Glutamate carboxypeptidase II levels in rodent brain using [125I]DCIT quantitative autoradiography.
The ability to visualize quantitatively glutamate carboxypeptidase II (GCPII) levels in vivo could advance our understanding of its function in health and disease. In the current study, we synthesized and evaluated a radiolabeled (iodine-125) analog of N-[N-[(S)-1,3-dicarboxypropyl]carbamoyl]-S-3-iodo-L-tyrosine (DCIT), a potent antagonist of GCPII activity. We examined the regional distribution of [125I]DCIT binding in the rodent brain using quantitative autoradiography in order to confirm the validity of this radioligand as a marker of GCPII in the brain. The ultimate goal is to develop an imaging agent for assessing GCPII levels in the living brain. The specific binding of [125I]DCIT to rat brain followed a regional distribution consistent with previous studies describing regional brain GCPII gene expression and activity. We found a modest rostrocaudal gradient in which specific binding of [125I]DCIT to GCPII was lowest in cortical regions, with increasing levels of binding in midbrain structures and high levels of binding in hindbrain and brainstem. Autoradiography of [125I]DCIT in GCPII knockout and wild type mouse brain showed a gene-dose dependency confirming the selectivity of this radioligand for GCPII. We propose that [125I]DCIT is a selective radioligand that can be used to quantify brain GCPII levels in vitro using quantitative autoradiography.
Guilarte
Tomás R
TR
Department of Environmental Health Sciences, Johns Hopkins School of Public Health, Baltimore, MD 21287-2182, USA.
McGlothan
Jennifer L
JL
Foss
Catherine A
CA
Zhou
Jia
J
Heston
Warren D
WD
Kozikowski
Alan P
AP
Pomper
Martin G
MG
D000818:Animals; D001667:Binding, Competitive / Q000502:physiology; D015415:Biomarkers / Q000737:chemistry / Q000378:metabolism; D001921:Brain / Q000033:anatomy & histology / Q000000981:diagnostic imaging / Q000201:enzymology*; D002219:Carbamates / Q000138:chemical synthesis / Q000378:metabolism; D003952:Diagnostic Imaging / Q000379:methods; D004151:Dipeptides / Q000378:metabolism*; D004791:Enzyme Inhibitors / Q000378:metabolism; D018628:Gene Dosage; D015971:Gene Expression Regulation, Enzymologic / Q000502:physiology; D043425:Glutamate Carboxypeptidase II / Q000235:genetics / Q000378:metabolism*; D007457:Iodine Radioisotopes; D051379:Mice; D018345:Mice, Knockout; D011869:Radioligand Assay / Q000379:methods; D011877:Radionuclide Imaging; D019275:Radiopharmaceuticals / Q000138:chemical synthesis / Q000378:metabolism; D051381:Rats; D020318:Rats, Long-Evans; D014443:Tyrosine / Q000031:analogs & derivatives* / Q000138:chemical synthesis / Q000378:metabolism
D015415:Biomarkers; D002219:Carbamates; D004151:Dipeptides; D004791:Enzyme Inhibitors; D007457:Iodine Radioisotopes; C506257:N-(N-((S)-1,3-dicarboxypropyl)carbamoyl)-S-3-iodo-L-tyrosine; D019275:Radiopharmaceuticals; C027172:isospaglumic acid; D014443:Tyrosine; D043425:Glutamate Carboxypeptidase II