• Title/Summary/Keyword: HSP 70

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Identification of heat shock protein70-2 and protamine-1 mRNA, proteins, and analyses of their association with fertility using frozen-thawed sperm in Madura bulls

  • Zulfi Nur Amrina Rosyada;Berlin Pandapotan Pardede;Ekayanti Mulyawati Kaiin;Ligaya I.T.A Tumbelaka;Dedy Duryadi Solihin;Bambang Purwantara;Mokhamad Fakhrul Ulum
    • Animal Bioscience
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    • v.36 no.12
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    • pp.1796-1805
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    • 2023
  • Objective: This study aims to identify heat shock protein70-2 (HSP70-2) and protamine-1 (PRM1) mRNA and protein in Madura bull sperm and demonstrate their relation as bull fertility biomarkers. Methods: The Madura bull fertility rates were grouped based on the percentage of first service conception rate (%FSCR) as high fertility (HF) (79.04%; n = 4), and low fertility (LF) (65.84%; n = 4). mRNA of HSP70-2 and PRM1 with peptidylprolyl isomerase A (PPIA) as a housekeeping gene were determined by quantitative real-time polymerase chain reaction, while enzyme-linked immunoassay was used to measure protein abundance. In the post-thawed semen samples, sperm motility, viability, acrosome integrity, and sperm DNA fragmentation index were analyzed. Data analysis was performed on the measured parameters of semen quality, relative mRNA expression, and protein abundance of HSP70-2 and PRM1, among the bulls with various fertility levels (HF and LF) in a one-way analysis of variance analysis. The Pearson correlation was used to analyze the relationship between semen quality, mRNA, proteins, and fertility rate. Results: Relative mRNA expression and protein abundance of HSP70-2 and PRM1 were detected and were found to be highly expressed in bulls with HF (p<0.05) and were associated with several parameters of semen quality. Conclusion: HSP70-2 and PRM1 mRNA and protein molecules have great potential to serve as molecular markers for determining bull fertility.

Constitutive Expression of Small Heat Shock Protein Increases Thermotolerance in Transgenic Plant (저 분자량 Heat Shock Protein의 항상적 발현에 의한 형질전판 식물체의 고온내성 증가)

  • 이병현
    • Korean Journal of Plant Tissue Culture
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    • v.27 no.1
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    • pp.13-18
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    • 2000
  • To investigate the function of chloroplast small HSP, transgenic tobacco plants (Nicotiana tabacum L. cv. Samsun) that constitutively overexpress the chloroplast small HSP (NtHSP21) from N. tabacum cv. Petit Havana SR1 were generated. Five homozygous lines of transformants showing different constitutive expression levels of the NtHSP21 were selected. To determine whether constitutive overexpression of NtHSP21 protein affects thermotolerance, wild-type and transformants were grown in Petri dishes, heat-stressed at 52$^{\circ}C$ for 45 min, and then incubated in normal growth condition. When heat-stressed wild-type plantlets were incubated at $25^{\circ}C$, leaf color gradually became white and all trio plantlets finally died within a week. As for the transformants, however, more than 70% of them remained green and survived under the conditions in which all the wild-type plants were dying. It was also found that the levels of NtHSP21 were correlated with the degree of thermotolerance. These results suggest that the NtHSP21 protein in transformants is responsible for the increase in thermotolerance.

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Effects of DBD-bio-plasma on the HSP70 of Fibroblasts: A New Approach on Change of Molecular Level by Heat Shock in the Cell (Fibroblasts 세포주의 HSP70에 대한 DBD-bio-plasma의 effects: Cell에서 Heat Shock에 의한 Molecular Level 변화로의 새로운 접근법)

  • Kim, Kyoung-Yeon;Yi, Junyeong;Nam, Min-Kyung;Choi, Eun Ha;Rhim, Hyangshuk
    • KSBB Journal
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    • v.30 no.1
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    • pp.21-26
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    • 2015
  • Plasma is an ionized gas mixture, consisting of neutral particles, positive ions, negative electrons, electronically excited atoms and molecules, radicals, UV photons, and various reactive species. Also, plasma has unique physical properties distinct from gases, liquids, and solids. Until now, non-thermal plasmas have been widely utilized in bio-medical applications (called bio-plasma) and have been developed for the plasma-related devices that are used in the medical field. Although numerous bio-plasma studies have been performed in biomedicine, there is no confirmation of the nonthermal effect induced by bio-plasma. Standardization of the biological application of plasma has not been evaluated at the molecular level in living cells. In this context, we investigated the biological effect of bio-plasma on living cells. Hence, we treated the fibroblasts with Dielectric Bauvier Discharge bio-plasma (DBD), and assessed the characteristic change at the molecular level, one of the typical cellular responses. Heat shock protein 70 (HSP70) regulates its own protein level in response to stimuli. HSP70 responds to heat shock by increasing its own expression at the molecular level in cells. Hence, we confirmed the level of HSP70 after treatment of mouse embryonic fibroblasts (MEFs) with DBD. Interestingly, DBD-plasma induced cell death, but there was no difference in the level of HSP70, which is induced by heat shock stimuli, in DBD-treated MEFs. Our data provide the basic information on the interaction between MEFs and DBD, and can help to design a molecular approach in this field.

Association of a Single Nucleotide Polymorphism in the 5'-Flanking Region of Porcine HSP70.2 with Backfat Thickness in Duroc Breed

  • Chen, Ming-Yu;Huang, San-Yuan;Lin, En-Chung;Hseu, Tzong-Hsiung;Lee, Wen-Chuan
    • Asian-Australasian Journal of Animal Sciences
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    • v.16 no.1
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    • pp.100-103
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    • 2003
  • Higher environmental temperature affects the economic performance of pigs. Heat shock protein 70 has been shown to play an important role in thermoresistance. The purpose of this study was to assess the effect of a single nucleotide polymorphism in the 5'-flanking region of porcine HSP70.2 on growth performance in Taiwanese Duroc. The genotype of this nt 393 polymorphic site could be verified by digestion with Bsa WI restriction enzyme of a PCR product. Pigs with TT and TC genotypes have thinner backfats than those with CC type (p<0.05). The result suggested that the polymorphic Bsa WI site in the 5'flanking region of porcine HSP70.2 may be used as a marker for the early selection of ultrasonic backfat thickness in Duroc pigs.

Immunohistochemical Localization of Heat Shock Protein 70 in the Central Nervous System of Nicotine-treated Rat Embryo (태서 중추신경계의 Heat Shock Protein 70 분포에 대한 Nicotine 영향)

  • 최병태;강호성
    • Journal of Life Science
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    • v.7 no.4
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    • pp.276-281
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    • 1997
  • This study was investigated to determine whether nicotine causes the morphological changes and expression of heat shock protein(HSP) 70 in the central nervous system of rat embryo. The pregnant rats were injected s.c. twice daily with 3 mg nicotine per 100g body weight from day 0 to 14 of gestation and embryos were removed on gestation day 15. As morphological changes, retardation of cell proliferaton was observed in the telencephalon of nicotine-treated groups and no changes in the other region were found. Minimal HSP 70 was expressed over chole central nervous system was similar between control and nicotine-treated group, the expression of blood cells in the meinges and chroid plexus was significantly greater in nicotine-treated group than in control.

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Pre-Exercise Protective Effects Against Renal Ischemic Reperfusion Injury in Hsp 70.1 Knockout Mice (Hsp70.1유전자결핍된 마우스에서 허혈 재관류 신장손상에 대한 전처치 운동의 보호효과)

  • Lee, Jin;Kim, Won-Kyu
    • Journal of Life Science
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    • v.20 no.4
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    • pp.555-560
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    • 2010
  • The objective of this study was to investigate levels of serum creatinine, CuSOD and MnSOD protein expression in the kidney after renal ischemic reperfusion with pre-exercise using heat shock protein 70.1 in knock-out mice (KO). The C57/BL6 strain (Wild type: WT) and KO were divided into 4 groups as follows: Sham control group (Sham), pre-exercise group (Ex), pre-exercise +ischemia group (Ex+IR), and ischemia group (IR). CuSOD and MnSOD expression were significantly decreased (p<0.01, p<0.05) and blood creatinine concentration was significantly increased (p<0.01) in the IR group of KO. In contrast, CuSOD and MnSOD expression in the Ex+IR group of KO were higher than the IR group, while creatinine concentration was significantly lower. These results suggest that Hsp70 is directly correlated to renal ischemic reperfusion injury. Pre-exercise in renal ischemia might prevent or inhibit positive oxidative stress inhibitory effects by increasing anti-oxidative enzymes (CuSOD, MnSOD) within the kidney and improve to prevent renal function. Thus, pre-exercise may have a protective role against renal injury after renal ischemia.

Physiological Studies on Acute Water-temperature Stress of Juvenile Abalone, Haliotis discus hannai (급격한 수온 스트레스에 따른 전복, Haliotis discus hannai 치패의 생리학적 연구)

  • Kim Tae-Hyung;Yang Moon-Hyu;Choe Mi-Kyung;Han Seok-Jung;Yeo In-Kyu
    • Journal of Aquaculture
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    • v.18 no.1
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    • pp.7-12
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    • 2005
  • This study was conducted to investigate antioxidant enzyme activity (catalase and superoxide dismutase) and Heat Shock Protein 70 (HSP70) mRNA variation in hepatopancreas of abalone (Haliotis discus hannai) cultured under several acute water temperatures. Abalones were cultured at 10, 15, 20, 25 and $30^{\circ}C$, for 0, 6, 12, 24 and 48 hours, respectively. The HSP70 mRNA expression in hepatopancreas was more increased at $30^{\circ}C$ compared to those at 10. 15, 20 (control) and $25^{\circ}C$. The superoxide dismutase (SOD) activity was increased in hepato-pancreas at all water temperature conditions compared to the control ($20^{\circ}C$). The SOD activity at high water temperature (25 and $30^{\circ}C$) tended to be increased after 12 hours, and was increased immediately after exposure to low water temperature (10 and $15^{\circ}C$). and then was recovered to starting level after the increase. Also, catalase (CAT) activity in hepatopancreas was increased in all the groups except for at $10^{\circ}C$ than the control ($20^{\circ}C$). Survival rate of abalone was $100\%$ at 10, 15, 20 and $25^{\circ}C$, but $92\%$ at $30^{\circ}C$. Thus, according to our study, when abalone is appeared at $20^{\circ}C$, defense mechanism against stress at low water temperature can be accelerated to be stabilized at about $5^{\circ}C$. In the case of exposure of abalone to high water temperature, antioxidant enzyme and HSP70 expression were increased due to elevated physiological stimulation factor, such as temperature.

Association between HSP70 Genotypes and Oocytes Development on In vitro Maturation/Fertilization in Pig

  • Wee, M.S.;Park, C.K.;Cho, S.R.;Lee, S.S.;Yeon, S.H.;Kim, C.D.;Cho, C.Y.;Choi, S.H.;Sang, B.D.;Son, D.S.;Li, Z.D.;Jin, H.J.
    • Asian-Australasian Journal of Animal Sciences
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    • v.21 no.10
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    • pp.1404-1410
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    • 2008
  • This study was performed to clarify whether the variation of stress related heat shock protein 70 (HSP70) (GenBank X68213) gene was associated with the nuclear morphological change of in vitro maturation and in vitro capacitation in oocytes of pig ovaries obtained at the slaughterhouse. The nucleic acid substitution of C to G at the 483rd position was found out in HSP70 K1 (290-512) from X68213. The ovaries were categorized into CC, CG, and GG genotypes using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) (BsiHKA I). After the second in vitro maturation of immature fresh oocytes, the relation of nuclear morphological change in oocytes with the genotype of HSP70 K1 gene was such that the MII ratios of the genotype GG and CG (46.93% and 42.20%, respectively) were significantly higher than that of the CC genotype (10.71%) (p<0.05). With respect to in vitro maturation of frozen-thawed oocytes by an open pulled straw (OPS) method, the percentage of oocytes matured to MII stage of the CG genotype showed a higher trend than CC and GG genotypes. After the in vitro maturation of immature fresh oocytes and frozen-thawed oocytes by the OPS method, the relation of the pronuclei change in oocytes matured in vitro with HSP70 genotype was assessed, and the result showed that the enlarged sperm heads (ESH) of matured fresh oocytes and frozen-thawed oocytes were 80.0% and 60.0% in the CC genotype, respectively. The CC genotype group had a significantly higher rate of ESH than the CG and the GG genotype group (p<0.05). The ratios of polyspermic invasion were not different among HSP70 of the three genotypes. It was considered that the rate of in vitro maturation of fertilized oocytes was expected to differ according to genotype of the stress related gene.

The effect of heat shock protein 70 on inducible nitric oxide synthase during sepsis in rats (백서 패혈증 모델에서 HSP70의 과도 발현이 iNOS의 발현에 미치는 효과에 관한 연구)

  • Lee, Yong-Keun;Ahn, Yung;Leem, Dae-Ho;Baek, Jin-A;Ko, Seung-O;Shin, Hyo-Keun
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.36 no.5
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    • pp.346-352
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    • 2010
  • Introduction: Heat shock protein70 (HSP70) is a highly conserved family of proteins produced after a variety of stresses. Many studies reported that the overexpression of HSP70 can improve the prognosis of the patients with sepsis through a reduction of the nitric oxide concentration. However, these results only revealed the effect of HSP70 and nitric oxide. No studies have examined the relationship between HSP70 and nitric oxide. The aim of this study was to evaluate the effect of the overexpression of HSP70 on the expression of inducible nitric oxide synthase and the nitric oxide concentration. In addition, the mechanism of the relationship of HSP70 and inducible nitric oxide synthase (iNOS) in sepsis was examined. Materials and Methods: The experiments were performed on male sprague-dawley rats. Sepsis was induced by a cecal ligation and puncture (CLP). Glutamine (GLN) or saline was administered 1 hour after the initiation of sepsis. Serum and lung tissues were acquired from the rats 12 hours or 24 hours after the initiation of sepsis. The nitric oxide concentration, the expression of HSP70 in lung, and the gene expression of iNOS in lung were analyzed. The three groups, sham operation, CLP and CLP+GLN, were compared. Results: Compared to the other groups, in CLP+GLN, GLN administered after the initiation of sepsis enhanced the expression of HSP70 in the lung at 12 hours ($47.19{\pm}10.04$ vs. $33.22{\pm}8.28$, P=0.025) and 24 hours ($47.06{\pm}10.60$ vs. $31.90{\pm}4.83$, P=0.004). In CLP+GLN, GLN attenuated the expression of iNOS messenger RNA (mRNA) in the lung at 12 hours ($5,513.73{\pm}1,051.60$ vs. $4,167.17{\pm}951.59$, P=0.025) and 24 hours ($18,740.27{\pm}8,241.20$ vs. $9,437.65{\pm}2,521.07$, P=0.016), and reduced the concentration of nitric oxide in the serum at 12 hours ($0.86{\pm}0.48$ vs. $3.82{\pm}2.53$, P=0.016) and 24 hours ($0.39{\pm}0.25$ vs. $1.85{\pm}1.70$, P=0.025). Conclusion: The overexpression of HSP70 induced by the administration of GLN in sepsis attenuates the expression of the iNOS gene but reduces the nitric oxide concentration.

Effects of Cadmium on Heat Shock Protein Induction and on Clinical Indices in Rats (카드뮴이 랫드의 Heat Shock Protein 발현에 미치는 영향과 독성학적 변화에 관한 연구)

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

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