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The mechanisms of latrophilin 1-mediated control of spontaneous exocytosis at the mouse neuromuscular junction

Petitto, Evelina (2015) The mechanisms of latrophilin 1-mediated control of spontaneous exocytosis at the mouse neuromuscular junction. Doctor of Philosophy (PhD) thesis, University of Kent,.

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Abstract

Latrophilin1 (LPHN1) is a presynaptic adhesion G protein-coupled receptor involved in the control of spontaneous exocytosis of neurotransmitters. The effects of LPHN1 activation on exocytosis have been described on several model systems, such as cultured hippocampal neurons and neuromuscular junction (NMJ) using its well known agonist LTXN4C, and include a massive increase in exocytosis characterised by periods of intense release (bursts) interspersed with periods of moderate activity (interburst intervals). However, the molecular mechanisms underlying these effects were yet to be determined. Based on previous observations that LPHN1 is associated to G proteins, and that its activation leads to activation of PLC and increased IP3, we hypothesize that LPHN1 controls exocytosis via the G?q protein pathway, whose activation ultimately results in the release of Ca2+ from IP3-sensitive Ca2+ stores. Using a pharmacological approach and the current clamp method at the mouse NMJ, we first used LPHN1 KO preparations to study the role of LPHN1 on spontaneous exocytosis in resting conditions, and to show that LPHN1 is the only receptor mediating the effects observed upon stimulation by LTXN4C. Then, we interfered with several molecules involved in the G?q pathway to test their involvement in LPHN1 activation, and we investigated the role of store-operated (SOCCs) and voltage-gated (VGCCs) Ca2+ channels in mediating the Ca2+ influx that is necessary for the development of LPHN1 effects. Our results support the hypothesis LPHN1 is involved in the regulation of spontaneous exocytosis at rest and that it is the receptor mediating the increased exocytosis following stimulation by LTXN4C; the suggestions that G?q and its intracellular pathway mediate the effects of LPHN1 activation on spontaneous exocytosis, and that SOCCs and VGCCs (particularly Cav2.1) mediate the Ca2+ influx necessary for the development of LPHN1 effects are also supported by our findings. Altogether, this work uncovered the mechanisms by which G protein-coupled receptors, in this case LPHN1, can regulate the rate of spontaneous neurotransmitter release at the mouse NMJ.

Item Type: Thesis (Doctor of Philosophy (PhD))
Thesis advisor: Ushkaryov, Yuri
Uncontrolled keywords: Spontaneous exocytosis; G protein-coupled receptors; latrophilin; neuroscience
Subjects: R Medicine > RS Pharmacy and materia medica
Divisions: Faculties > Sciences > Medway School of Pharmacy
Depositing User: Users 1 not found.
Date Deposited: 29 Feb 2016 14:37 UTC
Last Modified: 29 May 2019 17:03 UTC
Resource URI: https://kar.kent.ac.uk/id/eprint/54354 (The current URI for this page, for reference purposes)
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