Microtransplantation of cellular membranes from squid stellate ganglion reveals ionotropic GABA receptors

dc.contributor.author Conti, Luca
dc.contributor.author Limon, Agenor
dc.contributor.author Palma, Eleonora
dc.contributor.author Miledi, Ricardo
dc.date.accessioned 2013-03-19T15:21:06Z
dc.date.available 2013-03-19T15:21:06Z
dc.date.issued 2013-02-01
dc.description Author Posting. © Marine Biological Laboratory, 2013. This article is posted here by permission of Marine Biological Laboratory for personal use, not for redistribution. The definitive version was published in Biological Bulletin 224 (2013): 47-52. en_US
dc.description.abstract The squid has been the most studied cephalopod, and it has served as a very useful model for investigating the events associated with nerve impulse generation and synaptic transmission. While the physiology of squid giant axons has been extensively studied, very little is known about the distribution and function of the neurotransmitters and receptors that mediate inhibitory transmission at the synapses. In this study we investigated whether γ-aminobutyric acid (GABA) activates neurotransmitter receptors in stellate ganglia membranes. To overcome the low abundance of GABA-like mRNAs in invertebrates and the low expression of GABA in cephalopods, we used a two-electrode voltage clamp technique to determine if Xenopus laevis oocytes injected with cell membranes from squid stellate ganglia responded to GABA. Using this method, membrane patches containing proteins and ion channels from the squid's stellate ganglion were incorporated into the surface of oocytes. We demonstrated that GABA activates membrane receptors in cellular membranes isolated from squid stellate ganglia. Using the same approach, we were able to record native glutamate-evoked currents. The squid's GABA receptors showed an EC50 of 98 μmol l–1 to GABA and were inhibited by zinc (IC50 = 356 μmol l–1). Interestingly, GABA receptors from the squid were only partially blocked by bicuculline. These results indicate that the microtransplantation of native cell membranes is useful to identify and characterize scarce membrane proteins. Moreover, our data also support the role of GABA as an ionotropic neurotransmitter in cephalopods, acting through chloride-permeable membrane receptors. en_US
dc.description.sponsorship Grass Foundation Fellowships to L.C. and A.L. (www.grassfoundation.org). L.C. was additionally supported by the Ph.D. in Neurophysiology program of the University of Rome “La Sapienza.” All authors were Grass Fellows. This work was supported by Ministero della Sanita` Antidoping and PRIN project 2009 (to E.P.). en_US
dc.format.mimetype application/pdf
dc.identifier.citation Biological Bulletin 224 (2013): 47-52 en_US
dc.identifier.doi 10.1086/BBLv224n1p47
dc.identifier.uri https://hdl.handle.net/1912/5811
dc.language.iso en_US en_US
dc.publisher Marine Biological Laboratory en_US
dc.relation.uri https://doi.org/10.1086/BBLv224n1p47
dc.title Microtransplantation of cellular membranes from squid stellate ganglion reveals ionotropic GABA receptors en_US
dc.type Article en_US
dspace.entity.type Publication
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