Biomedical subjects

K W Cho

Publications and source records attributed to K W Cho.

At least 91 records · Page 5Linked to original sources

PubMed1995

Functional conservation of the Wnt signaling pathway revealed by ectopic expression of Drosophila dishevelled in Xenopus.

Wnt genes encode secreted growth factors that exhibit potent effects on both embryonic and postembryonic development in vertebrates and invertebrates. Recently, the dishevelled (dsh), shaggy/zeste-white 3, and armadillo genes have been shown to participate in Wnt (wingless; wg) signaling in Drosophila. Vertebrate genes that have sequence similarities to all of these Drosophila genes have been identified. To determine whether these structurally conserved components of insect wg signaling represent a functionally conserved Wnt signaling pathway in vertebrates, we investigated the role of Drosophila dsh in Xenopus Wnt signaling. Xenopus embryos ectopically injected with Drosophila dsh mRNA developed duplicated axes similar to those seen in embryos injected with Wnt mRNAs. The involvement of dsh function in the Wnt signaling pathway in Xenopus was demonstrated using two assays which are specifically sensitive to Wnt signaling: synergistic induction of dorsal mesoderm with bFGF and the specific induction of a Wnt-responsive reporter gene. These findings support the notion that the intracellular response to the Wnt signal has been conserved during evolution to such an extent that its components may be interchanged between distantly related species.

Animals↗

PubMed1995

Mechanical basis of ANP secretion in beating atria: atrial stroke volume and ECF translocation.

To investigate the mechanism by which an increase in pacing frequency or distension increases the secretion of atrial natriuretic peptide (ANP), the changes in atrial volume during contraction (atrial stroke volume), transmural transport of the extracellular fluid (ECF), and ANP secretion were quantified in the beating perfused rabbit atria. The atrium was stimulated by transmural field stimulation or by atrial distension induced by an increase in intraatrial pressure. Atrial stretch and incremental increases in pacing frequency up to 2 Hz activated the secretion of ANP coincident with an increase in atrial stroke volume and the transendocardial translocation of the ECF. These results showed positive relationships between changes in the secretion of ANP and the atrial stroke volume or the translocation of the ECF. The translocation of the ECF was also positively correlated with the change in atrial stroke volume. The accentuated secretion of ANP and translocation of the ECF waned at higher stimulating rates to show a peak value. Even under this condition, the secretion of ANP was a function of the translocation of the ECF. These data suggest that the increases in atrial stroke volume and translocation of ECF are fundamental factors in the ANP stimulation in response to atrial stretch and increases in atrial rate.

Animals↗

PubMed1995

Anterior neurectoderm is progressively induced during gastrulation: the role of the Xenopus homeobox gene orthodenticle.

In order to study the regional specification of neural tissue we isolated Xotx2, a Xenopus homolog of the Drosophila orthodenticle gene. Xotx2 is initially expressed in Spemann's organizer and its expression is absent in the ectoderm of early gastrulae. As gastrulation proceeds, Xotx2 expression is induced in the overlying ectoderm and this domain of expression moves anteriorly in register with underlying anterior mesoderm throughout the remainder of gastrulation. The expression pattern of Xotx2 suggests that a wave of Xotx2 expression (marking anterior neurectoderm) travels through the ectoderm of the gastrula with the movement of underlying anterior (prechordal plate) mesoderm. This expression of Xotx2 is reminiscent of the Eyal-Giladi model for neural induction. According to this model, anterior neural-inducing signals emanating from underlying anterior mesoderm transiently induce anterior neural tissues after vertical contact with the overlying ectoderm. Further patterning is achieved when the ectoderm receives caudalizing signals as it comes in contact with more posterior mesoderm during subsequent gastrulation movements. Functional characterization of the Xotx2 protein has revealed its involvement in differentiation of the anterior-most tissue, the cement gland. Ectopic expression of Xotx2 in embryos induces extra cement glands in the skin as well as inducing a cement gland marker (XAG1) in isolated animal cap ectoderm. Microinjection of RNA encoding the organizer-specific homeo-domain protein goosecoid into the ventral marginal zone results in induction of the Xotx2 gene. This result, taken in combination with the indistinguishable expression patterns of Xotx2 and goosecoid in the anterior mesoderm suggests that Xotx2 is a target of goosecoid regulation.

Animals↗

PubMed1994

Effect of extracellular calcium depletion on the two-step ANP secretion in perfused rabbit atria.

Employing isolated perfused rabbit atrial model we have found that stretch-activated atrial natriuretic peptide (ANP) secretion takes place in two steps: release of ANP from myocytes into surrounding intercellular space, and then translocation of the released ANP into atrial lumen along with the extracellular fluid (ECF) translocated upon releasing the stretch. Ca2+, one of the most important factors regulating secretory processes, has been reported by many workers to have influence on the ANP secretion, but at large variance. In the present study, therefore, we undertook to clarify the influence of Ca2+ depletion on ANP secretion and further to define which of the two steps is affected by Ca2+ removal. Extracellular Ca2+ depletion resulted in increased basal secretion and in accentuated secretion of immunoreactive (ir) ANP in response to mechanical stimuli. The translocation of ECF increased in response to atrial stretch-and-release, but it was not affected by Ca2+ depletion. The irANP concentration in the extracellular space calculated as the amount of irANP secreted in relation to the ECF translocated significantly increased when extracellular Ca2+ was depleted. These results indicate that extracellular Ca2+ depletion accentuates the stretch-induced ANP secretion through the augmentation of ANP release into the interstitium without changes in the ECF translocation.

Animals↗

PubMed1994

Mechanism of aldehyde inhibition of Vibrio harveyi luciferase. Identification of two aldehyde sites and relationship between aldehyde and flavin binding.

Vibrio harveyi luciferase is sensitive to aldehyde substrate inhibition, and two kinetic schemes have been previously postulated to account for such an inhibition. One scheme depicts a sequential binding of 2 aldehyde molecules, yielding an active enzyme-aldehyde binary complex and subsequently an inactive enzyme-(aldehyde)2 ternary complex (Holzman, T. F., and Baldwin, T. O. (1983) Biochemistry 22, 2838-2846). This two-aldehyde model was later withdrawn, and recently, a different scheme was proposed, following which the prior binding of one aldehyde to the native luciferase forms an inactive dead-end complex (Abu-Soud, H. M., Clark, A. C., Francisco, W. A., Baldwin, T. O., and Raushel, F. M. (1993) J. Biol. Chem. 268, 7699-7706). In this work, kinetic and equilibrium studies were carried out to elucidate further the mechanism of aldehyde inhibition. Two, presumably independent, aldehyde-binding sites were detected, with a higher affinity site for the aldehyde substrate and a weaker affinity site for the aldehyde inhibitor. Binding to and dissociation from the inhibitor site by decanal were revealed by chemical relaxation analysis to be slow processes. Furthermore, whereas the binding of the decanal substrate enhances the affinity of the reduced riboflavin 5'-phosphate (FMNH2) site, the binding of decanal to the inhibitor site competes against FMNH2 binding, thus resulting in inhibition of luciferase activity. These findings are not compatible with either of the two earlier schemes mentioned above. A new kinetic model is formulated for the mechanism of aldehyde inhibition. Theoretical kinetic behaviors predicted on the basis of this model are in excellent agreement with experimental observations. A particularly reactive cysteine (residue 106) on the alpha subunit has been previously demonstrated to be at or near an aldehyde site (Fried, A., and Tu, S.-C. (1984) J. Biol. Chem. 259, 10754-10759). Evidence is presented to indicate that this residue is at or near the aldehyde inhibitor site. Relative locations of this residue and binding sites for FMNH2, the aldehyde substrate, and the aldehyde inhibitor are proposed.

Aldehydes↗

PubMed1994

Identification of immunoreactive atrial natriuretic peptide in the gallbladder and bile juice of rabbit, pig and human.

The presence of immunoreactive atrial natriuretic peptide (irANP) in rabbit, pig and human gallbladders was investigated using radioimmunoassay and immunohistochemistry. Serial dilution curves of gallbladder tissue and bile juice extracts were paralleled to the standard curve of atriopeptin III. Gel filtration profiles of gallbladder tissue extracts showed a major peak corresponding to rat pro-ANP. The amounts of irANP in rabbit, pig and human gallbladders were 30.0 +/- 12.3 pg/mg (n = 7), 7.0 +/- 2.0 fg/mg (n = 7) and 17.7 +/- 2.0 fg/mg wet tissue (n = 8), respectively. Bile juice was also shown to contain irANP but with small molecular mass. The amounts of irANP in the rabbit, pig and human bile juice were 25.0 +/- 2.0 (n = 7) and 0.50 +/- 0.02 (n = 7), and 1.3 +/- 0.1 pg/ml (n = 8), respectively. The immunohistochemical staining of the rabbit gallbladder tissue revealed the presence of irANP in the luminal epithelium and smooth muscle layer. The amount of irANP was higher in the luminal epithelium than in the rest of the gallbladder tissue from rabbits (0.30 +/- 0.06 vs. 0.01 +/- 0.01 pg/microgram protein, P < 0.01). These findings suggest that ANP may be synthesized and stored in the gallbladder, and may have a role in the regulation of fluid balance and cystic motility.

Animals↗

PubMed1994

Atrial natriuretic peptide immunoreactivity in the eggs of the silkworm Bombyx mori.

Synthesis and secretion of atrial natriuretic peptides (ANPs) are not confined to the atrium, but are also present in other tissues. Recently, we have found synthesis of ANP in the eggs of several vertebrate animals. The present study was undertaken to determine whether immunoreactive ANP (irANP) is present in the egg of an invertebrate, the silkworm (Bombyx mori L.). The serial dilution curve of egg extracts of silkworm was parallel to the standard curve of atriopeptin III. Analysis of ANP immunoreactivity by gel filtration chromatography and reverse-phase HPLC showed that the major immunoreactivity corresponded to rat proANP. The semipurified irANP of egg extracts produced a dose-dependent relaxation on rat aortic strips, which was blocked by preincubation with anti-ANP antiserum. Therefore, we suggest that ANP is synthesized in the silkworm egg.

Animals↗

PubMed1994

Expression of zebrafish goosecoid and no tail gene products in wild-type and mutant no tail embryos.

goosecoid is an immediate early gene expressed at the dorsal blastoporal lip of the Xenopus gastrula. Microinjection experiments have suggested a direct role for goosecoid in organizing the dorsoventral axis of the frog embryo. Here we characterize the zebrafish homologue of goosecoid (gsc) and compare its expression to that of Brachyury or no tail (ntl), another immediate early gene required in developing mesoderm. We show that gsc exhibits two independent phases of expression: an early one in cells anterior to the presumptive notochord, but not in cells of the notochord itself, and a later one in neural crest derivatives in the larval head. Zygotic gsc transcripts are detected soon after the midblastula transition, and at the blastula stage form a gradient with a maximum at the dorsal side. Use of gsc as a dorsal marker allowed us to demonstrate that ntl expression is initially activated at the dorsal side of the blastula. At this early stage, gsc and ntl show overlapping domains of expression and are co-expressed in cells at the dorsal midline of the early gastrula. However, gsc- and ntl-expressing cells become separated in the course of gastrulation, with gsc being expressed in the axial hypoblast (prechordal plate) anterior to the ntl-expressing presumptive notochord cells. Studies with mutant embryos suggest that gsc is independent of ntl function in vivo.

Amino Acid Sequence↗

PubMed1994

Effects of positive acceleration on atrial natriuretic peptide in humans.

To study the effects of positive acceleration on the plasma atrial natriuretic peptide (ANP) and renin concentration (PRC), students of the Korean Air Force Academy and pilots were exposed to +6 Gz for 30 s in a human centrifuge without anti-G suits. An acceleration to +6 Gz caused a significant increase (p < 0.01) in the heart rate of both the student and pilot groups. We performed blood collection within 2 min after terminating centrifugation. Systolic blood pressure was increased significantly (p < 0.01), and heart rates were elevated and did not return to control levels by 2 min after centrifugation. Plasma ANP decreased significantly in both groups (p < 0.05). The difference in the percentage of ANP decrement was not significant between student and pilot groups. PRC increased significantly (p < 0.05) in both groups. This study demonstrated that high +Gz exposure in man caused a significant reduction of plasma ANP concentration, although PRC, heart rate, and blood pressure increase.

Acceleration↗

PubMed1993

The homeobox gene goosecoid controls cell migration in Xenopus embryos.

Goosecoid (gsc), a homeobox gene expressed specifically in the dorsal blastopore lip of the Xenopus gastrula, is considered to play an important role in Spemann's organizer phenomenon. Lineage tracing and time-lapse microscopy were used to follow the fate of embryonic cells microinjected with gsc mRNA. Microinjected gsc has non-cell autonomous effects, recruiting neighboring uninjected cells into a twinned dorsal axis. Ectopic expression of gsc mRNA in ventral blastomeres as well as overexpression of gsc in dorsal blastomeres leads to cell movement toward the anterior of the embryo. The results suggest a function for gsc in the control of gastrulation movements in groups of cells, but not in dissociated cells, and demonstrate that a vertebrate homeobox gene can regulate region-specific cell migration.

Animals↗

PubMed1993

Characterization of the Xenopus Hox 2.4 gene and identification of control elements in its intron.

We report on the Xenopus homolog of the Hox 2.4 gene. This gene occupies the next to 5'-most position in the Xenopus Hox2 complex. Hox 2.4 RNA is first detected at the early neurula stage, reaching a peak at the early tailbud stage, and is localized in the middle and posterior portions of the embryos. Antibodies raised against a fusion protein show expression of Hox 2.4 protein in Xenopus embryos in a band located in the mid spinal cord. Thus, the protein is expressed in a narrower domain than that of Hox 2.4 mRNA. The Xenopus Hox 2.4 antibody cross-reacts readily with mouse embryonic tissue, where the protein is detected in migrating neural crest cells, the dorsal portion of the spinal cord, somites, lateral plate mesoderm, and in the forelimb bud. The Xenopus Hox 2.4 intron shares considerable sequence identity with the intron in the mouse homolog. A reporter gene containing an element from this intron which can bind homeodomain proteins is activated following microinjection into Xenopus embryos. The short distance between the end of the Hox 2.4 cDNA and the start site of the neighboring gene in the complex raises the possibility that this transcriptional element might be shared by two Hox genes.

Amino Acid Sequence↗

PubMed1993

Single injection of pentobarbital induces long-lasting effects on ANP synthesis and gene expression in the rat atria.

To define the long-term effects of pentobarbital sodium on the plasma and atrial atrial natriuretic peptide (ANP) system, experiments were performed in female Sprague-Dawley rats. The plasma levels of immunoreactive (ir) ANP showed chronic as well as acute response to pentobarbital sodium administration. A single dose (30 mg/kg, i.p.) of pentobarbital sodium resulted in a suppression in the plasma levels of irANP up to 1 week of administration. The suppressive effect on plasma irANP concentrations was dose-dependent. Right but not left atrial contents of irANP increased by an administration of pentobarbital sodium up to 4 weeks. ANP mRNA contents of the atrial exposed to pentobarbital sodium began to increase after 2 days, reached to the peak after 2 weeks, and began to return to control values after 6 weeks. Surgical stress accentuated these patterns of plasma and atrial ANP responses to pentobarbital sodium treatment. The present data, therefore, suggest that even a single anesthetic dose of pentobarbital could elicit long-lasting profound changes in ANP system, i.e., changes in gene expression, synthesis and the secretion of ANP.

Animals↗

PubMed1993

Immunoreactive atrial natriuretic peptides in the oocyte.

1. The presence and partial characterization of immunoreactive ANP (irANP) in eggs were investigated using high performance liquid chromatography (HPLC), immunohistochemistry and Northern-blot hybridization in mammalian and nonmammalian vertebrates. 2. Serial dilution curves of egg extracts from frogs and freshwater teleostean fishes (silver crusian carp and loach) were parallel to the standard curve of atriopeptin III. Rat oocytes also contained irANP (25-30 pg/egg). 3. The HPLC profile of irANP showed two main peaks corresponding to low and high molecular weight of irANP. 4. ANP mRNA was detected in the fish eggs. With immunohistochemical analysis of the rat ovarian follicles, this ANP-like material was localized mainly at the oocytes. 5. We, therefore, suggest that the vertebrate oocytes synthesize ANP and that the presence of ANP in the oocytes may be linked with the cell differentiation.

Animals↗

PubMed1993

Fluorescent polyene aliphatics as spectroscopic and mechanistic probes for bacterial luciferase: evidence against carbonyl product from aldehyde as the primary excited species.

The fluorescent alpha-parinaric acid (alpha-PAC) and beta-parinaric acid (beta-PAC) were converted to the corresponding aldehydes and alcohols all of which exhibited absorption and fluorescence properties closely resembling those of the parent acids. alpha-PAC and beta-PAC each binds to luciferase in competition with aldehyde. The hydrophobic nature of the aldehyde site was indicated by the enhanced fluorescence quantum yields of the bound alpha-PAC and beta-PAC. These two polyene acids and the beta-parinaryl alcohol were shown to stabilize the luciferase flavin-peroxide intermediate. alpha-Parinaraldehyde (alpha-PAD) and beta-parainaraldehyde (beta-PAD) were active substrates for Vibrio harveyi and Vibrio fischeri luciferases and, for the former enzyme, exhibited Km values similar to and quantum yields about 20-30% as those for decanal and dodecanal. For the V. harveyi luciferase with reduced FMN as a co-substrate, the alpha-PAD- or beta-PAD-initiated luminescence was indistinguishable from the normal emission obtained with octanal (lambda max 495 nm) showing no additional 430-nm component correlatable with emission from excited alpha-PAC or beta-PAC. In reactions using reduced 2-thioFMN for V. harveyi luciferase or reduced FMN for V. fischeri luciferase plus yellow fluorescent protein, the replacement of octanal by beta-PAD again resulted in no additional 430-nm emission. The lack of any emission correlatable with excited alpha-PAC, beta-PAC, or equivalent carbonyl product was not due to the quenching of the polyene moiety by chemical transformation, binding to luciferase, or a 100% energy transfer to the flavin 4a-hydroxide emitter.(ABSTRACT TRUNCATED AT 250 WORDS)

Aldehydes↗

PubMed1993

Extracellular fluid translocation in perfused rabbit atria: implication in control of atrial natriuretic peptide secretion.

1. Transmural transport of 22Na+, 51Cr-EDTA, [3H]inulin and [14C]Dextran (57 kDa) was measured in perfused rabbit atria. The radiolabelled extracellular space (ECS) markers and [14C]Dextran were introduced into the pericardial space or atrial lumen. Atrial volume changes were induced by steps up and down in atrial pressure. 2. Basal rates of transmural transport of radiolabelled ECS markers across the atrial wall were relatively stable up to 70 min. Atrial stretch and release resulted in a rapid but transient, and reversible increase in the ECS fluid (ECF) translocation. The increased translocation of the ECF into the atrial lumen occurred within 15 s of the reduction of atrial distension and returned to the baseline level within 60 s. 3. Transmural transport of [3H]inulin across the atrial wall was bidirectional. 4. The clearance of radiolabelled ECS markers was molecular-size dependent. The transmural clearance of [3H]inulin was dependent on the distension-reduction volume changes induced by atrial stretch and release. Little transport of [14C]Dextran across the atrial wall was observed. 5. The ECF translocation across the atrial wall was not influenced by changes in external Ca2+ but was suppressed by low temperature. 6. Dynamic changes in the ECS of the atrium were observed in response to atrial distension and reduction. The ECS of the atrium increased on distension and decreased on reduction of atrial distension. 7. Reduction in atrial distension resulted in an increase in the secretion of immunoreactive atrial natriuretic peptide (ANP) which coincided with an increase in the translocation of the ECF. The secretion of immunoreactive ANP was a function of the translocation of the ECF. 8. It is suggested that atrial stretch and release may play a role in driving fluid flow within the interstitium and fluid translocation out of the interstitium. This fluid movement presumably leads to convective transport of released ANP into the atrial lumen.

Animals↗

PubMed1992

Presence and release of immunoreactive atrial natriuretic peptide in granulosa cells of the pig ovarian follicle.

Atrial natriuretic peptide (ANP) has been reported to be locally synthesized in the ovary although its physiological roles are still unknown. To define the origin of ovarian ANP, we demonstrated the presence and release of immunoreactive (ir) ANP in pig granulosa cells and characterized its biochemical properties. Serial dilution curves made with the extracts of pig granulosa cells, their perfusates and follicular fluid were paralleled to the standard curve of ANP. The amount of irANP in the granulosa cell was 2 fg/cell. The total amount of irANP in granulosa cells significantly correlated with the levels of irANP in follicular fluid. Additionally, the total content of irANP in the follicle negatively correlated with the follicular size. On reverse phase HPLC, the major form of irANP in granulosa cells and follicular fluid was high molecular weight but that in perfusate was low molecular weight. In Northern blot analysis, ANP mRNA was detected in the pig granulosa cells. Immunohistochemistry showed ANP prohormone location in granulosa cells of rat ovary. These data strongly suggest that the granulosa cells synthesize and secrete ANP.

Animals↗

PubMed1992

Gastrulation in the mouse: the role of the homeobox gene goosecoid.

Mouse goosecoid is a homeobox gene expressed briefly during early gastrulation. Its mRNA accumulates as a patch on the side of the epiblast at the site where the primitive streak is first formed. goosecoid-expressing cells are then found at the anterior end of the developing primitive streak, and finally in the anteriormost mesoderm at the tip of the early mouse gastrula, a region that gives rise to the head process. Treatment of early mouse embryos with activin results in goosecoid mRNA accumulation in the entire epiblast, suggesting that a localized signal induces goosecoid expression during development. Transplantation experiments indicate that the tip of the murine early gastrula is the equivalent of the organizer of the amphibian gastrula.

Activins↗

PubMed1992

Right atrial predominance of atrial natriuretic peptide secretion in isolated perfused rat atria.

In order to investigate the regulatory mechanism for the atrial release of atrial natriuretic peptide (ANP), a perfused rabbit atrial model was devised. In the present experiments, the effect of a reduction in atrial distension on the immunoreactive ANP (irANP) secretion was investigated and compared in the perfused right and left atria of rats. Elevations in right and left atrial pressure resulted in proportional increases in the volume of atrial distension-reduction which was larger in the right than in the left atria. The basal rate of irANP secretion was higher in the right than in the left atria. Increases in the volume of atrial distension-reduction resulted in proportional increases in irANP secretion in both atria. Increment in irANP secretion in response to a reduction in atrial distension was significantly higher in the right than in the left atria. Higher rate of irANP secretion in response to unit volume change was observed in the right atria. Increases in the volume of atrial distension-reduction resulted in accentuated irANP responses in the right atrium. IrANP content was significantly higher in the right than in the left atria. The results suggest that the right atrium is a predominant site in ANP secretion in rats.

Animals↗