Brain research. Molecular brain research
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Brain Res. Mol. Brain Res. · Jan 2000
Cell death in regenerating populations of neurons in BDNF mutant mice.
There are two populations of neurons which are continually renewed in the adult, the dentate gyrus granule neurons and the olfactory bulb granule and periglomerular neurons. In the dentate gyrus, a secondary proliferative zone termed the subgranular zone is established along the interface between the dentate gyrus and the hilus where granule cells are born throughout life. Olfactory bulb neurons are generated in the anterior subventricular zone of the lateral ventricle and migrate via the rostral migratory stream to the olfactory bulb. ⋯ However, increased apoptosis was not limited to precursors, as apoptotic cells were also found in the granule cell layer of the dentate gyrus and in the granule and periglomerular layers of the olfactory bulb. The excessive cell death was limited to these populations of neurons as no excessive cell death was detected in other forebrain areas. We conclude that BDNF is essential for the survival of neurons specifically in populations which are continuously being regenerated in the brain.
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Brain Res. Mol. Brain Res. · Dec 1999
Developmental changes in the expression of Shaker- and Shab-related K(+) channels in neurons of the rat trigeminal ganglion.
We have investigated properties of voltage-gated K(+) channels in neurons of the pre- and postnatal rat trigeminal ganglion (TG). To correlate functional data with information on gene expression of Shaker- and Shab-related channels in these pseudo-unipolar neurons, the patch-clamp technique was combined with the single-cell reverse transcription-polymerase chain reaction (RT-PCR). A majority (80%) of prenatal TG neurons possessed only sustained delayed rectifier currents with half-maximal current inactivation at -30 mV. ⋯ Most cells simultaneously expressed several different Shaker- and Shab-like transcripts. At postnatal day 14, the frequency of cells carrying transcripts encoding Kv1.1 decreased. Detailed analysis revealed a higher 4-AP sensitivity of TG neurons expressing Kv1.1 transcripts.
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Brain Res. Mol. Brain Res. · Dec 1999
Vascular endothelial growth factor upregulation in transient global ischemia induced by cardiac arrest and resuscitation in rat brain.
This study examined vascular endothelial growth factor (VEGF) expression in rat brain after reversible global cerebral ischemia produced by cardiac arrest and resuscitation. Three alternative splicing forms, VEGF(188), VEGF(164) and VEGF(120), were observed in cortex, hippocampus and brainstem by RT-PCR analysis. After 24 h of recovery from cardiac arrest, mRNA levels corresponding to VEGF(188) and VEGF(164) were significantly increased by about double in all the regions analyzed. ⋯ VEGF protein expression measured by Western blot was also increased by about double at 24 and 48 h of recovery but returned to control levels after 7 days of recovery. VEGF immunohistochemistry localized this increased expression mostly associated with astrocytes. Considering its biological activity, VEGF induction after cardiac arrest and resuscitation may be responsible for the increased vascular permeability and the resultant vasogenic edema, found 24-48 h after reversible global ischemia.
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Brain Res. Mol. Brain Res. · Oct 1999
Mutation of human mu opioid receptor extracellular "disulfide cysteine" residues alters ligand binding but does not prevent receptor targeting to the cell plasma membrane.
The mu opioid receptor, a primary site of action in the brain for opioid neuropeptides and opiate drugs of abuse, is a member of the seven transmembrane, G protein-coupled receptor (GPCR) superfamily. Two cysteine residues, one in each of the first two of three extracellular loops (ECLs), are highly conserved among GPCRs, and there is direct or circumstantial evidence that the residues form a disulfide bond in many of these receptors. Such a bond would dramatically govern the topology of the ECLs, and possibly affect the position of the membrane-spanning domains. ⋯ The two point mutants possessing serine-for-cysteine substitutions were also observed to successfully reach the cell plasma membrane, as evidenced by electron microscopy. Consistent with related work with other GPCRs, the mu opioid receptor apparently also employs the extracellular disulfide bond. This information now permits accurate molecular modeling of extracellular aspects of the receptor, including plausible scenarios of mu receptor docking of opioid ligands known to require specific extracellular loop features for high affinity binding.
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Brain Res. Mol. Brain Res. · Jun 1999
Localization of promoter elements in the human mu-opioid receptor gene and regulation by DNA methylation.
The regulation of mu-opioid receptor gene expression was investigated using several molecular techniques. Genomic clones containing portions of the human mu-opioid receptor gene were sequenced. 5'-RACE analysis of human brain cDNA confirmed the presence of mRNAs up to -313 from the start codon. As was found for the mouse and rat genes, transcription apparently initiates in the absence of a discernable TATA box. ⋯ In contrast, two neural derived cell lines that do not express the mu-opioid receptor were nearly totally methylated while non-neural cell lines had intermediate levels of CpG methylation. Additional transient transfection experiments revealed that CpG methylation of the 5'-flanking region suppressed reporter gene expression. These results indicate that CpG methylation plays an important role in regulating mu-opioid receptor expression in neural cells; however, no association was found with regulation of expression in non-neural cells.