• Bulletin of the Korean Chemical Society
• /
• v.22 no.5
• /
• pp.514-518
• /
• 2001

We have explored the importance of two ATP binding domains of Hsp104 protein in protection of yeast cells from cadmium exposure. In the previous study we have discovered that the presence of two ATP binding sites was essential in providing heat sh ock protection as well as rescuing cells from oxidative stress. In this paper we first report wild type cell with functional hsp104 gene is more resistant to cadmium stress than hsp104-deleted mutant cell, judging from decrease in survival rates as a result of cadmium exposure. In order to demonstrate functional role of two ATP binding sites in cadmium defense, we have transformed both wild type (SP1) and hyperactivated ras mutant (IR2.5) strains with several plasmids differing in the presence of ATP binding sites. When an extra copy of functional hsp104 gene with both ATP binding sites was overexpressed with GPD-promoter, cells showed increased survival rate against cadmium stress than mutants with ATP binding sites changed. The degree of protection in the presence of two ATP binding sites was similarly observed in ira2-deleted hyperactivated ras mutant, which was more sensitive to oxidative stress than wild type cell. We have concluded that the greater sensitivity to cadmium stress in the absence of two ATP binding sites is attributed to the higher concentration of reactive oxygen species (ROS) produced by cadmium exposure based on the fluorescence tests. These findings, taken all together, imply that the mechanism by which cadmium put forth toxic effects may be closely associated with the oxidative stress, which is regulated independently of the Ras-cAMP pathway. Our study provides a better understanding of cadmium defense itself and cross-talks between oxidative stress and metal stress, which can be applied to control human diseases due to similar toxic environments.

• Journal of Environmental Health Sciences
• /
• v.22 no.4
• /
• pp.91-101
• /
• 1996

Exposure indices are important tools which enable scientists to reliably predict and detect exposures to xenobiotics and resultant cell injury. Since the de novo synthesis of stress proteins can be detected early after exposure to some agents, analysis of toxicant-induced changes in gene expression, i.e. alterations in patterns of protein synthesis, may be useful to develop as biomarkers of exposure and toxicity. The acute and chronic effects of cadmium(Cd, $CdCl_2$ 20 mg/kg) on Wistar male rats were evaluated concerning cadmium contents, tissues enzyme activity, HSP expression. The results of the study were as follows: 1. Less cadmium was absorbed through the digestive tracts, but the ratio of contents in renal to hepatic cadmium was higher at 8 weeks after treatment. 2. ALT(alanine aminotransferase), AST(aspartate aminotransferase), glucose, BUN(blood urea nitrogen), creatinine, the key indices of the clinical changes in hepatic and renal function were significantly changed by the cadmium treatment after 1 week in liver, after 4 weeks in kidney. 3. Enhanced synthesis of 70 KDa relative molecular mass proteins were detected in 2 hours after cadmium exposure, with maximum activity occurring at 8~48 hours. Induction of $HSP_{70}$ was evident at proximal tubules and glomeruli in kidney. Testicular cells produced enough HSP to be detected normally. From the above results, it could be concluded that $HSP_{70}$ induction by the cadmium treatment was a rapid reaction to indicate the exposure of xenobiotics.

• Reproductive and Developmental Biology
• /
• v.35 no.3
• /
• pp.307-311
• /
• 2011

Heat shock protein 90 (Hsp90) is ATPase-directed molecular chaperon and affects survival of several cells. In our previous study, inhibitory effect of Hsp90 by inducing cell cycle arrest and apoptosis in the pig embryonic and primary cells was reported. However, its role during early bovine embryonic development is not sufficient. In this study, we traced the effects of Hsp90 inhibitor, 17-allylamino-17-demethoxygeldanamycin (17-AAG), on early bovine embryonic development. We also investigated several indicators of developmental potential, including structural integrity, gene expression (apoptosis-related genes), and apoptosis, which are affected by 17-AAG. Bovine embryos were cultured in the CR1-aa medium with or without 17-AAG for 7 days. In result, significant differences in developmental potential were detected between the embryos that were cultured with or without 17-AAG ($33.1{\pm}9.6$ vs $21.7{\pm}8.3%$). The structural integrity of the blastocysts was examined by differential staining. Blastocysts from the dbcAMP-treated group had higher numbers of ICM, TE, and total cells than those from the untreated group. Terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) showed that the number of containing fragmented DNA at the blastocyst stage increased in the 17-AAG treated group compared with control (11.2 vs 3.9, respectively). Blastocysts that developed in the 17-AAG treated group had low structural integrity and high apoptotic nuclei than those of the untreated control, resulting in decrease the embryonic qualities of preimplantation bovine blastocysts. The mRNA expression of the pro-apoptotic gene (Bax) increased in 17-AAG treated group, whereas expression of the antiapoptotic gene (Bcl-XL) decreased. In conclusion, Hsp90 also appears to play a direct role in bovine early embryo developmental competence including structural integrity of blastocysts. Also, these results indicate that Hsp90 is closely associated with apoptosis-related genes expression in developing bovine embryos.

• Food Science and Preservation
• /
• v.15 no.4
• /
• pp.562-567
• /
• 2008

To develop human risk assessment protocols, we explored housekeeping gene and cytokine expression in mouse spleen cells using Rt-PCR. We normalized housekeeping gene expression by RT-PCR; gene expression was highly uniform in potato leafs and mouse spleen cells. We measured the expression of frequently used housekeeping genes, such as those encoding APRT, $\beta$-tubulin, Actin, Hsp 20.2, Cyclophilin, 18S RNA, Efla, Tbp, GAPDH, $\beta$-actin, Tuba2, Hprt, Cyclophlin A, Tfrc, and RPL13A in mice fed GM or non-GM potatoes. Housekeeping gene expression did not show any significant differences between GM and non-GM potato-fed mice. The murine model of potato-fed mice did not express IL-4 and IL-13 at a significant levels.

• Journal of the Korea Academia-Industrial cooperation Society
• /
• v.13 no.8
• /
• pp.3785-3791
• /
• 2012

Dissolved oxygen is one of the most important factors controlling growth in aquatic organisms. Hypoxia is generally defined as dissolved oxygen less than 2.8 mg $O_2/L$ (equivalent to 2 mL $O_2/L$ or 91.4 mM). Therefore, hypoxia zone can cause a serious problem in marine ecosystem. In this study, to investigate embryotoxic (fertilization and embryo development rates) effects of hypoxia on sea urchin Strongylocentrotus nudus were exposed to dissolved oxygen levels of 7.6 mg $O_2/L$ (normoxia) and 1.8 mg $O_2/L$ (hypoxia) for 2 days at $15^{\circ}C$ and 33 ‰. Also, Expression levels of stress related gene (HSP70) and antioxidant related gene (glutathione reductase) in the sea urchins exposed to hypoxia were confirmed by Immunoblotting and RT-PCR analysis. In results, we showed that developmental rates were dramatically reduced in hypoxia condition. Molecular analysis demonstrated that higher HSP70 (5.5 fold) and glutathione reductase gene (2.79 fold) were present in the sea urchin exposed to hypoxia. Our results suggested that hypoxia can cause the abnormal development and elicits a stress and antioxidant response on sea urchin.

• Annals of Clinical Neurophysiology
• /
• v.22 no.2
• /
• pp.121-124
• /
• 2020

The most common form of autosomal recessive hereditary spastic paraplegia (HSP) is caused by mutations in SPG11/KIAA1840 gene, which encodes for spatacsin. The clinical presentation of SPG11 is characterized by cognitive impairment, peripheral neuropathy and a thin corpus callosum in brain magnetic resonance imaging. We identified a novel homozygous nonsense mutation (c.6082C>T [p.Q2028]) in exon 32 of SPG11 in Korean siblings. Our findings suggest that this novel homozygous mutation in SPG11 is associated with HSP and with dysgenesis of the corpus callosum.

• Journal of Life Science
• /
• v.24 no.2
• /
• pp.105-111
• /
• 2014

The purpose of this study was to characterize equine heat-shock protein (Hsp) genes and analyze their expression pattern in various horse tissues and blood leukocytes after exercise. In a previous study, RNA sequencing of blood and skeletal muscles of thoroughbreds before and after exercise was performed using differently expressed gene (DEG) analysis. Three Hsp genes (HspH1, Hsp90${\alpha}$ and Hsp70) were selected by DEG analysis and were found to be differentially expressed in either blood or muscle. To validate and extend previous observations on these genes, we performed RT-PCR analyses of horse tissue as well as real-time qPCR analyses of blood leukocytes after exercise. mRNA expression of these Hsp genes was found to be ubiquitous in the analyzed tissues (including thyroid, colon, skeletal muscle, cecum, kidney, spinal cord, heart, and lung). In addition, Hsp mRNA expression of these genes in extracted whole blood increased after 120 minutes of exercise compared to the baseline condition. These results are in agreement with the results of human and other experimental animals, suggesting that regulatory mechanisms that are responsible for upregulation of Hsp gene transcription may be conserved among species. Further investigations to correlate Hsp gene expression patterns with athletic performance or recovery processes after exercise are warranted.

• Korean Journal of Poultry Science
• /
• v.49 no.2
• /
• pp.89-98
• /
• 2022

This study compared the production and physiological characteristics of male and female Korean native commercial chickens. We investigated the growth performance, vitality, and stress response of 479 male and 608 female chickens from 1 d to 12 weeks of age. The body weight, feed consumption and feed conversion ratio were measured as growth performance. The survival rate, amount of telomeric DNA, DNA damage rate, heterophil-lymphocyte ratio (H/L ratio), and heat shock protein (HSP)-70, HSP-90α, and HSP-90β gene expression levels were analyzed as indicators of vitality and stress response. Body weight was significantly higher in male chickens than in female chickens after 2 weeks of age (P<0.01). Feed intake was higher in male chickens than in female chickens, whereas the feed conversion ratio showed the opposite trend. The survival rate was significantly higher in female chickens than in male chickens (82.8% vs. 73.8%, P<0.05). Stress response analysis revealed no differences between male and female chickens in terms of telomeric DNA content, DNA damage rate, H/L ratio, and HSP gene expression levels. Taken together, it was concluded that there was a significant difference in the growth performance and survival rate between male and female Korean native commercial chickens, whereas there was no difference in the degree of stress response between them. Therefore, in terms of the productivity of Korean native commercial chickens, it is judged that separate-sex rearing is much more advantageous than mixed rearing, regardless of the physiological characteristics of males and females.

• Korean Journal of Plant Tissue Culture
• /
• v.22 no.1
• /
• pp.19-23
• /
• 1995

Carrot cultured cells are able to respond to a temperature increase by inducing a set of new proteins, heat shock proteins (HSP). Such an induction of the HS gene was known to be achieved mainly at the level of transcription. However there has been an increasing number of evidences showing that a translational control was involved in the regulation of the HS gene expression. A comparison of HSP synthesized in vivo to in vivo (represent for mRNA level since the amount of the proteins produced by in vivo translation system will be proportional to an amount of the corresponding mRNA)showed no correlation between the amount of HS mRNA and the amount of the corresponding HSP at $30^{\circ}C$, It appears that a translational control may exert a major role in the expression of HS gene in carrot callus cells at $30^{\circ}C$.

• Journal of Korean Society on Water Environment
• /
• v.29 no.2
• /
• pp.232-238
• /
• 2013

Water temperature is key factor influencing growth and reproduction of fish and its increase give rise to various physiological changes including gene expression. Heat shock protein (Hsp), one of the molecular chaperones, is highly conserved throughout evolution and its expression is induced by various stressors such as temperature, oxidative, physical and chemical stresses. Here, we isolated partial cDNA clones encoding 70-kDa Hsp (Hsp70) and $\beta$-actin using reverse transcriptase-PCR (RT-PCR) from gut of Rhynchocypris kumgangensis, a Korean indigenous species and cold-water fish, and investigated expression profiles of Hsp70 under an increase of water temperature using $\beta$-actin as an internal control for RT-PCR. Cloned Hsp70 cDNA of R. kumgangensis showed homology to Ctenopharyngodon idella (96%), Hypophthalmichthys molitrix (96%), Danio rerio (93%) and Oncorhynchus mykiss (81%) Hsp70. Cloned $\beta$-actin cDNA of R. kumgangensis showed homology to D. rerio (98%), H. molitrix (97%), C. idella (97%) and O. mykiss (90%) $\beta$-actin. Both mRNA of Hsp70 and $\beta$-actin were expressed in gut, brain, and liver in R. kumgangensis. Futhermore, expression of Hsp70, in brain, was highly augmented by an increase of water temperature. These results suggest that Hsp70 mRNA expression level in brain can be used as a biological molecular marker to represent physiological stress against an increase of water temperature.