Cloning and characterization of a Caenorhabditis elegans D2‐like dopamine receptor

The neurotransmitter dopamine plays an important role in the regulation of behavior in both vertebrates and invertebrates. In mammals, dopamine binds and activates two classes of dopamine receptors, D1‐like and D2‐like receptors. However, D2‐like dopamine receptors in Caenorhabditis elegans have not yet been characterized. We have cloned a cDNA encoding a putative C. elegans D2‐like dopamine receptor. The deduced amino acid sequence of the cloned cDNA shows higher sequence similarities to vertebrate D2‐like dopamine receptors than to D1‐like receptors. Two splice variants that differ in the length of their predicted third intracellular loops were identified. The receptor heterologously expressed in cultured cells showed high affinity binding to [125I]iodo‐lysergic acid diethylamide. Dopamine showed the highest affinity for this receptor among several amine neurotransmitters tested. Activation of the heterologously expressed receptor led to the inhibition of cyclic AMP production, confirming that this receptor has the functional property of a D2‐like receptor. We have also analyzed the expression pattern of this receptor and found that the receptor is expressed in several neurons including all the dopaminergic neurons in C. elegans.

[1]  Rajesh Ranganathan,et al.  C. elegans Locomotory Rate Is Modulated by the Environment through a Dopaminergic Pathway and by Experience through a Serotonergic Pathway , 2000, Neuron.

[2]  Ronald L. Davis,et al.  DAMB, a Novel Dopamine Receptor Expressed Specifically in Drosophila Mushroom Bodies , 1996, Neuron.

[3]  C. Gerfen,et al.  Multiple D2 dopamine receptors produced by alternative RNA splicing , 1989, Nature.

[4]  P. Strange,et al.  Differences in the Ligand Binding Properties of the Short and Long Versions of the D2 Dopamine Receptor , 1993, Journal of neurochemistry.

[5]  M. Martres,et al.  Alternative splicing directs the expression of two D2 dopamine receptor isoforms , 1989, Nature.

[6]  W. Schafer,et al.  A calcium-channel homologue required for adaptation to dopamine and serotonin in Caenorhabditis elegans , 1995, Nature.

[7]  J. Glowinski,et al.  Lack of autoreceptor-mediated inhibitory control of dopamine release in striatal synaptosomes of D2 receptor-deficient mice , 1998, Brain Research.

[8]  C. D. Livingstone,et al.  Molecular modelling of D2-like dopamine receptors. , 1992, The Biochemical journal.

[9]  A. Kopin,et al.  A Drosophila dopamine 2-like receptor: Molecular characterization and identification of multiple alternatively spliced variants , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[10]  S. R. Nash,et al.  Dopamine receptors: from structure to function. , 1998, Physiological reviews.

[11]  B. Sommer,et al.  The dopamine D2 receptor: two molecular forms generated by alternative splicing. , 1989, The EMBO journal.

[12]  E. Borrelli,et al.  Structure and function of dopamine receptors , 2000, Neuroscience & Biobehavioral Reviews.

[13]  L. Ségalat,et al.  Modulation of serotonin-controlled behaviors by Go in Caenorhabditis elegans , 1995, Science.

[14]  H. V. Van Tol,et al.  Dopamine signaling in Caenorhabditis elegans-potential for parkinsonism research. , 2001, Parkinsonism & related disorders.

[15]  David Hirsh,et al.  A trans-spliced leader sequence on actin mRNA in C. elegans , 1987, Cell.

[16]  G. Gerhardt,et al.  Direct in vivo evidence that D2 dopamine receptors can modulate dopamine uptake , 1994, Neuroscience Letters.

[17]  S. Brenner,et al.  The structure of the nervous system of the nematode Caenorhabditis elegans. , 1986, Philosophical transactions of the Royal Society of London. Series B, Biological sciences.

[18]  P. Calabresi,et al.  Loss of autoreceptor function in dopaminergic neurons from dopamine D2 receptor deficient mice. , 1997, Neuroscience.

[19]  Jeremy Mendel,et al.  Participation of the protein Go in multiple aspects of behavior in C. elegans , 1995, Science.

[20]  P S Goldman-Rakic,et al.  Prominence of the dopamine D2 short isoform in dopaminergic pathways. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[21]  A. Baumann,et al.  Characterization of a Dopamine D1 Receptor from Apis mellifera: Cloning, Functional Expression, Pharmacology, and mRNA Localization in the Brain , 1998, Journal of neurochemistry.

[22]  R. Blakely,et al.  The Caenorhabditis elegans gene T23G5.5 encodes an antidepressant- and cocaine-sensitive dopamine transporter. , 1998, Molecular pharmacology.

[23]  Janet S. Duerr,et al.  The cat-1 Gene of Caenorhabditis elegansEncodes a Vesicular Monoamine Transporter Required for Specific Monoamine-Dependent Behaviors , 1999, The Journal of Neuroscience.

[24]  J. O. Schenk,et al.  D2 Receptors May Modulate the Function of the Striatal Transporter for Dopamine: Kinetic Evidence from Studies In Vitro and In Vivo , 1993, Journal of neurochemistry.

[25]  S. Ishiura,et al.  Identification of a dopamine receptor from Caenorhabditis elegans , 2002, Neuroscience Letters.

[26]  S. W. Emmons,et al.  Patterning of dopaminergic neurotransmitter identity among Caenorhabditis elegans ray sensory neurons by a TGFbeta family signaling pathway and a Hox gene. , 1999, Development.

[27]  H. Niznik,et al.  A primordial dopamine D1‐like adenylyl cyclase‐linked receptor from Drosophila melanogaster displaying poor affinity for benzazepines , 1995, FEBS letters.

[28]  D. Jackson,et al.  Unique binding characteristics of antipsychotic agents interacting with human dopamine D2A, D2B, and D3 receptors. , 1993, Molecular pharmacology.

[29]  P. Evans,et al.  Cloning and Functional Characterization of a Novel Dopamine Receptor from Drosophila melanogaster , 1996, The Journal of Neuroscience.

[30]  J. Sulston,et al.  Dopaminergic neurons in the nematode Caenorhabditis elegans , 1975, The Journal of comparative neurology.

[31]  A. Baumann,et al.  Primary structure and functional characterization of a Drosophila dopamine receptor with high homology to human D1/5 receptors. , 1994, Receptors & channels.

[32]  David H. Hall,et al.  Neurotoxin-induced degeneration of dopamine neurons in Caenorhabditis elegans , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[33]  Susumu Tonegawa,et al.  Dopamine D2 Long Receptor-Deficient Mice Display Alterations in Striatum-Dependent Functions , 2000, The Journal of Neuroscience.

[34]  N. Pollock,et al.  Serine mutations in transmembrane V of the dopamine D1 receptor affect ligand interactions and receptor activation. , 1992, The Journal of biological chemistry.

[35]  D. Weinshenker,et al.  Genetic and pharmacological analysis of neurotransmitters controlling egg laying in C. elegans , 1995, The Journal of neuroscience : the official journal of the Society for Neuroscience.

[36]  Alessandro Usiello,et al.  Distinct functions of the two isoforms of dopamine D2 receptors , 2000, Nature.

[37]  V. Ambros,et al.  Efficient gene transfer in C.elegans: extrachromosomal maintenance and integration of transforming sequences. , 1991, The EMBO journal.

[38]  INTERNATIONAL SOCIETY FOR NEUROCHEMISTRY , 1976 .

[39]  S. George,et al.  Functional analysis of the D2L dopamine receptor expressed in a cAMP-responsive luciferase reporter cell line. , 1998, Biochemical pharmacology.

[40]  L. Avery,et al.  Mutations in a C. elegans Gqα Gene Disrupt Movement, Egg Laying, and Viability , 1996, Neuron.