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J Zuo

Publications and source records attributed to J Zuo.

At least 73 records · Page 4Linked to original sources
PubMed1995

Cloning of rat grp75, an hsp70-family member, and its expression in normal and ischemic brain.

Following metabolic stress a variety of gene products are induced in cells in the brain, some of which may protect the tissue from subsequent stresses. The heat shock proteins (hsps), in particular hsp70, have been widely studied in this context, but evidence for the involvement of known hsps in protection of the CNS is inconclusive. We have therefore undertaken the search for other stress-induced proteins which may mitigate ischemic injury. Beginning with degenerate RT-PCR, we have isolated a rat-brain cDNA encoding a protein highly similar to human grp75, a mitochondrial member of the hsp70-family of stress proteins. It is also highly similar to two non-mitochondrial proteins; mortalin, a senescence-related gene product, and pbp74, a protein implicated in B-cell peptide processing. Sequence structure and phylogenetic analyses predict mitochondrial localization and induction by a calcium ionophore and glucose deprivation in PC12 cells support its identification as rat grp75. In situ analysis of normal brain reveals an unusual distribution, with very high expression in neurons of the basal forebrain, reticular and subthalamic nuclei, globus pallidus, amygdala and elsewhere. grp75-mRNA is upregulated following focal brain ischemia in a distinctive fashion. When the degree of injury is small, induction occurs in the area of injury, similar to the pattern observed for hsp70. However, when the injury is extensive, hsr is upregulated in neurons outside the ischemic area. The induction of grp75 may represent a sensitive marker of metabolically compromised tissue.

Amino Acid Sequence↗
PubMed1995

Scrapie prions selectively modify the stress response in neuroblastoma cells.

The fundamental event underlying scrapie infection seems to be a conformational change in the prion protein. To investigate proteins that might feature in the conversion of the cellular prion protein (PrPC) into the scrapie isoform (PrPSc), we examined mouse neuroblastoma N2a cells for the expression and cellular distribution of heat shock proteins (Hsps), some of which function as molecular chaperones. In scrapie-infected N2a (ScN2a) cells, Hsp72 and Hsp28 were not induced by heat shock, sodium arsenite, or an amino acid analog, in contrast to uninfected control N2a cells, while other inducible Hsps were increased by these treatments. Following heat shock of the N2a cells, constitutively expressed Hsp73 was translocated from the cytoplasm into the nucleus and nucleolus. In contrast, the distribution of Hsp73 in ScN2a cells was not altered by heat shock; the discrete cytoplasmic structures containing Hsp73 were largely resistant to detergent extraction. These alterations in the expression and subcellular translocation of specific Hsps in ScN2a cells may reflect the cellular response to the accumulation of PrPSc. Whether any of these Hsps feature in the conversion of PrPC into PrPSc or the pathogenesis of prion diseases remains to be established.

Animals↗
PubMed1995

Short circuiting stress protein expression via a tyrosine kinase inhibitor, herbimycin A.

We set out to identify pharmacological means by which to activate the so-called heat shock or stress response and thereby harness the protective effect afforded to the cell by its acquisition of a thermotolerant phenotype. An earlier report by Murakami et al. (1991, Exp. Cell Res., 195: 338-344) described the increased expression of the 70 kDa heat shock proteins in human A431 cells exposed to Herbimycin A (HA), a benzoquinoid ansamycin antibiotic. We show here that treatment of cells with HA results in the increased expression of all of the constitutively expressed stress proteins and confers upon the cells a thermotolerant-like phenotype. Increases in the expression of the stress proteins continued for as long as the cells were exposed to the drug and was independent of the pre-existing levels of the stress proteins. Unlike heat shock or other metabolic stressors, we did not observe any adverse cellular effects following HA exposure. For example, unlike most agents/treatments that elicit the stress response HA-treated cells exhibited no obvious abnormalities with respect to protein maturation, protein insolubility, the integrity of the intermediate filament cytoskeleton, or overall cell viability. In addition, unlike other metabolic stressors, HA treatment did not result in the translocation of hsp 73 into the nucleus/nucleolus. Finally, for at least rodent cells, HA exposure did not result in any obvious activation of the heat shock transcription factor. Based on these findings, we suggest that HA treatment of cells results in a "short-circuiting" of the pathway(s) that normally regulates the expression of the stress proteins. These results are discussed as they pertain to the potential use of HA in animals as a way to harness the protective effects afforded by the stress response.

Animals↗
PubMed1995

Myometrial estradiol and progesterone receptor changes in preterm and term pregnancies.

OBJECTIVE: To determine if labor is associated with changes in myometrial estradiol (E2) and progesterone receptors. METHODS: Lower myometrial segments were obtained from women undergoing cesarean deliveries at term in labor (n = 10), term not in labor (n = 10), preterm in labor (n = 9), and preterm not in labor (n = 11). Western immunoblotting was used to determine the presence and molecular size of E2 and progesterone receptor proteins. Immunocytochemistry was used to determine E2 and progesterone receptor changes in preterm and term pregnancies. RESULTS: Myometrium from pregnant women contained 74-kilodalton (kDa) E2 receptor and 94- and 110-kDa progesterone receptor proteins. These receptors are present in both myometrial smooth muscle and myometrial blood vessels. The nuclei of myometrial smooth muscle cells primarily contain both receptors. The immunostaining for progesterone receptors was less in patients in labor compared with those not in labor in preterm and term pregnancies. In comparing patients not in labor, the immunostaining for progesterone receptors was less at term compared with preterm pregnancy. Unlike the differences in progesterone receptors, there are no obvious differences in E2 receptor immunostaining in myometrial samples from all four groups of women. CONCLUSION: A myometrial decrease in progesterone receptors, rather than an increase in E2 receptors, may play a role in the onset of labor in women with term or preterm pregnancies.

Adult↗
PubMed1995

Glucocorticoid receptor expression in the postnatal rat cochlea.

The glucocorticoid receptor (GR) expression in the neonatal rat cochlea was investigated by utilization of a polyclonal antibody against GR, the immunoreactivity of which exhibited a distinct, age-dependent developmental pattern in tissues of the spiral ligament (SL). Immunostaining of GR appeared initially at the 7th postnatal day (PND), increased rapidly between the 14th and 21st PND, and reached adult-like expression levels by the 21st PND. Less pronounced, developmentally regulated expression patterns of GR were observed in cells of the spiral limbus (SLi), spiral ganglion (SG), organ of Corti (OC), and cochlear nerve (CN). For example, high expression levels of GR were observed in the SLi, SG and OC at 3 PND; subsequently, GR immunoreactivity levels decreased from 7 to 14 PND, and then GR immunoreactivity intensified in these regions by 21 PND. No remarkable changes in GR expression were observed in stria vascularis (SV). These data indicate that GR expression in the inner ear is tissue and age-specific, and that GR expression parallels both Na,K-ATPase expression and endocochlear potential development.

Aging↗
PubMed1995

Decreased expression of functional human chorionic gonadotropin/luteinizing hormone receptor gene in human uterine leiomyomas.

Human myometrium contains functional hCG/LH receptors. The purpose of this study was to determine whether or not leiomyomas that arise from myometrium also contain these receptors. Northern blotting demonstrated that leiomyomas and normal adjacent myometria contained hCG/LH receptor mRNA transcripts. Western immunoblotting showed that leiomyomas and corresponding normal myometria also contained 60- and 50-kDa receptor proteins. Ligand blotting revealed that only the 50-kDa receptor protein in leiomyomas and corresponding normal myometria could bind 125I-hCG and that this binding inhibited by excess unlabeled hCG. In situ hybridization and immunocytochemistry revealed that smooth muscle cells in leiomyomas and corresponding normal myometria contained the hCG/LH receptor mRNA transcripts and receptor proteins. The receptor levels were lower in leiomyomas than in corresponding normal myometria. However, the receptors were functional, as treatment with hCG resulted in a decrease of connexin-43 protein levels in leiomyomas as in normal myometria. In summary, human uterine leiomyomas express a functional hCG/LH receptor gene at a reduced level compared with that for normal corresponding myometria. This finding could be relevant to an understanding of the growth control mechanisms in leiomyomas and the manner in which medical therapy for leiomyomas might work.

Adult↗
PubMed1995

Helicobacter pylori infection and gastric metaplasia in the duodenum in China.

Biopsy specimens from antral and duodenal mucosa were obtained from 64 patients with active duodenal ulcer and from 78 with nonulcer dyspepsia. Gastric metaplasia in the duodenal bulb was more frequent in patients with duodenal ulcer (82.5%) than in patients with dyspepsia and duodenitis (53.1%) or in patients with dyspepsia without duodenitis (34.5%). Helicobacter pylori infection in the duodenal bulb was found more often in patients with moderate to severe gastric metaplasia (62.3%) than in patients with mild gastric metaplasia (20%). Therefore, patients from a developing country, China, showed the same relationship between duodenitis, gastric metaplasia, duodenal colonization with H. pylori, and duodenal ulcer previously demonstrated in developed countries.

Adolescent↗
PubMed1995

Generation of a high-resolution genetic map and a YAC contig of the Lurcher locus on mouse chromosome 6.

Lurcher (Lc) is a semidominant mouse mutant that displays progressive neurodegeneration during perinatal development. This genetic lesion results in apoptotic neuronal death in a dosage dependent and cell autonomous manner in specific neurons during their terminal differentiation. To understand the molecular basis of the Lc mutation, we have adopted a positional cloning approach based on its location on mouse chromosome 6. To define the Lc locus, we have extended our previous analysis of an intersubspecific backcross between Mus m. castaneus and B6CBACa-Aw-j/A-Lc consisting of 504 animals (Norman et al. 1991). In addition, 580 animals of a generic backcross between Mus spretus and C57BL/6 (The European Collaborative Interspecific Backcross) were utilized for the fine genetic mapping of the Lc locus. Using three RFLP markers and nine microsatellite markers in the vicinity of the Lc locus, we determined the order and relative genetic distances of these markers at a resolution of 0.1 cM. The Lc mutation was mapped between two flanking markers, D6Mit121 and D6Mit175, separated by a genetic distance of 0.5 cM. We then initiated the cloning of the genomic region surrounding these two markers by screening a YAC library and characterizing YAC end sequences for further screening. This effort has resulted in the construction of a YAC contig consisting of 14 YACs and spanning a 3-Mb region. Markers isolated from these YACs were used to further define the Lc locus, resulting in a physical map that places the Lc gene within an estimated 300-kb interval. This set of YACs and markers will serve as DNA sources for the identification of the Lc gene.

Animals↗
PubMed1995

Multiple layers of regulation of human heat shock transcription factor 1.

Upon heat stress, monomeric human heat shock transcription factor 1 (hHSF1) is converted to a trimer, acquires DNA-binding ability, is transported to the nucleus, and becomes transcriptionally competent. It was not known previously whether these regulatory changes are caused by a single activation event or whether they occur independently from one another, providing a multilayered control that may prevent inadvertant activation of hHSF1. Comparison of wild-type and mutant hHSF1 expressed in Xenopus oocytes and human HeLa cells suggested that retention of hHSF1 in the monomeric form depends on hydrophobic repeats (LZ1 to LZ3) and a carboxy-terminal sequence element in hHSF1 as well as on the presence of a titratable factor in the cell. Oligomerization of hHSF1 appears to induce DNA-binding activity as well as to uncover an amino-terminally located nuclear localization signal. A mechanism distinct from that controlling oligomerization regulates the transcriptional competence of hHSF1. Components of this mechanism were mapped to a region, including LZ2 and nearby sequences downstream from LZ2, that is clearly separated from the carboxy-terminally located transcription activation domain(s). We propose the existence of a fold-back structure that masks the transcription activation domain in the unstressed cell but is opened up by modification of hHSF1 and/or binding of a factor facilitating hHSF1 unfolding in the stressed cell. Activation of hHSF1 appears to involve at least two independently regulated structural transitions.

Animals↗
PubMed1995

Expression of Na, K-ATPase alpha and beta isoforms in the neonatal rat cochlea.

The postnatal expression of five Na, K-ATPase alpha (alpha 1, alpha 2, alpha 3) and beta (beta 1, beta 2) subunit isoforms in the rat cochlea was investigated by immunocytochemistry. High levels of expression of the alpha 1 and beta 2 isoforms were observed in stria vascularis (SV) at all developmental stages. alpha 1 and beta 1 isoforms showed a distinct time-dependent developmental expression pattern in tissues of the spiral ligament (SL) and spiral limbus (SLi). Limited, temporary expression of alpha 2 and alpha 3 subunit isoforms were found in SV and SL. Expression of each isoform was also seen in organ of Corti (OC), spiral ganglion (SG), cochlear nerve (CN) and Kölliker's Organ (KO). These observations suggest that individual isoforms may exert specific actions postnatally during final cochlear maturation.

Animals↗
PubMed1995

[Proto-oncogene point mutation of c-Ha-ras in bronchogenic carcinoma].

11 bronchogenic carcinoma and 8 corresponding paracarcinoma specimens were detected for activating point mutation in codon 12 of c-Ha-ras gene by Southern blotting hybridization technique. The c-Ha-ras gene was found to be activated by point mutation in codon 12 in 8 of bronchogenic carcinoma and 1 of paracarcinoma tissues. The mutative and unmutative hybrid band was both discovered in 2 of paracarcinoma tissues. Analysis of clinical and pathological features, we conclude that: (1) The point mutation of c-Ha-ras gene codon 12 is found in 77.8% bronchogenic carcinoma among Chinese; (2) At least in part cases, mutation of c-Ha-ras gene codon 12 is not the "initiation" event, but may be a early-stage event in the pathogenesis; (3) No apparent association between the c-Ha-ras gene codon 12 mutation and pathological types, disease stages and history of smoking is present.

Adenocarcinoma↗
PubMed1994

Activation of the DNA-binding ability of human heat shock transcription factor 1 may involve the transition from an intramolecular to an intermolecular triple-stranded coiled-coil structure.

Heat stress regulation of human heat shock genes is mediated by human heat shock transcription factor hHSF1, which contains three 4-3 hydrophobic repeats (LZ1 to LZ3). In unstressed human cells (37 degrees C), hHSF1 appears to be in an inactive, monomeric state that may be maintained through intramolecular interactions stabilized by transient interaction with hsp70. Heat stress (39 to 42 degrees C) disrupts these interactions, and hHSF1 homotrimerizes and acquires heat shock element DNA-binding ability. hHSF1 expressed in Xenopus oocytes also assumes a monomeric, non-DNA-binding state and is converted to a trimeric, DNA-binding form upon exposure of the oocytes to heat shock (35 to 37 degrees C in this organism). Because endogenous HSF DNA-binding activity is low and anti-hHSF1 antibody does not recognize Xenopus HSF, we employed this system for mapping regions in hHSF1 that are required for the maintenance of the monomeric state. The results of mutagenesis analyses strongly suggest that the inactive hHSF1 monomer is stabilized by hydrophobic interactions involving all three leucine zippers which may form a triple-stranded coiled coil. Trimerization may enable the DNA-binding function of hHSF1 by facilitating cooperative binding of monomeric DNA-binding domains to the heat shock element motif. This view is supported by observations that several different LexA DNA-binding domain-hHSF1 chimeras bind to a LexA-binding site in a heat-regulated fashion, that single amino acid replacements disrupting the integrity of hydrophobic repeats render these chimeras constitutively trimeric and DNA binding, and that LexA itself binds stably to DNA only as a dimer but not as a monomer in our assays.

Amino Acid Sequence↗
PubMed1994

Lack of self-regulation of human chorionic gonadotropin biosynthesis in human choriocarcinoma cells.

Human gestational trophoblastic neoplasms overexpress hCG/LH receptors. Whether this overexpression is a reflection of a loss of self-regulation of hCG biosynthesis was investigated using JAR human choriocarcinoma cells. The results show that exogenous hCG did not affect steady state hCG alpha and hCG beta mRNA or dimer hCG protein levels in JAR cells. The JAR cells, however, responded to 8-bromo-cAMP with an increase in hCG alpha mRNA levels, suggesting that cAMP-mediated regulation of the hCG subunit genes was intact in the cells. Disruption of receptor function by a receptor antibody, which resulted in an increase in hCG alpha mRNA levels and hCG secretion in normal trophoblasts, had no effect on JAR cells. Unlike normal trophoblasts, which contain a predominant receptor transcript of 1.8 kilobases (kb), with minor higher molecular size (7.5 and 5.4 kb) transcripts occasionally seen, JAR cells contain a higher abundance of multiple transcripts (7.5, 5.4, 3.5, and 1.8 kb), with the predominant transcript being 5.4 kb. In addition, although normal trophoblasts contain an 80-kilodalton receptor protein, JAR cells contain only a 50-kilodalton hCG/LH receptor isoform. In contrast to the effects of exogenous hCG on normal placental tissue in vitro, it was unable to down-regulate receptor transcripts or receptor protein in JAR cells. In summary, JAR cells lack the ability to self-regulate hCG biosynthesis. This loss could explain how hCG can reach very high levels in gestational trophoblastic disease compared to those in normal pregnancy.

Choriocarcinoma↗
PubMed1994

Differential cyclooxygenase-1 and -2 gene expression in human myometria from preterm and term deliveries.

The expression of cyclooxygenase-1 (COX-1) and COX-2 genes in myometria from preterm and term pregnancies was investigated by in situ hybridization, immunoblotting, and immunocytochemistry. In situ hybridization revealed that myometrial smooth muscle contained both COX-1 and COX-2 messenger ribonucleic acid transcripts. Immunoblotting revealed that myometria contained 55- and 67-kilodalton COX-1 proteins as well as 72- and 56-kilodalton COX-2 proteins. Immunocytochemistry showed that COX-1 and COX-2 proteins are present in the myometrial smooth muscle. The expression of COX-1 and COX-2 genes differed in myometria. For example, although COX-1 expression was lower, COX-2 expression was higher at term compared to preterm pregnancy not in labor. In addition, although COX-1 expression was not dependent on labor, COX-2 expression was lower with labor compared to not in labor for both preterm and term pregnancy. The myometrial smooth muscle also immunostained for prostaglandin E2 (PGE2) and PGF2 alpha, suggesting that the cyclooxygenases are catalytically active. The changes in PGE2 and PGF2 alpha paralleled the changes in COX-1 enzyme in terms of being lower at term compared to preterm pregnancy and the changes in COX-2 enzyme in terms of being lower during labor at preterm and term pregnancy. In summary, our results demonstrate that pregnant human myometria express both COX-1 and COX-2 genes. The expression of these genes differed in preterm or term pregnancy myometria from in labor and not in labor.

Dinoprost↗
PubMed1994

Human myometrial chorionic gonadotropin/luteinizing hormone receptors in preterm and term deliveries.

Nonpregnant human myometrium contains functional hCG/LH receptors. The present study investigated whether pregnant human myometrium also contains these receptors and whether they vary as a function of delivery in preterm or term pregnancies. Northern blotting revealed that pregnant human myometrium contains 4.3- and 2.2-kilobase receptor messenger ribonucleic acid transcripts. Immunoblotting with a specific hCG/LH receptor antibody showed that myometrium contains 70- and 50-kilodalton immunoreactive proteins. Ligand blotting demonstrated that only the 50-kilodalton protein could bind [125I]hCG, and this binding was inhibited by excess unlabeled hCG. In situ hybridization and immunocytochemistry revealed that the receptor messenger ribonucleic acid and receptor protein are present in myometrial and vascular smooth muscle. The myometrial smooth muscle receptor levels were lower during labor compared to those before labor at preterm and term pregnancy. In summary, our study demonstrates that pregnant human myometrium express hCG/LH receptor gene. The receptor levels were lower during labor compared to those before labor in preterm or term pregnancy. These data suggest that hCG, via its receptors, may contribute to myometrial quiescence until labor begins at the end of pregnancy.

Blotting, Northern↗
PubMed1994

[An approach for PCR amplification of long DNA fragments].

The polymerase chain reaction (PCR) technique, evolved as a standard laboratory technique, has a major limitation: its inability to efficiently amplify fragments higher than 1.5 kb. In this paper, PCR conditions allowing the efficient amplification of long DNA fragments, especially a new proper buffer system, have been developed. The difference between the new PCR reaction buffer system and standard reaction system is that no KCl is included in the former. The results indicated that specificity and reproducibility for amplifying long DNA fragments by using new buffer system are satisfactory.

Animals↗
PubMed1994

Spectrum of glycidyl methacrylate-induced mutation in plasmid-Escherichia coli system.

In order to characterize the spectrum of mutation induced by glycidyl methacrylate (GMA), the plasmid pBR322 was modified with this mutagen in vitro, transfected into appropriate Escherichia coli host HB101. The mutants were then screened and defined by DNA sequencing. Sequence analysis reveals that GMA induces two classes of mutations: deletion of the mono-, di- or tetra-base or the insertion of mono- or di-base. Both types of mutations, with about 10% frequency, occur predominantly at C-G runs and at 5'-CNCCN-3' sequence, which are hotspots for GMA damage and may cause frameshift mutation.

Amino Acid Sequence↗