Cloning and characterization of an ionotropic glutamate receptor subunit expressed in the squid nervous system.
In this paper we describe the cloning of a putative ionotropic glutamate receptor subunit, SqGluR, and its distribution in the nervous system of the squid. A full-length cDNA was assembled from a cDNA library of the stellate ganglion/giant fibre lobe complex of Loligo opalescens. The deduced amino acid sequence of the mature SqGluR displayed 44-46% amino acid identity with mammalian GluR1-GluR4 and 53% with Lym-eGluR1 from Lymnaea stagnalis. In situ hybridizations in adult squid confirmed that the SqGluR mRNA is abundant in giant fibre lobe neurons, in large, presumptive motor neurons of the stellate ganglion proper and in the supraoesophageal and optic lobes of the central nervous system. In newborn squid, SqGluR mRNA expression was detected throughout the nervous system but not elsewhere. A synthetic peptide corresponding to the last 15 amino acids of the SqGluR C-terminus was used to generate polyclonal antibodies, which were used for immunoblot analysis to demonstrate widespread expression in the squid central and peripheral nervous systems. Injection of the synthetic peptide into the postsynaptic side of the giant synapse inhibited synaptic transmission.
Battaglia
A A
AA
King's College London, Centre for Neuroscience Research, Sensory Function Group, Guy's Campus, Hodgkin Building, London Bridge, London SE1 1UL, UK.
Nardi
G
G
Steinhardt
A
A
Novakovic
A
A
Gentile
S
S
Iaccarino Idelson
P
P
Gilly
W F
WF
de Santis
A
A
D000595:Amino Acid Sequence; D000818:Animals; D000831:Animals, Newborn / Q000254:growth & development / Q000378:metabolism; D003001:Cloning, Molecular; D049832:Decapodiformes; D004558:Electric Stimulation; D004594:Electrophysiology; D019706:Excitatory Postsynaptic Potentials / Q000187:drug effects / Q000502:physiology; D015723:Gene Library; D007987:Gonadotropin-Releasing Hormone / Q000276:immunology / Q000378:metabolism; D015151:Immunoblotting; D017403:In Situ Hybridization; D066298:In Vitro Techniques; D008845:Microinjections; D008969:Molecular Sequence Data; D009420:Nervous System / Q000254:growth & development / Q000378:metabolism*; D009474:Neurons / Q000378:metabolism*; D010455:Peptides / Q000276:immunology / Q000378:metabolism / Q000494:pharmacology; D021122:Protein Subunits / Q000737:chemistry / Q000235:genetics*; D016372:RNA, Antisense / Q000378:metabolism; D012333:RNA, Messenger / Q000096:biosynthesis / Q000302:isolation & purification; D017470:Receptors, Glutamate / Q000737:chemistry / Q000235:genetics*; D017386:Sequence Homology, Amino Acid; D013233:Stellate Ganglion / Q000187:drug effects / Q000502:physiology; D014158:Transcription, Genetic
D010455:Peptides; D021122:Protein Subunits; D016372:RNA, Antisense; D012333:RNA, Messenger; D017470:Receptors, Glutamate; D007987:Gonadotropin-Releasing Hormone