Calcium Transient Registration in Response to Single Stimulation and During Train of Pulses in Mouse Neuromuscular Junction

Samigullin, Dmitry; Khaziev, Eduard; Zhilyakov, Nikita; Sudakov, Igor A.; Bukharaeva, É. A.; Nikolsky, E. E.

doi:10.1007/s12668-016-0318-6

Cited by 7 publications

(3 citation statements)

References 16 publications

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“…It is noteworthy that this loading method is suitable only for preparations that can withstand long incubations. To reduce the dye-loading time when studies are being conducted on more fragile tissues ( e.g., synapses of warm-blooded animals), it is necessary to downscale the nerve stump length and to use micropipettes for loading 29 39 .…”

Section: Discussionmentioning

confidence: 99%

Loading a Calcium Dye into Frog Nerve Endings Through the Nerve Stump: Calcium Transient Registration in the Frog Neuromuscular Junction

Samigullin

Khaziev

Zhilyakov

et al. 2017

JoVE

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One of the most feasible methods of measuring presynaptic calcium levels in presynaptic nerve terminals is optical recording. It is based on using calcium-sensitive fluorescent dyes that change their emission intensity or wavelength depending on the concentration of free calcium in the cell. There are several methods used to stain cells with calcium dyes. Most common are the processes of loading the dyes through a micropipette or pre-incubating with the acetoxymethyl ester forms of the dyes. However, these methods are not quite applicable to neuromuscular junctions (NMJs) due to methodological issues that arise. In this article, we present a method for loading a calcium-sensitive dye through the frog nerve stump of the frog nerve into the nerve endings. Since entry of external calcium into nerve terminals and the subsequent binding to the calcium dye occur within the millisecond time-scale, it is necessary to use a fast imaging system to record these interactions. Here, we describe a protocol for recording the calcium transient with a fast CCD camera.

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Section: Discussionmentioning

confidence: 99%

Loading a Calcium Dye into Frog Nerve Endings Through the Nerve Stump: Calcium Transient Registration in the Frog Neuromuscular Junction

Samigullin

Khaziev

Zhilyakov

et al. 2017

JoVE

Self Cite

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“…The last suggestion is very important, because it has been shown earlier that nNAChRs are more permeable to Ca 2+ as compared to permeability of their muscle-type counterparts [40,41]. At the same time, it was shown that cadmium up to a concentration of 200 µM does not block currents through nNAChRs [42], but significantly blocks currents through VGCCs [43,44].…”

Section: Presynaptic Cholinergic Receptors and The Role Of Calcium Influx In The Mechanism Of Ach Release Autoinhibitionmentioning

confidence: 97%

“…Nerve motor endings were loaded with Oregon Green 488 BAPTA-1 Hexapotassium Salt 1 mM high-affinity calcium-sensitive dye (Molecular Probes, Eugene, OR, USA) through the nerve stump, as described previously [44]. The fluorescence signal was recorded using an imaging setup based on an Olympus BX-51 microscope with a ×40 water-immersion objective (Olympus, Tokyo, Japan).…”

Section: Calcium Transient Recordingmentioning

confidence: 99%

Activation of Neuronal Nicotinic Receptors Inhibits Acetylcholine Release in the Neuromuscular Junction by Increasing Ca2+ Flux through Cav1 Channels

Zhilyakov

Arkhipov

Malomouzh

et al. 2021

IJMS

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Cholinergic neurotransmission is a key signal pathway in the peripheral nervous system and in several branches of the central nervous system. Despite the fact that it has been studied extensively for a long period of time, some aspects of its regulation still have not yet been established. One is the relationship between the nicotine-induced autoregulation of acetylcholine (ACh) release with changes in the concentration of presynaptic calcium levels. The mouse neuromuscular junction of m. Levator Auris Longus was chosen as the model of the cholinergic synapse. ACh release was assessed by electrophysiological methods. Changes in calcium transients were recorded using a calcium-sensitive dye. Nicotine hydrogen tartrate salt application (10 μM) decreased the amount of evoked ACh release, while the calcium transient increased in the motor nerve terminal. Both of these effects of nicotine were abolished by the neuronal ACh receptor antagonist dihydro-beta-erythroidine and Cav1 blockers, verapamil, and nitrendipine. These data allow us to suggest that neuronal nicotinic ACh receptor activation decreases the number of ACh quanta released by boosting calcium influx through Cav1 channels.

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Activation of TRPV1 Channels Inhibits the Release of Acetylcholine and Improves Muscle Contractility in Mice

Arkhipov,

Fedorov,

Nurullin

et al. 2023

Cell Mol Neurobiol

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Calcium Transient Registration in Response to Single Stimulation and During Train of Pulses in Mouse Neuromuscular Junction

Cited by 7 publications

References 16 publications

Loading a Calcium Dye into Frog Nerve Endings Through the Nerve Stump: Calcium Transient Registration in the Frog Neuromuscular Junction

Loading a Calcium Dye into Frog Nerve Endings Through the Nerve Stump: Calcium Transient Registration in the Frog Neuromuscular Junction

Activation of Neuronal Nicotinic Receptors Inhibits Acetylcholine Release in the Neuromuscular Junction by Increasing Ca2+ Flux through Cav1 Channels

Activation of TRPV1 Channels Inhibits the Release of Acetylcholine and Improves Muscle Contractility in Mice

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