Molecular characterization and localization of human metabotropic glutamate receptor type 3.
A human brain cDNA library was screened using amplified human metabotropic glutamate receptor (mGluR) cDNA sequences as probes. The resulting clones included one containing the complete coding sequence of mGluR3. This sequence has 90% DNA sequence identity with rat mGluR3 and the predicted protein sequence has 97% identity. The mGluR3 cDNA was transfected in Chinese hamster ovary (CHO) cells. Stimulation of the expressed receptor by (2S,3S,4S)-alpha-(carboxycyclopropyl) glycine (L-CCG-I) resulted in a reduction of forskolin-stimulated cyclic AMP (cAMP) with EC50 values of 0.15-0.3 microM. A specific probe from the human mGluR3 clone was used to hybridise to Northern blots of mRNA from various human tissues and different brain regions. The mGluR3 mRNA is brain-specific, and is expressed in all the brain regions represented on the blot. In-situ hybridization studies on human brain sections confirmed this widespread distribution with expression in neurones in the cerebral cortex, caudate-putamen, thalamus and cerebellum.
Makoff
A
A
Biology Division, Wellcome Foundation, Beckenham, Kent, UK.
Volpe
F
F
Lelchuk
R
R
Harrington
K
K
Emson
P
P
D000595:Amino Acid Sequence; D000679:Amygdala / Q000378:metabolism; D000818:Animals; D001483:Base Sequence; D001921:Brain / Q000378:metabolism*; D016466:CHO Cells; D006224:Cricetinae; D015342:DNA Probes; D018076:DNA, Complementary; D005625:Frontal Lobe / Q000378:metabolism; D006801:Humans; D008969:Molecular Sequence Data; D009474:Neurons / Q000378:metabolism; D009928:Organ Specificity; D051381:Rats; D018094:Receptors, Metabotropic Glutamate / Q000032:analysis / Q000096:biosynthesis* / Q000737:chemistry; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry; D017386:Sequence Homology, Amino Acid; D014158:Transcription, Genetic; D014162:Transfection
D015342:DNA Probes; D018076:DNA, Complementary; D018094:Receptors, Metabotropic Glutamate; D011994:Recombinant Proteins; C102718:metabotropic glutamate receptor 3