Distribution of the neurons containing inositol 1,4,5-trisphosphate 3-kinase and its messenger RNA in the developing rat brain.
As a result of its interaction with a specific receptor, inositol 1,4,5-trisphosphate (InsP3) mobilizes intracellular calcium. The metabolism of InsP3 is rather complex: InsP3 3-kinase produces Inositol 1,3,4,5-tetrakisphosphate (InsP4), a putative second messenger also involved in the intraneuronal calcium homeostasis. The distribution of the messenger RNA coding for the recently cloned InsP3 3-kinase was studied in the developing rat brain by using oligonucleotides derived from the rat cDNA sequence and in situ hybridization combined with Northern blot analysis. In addition, the locations of the enzyme were determined by immunohistochemistry in combination with Western blot analysis. By Northern blot and Western blot analyses on rat brain, the kinase was not detected in the embryo, was first found slightly at birth, and reached adult levels around 2-3 postnatal weeks. These findings were confirmed in the different positive regions by in situ hybridization conducted at the macroscopic level. At the cellular level, the mRNA was found exclusively in the neuronal populations previously demonstrated in the adult. The levels of transcripts per neuron were however higher in the adult than in the neonate brain. The enzyme mRNA could be detected first at postnatal day 0, (birth, P0) in the perikarya of the cerebellar Purkinje cells, followed at P4 by the hippocampal CA1 pyramidal cells and granule cells of the dentate gyrus and finally, at P9, by a majority of the neurons in the cortical layers II-III and V, especially in the frontal cortex and cingulate cortex; claustrum; caudate, putamen, accumbens, olfactory tubercle and calleja islets; anterior olfactory nucleus; taenia tecta; piriform piriform cortex; dorsolateral septum; bed nucleus stria terminalis; amygdala; hippocampal CA2-4 sectors and subiculum. By immunohistochemistry, the enzyme was initially found in the periphery of the cell bodies of the neonatal neurons; was progressively enriched in the developing dendritic arborization during the first postnatal weeks where it remained exclusively localized in the adult. In conclusion, in the developing brain, InsP3 3-kinase was first detected at birth, and thereafter its concentrations increased to reach adult levels around 2-3 postnatal weeks. At the cellular level, the kinase was exclusively found in the neurons. The small amounts of transcripts found per neuron in the neonate increase during synaptogenesis and the protein became progressively enriched in the developing dendritic arborization, where it is localized in the adult.
Laboratory of Neurophysiology and Neuropeptide Research, Faculty of Medicine, Université Libre de Bruxelles, Belgium.
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Mailleux
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Takazawa
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Erneux
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Vanderhaeghen
D000818:Animals; D015152:Blotting, Northern; D015153:Blotting, Western; D001921:Brain / Q000196:embryology / Q000201:enzymology* / Q000254:growth & development*; D001923:Brain Chemistry / Q000502:physiology; D005314:Embryonic and Fetal Development / Q000502:physiology; D007150:Immunohistochemistry; D017403:In Situ Hybridization; D009474:Neurons / Q000737:chemistry / Q000201:enzymology*; D010770:Phosphotransferases / Q000032:analysis* / Q000235:genetics; D017853:Phosphotransferases (Alcohol Group Acceptor); D012333:RNA, Messenger / Q000032:analysis*; D051381:Rats; D017208:Rats, Wistar
D012333:RNA, Messenger; D010770:Phosphotransferases; D017853:Phosphotransferases (Alcohol Group Acceptor); C051038:Inositol 1,4,5-trisphosphate 3-kinase