia of the gallbladder. In addition to staining the ganglia, the ... More
Characterization of Merkel cells and mechanosensory axons of the rat by styryl pyridinium dyes.
AuthorsNurse CA, Farraway L
JournalCell Tissue Res
PubMed ID2472215
The epidermal Merkel cells and their sensory innervation serve tactile sensation in vertebrates. In this study the fluorescent cationic mitochondrial dye, 4-(4-diethylaminostyryl)-N-methylpyridinium iodide (4-Di-2-ASP), which has recently been used as a vital stain for motor and autonomic nerve terminals, was tested for its ability to stain Merkel cells and sensory ... More
Visualization of human choroidal ganglion cells with the supravital fluorescent dye 4-(4-diethylaminostyryl)-N-methylpyridium iodide.
AuthorsBergua A, Neuhuber WL, Naumann GO
JournalOphthalmic Res
PubMed ID7877799
The distribution and morphology of ganglion cell bodies and nerve fibers in supravital human choroidea is studied by staining with the fluorescent dye 4-(4-diethylaminostyryl)-N-methylpyridium iodide (4-Di-2-ASP). We have used the choroids from human donor eyes donated for cornea transplantation. The isolated tissues were rinsed in ringer solution and processed within ... More
Properties of "reconstructed" motor synapses of the garter snake.
AuthorsWilkinson RS, Lunin SD
JournalJ Neurosci
PubMed ID8182475
We have developed a technique, called synaptic reconstruction, that permits nerve terminals of living vertebrate neuromuscular junctions (NMJs) to be isolated and then manually recombined with vacant endplate sites to form functional synapses. By reconstructing NMJs with various combinations of pre- and postsynaptic partners, or with varying degrees of pre- ... More
The use and effects of vital fluorescent dyes: observation of motor nerve terminals and satellite cells in living frog muscles.
AuthorsHerrera AA, Banner LR
JournalJ Neurocytol
PubMed ID2351996
Several different fluorescent mitochondrial dyes were tested as vital stains for motor nerve terminals and other cells in frog skeletal muscles. It was found that 3,3' diethyloxadicarbocyanine iodide and 4-(4-diethylaminostyryl)-N-methylpyridinium iodide were most useful. Both dyes labelled motor nerve terminals with high reliability. Electrophysiological and morphological control experiments showed that ... More
Functional repair of motor endplates after botulinum neurotoxin type A poisoning: biphasic switch of synaptic activity between nerve sprouts and their parent terminals.
Authorsde Paiva A, Meunier FA, Molgó J, Aoki KR, Dolly JO
JournalProc Natl Acad Sci U S A
PubMed ID10077661
Blockade of acetylcholine release by botulinum neurotoxin type A at the neuromuscular junction induces the formation of an extensive network of nerve-terminal sprouts. By repeated in vivo imaging of N-(3-triethyl ammonium propyl)-4-(4-(dibutylamino)styryl) pyridinium dibromide uptake into identified nerve endings of the mouse sternomastoid muscle after a single intramuscular injection of ... More
Repeated, in vivo observation of frog neuromuscular junctions: remodelling involves concurrent growth and retraction.
AuthorsHerrera AA, Banner LR, Nagaya N
JournalJ Neurocytol
PubMed ID2351997
The fluorescent dye 4-(4-diethylaminostyryl)-N-methylpyridinium iodide was used as a vital stain to study remodelling of motor nerve terminals in sartorius muscles of living frogs (Rana pipiens). Identified terminals were observed twice in vivo at intervals of 87-192 days. After the second observation, muscles were fixed and stained with the nitroblue ... More
"Satellite cells" and nerve terminals in the crayfish opener muscle visualized with fluorescent dyes.
AuthorsHarrington CC, Atwood HL
JournalJ Comp Neurol
PubMed ID8550891
Nerve terminals and associated cells on the muscle's surface were visualized in the crayfish opener muscle with several fluorescent dyes in conjunction with confocal microscopy and conventional fluorescence microscopy. The nerve terminals of the excitatory and inhibitory axons were best seen with 4-diethylaminostyryl-N-methylpyridinium iodide (4-Di-2-Asp). This dye is selectively accumulated ... More
Structural and functional specialization of A delta and C fiber free nerve endings innervating rabbit corneal epithelium.
AuthorsMacIver MB, Tanelian DL
JournalJ Neurosci
PubMed ID8410200
An in vitro preparation of rabbit cornea was used to compare anatomical specialization and electrophysiological responses of A delta and C fiber sensory afferents which terminate as free nerve endings. Living nerve endings were visualized using epifluorescence microscopy and the vital dye 4-di2-ASP, and response properties were determined using microstimulation ... More
Localization of L-type Ca2+ channels at perisynaptic glial cells of the frog neuromuscular junction.
AuthorsRobitaille R, Bourque MJ, Vandaele S
JournalJ Neurosci
PubMed ID8613781
The presence of L-type Ca2+ channels at the frog neuromuscular junction (nmj) was studied by monitoring changes in intracellular Ca2+ evoked in presynaptic terminals and perisynaptic Schwann cells (PSCs) and by studying the distribution of Ca2+ channels using a monoclonal antibody directed against the alpha 2/delta subunit of L channels. ... More
Muscle length alters geometry of arterioles and venules in hamster retractor.
AuthorsNakao M, Segal SS
JournalAm J Physiol
PubMed ID7840281
We investigated how changes in muscle length (Lm) would alter the geometry of arterioles and venules and whether such an effect would depend on the orientation of microvessels to muscle fibers. The parallel-fibered retractor muscle of anesthetized (pentobarbital sodium, 65 mg/kg) male hamsters (n = 20, 105 +/- 4 g) ... More
Sarcomere length and capillary curvature of rat hindlimb muscles in vivo.
AuthorsLedvina MA, Segal SS
JournalJ Appl Physiol
PubMed ID7665398
Mammalian skeletal muscle fibers have been reported to develop maximum force at a sarcomere length (Ls) of approximately 2.5 microns. However, the functional range of muscle length (Lm) and Ls encountered by skeletal muscle in vivo is not well defined. Changes in Ls markedly influence capillary geometry, but this effect ... More
Imaging neuromuscular junctions by confocal fluorescence microscopy: individual endplates seen in whole muscles with vital intracellular staining of the nerve terminals.
AuthorsMarques MJ, Santo Neto H
JournalJ Anat
PubMed ID9688508
The mammalian neuromuscular junction has been extensively studied by different methods to understand better the biological aspects of its normal development, ageing and pathological conditions, such as disorders of neuromuscular transmission. In the present report, a new technique is described that combines confocal microscopy with the use of a vital ... More
Repeated in vivo visualization of neuromuscular junctions in adult mouse lateral gastrocnemius.
AuthorsWigston DJ
JournalJ Neurosci
PubMed ID2355249
The structure of individually identified neuromuscular junctions (NMJs) in mouse lateral gastrocnemius (LG) muscles was studied on 2 or more occasions over 3-6 months. Presynaptic motor nerve terminals and their underlying acetylcholine receptors were stained in living animals with the fluorescent dye 4-(4-diethylaminostyryl)-N-methylpyridinium iodide) and tetramethylrhodamine isothiocyanate-conjugated alpha-bungarotoxin (R alpha ... More
Extension of synaptic extracellular matrix during nerve terminal sprouting in living frog neuromuscular junctions.
AuthorsChen L, Ko CP
JournalJ Neurosci
PubMed ID8301362
Remodeling of the synaptic extracellular matrix (ECM) and its dynamic relationship with nerve terminal plasticity have been demonstrated in normal frog neuromuscular junctions (NMJs) in vivo (Chen et al., 1991). Our previous work has led to a hypothesis that extension of synaptic ECM precedes nerve terminal growth during synaptic remodeling. ... More
D2 receptors regulate dopamine transporter function via an extracellular signal-regulated kinases 1 and 2-dependent and phosphoinositide 3 kinase-independent mechanism.
AuthorsBolan EA, Kivell B, Jaligam V, Oz M, Jayanthi LD, Han Y, Sen N, Urizar E, Gomes I, Devi LA, Ramamoorthy S, Javitch JA, Zapata A, Shippenberg TS
JournalMol Pharmacol
PubMed ID17267664
The dopamine transporter (DAT) terminates dopamine (DA) neurotransmission by reuptake of DA into presynaptic neurons. Regulation of DA uptake by D(2) dopamine receptors (D(2)R) has been reported. The high affinity of DA and other DAT substrates for the D(2)R, however, has complicated investigation of the intracellular mechanisms mediating this effect. ... More
Synaptic differentiation of a single motor neuron: conjoint definition of transmitter release, presynaptic calcium signals, and ultrastructure.
AuthorsCooper RL, Marin L, Atwood HL
JournalJ Neurosci
PubMed ID7790906
The opener muscle in the walking legs of the crayfish (Procambarus clarkii) is innervated by only one excitatory motor neuron, yet excitatory postsynaptic potentials (EPSPs) of proximal fibers are eightfold larger than those of central muscle fibers at low frequencies of activation, due in large measure to differences in presynaptic ... More
Vital confocal microscopy in bone.
AuthorsBoyde A, Wolfe LA, Maly M, Jones SJ
JournalScanning
PubMed ID7767496
We wished to exploit confocal microscopy for high spatial and temporal resolution vital microscopy in bone. To this end, we evolved implants with glass windows supported in titanium, which were placed in the medial proximal tibial plateau of the rabbit, and special small, self-focussing objectives (dry 10/0.25, water immersion 20/0.45, ... More
Novel fluorescence-based approaches for the study of biogenic amine transporter localization, activity, and regulation.
Pre-synaptic norepinephrine (NE) and dopamine (DA) transporters (NET and DAT) terminate catecholamine synaptic transmission through reuptake of released neurotransmitter. Recent studies reveal that NET and DAT are tightly regulated by receptor and second messenger-linked signaling pathways. Common approaches for studying these transporters involve use of radiolabeled substrates or antagonists, methods ... More