Metabotropic glutamate receptor activation in vivo induces intraneuronal amyloid immunoreactivity in guinea pig hippocampus.
Stimulation of metabotropic glutamate receptors in vitro has been shown to accelerate the breakdown of amyloid precursor protein (APP) to form increased production of non-amyloidogenic secreted APP (sAPP). The mechanism whereby this occurs is not entirely clear but it is presumed to be linked to generation of diacylglycerol and activation of protein kinase C because other neurotransmitter receptors such as m1 and m3 muscarinic receptors, known to be coupled to this second messenger cascade, likewise increase sAPP production. Although it is presumed that a reciprocal relationship exists between the formation of amyloid beta protein (Abeta) and the production of sAPP, recent evidence suggests alternative processing can occur. Given the fact that much of the observations on APP metabolism have been made in vitro we sought to investigate the effect of metabotropic receptor activation on Abeta in vivo in a species known to contain the same amino acid sequence of Abeta as found in humans. Intrahippocampal injection of the mGluR agonist 1S,3R-ACPD in guinea pigs produced neurodegeneration of CA1 hippocampal pyramidal neurons at 12 h postinjection. Immunocytochemistry of sections from ACPD injected animals using selective antibodies to Abeta revealed the presence of punctate intraneuronal granules in pyramidal neurons of the hippocampus. These structures appeared to be localized within the nucleus and were particularly prominent in neurons within the region of neurodegeneration. Immunoreactivity was not observed in vehicle injected controls nor in sections from ACPD injected animals stained with preadsorbed antiserum. Abeta immunodetection was correlated with the onset of neurodegeneration since animals evaluated at 1 h and 4 h postinjection lacked both Abeta immunoreactivity as well as neurodegeneration. Evaluation of animals injected with NMDA revealed neurodegeneration but no Abeta immunoreactivity suggesting Abeta formation did not appear to be due to non-selective excitotoxicity. Staining of sections with antibodies directed to various regions of APP demonstrated increased C-terminal APP immunoreactivity in pyramidal neurons in the vicinity of degeneration. These data support recent in vitro studies illustrating that Abeta can be found intracellularly within neurons.
Eli Lilly and Company, CNS Division, Lilly Corporate Center, Indianapolis, IN 46285, USA.
D T
DT
Stephenson
J A
JA
Clemens
D000682:Amyloid / Q000378:metabolism*; D016564:Amyloid beta-Protein Precursor / Q000378:metabolism; D000818:Animals; D003515:Cycloleucine / Q000031:analogs & derivatives / Q000494:pharmacology; D006168:Guinea Pigs; D006624:Hippocampus / Q000378:metabolism* / Q000473:pathology; D007150:Immunohistochemistry; D008297:Male; D008845:Microinjections; D009474:Neurons / Q000378:metabolism / Q000473:pathology; D018094:Receptors, Metabotropic Glutamate / Q000819:agonists*
D000682:Amyloid; D016564:Amyloid beta-Protein Precursor; D018094:Receptors, Metabotropic Glutamate; D003515:Cycloleucine; C036361:1-amino-1,3-dicarboxycyclopentane