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Title: The mechanisms of latrophilin 1-mediated control of spontaneous exocytosis at the mouse neuromuscular junction
Author: Petitto, Evelina
ISNI:       0000 0004 5922 9248
Awarding Body: University of Kent
Current Institution: University of Kent
Date of Award: 2015
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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 Gαq 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 Gαq 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 Gαq 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.
Supervisor: Ushkaryov, Yuri Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID:  DOI: Not available
Keywords: RS Pharmacy and materia medica