Quinolinic acid-induced increases in calbindin D28k immunoreactivity in rat striatal neurons in vivo and in vitro mimic the pattern seen in Huntington's disease.
In Huntington's disease striatal neurons undergo marked changes in dendritic morphology and coincidently exhibit an increase in immunoreactive calbindin D28k (calbindin), a cytosolic calcium-binding protein which is highly abundant in these neurons. Previous studies in the rat striatum have shown that excitotoxic injury, which is linked to a rise in intracellular Ca2+, mimics many of the neurochemical and neuropathological characteristics of Huntington's disease. We speculated, therefore, that the apparent increase in calbindin labeling in Huntington's disease spiny neurons may signal the response to an excitotoxic process. To investigate this possibility, we compared the cellular features of calbindin immunoreactivity in grade 1-4 Huntington's disease cases with those seen in rat striatal neurons in vivo and in vitro following treatment with N-methyl-D-aspartate (NMDA) receptor agonist, quinolinic acid. In human post mortem control cases calbindin immunoreactivity was seen primarily in the somata and proximal dendrites of striatal neurons. In the Huntington's disease cases, calbindin labeling was markedly increased throughout the second and third order dendrites and in spines, and this change was more prevalent in advanced cases (grades 3-4). In the rat brain, two weeks after intrastriatal injection of quinolinic acid (6-20 ng), surviving medium-spiny neurons in the transition zone around the lesion core exhibited a marked increase in calbindin immunoreactivity similar to that seen in Huntington's disease spiny neurons. In more peripheral areas away from the lesion and on the contralateral unlesioned side, calbindin immunostaining was confirmed to somata and proximal dendrites. In situ hybridization histochemistry with an 35S-labeled oligonucleotide probe showed no change or a decrease in calbindin mRNA levels in neurons within the transition zone, suggesting that the observed increase in calbindin staining was not the result of increased transcription. In 12 day old postnatal striatal cultures, 2-6 h exposures to quinolinic acid (0.5 mM) significantly increased the length of neurites exhibiting calbindin immunoreactivity when compared to untreated controls. This effect was blocked by the selective NMDA receptor blocker (+/-)-2-amino-5-phosphonopentanoic acid (AP-5), indicating that an NMDA receptor-mediated mechanism contributed to the change in staining pattern. Results in rats suggest that the subcellular redistribution of calbindin immunoreactivity observed in Huntington's disease spiny neurons may be related to an NMDA receptor-induced excitotoxic process. An increased availability of calbindin protein at dendrites and spines may reflect a greater demand for Ca2+ buffering precipitated by an abnormal rise in in intracellular Ca2+.
Huang
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Laboratory of Cellular Neurobiology, Massachusetts General Hospital, Boston 02114, USA.
Zhou
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D
Sapp
E
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Aizawa
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H
Ge
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Bird
E D
ED
Vonsattel
J P
JP
DiFiglia
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M
D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D000818:Animals; D001483:Base Sequence; D064092:Calbindin 1; D064026:Calbindins; D002478:Cells, Cultured; D006801:Humans; D006816:Huntington Disease / Q000378:metabolism* / Q000473:pathology; D007150:Immunohistochemistry; D017403:In Situ Hybridization; D008297:Male; D008875:Middle Aged; D008969:Molecular Sequence Data; D017072:Neostriatum / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D009419:Nerve Tissue Proteins / Q000096:biosynthesis*; D016501:Neurites / Q000187:drug effects / Q000378:metabolism / Q000648:ultrastructure; D017378:Quinolinic Acid / Q000494:pharmacology*; D012333:RNA, Messenger / Q000096:biosynthesis; D051381:Rats; D017207:Rats, Sprague-Dawley; D064030:S100 Calcium Binding Protein G / Q000096:biosynthesis*
C576498:CALB1 protein, human; C576500:Calb1 protein, rat; D064092:Calbindin 1; D064026:Calbindins; D009419:Nerve Tissue Proteins; D012333:RNA, Messenger; D064030:S100 Calcium Binding Protein G; D017378:Quinolinic Acid