Phosphoinositide hydrolysis in vivo with group I metabotropic glutamate receptor agonists

The present report describes the effect of mGluR agonists and antagonists administration on phospholipase C activation by measuring accumulation of [ 3H] inositol monophosphates (IP) in rats pre-labeled with [ 3H] myo-inositol (i.c.v. 24 h pre-treatment). The levels of accumulated [ 3H]IP were then determined from clarified tissue homogenates using ion-exchange chromotography. Following lithium chloride treatment (10 mg/kg, s.c.), (R/S)-3,5-dihydroxyphenylglycine (DHPG), a selective group I mGluR agonist was found to dose-dependently cause a maximal increase in the levels of [ 3H]IP at 0.3 to 3 μmol/8 μl i.c.v. with lower doses resulting in less efficacious or no responses. This effect was temporal-dependent reaching a plateau at 2 h. The DHPG-induced increases in [ 3H]IP were most pronounced in the hippocampus where a 3- to 5-fold increase above vehicle was consistently found, but significant approximately 2-fold increases were also seen in the cerebellum, striatum and frontal cortex. The mixed group I and II agonist, (1S,3R)-1-aminocyclopentane- trans-1,3-dicarboxylic acid (1S,3R- t-ACPD), similarly resulted in dose-dependent increases in [ 3H]IP levels with doses of 1 to 3 μmol i.c.v. Furthermore, this effect was enantiomer specific since the less active 1R,3S- t-ACPD failed to alter phosphoinositol hydrolysis. Administration of the selective mGluR5 agonist (R/S)-2-chloro-5-hydroxyphenyl-glycine (CHPG) resulted in a dose-dependent increase in hippocampal but not cerebellar levels of [ 3H]IP, consistent with the receptor distribution of the two group I mGluRs. The Group II agonist LY354740 (1S,2S,5R,6S-2-aminobicycl[3.1.0]hexane-2,6-dicarboxylate monohydrate) and the group III agonist L-AP 4 (L-(+)-2-amino-4-phosphonobutyric acid) failed to alter the levels of [ 3H]IP. LY341495 (2S-2-amino-2-(1S,2S-2-carboxycycloprop-1-yl)-3-(xanth-9-yl)propanoic acid) is a nM potent Group II antagonist. However, LY341495 has also been found to have μM potency in inhibiting mGluR1 and 5. The stimulation of [ 3H]PI hydrolysis by 1 μmol DHPG was dose-dependently blocked by co-administration of the mGluR antagonists, LY341495 at doses that are constant with an interaction at Group I mGluR's. Taken together these results suggest that stimulation of group I mGluRs results in measurable increases in PI hydrolysis in vivo. This method could be quite useful in determining the doses and routes of administration of agonists and antagonists that are required to interact with group