Fusion pores with low conductance are cation selective

Delacruz, Joannalyn; Sharma, Satyan; Rathore, Shailendra; Huang, Meng; Lenz, Joan Sesing; Lindau, Manfred

doi:10.1016/j.celrep.2021.109580

Cited by 3 publications

(3 citation statements)

References 56 publications

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“…Alternatively, it has been reported that the fusion pore is cation-selective ( 31 ). Since both Alexa 488 and CF488A are anionic, it is possible that the delay in external dye entry may be due to electrostatic repulsion.…”

Section: Resultsmentioning

confidence: 99%

High-speed imaging reveals the bimodal nature of dense core vesicle exocytosis

Zhang

Rumschitzki

Edwards

2022

Proc. Natl. Acad. Sci. U.S.A.

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During exocytosis, the fusion of secretory vesicle with plasma membrane forms a pore that regulates release of neurotransmitter and peptide. Heterogeneity of fusion pore behavior has been attributed to stochastic variation in a common exocytic mechanism, implying a lack of biological control. Using a fluorescent false neurotransmitter (FFN), we imaged dense core vesicle (DCV) exocytosis in primary mouse adrenal chromaffin cells by total internal reflection fluorescence microscopy at millisecond resolution and observed strikingly divergent modes of release, with fast events lasting <30 ms and slow events persisting for seconds. Dual imaging of slow events shows a delay in the entry of external dye relative to FFN release, suggesting exclusion by an extremely narrow pore <1 nm in diameter. Unbiased comprehensive analysis shows that the observed variation cannot be explained by stochasticity alone, but rather involves distinct mechanisms, revealing the bimodal nature of DCV exocytosis. Further, loss of calcium sensor synaptotagmin 7 increases the proportion of slow events without changing the intrinsic properties of either class, indicating the potential for independent regulation. The identification of two distinct mechanisms for release capable of independent regulation suggests a biological basis for the diversity of fusion pore behavior.

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Section: Resultsmentioning

confidence: 99%

High-speed imaging reveals the bimodal nature of dense core vesicle exocytosis

Zhang

Rumschitzki

Edwards

2022

Proc. Natl. Acad. Sci. U.S.A.

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“…However, surprisingly, in many other cases, apparent fluctuations of a foot signal preceding a spike were the result of overlapping catecholamine release activity at different locations, suggesting that distant exocytotic events and foot signals may occur in a synchronized manner. Thus, the interpretation of amperometric foot signals may seem less straightforward than typically assumed [ 43 ].…”

Section: Frontier Advances Reported At the Isccb-21 Meeting In Hamburgmentioning

confidence: 99%

Forty years of the adrenal chromaffin cell through ISCCB meetings around the world

Maneu

Borges

Gandı́a

et al. 2023

Pflugers Arch - Eur J Physiol

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This historical review focuses on the evolution of the knowledge accumulated during the last two centuries on the biology of the adrenal medulla gland and its chromaffin cells (CCs). The review emerged in the context of a series of meetings that started on the Spanish island of Ibiza in 1982 with the name of the International Symposium on Chromaffin Cell Biology (ISCCB). Hence, the review is divided into two periods namely, before 1982 and from this year to 2022, when the 21st ISCCB meeting was just held in Hamburg, Germany. The first historical period extends back to 1852 when Albert Kölliker first described the fine structure and function of the adrenal medulla. Subsequently, the adrenal staining with chromate salts identified the CCs; this was followed by the establishment of the embryological origin of the adrenal medulla, and the identification of adrenaline-storing vesicles. By the end of the nineteenth century, the basic morphology, histochemistry, and embryology of the adrenal gland were known. The twentieth century began with breakthrough findings namely, the experiment of Elliott suggesting that adrenaline was the sympathetic neurotransmitter, the isolation of pure adrenaline, and the deciphering of its molecular structure and chemical synthesis in the laboratory. In the 1950s, Blaschko isolated the catecholamine-storing vesicles from adrenal medullary extracts. This switched the interest in CCs as models of sympathetic neurons with an explosion of studies concerning their functions, i.e., uptake of catecholamines by chromaffin vesicles through a specific coupled transport system; the identification of several vesicle components in addition to catecholamines including chromogranins, ATP, opioids, and other neuropeptides; the calcium-dependence of the release of catecholamines; the underlying mechanism of exocytosis of this release, as indicated by the co-release of proteins; the cross-talk between the adrenal cortex and the medulla; and the emission of neurite-like processes by CCs in culture, among other numerous findings. The 1980s began with the introduction of new high-resolution techniques such as patch-clamp, calcium probes, marine toxins-targeting ion channels and receptors, confocal microscopy, or amperometry. In this frame of technological advances at the Ibiza ISCCB meeting in 1982, 11 senior researchers in the field predicted a notable increase in our knowledge in the field of CCs and the adrenal medulla; this cumulative knowledge that occurred in the last 40 years of history of the CC is succinctly described in the second part of this historical review. It deals with cell excitability, ion channel currents, the exocytotic fusion pore, the handling of calcium ions by CCs, the kinetics of exocytosis and endocytosis, the exocytotic machinery, and the life cycle of secretory vesicles. These concepts together with studies on the dynamics of membrane fusion with super-resolution imaging techniques at the single-protein level were extensively reviewed by top scientists in the field at the 21st ISCCB meeting in Hamburg in the summer of 2022; this frontier topic is also briefly reviewed here. Many of the concepts arising from those studies contributed to our present understanding of synaptic transmission. This has been studied in physiological or pathophysiological conditions, in CCs from animal disease models. In conclusion, the lessons we have learned from CC biology as a peripheral model for brain and brain disease pertain more than ever to cutting-edge research in neurobiology. In the 22nd ISCCB meeting in Israel in 2024 that Uri Asheri is organizing, we will have the opportunity of seeing the progress of the questions posed in Ibiza, and on other questions that undoubtedly will arise.

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“…Alternatively, it has been reported that the fusion pore is cation-selective (Delacruz et al, 2021). Since both Alexa 488 and CF488A are anionic, it is possible that the delay in external dye entry may be due to electrostatic repulsion.…”

Section: The Fusion Pore Can Limit Entry Of External Dyementioning

confidence: 99%

High-Speed Imaging Reveals the Bimodal Nature of Dense Core Vesicle Exocytosis

Zhang

Rumschitzki

Edwards

2021

Preprint

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During exocytosis, the fusion of secretory vesicle with plasma membrane forms a pore that regulates release of neurotransmitter and peptide. Osmotic forces contribute to exocytosis but release through the pore is thought to occur by diffusion. Heterogeneity of fusion pore behavior has also suggested stochastic variation in a common exocytic mechanism, implying a lack of biological control. Imaging at millisecond resolution to observe the first events in exocytosis, we find that fusion pore duration is bimodal rather than stochastic. Loss of calcium sensor synaptotagmin 7 increases the proportion of slow events without changing the intrinsic properties of either class, indicating the potential for independent regulation. In addition, dual imaging shows a delay in the entry of external dye relative to release that indicates discharge at high velocity rather than strictly by diffusion.

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Fusion pores with low conductance are cation selective

Abstract: Highlights d Exocytotic transmitter release is an electrodiffusion process d Narrow fusion pores are cation selective d Fusion pore ion selectivity decreases as the fusion pore expands

Cited by 3 publications

References 56 publications

High-speed imaging reveals the bimodal nature of dense core vesicle exocytosis

High-speed imaging reveals the bimodal nature of dense core vesicle exocytosis

Forty years of the adrenal chromaffin cell through ISCCB meetings around the world

High-Speed Imaging Reveals the Bimodal Nature of Dense Core Vesicle Exocytosis

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