98%
921
2 minutes
20
Neurons and endocrine cells secrete neurotransmitter and hormones in discrete packets in a process called quantal exocytosis. Electrochemical microelectrodes can detect spikes in current resulting from the oxidation of individual quanta of transmitter only if the electrodes are small and directly adjacent to release sites on the cell. Here we report development of a microchip device that uses microfluidic traps to automatically target individual or small groups of cells to small electrochemical electrodes. Microfluidic channels and traps were fabricated by multi-step wet etch of a silicon wafer whereas Pt electrodes were patterned in register with the trap sites. We demonstrate high-resolution amperometric measurement of quantal exocytosis of catecholamines from chromaffin cells on the device. This reusable device is a step towards developing high-throughput lab-on-a-chip instruments for recording quantal exocytosis to increase the pace of basic neuroscience research and to enable screening of drugs that target exocytosis.
Download full-text PDF |
Source |
---|---|
http://dx.doi.org/10.1039/b913216c | DOI Listing |
Front Cell Dev Biol
May 2025
Department of Cell and Developmental Biology, Gray Faculty of Medical and Health Sciences, Tel Aviv University, Tel Aviv, Israel.
Mast cells (MCs) are sentinel cells of the immune system that play important protective roles in innate host defenses but are also key effectors of allergic responses and chronic inflammatory diseases. Both physiological and pathophysiological responses of MCs are mediated by the release of inflammatory mediators, many of which are stored, preformed, in secretory granules (SGs), and released by regulated exocytosis in response to multiple stimuli. MC SGs belong to the family of lysosome related organelles (LROs), as indicated by their content of lysosomal hydrolases, lysosomal membrane proteins and acidic pH.
View Article and Find Full Text PDFJ Physiol
June 2025
Institute of Molecular and Cellular Biology, National Taiwan University, Taipei, Taiwan.
Ca-dependent exocytosis initiates with the formation of fusion pores comprising the soluble N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE) complex. Although cellular signalling typically occurs in transient oscillations on the order of tens of seconds, it remains unclear how such rapid SNARE phosphorylation influences fusion pore kinetics, analogous to transient regulation observed in ion channels. Here we demonstrate that protein kinase A (PKA)-mediated phosphorylation of SN25b (the neuronal isoform of synaptosome-associated protein of 25 kD) modulates secretion rate and fusion pore kinetics in PC12 cells (rat pheochromocytoma derivatives).
View Article and Find Full Text PDFNat Struct Mol Biol
August 2025
Departments of Neurology and Physiology, UCSF School of Medicine, San Francisco, CA, USA.
The concentration of neurotransmitters inside synaptic vesicles (SVs) underlies the quantal nature of synaptic transmission. Uptake of glutamate, the principal excitatory neurotransmitter, is driven by membrane potential. To prevent nonquantal efflux of glutamate after SV exocytosis, the vesicular glutamate transporters (VGLUTs) are allosterically inhibited by the neutral pH of the synaptic cleft.
View Article and Find Full Text PDFBiophys Rep (N Y)
June 2025
National Institute of Neurological Disorders and Stroke, 35 Convent Dr., Bldg. 35, Bethesda, MD 20892, USA. Electronic address:
Exocytosis, which mediates important functions like synaptic transmission and stress responses, has been postulated to release all transmitter molecules in the vesicle in the "all-or-none" quantal hypothesis. Challenging this hypothesis, amperometric current recordings of catecholamine release propose that sub-quantal or partial transmitter release is dominant in various cell types, particularly chromaffin cells. The sub-quantal hypothesis predicts that fusion pore closure (kiss-and-run fusion), the cause of sub-quantal release, is dominant, and blocking pore closure increases quantal size.
View Article and Find Full Text PDFNeurochem Res
November 2024
Laboratory of Biophysics of Synaptic Processes, Kazan Institute of Biochemistry and Biophysics, FRC Kazan Scientific Center of RAS, 2/31 Lobachevsky St, Kazan, 420111, Russia.