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J H Cha

Publications and source records attributed to J H Cha.

At least 37 records · Page 2Linked to original sources
PubMed2000

Decreased osteopontin expression in the rat kidney on a sodium deficient diet.

Osteopontin (OPN) is a secreted phosphoprotein that is constitutively expressed in the normal kidney and is induced by various experimental and pathologic conditions. Several possible functions of OPN have been suggested, however the mechanism and significance of OPN expression are still uncertain. Since high salt concentration or salt crystal have been known to enhance OPN expression in intact kidney or cultured renal cells, in the present study we examined whether or not a low salt condition had an effect on OPN expression in the kidney. Adult male Sprague-Dawley rats were fed either a normal sodium or a sodium deficient diet for 1 week. Kidneys were processed for in situ hybridization using a digoxigenin-labeled riboprobe and for immunohistochemistry using antibodies to OPN, renin, and Na-K-ATPase. In rats fed a normal sodium diet, OPN mRNA and protein were expressed only in the descending thin limbs of Henle's loop (DTL) and in the papillary and pelvic surface epithelium (PSE). In rats fed a sodium deficient diet, there was a marked decrease in OPN immunoreactivity in the DTL, but no changes in PSE. In contrast, no changes were observed in OPN mRNA expression in the DTL by in situ hybridization, indicating that decreased OPN protein expression was a result of translational regulation. As expected, rats fed a sodium deficient diet were associated with increased immunoreactivity for Na-K-ATPase and renin compatible with activation of the renin-angiotensin system. These results suggest that dietary sodium may be involved in the regulation of OPN expression in the DTL of the rat kidney.

Animals↗
PubMed1999

Intranuclear inclusions in subtypes of striatal neurons in Huntington's disease transgenic mice.

R6/2 transgenic mice express exon 1 of an abnormal human Huntington's disease (HD) gene and develop a neurological phenotype similar to HD. These mice develop ubiquitinated neuronal intranuclear inclusions (NII) which might play a central role in the pathophysiology of HD. We studied the distribution of NII in subpopulations of striatal neurons in 12-week-old R6/2 transgenic mice using fluorescent double label immunohistochemistry. We observed that most of the Calbindin-D28K positive projection neurons (89%) and the Parvalbumin positive interneurons (86%) showed ubiquitinated NII. In interneurons, however, which contain either choline acetyltransferase, neuronal nitric oxide synthase, or Calretinin, the frequency of NII was much lower (22%, 8%, 9%, respectively). Our data suggest that subpopulations of striatal neurons differ remarkably in their capability of forming ubiquitinated NII. Interneurons which are known to resist neurodegeneration in HD show less NII.

Animals↗
PubMed1999

Expression of osteopontin mRNA in the adult rat brain.

Examining the distribution of osteopontin (OPN) in the rat brain using in situ hybridization, it was found that OPN mRNA was restricted to likely neurons in the olfactory bulb and the brain stem; it was not detected in the telencephalon and the diencephalon. In addition, Northern blot analysis showed a confined expression only in the brain stem with higher level in the pons and the medulla than in the midbrain. In the brain stem, it was found in functionally diverse areas including motor-related areas, sensory system and reticular formation. This specific but widespread distribution suggests that OPN may play an important role in the adult rat brain.

Animals↗
PubMed1999

Transient upregulation of osteopontin mRNA in hippocampus and striatum following global forebrain ischemia in rats.

We investigated the spatial and temporal expression of osteopontin (OPN) mRNA following transient forebrain ischemia in rats. Experiments were carried out using a four-vessel occlusion model for forebrain ischemia. The transient induction of OPN mRNA after global ischemia occurred earlier in the striatum than in the hippocampus. It was pronounced in the dorsomedial striatum close to the lateral ventricle and in the CA1 subfield and the subiculum of the hippocampus before microglial cells became more reactive. It also could be detected in the dentate hilus and to a marginal extent in the CA3. Our results suggest that the hippocampus and the striatum following global forebrain ischemia upregulate OPN mRNA in different spatiotemporal profiles.

Animals↗
PubMed1999

Molecular cloning and expression analysis of a mouse phospholipase C-delta1.

We describe here the molecular cloning and expression analysis of mouse PLC-delta1 (mPLC-delta1), a key enzyme in cell signal transduction. A mouse brain cDNA library was screened in order to isolate the mPLC-delta1 cDNA. The mPLC-delta1 cDNA was 2660 bp in length. The predicted open reading frame encodes a protein of 756 amino acids with an estimated molecular mass of 85 kDa. The deduced amino acid sequence exhibits 96.9% and 92.7% identity with the sequence of rat and human PLC-delta1, respectively. The mPLC-delta1 mRNA was highly expressed in brain, heart, lung, and testis. We found that transcripts of mPLC-delta1 are present in almost all regions of mouse brain examined, implying that the enzyme may play a role in some fundamental cellular process in brain. In male reproductive tract, mPLC-delta1 mRNA was widely expressed in the epididymis as well as in the testis. In situ hybridization studies indicate that distribution of mPLC-delta1 mRNA in mouse testis is discrete and unique. The expression of mPLC-delta1 mRNA was defined in the periphery of each seminiferous tubule, especially in spermatogonia, which might imply that mPLC-delta1 plays a role in proliferation of spermatogonia. To the best our knowledge, this is the first report to demonstrate the high expression of mPLC-delta1 mRNA in spermatogonia of testis. Taken together, these results suggest that mPLC-delta1 may carry out fundamental roles in almost all of mouse tissues, especially in brain and specific roles in testis.

Amino Acid Sequence↗
PubMed1999

Altered neurotransmitter receptor expression in transgenic mouse models of Huntington's disease.

Alterations in neurotransmitter receptors are a pathological hallmark of the neurodegeneration seen in Huntington's disease (HD). However, the significance of these alterations has been uncertain, possibly reflecting simply the loss of brain cells. It is not known for certain whether the alteration of neurotransmitter receptors occurs before the onset of symptoms in human HD. Recently we developed transgenic mice that contain a portion of a human HD gene and develop a progressive abnormal neurological phenotype. Neurotransmitter receptors that are altered in HD (receptors for glutamate, dopamine, acetylcholine and adenosine) are decreased in the brain transgenic mice, in some cases before the onset of behavioural or motor symptoms. In transgenic mice, neurotransmitter receptor alterations occur before neuronal death. Further, receptor alterations are selective in that certain receptors, namely N-methyl-D-aspartate and gamma-aminobutyric acid receptors, are unaltered. Finally, receptor decreases are preceded by selective decreases in the corresponding mRNA species, suggesting the altered transcription of specific genes. These results suggest that (i) receptor decreases precede, and therefore might contribute to, the development of clinical symptoms, and (ii) altered transcription of specific genes might be a key pathological mechanism in HD.

Animals↗
PubMed1999

Inhibition of caspase-1 slows disease progression in a mouse model of Huntington's disease.

Huntington's disease is an autosomal-dominant progressive neurodegenerative disorder resulting in specific neuronal loss and dysfunction in the striatum and cortex. The disease is universally fatal, with a mean survival following onset of 15-20 years and, at present, there is no effective treatment. The mutation in patients with Huntington's disease is an expanded CAG/polyglutamine repeat in huntingtin, a protein of unknown function with a relative molecular mass of 350,000 (M(r) 350K). The length of the CAG/polyglutamine repeat is inversely correlated with the age of disease onset. The molecular pathways mediating the neuropathology of Huntington's disease are poorly understood. Transgenic mice expressing exon 1 of the human huntingtin gene with an expanded CAG/polyglutamine repeat develop a progressive syndrome with many of the characteristics of human Huntington's disease. Here we demonstrate evidence of caspase-1 activation in the brains of mice and humans with the disease. In this transgenic mouse model of Huntington's disease, expression of a dominant-negative caspase-1 mutant extends survival and delays the appearance of neuronal inclusions, neurotransmitter receptor alterations and onset of symptoms, indicating that caspase-1 is important in the pathogenesis of the disease. In addition, we demonstrate that intracerebroventricular administration of a caspase inhibitor delays disease progression and mortality in the mouse model of Huntington's disease.

Animals↗
PubMed1999

Expression of vascular endothelial growth factor mRNA following transient forebrain ischemia in rats.

Using a reproducible two-vessel occlusion model for forebrain ischemia in rats, we investigated the temporal and spatial changes of vascular endothelial growth factor (VEGF) expression after transient forebrain ischemia with Northern blot analysis and in situ hybridization. Northern blot analysis revealed that VEGF mRNA of the hippocampus was increased from 12 h after reperfusion, with a peak at 1 day. In situ hybridization and double labeling for VEGF mRNA and glial fibrillary acidic protein showed a transient induction of VEGF mRNA in the neurons of the hippocampus from 12 h of reperfusion with a peak at 1 day, and in the astrocytes of the hippocampus, caudoputamen, thalamus and cortical regions at 1 day. After 3 days, no more VEGF signal was detected. Our results demonstrate that astrocytes and neurons each upregulate VEGF mRNA in different temporal and spatial patterns after transient forebrain ischemia in the rat, and these patterns appear to be different from those in transient focal cerebral ischemia.

Animals↗
PubMed1999

Hamster diphtheria toxin receptor: a naturally occurring chimera of monkey and mouse HB-EGF precursors.

The sensitivity of mammalian cell lines to diphtheria toxin (DT) varies between species. Monkey (Mk) Vero cells are highly sensitive to DT, whereas rat and mouse (Ms) cells are resistant; hamster (Hm) cells display moderate DT sensitivity. The precursor of the Mk heparin-binding epidermal growth factor-like growth factor (proHB-EGF) functions as a DT receptor but the Ms proHB-EGF does not. In this study we have cloned, expressed, and characterized the Hm proHB-EGF/DT receptor. The expression of Hm proHB-EGF confers moderate DT sensitivity to normally DT-resistant mouse cells. The amino acid sequence of Hm preproHB-EGF shows that, overall, it more closely resembles the Ms preproHB-EGF sequence, except in the DT-binding region where it more closely resembles the Mk sequence. In the DT-binding region the Hm proHB-EGF sequence differs from the Mk proHB-EGF in only four amino acid residues (124, 126, 133, and 147); one of these residues, Ile133 in Mk proHB-EGF, has been previously reported to be important for DT binding and sensitivity. Analysis of Mk proHB-EGF mutants with residues substituted for Ile133 suggests that Asn133 in Hm proHB-EGF may be responsible for the moderate DT sensitivity of Hm proHB-EGF-expressing cells.

Amino Acid Sequence↗
PubMed1999

Intracisternal antisense oligonucleotide to growth associated protein-43 blocks the recovery-promoting effects of basic fibroblast growth factor after focal stroke.

Focal infarction (stroke) of the lateral cerebral cortex of rats (including the sensorimotor cortex) produces deficits in sensorimotor function of the contralateral limbs that recover partially over time. In previous studies, we found that the intracisternal injection of basic fibroblast growth factor (bFGF), a potent neurotrophic growth factor, starting at 1 day after stroke, significantly enhanced recovery of sensorimotor function of the contralateral forelimb and hindlimb. Moreover, immunoreactivity (IR) for growth-associated protein-43 (GAP-43), a molecular marker of new axonal growth, was increased in the intact contralateral sensorimotor cortex following bFGF treatment. In the current study, we found that the intracisternal administration of antisense, but not missense, oligonucleotide to GAP-43 blocked the recovery-enhancing effects of bFGF and blocked the increase in GAP-43 IR in the contralateral cortex. These results suggest that upregulation of GAP-43 expression and consequent enhanced axonal sprouting in intact uninjured parts of the brain are likely mechanisms for the recovery-promoting effects of bFGF.

Animals↗
PubMed1999

[Huntington chorea. Animal models reveal new hypotheses for pathophysiology and therapy].

Huntington's disease (HD) is member of a growing family of neurodegenerative diseases which are caused by a CAG-Triplet expansion in the coding region of their respective genes. The results of the research of the last years is very suggestive of a common pathomechanisms of all these diseases even though their clinical appearance may be quite different. The development of new animal models by transferring the human gene defect into the mouse genome has led to the finding of so-called intranuclear inclusion bodies. This new observation allowed to come closer to solving the problem how this genetic defect causes neurodegeneration. Recent studies on transgenic HD mice could also demonstrate a possible connection between the genetic defect and glutamate exitotoxicity in the neurodegenerative process of HD which had been emphasized by earlier animal models of the disease. Transgenic animal models of HD will have an important impact on the understanding of the disease mechanisms and may contribute to a faster development and testing of new therapeutic approaches.

Animals↗
PubMed1999

Intercalated cell subtypes in connecting tubule and cortical collecting duct of rat and mouse.

At least two populations of intercalated cells, type A and type B, exist in the connecting tubule (CNT), initial collecting tubule (ICT), and cortical collecting duct (CCD). Type A intercalated cells secrete protons via an apical H+-ATPase and reabsorb bicarbonate by a band 3-like Cl-/HCO3-exchanger, AE1, located in the basolateral plasma membrane. Type B intercalated cells secrete bicarbonate by an apical Cl-/HCO3- exchanger that is distinct from AE1 and remains to be identified. They express H+-ATPase in the basolateral plasma membrane and in vesicles throughout the cytoplasm. A third type of intercalated cell with apical H+-ATPase, but no AE1, has been described in the CNT and CCD of both rat and mouse. The prevalence of the third cell type is not known. The aim of this study was to characterize and quantify intercalated cell subtypes, including the newly described third non A-non B cell, in the CNT, ICT, and CCD of the rat and mouse. A triple immunolabeling procedure was developed in which antibodies to H+-ATPase and band 3 protein were used to identify subpopulations of intercalated cells, and segment-specific antibodies were used to identify distal tubule and collecting duct segments. In both rat and mouse, intercalated cells constituted approximately 40% of the cells in the CNT, ICT, and CCD. Type A, type B, and non A-non B intercalated cells were observed in all of the three segments, with type A cells being the most prevalent in both species. In the mouse, however, non A-non B cells constituted more than half of the intercalated cells in the CNT, 39% in the ICT, and 22% in the CCD, compared with 14, 7, and 5%, respectively, in the rat. In contrast, type B intercalated cells accounted for only 8 to 16% of the intercalated cells in the three segments in the mouse compared with 26 to 39% in the rat. It is concluded that striking differences exist in the prevalence and distribution of the different types of intercalated cells in the CNT, ICT, and CCD of rat and mouse. In the rat, the non A-non B cells are fairly rare, whereas in the mouse, they constitute a major fraction of the intercalated cells, primarily at the expense of the type B intercalated cells.

Animals↗
PubMed1998

Diphtheria toxin:receptor interaction: association, dissociation, and effect of pH.

Diphtheria toxin (DT) binds to a specific heparin-binding epidermal growth factor-like growth factor (HB-EGF) precursor that is expressed in DT-sensitive cells. DT binds to the cell-surface HB-EGF precursor with an apparent dissociation constant (KD) of approximately 1 x 10(-8) - 10(-9) M at 4 degreesC, a temperature at which toxin binds but is not internalized. The interaction of DT with the cell-surface receptor, however, may be influenced by other cell-surface components. We used a biosensor method to measure the binding of DT to immobilized recombinant human HB-EGF (hHB-EGF) at 25 degreesC with no other cellular components present. We observed that at pH 7.4, using this in vitro two component system, DT binds to hHB-EGF with an apparent KD of 2.7 x 10(-8) M. We also observed that the dissociation of DT from hHB-EGF at pH values that approach those of the endosome occurs at a faster rate as the pH is decreased. These results suggest that the low pH of the endosome is sufficient to allow DT to dissociate from the HB-EGF precursor, prior to the translocation of the enzymatically active fragment of DT into the cytosol.

Biosensing Techniques↗
PubMed1998

Altered brain neurotransmitter receptors in transgenic mice expressing a portion of an abnormal human huntington disease gene.

Loss of neurotransmitter receptors, especially glutamate and dopamine receptors, is one of the pathologic hallmarks of brains of patients with Huntington disease (HD). Transgenic mice that express exon 1 of an abnormal human HD gene (line R6/2) develop neurologic symptoms at 9-11 weeks of age through an unknown mechanism. Analysis of glutamate receptors (GluRs) in symptomatic 12-week-old R6/2 mice revealed decreases compared with age-matched littermate controls in the type 1 metabotropic GluR (mGluR1), mGluR2, mGluR3, but not the mGluR5 subtype of G protein-linked mGluR, as determined by [3H]glutamate receptor binding, protein immunoblotting, and in situ hybridization. Ionotropic alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid and kainate receptors were also decreased, while N-methyl-D-aspartic acid receptors were not different compared with controls. Other neurotransmitter receptors known to be affected in HD were also decreased in R6/2 mice, including dopamine and muscarinic cholinergic, but not gamma-aminobutyric acid receptors. D1-like and D2-like dopamine receptor binding was drastically reduced to one-third of control in the brains of 8- and 12-week-old R6/2 mice. In situ hybridization indicated that mGluR and D1 dopamine receptor mRNA were altered as early as 4 weeks of age, long prior to the onset of clinical symptoms. Thus, altered expression of neurotransmitter receptors precedes clinical symptoms in R6/2 mice and may contribute to subsequent pathology.

Animals↗
PubMed1998

Toxin binding site of the diphtheria toxin receptor: loss and gain of diphtheria toxin binding of monkey and mouse heparin-binding, epidermal growth factor-like growth factor precursors by reciprocal site-directed mutagenesis.

The transmembrane precursor of the monkey (Mk) heparin-binding, epidermal growth factor-like growth factor (proHB-EGF) functions as a diphtheria toxin (DT) receptor, whereas the mouse (Ms) precursor does not. Previously, using chimeric Ms/Mk precursors, we have shown that DT resistance of cells bearing Ms proHB-EGF may be accounted for by several amino acid substitutions between residues 122 and 148 within the EGF-like domain and that Glu-141 is an important amino acid residue for DT binding. In this study, reciprocal site-directed mutagenesis was performed on the major non-conserved residues in the region of 122-148, alone or in combination, between Mk and Ms precursors to identify more precisely which amino acid residues are important for DT binding. Two approaches were used. The first, more traditional approach was to destroy DT sensitivity and binding of Mk proHB-EGF by substitution(s) with the corresponding Ms residue(s). From the single mutations, the greatest loss of DT sensitivity was observed with Mk/Glu-141His (approximately 4000-fold) and the next greatest with Mk/Ile-133Lys (approximately fourfold). The double mutations Mk/Leu-127Phe/Glu-141His, Mk/Ile-133Lys/Glu-141His and Mk/His-135Leu/Glu-141His resulted in complete toxin resistance (> 100000-fold). The second approach, both novel and complementary, was to gain DT binding and sensitivity of Ms proHB-EGF by substitution(s) with the corresponding Mk residue(s). Surprisingly, the single mutation Ms/His-141Glu resulted in the gain of moderate DT sensitivity (> 260-fold). The double mutation Ms/Lys-133Ile/His-141Glu and the triple mutation Ms/Lys-133Ile/Leu-135His/His-141Glu resulted in a progressive gain in toxin sensitivity (> 4700-fold and >16000-fold respectively) and affinity. This triple mutant cell line is essentially as sensitive (IC50 = 3.1 ng ml(-1)) as the highly toxin-sensitive monkey Vero cell line (IC50 = 4 ng ml(-1)), indicating that these three Mk residues enable the Ms proHB-EGF to act as a fully functional DT receptor. Taken together, these results indicate that Glu-141 plays the most critical role in DT binding and sensitivity and that two additional amino acid residues, Ile-133 and His-135, also play significant roles.

Amino Acid Sequence↗
PubMed1998

Distributional patterns of phospholipase C isozymes in rat kidney.

As the first step to investigate the physiological function of phospholipase C (PLC), we determined the distribution patterns of PLC isozymes in normal rat kidneys using Western blotting analysis and immunohistochemistry. Western blotting analysis was performed in four regions (cortex, outer stripe and inner stripe of the outer medulla, and the inner medulla). PLC-beta1 and beta3 were detected in the inner stripe of the outer medulla and the inner medulla. PLC-gamma1 was distributed homogeneously along the corticomedullary axis. PLC-gamma2 was observed in the medulla and PLC-delta1 showed a gradual increase from the cortex to the inner medulla. In contrast, no PLC-beta4 was detected in all regions. On immunohistochemistry, the immunoreactivities to PLC antibodies were observed as follows: PLC-beta1, from the thick ascending limb (TAL) to the inner medullary collecting tubule (IMCT); PLC-beta3, in the glomerulus (Glm), the ascending thin limb (ATL) and the collecting tubule; PLC-beta4, Glm, the proximal convoluted tubule (PCT), ATL, the distal convoluted tubule, the connecting tubule, and the collecting tubules; PLC-gamma1, PCT, TAL and IMCT; PLC-delta1, homogeneously from PCT to IMCT. PLC-beta3 immunoreactivities were detected in the nuclei of the TAL, ATL, outer medullary collecting tubule (OMCT) and IMCT. PLC-beta4 and gamma2 were observed in Glm, MTAL, ATL, OMCT and IMCT. These results suggest the intrarenal site-specific existence of PLC isozymes that may regulate kidney functions through the PLC-mediated signal transductions.

Animals↗
PubMed1998

Inhibition of calbindin D28K expression by cyclosporin A in rat kidney: the possible pathogenesis of cyclosporin A-induced hypercalciuria.

A recent study by Steiner et al. (Biochem Pharmacol 51: 253-258, 1996) demonstrated a decreased calbindin D28K expression in the kidneys of cyclosporin A (CsA)-treated rats. To evaluate the association of renal calcium handling with calbindin D28K expression in CsA-treated rats, two separate experiments (vehicle [VH] versus CsA groups, 1,25-dihydroxyvitamin D3 [VitD] versus VitD + CsA groups) were done simultaneously. CsA (25 mg/kg per d, subcutaneously) and VitD (0.5 microg/kg per d, subcutaneously) were given for 7 d. The CsA group showed decreased serum calcium, increased urine calcium excretion, and decreased calbindin D28K protein level and immunoreactivity compared with the VH group. The VitD + CsA treatment decreased serum calcium, increased urine calcium excretion, and decreased calbindin D28K protein level and immunoreactivity compared with the VitD alone. CsA treatment did not affect the serum parathyroid hormone and VitD levels. This study demonstrates an association of calbindin D28K expression with the urinary calcium excretion in CsA-treated rats, and suggests that decreased calbindin D28K expression may play a role in renal calcium wasting.

Animals↗