Hsp70 Gene Expression Research Articles

The role of regulator FucP in Edwardsiella tarda pathogenesis and the inflammatory cytokine response in tilapia

The animal intestine provides a competitive environment for the microbiota. Successful colonization by pathogens requires sensing chemical signals to regulate the expression of virulence genes. Some bacteria rely on a two-component chemical signal transduction system, named FusKR, to regulate virulence genes expression by intestinal fucose. Here we construct the fucP gene deletion strain prove FucP regulation of the T3SS in E. tarda. The result showed that the mutant strain had down-regulated significantly the gene expression of FusKR and T3SS compared to the wild-type strain (P < 0.05). This mutant strain significantly increased LD50 in zebrafish compared to the wild-type strain (P < 0.05), and significantly decreased penetration and motility in mucin than the wild-type strain (P < 0.05). Meanwhile, tilapia infected with mutant strain show significantly reduced E. tarda adherence and colonization than those infected with the wild-type strain (P < 0.05). Fish infected with EIB202 and ΔfucP showed significantly higher (P < 0.05) gene expression of IL-1β, TNF-α, IFN-γ, TGF-β and HSP-70 in head kidney than fish treated with PBS in the whole observed period; however CPP-3 did not show significant differences (P > 0.05) in all groups. Fish infected with EIB202 showed significantly higher (P < 0.05) gene expression of TGF-β in head kidney than fish treated with ΔfucP in the whole observed period; however other cytokines did not show significant differences (P > 0.05) in the whole observed period. In addition, Fish infected with EIB202 showed significantly higher (P < 0.05) gene expression of IL-1β, TNF-α and TGF-β in spleen than fish treated with ΔfucP in the whole observed period, however IFN-γ, CPP-3, and HSP-70 did not show significant differences (P > 0.05) in the whole observed period. Although the gene expression of cytokines was induced similarly by both strains, all results indicate that the fucP gene deletion down-regulates the key gene expression of FucKR and T3SS, reduces the pathogenicity of E. tarda in fish, particularly decreases inducing the gene expression of TGF-β in the head kidney and IL-1β, TNF-α and TGF-β in the spleen.

Rosmarinic acid and siRNA combined therapy represses Hsp27 (HSPB1) expression and induces apoptosis in human glioma cells.

High expression of Hsp27 in glioma cells has been closely associated with tumor cell proliferation and apoptosis inhibition. The aim of the present study was to asses the effects of rosmarinic acid (RA) on Hsp27 expression and apoptosis in non-transfected and transfected human U-87 MG cells. The effect of rosmarinic acid was compared to quercetin, which is known to be a good Hsp27 inhibitor. In order to block the expression of Hsp27 gene (HSPB1), transfection with specific siRNAs was performed. Western blotting technique was used to assess the Hsp27 expression, and caspase-3 colorimetric activity assay was performed to determine apoptosis induction. According to the results, it was found that RA and quercetin effectively silenced Hsp27 and both agents induced apoptosis by activating the caspase-3 pathway. Eighty and 215μM RA decreased the level of Hsp27 by 28.8 and 46.7% and induced apoptosis by 30 and 54%, respectively. For the first time, we reported that rosmarinic acid has the ability to trigger caspase-3 induced apoptosis in human glioma cells. As a result of siRNA transfection, the Hsp27 gene was silenced by ~ 50% but did not cause a statistically significant change in caspase-3 activation. It was also observed that apoptosis was induced at a higher level as a result of Hsp27 siRNA and subsequent quercetin or RA treatment. siRNA transfection and 215μM RA treatment suppressed Hsp27 expression level by 90.5% and increased caspase-3 activity by 58%. Herein, we demonstrated that RA administered with siRNA seems to be a potent combination for glioblastoma therapy.

Dietary effects of succinic acid on the growth, digestive enzymes, immune response and resistance to ammonia stress of Litopenaeus vannamei

Organic acids acts as an growth promoter and antimicrobial agent in aquaculture. The present study investigated the effects of a natural organic acid - succinic acid (SA) on the growth, digestive enzymes, immune response and resistance to ammonia stress of Litopenaeus vannamei. The shrimps were firstly fed with diets containing different levels of SA: 0% (Control), 0.25% (SA1), 0.50% (SA2), and 1.0% (SA3) (w/w) for 56 days, followed by an acute ammonia stress for 48 h. The results indicated that dietary of SA improved the growth of shrimp, and increased the survival rate of shrimp after ammonia stress for 48 h. The amylase, lipase and pepsin activity increased in hepatopancreas in three SA group, while trypsin activity was only increased in the SA1 and SA2 groups. At 56 d, T-NOS activity, proPO and HSP70 gene expression level increased in the three SA group, PO activity increased in the SA1 and SA2 groups, T-AOC content and Toll gene expression level increased in the SA2 and SA3 groups, Trx and SOD gene expression level increased in the SA2 group, while Imd, GS and GDH gene expression level was no changes. After exposure to ammonia stress for 48 h, immune biochemical parameters (T-AOC and PO) and genes (proPO, HSP70, Trx and GDH) expression level increased in the three SA group, T-NOS activity, Toll, Imd and GS gene expression level increased in the SA2 and SA3 groups, while SOD gene expression level increased in the SA1 and SA2 groups. These results indicated that SA improved growth, enhanced digestive and immune capacities of L. vannamei against ammonia stress, and may be a potential feed additive for shrimp. The optimal dietary supplementation dosage is 0.50% (w/w) in diet.

Quantifying Environmental Multi‐Stress Response in Mussels via Respirometry and qPCR

Rocky intertidal organisms, such as the sessile West Coast mussel Mytilus californianus (M.c.), experience a dramatic set of acute and chronic environmental stressors. Mussels transition daily from submerged to emerged conditions, experiencing seasonal temperature changes in water, daily shifts in air temperature, and fluctuating durations of exposure based on tidal periods. M.c. must therefore tolerate both gradual and rapid shifts in environmental conditions. To better quantify differential individual outcomes of M.c. to simultaneous acute and chronic stressors, we employ both phenotypic and genotypic methods. We couple respirometric rates to differential gene expression on a suite of genes implicated in stress response: Heat Shock Protein 70 (HSP70; chaperone), Hypoxia‐Induced Factor 1α (HIF‐1α; hypoxia), Glutathione Peroxidase (GPX; oxidative stress), Thioredoxin Reductase (TRR; oxidative stress), and Caspase‐8 (CASP8; apoptosis), referenced to Elongation Factor 1α (EF‐1α; reference).Mussels were collected from Ventura, CA and acclimated for three weeks prior to study. Our full‐factorial experiment subjected M.c. to a range of conditions: water temperatures (14°C, 19°C, 24°C), air temperatures (7°C, 18°C, 35°C), and emersion times (4 hours, 8 hours) using 12 mussels per treatment. Two‐hour in vivo respirometry trials were completed using lab‐designed Chambers for Organismal Response to Environmental Stressors (CORES) and a NeoFox spectrophotometer (Ocean Optics, Dunedin, FL). Gills were excised and divided evenly for dry‐weight respirometry measurements and RNA extraction. Extracted RNA was converted to cDNA for quantitative PCR in technical triplicate using a CFX96 RT‐PCR System (BioRad, Hercules, CA). We quantify gene expression of HSP70, HIF‐1α, GPX, TRR, and CASP8 across all 18 experimental conditions and pair these results with corresponding respirometric rates.Mytilus californianus show a strong acute response when exposed to high air temperature (35°C) across all water conditions, with HSP70 expression elevated by 750x −2460x over 18°C air treatments. However, HSP70 is slightly downregulated (2.5–6.3x) as water temperature increases with highest expression found at 14°C. This may be due to the differential response to acute (air) versus chronic (water) stressors. When considering genes implicated in longer term organismal response, such as CASP8 apoptotic repair mechanisms, the response rate is different, however. We see slight rate changes (1–4x) between air temperature conditions, but a significant downregulation (11–59x) of CASP8 at 19°C water temperature when compared to 14°C and 24°C treatments. We also see a characteristic increase in respiration rates (2–3x) with increasing water temperature (14 to 24°C) which is broadly in line with the Q10 hypothesis. Notably, M.c. exhibit a wide range of individual respirometric responses to the most stressful condition set (24°C water, 35°C air, 8‐hour emersion). Variability also increases in larger deviations of HSP70 expression rates at higher temperatures, suggesting that individual variation may play a large role in determining outcomes under extreme multi‐stress conditions.Support or Funding InformationThis work is supported by CSU Channel Islands and the Santa Rosa Island Research Station.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

HSP70 from the Antarctic sea urchin Sterechinus neumayeri: molecular characterization and expression in response to heat stress

BackgroundHeat stress proteins are implicated in stabilizing and refolding denatured proteins in vertebrates and invertebrates. Members of the Hsp70 gene family comprise the cognate heat shock protein (Hsc70) and inducible heat shock protein (Hsp70). However, the cDNA sequence and the expression of Hsp70 in the Antarctic sea urchin are unknown.MethodsWe amplified and cloned a transcript sequence of 1991 bp from the Antarctic sea urchin Sterechinus neumayeri, experimentally exposed to heat stress (5 and 10 °C for 1, 24 and 48 h). RACE-PCR and qPCR were employed to determine Hsp70 gene expression, while western blot and ELISA methods were used to determine protein expression.ResultsThe sequence obtained from S. neumayeri showed high identity with Hsp70 members. Several Hsp70 family features were identified in the deduced amino acid sequence and they indicate that the isolated Hsp70 is related to the cognate heat shock protein type. The corresponding 70 kDa protein, called Sn-Hsp70, was immune detected in the coelomocytes and the digestive tract of S. neumayeri using a monospecific polyclonal antibody. We showed that S. neumayeri do not respond to acute heat stress by up-regulation of Sn-Hsp70 at transcript and protein level. Furthermore, the Sn-Hsp70 protein expression was not induced in the digestive tract.ConclusionsOur results provide the first molecular evidence that Sn-Hsp70 is expressed constitutively and is non-induced by heat stress in S. neumayeri.

Effect of liposome‑mediated HSP27 transfection on collagen synthesis in alveolar type II epithelial cells.

To investigate the effect of liposome Lipofectamine® 2000‑mediated HSP27 plasmid transfection in A549 human alveolar typeII epithelial cell line on collagen synthesis during transforming growth factor‑β1 (TGF‑β1)‑induced typeII epithelial cell transition to myofibroblasts. Cells were transfected with varying ratios of the Lipofectamine® 2000‑mediated heat shock protein27 (HSP27) plasmid and the transfection efficiency was determined using flow cytometry. The maximum transfection efficacy was confirmed by laser confocal microscopy. HSP gene expression and the most efficient HSP27 plasmid were determined using reverse transcription‑quantitative polymerase chain reaction. Western blot analysis was used to examine HSP27 and collagen expression levels. With a transfection efficiency of 83%, the 8µg:20µl ratio of liposome: Plasmid had the highest transfection levels. Among the four different interference sequences in the HSP27 plasmid, the Dsequence had the highest interference effect with 70% silencing of the HSP27 gene. The expression of typeIandIII collagen in TGF‑β1‑induced transition of A549 human alveolar typeII epithelial cell line to myofibroblasts was significantly downregulated by the successful transfection with HSP27‑interfering plasmid. The expression of typeI andIII collagen in the TGF‑β1‑induced transition of A549 cells to myofibroblasts was significantly downregulated by transfection of A549 cells with HSP27 plasmid D‑interfering sequence and optimal ratio of Lipofectamine®2000 and HSP27 plasmid.

Multibiomarker response shows how native and non‐native freshwater bivalves differentially cope with heat‐wave events

Abstract Temperature has a key impact on the physiology, food acquisition, metabolic rate, growth, and reproduction of organisms. Thus, exploring the effects of climate change on species physiology is important for the conservation and management of native and non‐native bivalve species. In this work, an integrative laboratory approach was used to compare the physiological, cellular, and molecular responses of the native Unio delphinus and the invasive non‐native Corbicula fluminea under a natural heat‐wave event. Whereas the filtration rate in C. fluminea was clearly affected throughout the whole heat‐wave event (from the heat peak onwards), it was only significantly affected during the heat‐wave climax in U. delphinus, and recovered afterwards. At the cellular level, lysosomal membrane stability was affected during the heat‐wave climax, but recovered afterwards, in both native and non‐native bivalve species. At the molecular level, hsp70 and hsp90 gene expression were increased and decreased (respectively) in C. fluminea after the heat wave. In contrast, only hsp90 gene expression decreased in U. delphinus during the climax and after the heat wave. The present work demonstrates that accurate monitoring of native and non‐native species using an integrated biomarker approach provides new insights into how both species might cope with the expected increase in frequency and intensity of heat events in a global warming context. Direct and indirect impacts of heat waves have been associated with massive mortalities of both native and non‐native species. Despite present results showing that C. fluminea are more sensitive to heat waves than U. delphinus, massive mortality events might act in favour of C. fluminea as it has a higher reproductive capacity. Although native freshwater mussel species conservation is already threatened by overexploitation, flow regulation and diversions, habitat destruction, and pollution, it is probably also adversely affected by global warming and the presence of invasive non‐native species, such as C. fluminea.

Mytilus galloprovincialis (Lamarck, 1819) spermatozoa: hsp70 expression and protamine-like protein property studies.

In this work, we describe results of the reproductive health monitoring studies in Mytilus galloprovincialis following spermatozoa hsp70 expression and protamine-like protein properties. Mussels control (ctr) were released within cages for 30days in three different marine sites near Naples (Campania, Italy): Bagnoli south (BAs) and Bagnoli north (BAn), both close to a disposal metallurgical factory and in Capo Miseno (CM). Studies of hsp70 gene expression carried out, by RT-qPCR, in mussel spermatozoa have shown varied expression levels, particularly 5, 13, and 15-fold more than ctr in CM, BAs, and BAn, respectively, indicating highest involvement of stress proteins in spermatozoa of mussels in Bagnoli. In order to evaluate the possible risk on Mytilus galloprovincialis sustainability loss, electrophoretic analyses were performed on protamine-like proteins (PL) of collected spermatozoa. The results showed that CM PL were apparently unaltered with respect to ctr PL, while BAs and BAn PL appeared in part in the form of peptides and in part as bands with low mobility. Further, CM and BAs PL showed, by electrophoretic mobility shift assay, a decrease in DNA binding ability and a change in their DNA binding mode. The results of this investigation show the usefulness of the study of alterations of spermatozoa hsp70 expression and protamine-like protein properties for eco-toxicological evaluation using Mytilus galloprovincialis as a bioindicator.

Effects of age and exercise on inflammatory cytokines, HSP70 and HSP90 gene expression and protein content in Standardbred horses

We hypothesised that the cortisol response to acute exercise, markers of oxidative stress, expression of inflammatory cytokines, heat shock protein (HSP)70 and HSP90 expression in whole blood and skeletal muscle, and HSP70 and HSP90 protein concentrations in skeletal muscle are altered by age and in response to acute submaximal exercise in horses. Young (n=6; 5.5±2.8 year) and aged (n=6; 22.6±2.25 year) unconditioned Standardbred mares underwent an acute submaximal exercise test. Blood samples were collected and analysed for plasma cortisol and malondialdehyde (MDA) concentrations, and for cytokine and HSP gene expression pre- and post-exercise. Gluteus medius biopsies were obtained for analysis of cytokine and HSP gene expression pre- and at 0, 4, 24 and 48 h post-exercise. Data were analysed for main effects using a two-way ANOVA for repeated measures. Post-hoc comparisons of means were conducted using Student-Neuman-Keuls for pair-wise multiple comparisons where appropriate. Acute submaximal exercise increased plasma cortisol concentration in both young and aged mares, and the duration of the post-exercise rise in cortisol was altered in aged horses. Plasma MDA concentration and expression of tumour necrosis factor-α (TNF-α) and interleukin (IL)-6 were unchanged in blood and muscle. Exercise increased IL-1β expression in whole blood of young and aged mares, with young mares having greater exercise-induced expression at 2 (P&lt;0.001) and 4 (P=0.019) h post-exercise. Both young and aged horses had increased HSP70 expression in whole blood following acute exercise, with young horses exhibiting 3-fold greater HSP70 expression than aged mares at 2 h post-exercise. HSP90 expression in whole blood following exercise was increased only in young horses. Both young and aged horses had increased HSP90 expression in skeletal muscle following exercise, but there was no difference due to age. However, the timing of HSP70 expression was different between young and aged horses. The age-related changes in cortisol and IL-1β expression following acute submaximal exercise can have implications for energy homeostasis and the adaption to such disturbances at a cellular and whole animal level. Quantification of HSP expression in whole blood may be a useful biomarker, with implications for cellular adaptation and survival in aged horses.

Seasonal plasticity in photoprotection modulates UV-induced hsp gene expression in copepods from a clear lake.

Zooplankton from clear alpine lakes is exposed to stressful levels of solar UV radiation (UVR). As these pelagic organisms experience high UVR and large changes in solar radiation conditions between ice‐free and ice‐cover periods, they have evolved various strategies to minimize UVR exposure and damage. Here, we studied the relation between photoprotection levels (mycosporine‐like amino acids, carotenoids), antioxidant capacities, and gene expression of heat shock proteins (hsps) as indicator of stress in the copepod Cyclops abyssorum tatricus during the course of a year. Expression of hsp60, hsp70, and hsp90 was measured in the field (baseline expression [BE]) and after UVR exposure in the laboratory. The BE differed among genes and seasons (hsp60: high during summer, hsp70 and hsp90: high during the ice‐cover period). The gene expression of hsp70 was upregulated after exposure to UVR (up to 5.2‐fold change), while hsp60 and hsp90 were only constitutively expressed. A strong seasonal pattern was found in the photoprotective compounds and antioxidant capacities, with highest levels during the ice‐free period. The extent of upregulation of hsp70 gene expression increased with decreasing photoprotection levels and peaked 24 h post UVR exposure (9.6‐fold change) at the time of lowest photoprotection (February). Our data suggest that hsp70 gene expression is modulated by seasonal plasticity in photoprotection. This ability of adequate stress response is essential for survival in highly variable ecosystems such as alpine lakes.

Effects of high temperature on biochemical parameters, oxidative stress, DNA damage and apoptosis of pufferfish (Takifugu obscurus)

Water temperature is an important environmental factor that affects physiology and biochemical activities of fish. In this study, we investigated of high temperature on biochemical parameters, oxidative stress, DNA damage and apoptosis of pufferfish. Thermal stress could significantly increase the levels of AST, ALT, LDH, GLU and TG, whereas the levels of ALP and TP decrease significantly. In addition, thermal stress also decreased total blood cell count, inhibited cell viability, and subsequently lead to DNA damage and apoptosis. The mRNA levels of p53, caspase-9 and caspase-3 were up-regulated under thermal stress. These results suggested that caspase-dependent and p53 signaling pathways could play important roles in thermal stress-induced apoptosis in fish. Furthermore, the gene expression of SOD, CAT, HSP90 and C3 were induced by thermal stress. This study provides new insights into the mechanism whereby thermal stress affects physiological responses and apoptosis in pufferfish.

3′-epi-12β-hydroxyfroside, a new cardenolide, induces cytoprotective autophagy via blocking the Hsp90/Akt/mTOR axis in lung cancer cells

Rationale: Cardenolides have potential as anticancer drugs. 3′-epi-12β-hydroxyfroside (HyFS) is a new cardenolide structure isolated by our research group, but its molecular mechanisms remain poorly understood. This study investigates the relationship between its antitumor activities and autophagy in lung cancer cells.Methods: Cell growth and proliferation were detected by MTT, lactate dehydrogenase (LDH) release, 5-ethynyl-20-deoxyuridine (EDU) and colony formation assays. Cell apoptosis was detected by flow cytometry. Autophagic and signal proteins were detected by Western blotting. Markers of autophagy and autophagy flux were also detected by immunofluorescence, transmission electron microscopy and acridine orange staining. Real time RT-PCR was used to analyze the gene expression of Hsp90. Hsp90 ubiquitination was detected by coimmunoprecipitation. The antitumore activities of HyFS were observed in nude mice.Results: HyFS treatment inhibited cell proliferation and induced autophagy in A549 and H460 lung cancer cells, but stronger inhibition of cell proliferation and induction of cell apoptosis were shown when HyFS-mediated autophagy was blocked. The Hsp90/Akt/mTOR axis was found to be involved in the activation of HyFS-mediated autophagy. Evidence of direct interaction between Hsp90 and Akt was observed. HyFS treatment resulted in decreased levels of heat shock protein 90 (Hsp90) and phosphorylated Akt, overexpression of Hsp90 increased activation of autophagy, and inhibition of Hsp90 expression decreased autophagy. In addition, ubiquitin-mediated degradation of Hsp90 and subsequent dephosphorylation of its client protein Akt were also found in HyFS-treated lung cancer cells. Moreover, combination treatment with HyFS and chloroquine showed remarkably increased tumor inhibition in both A549- and H460-bearing mice.Conclusion: Our results demonstrate that HyFS induced cytoprotective autophagy through ubiquitin-mediated degradation of Hsp90, which further blocked the Akt/mTOR pathway in lung cancer cells. Thus, a combination of a HyFS-like cardenolide and an autophagic inhibitor is a potential alternative approach for the treatment of lung cancer.

Factors governing development of nonalcoholic fatty liver disease and insulin resistance in obesity

The factors promoting development of non-alcoholic fatty liver disease in patients with obesity and different state of carbohydrate metabolism have been studied. 43 patients were examined; these included 26 patients with abdominal obesity (BMI=52.9±7.9 kg/m2). The control group consisted of 17 conditionally healthy donors without obesity (BMI=18.9-24.9 kg/m2), seven of them formed a comparison group that was included to compare the results of study on the levels of tissue-specific expression of HSP70 mRNA. The study of mRNA expression was performed by real-time PCR. The concentration of IL-6 and TNF-a was measured in blood serum by the ELISA method. In patients with obesity with diabetes mellitus type 2 (DM2), a significant increase in the serum level of proinflammatory cytokines was found in comparison with the group of patients without DM2 and control. The results of histological examination of liver biopsy specimens in obese patients revealed the most pronounced changes in the group of DM2 patients. Regardless of the stage of nonalcoholic fatty liver disease in obese DM2 patients, an increase in the area of fatty inclusions (relative to the group without type 2 diabetes) was recorded. The study of the HSP70 gene expression in peripheral blood mononuclear cells allowed its significant increase relative to the comparison group. The relationship between the level of expression of the HSP70 gene in metabolically active tissues (visceral, subcutaneous adipose tissue and liver) established in all obese patients with the serum content of proinflammatory cytokines (IL-6 and TNF-a) may indicate suppression of HSP70 expression in these tissues, background of systemic and local inflammation in obesity.

Essential oil from Cymbopogon flexuosus as the potential inhibitor for HSP90.

The essential oil of Cymbopogon flexuosus or lemongrass oil (LO) is reported to have antibacterial, antifungal and anticancerous effects. HSP90 is one of the major chaperones responsible for the proper folding of cancer proteins. In this paper we show that the essential oil of C. flexuosus significantly suppresses the HSP90 gene expression. The cytotoxicity of the compounds was tested by MTT assay and the gene expression studies were carried out using HEK-293 and MCF-7 cells. Also we tested the efficacy of the major component of this essential oil viz. citral and geraniol in inhibiting the HSP90 expression. The oil was found to be more cytotoxic to MCF-7 cells with different IC50 values for the oil (69.33 μg/mL), citral (140.7 μg/mL) and geraniol (117 μg/mL). The fold change of expression was calculated by RT-qPCR using ΔΔCt (2^-ΔΔCt) method and it was 0.1 and 0.03 in MCF-7 cells at 80 μg/mL and 160 μg/mL of LO. Western blot results showed suppression of HSP90 protein expression and HSP90 - ATPase activity inhibition was also observed using LO. This study shows the anticancer mechanism exhibited by the essential oil of C. flexuosus is by the inhibition of the important chaperone protein HSP90.

Ectopic overexpression of maize heat shock transcription factor gene ZmHsf04 confers increased thermo and salt-stress tolerance in transgenic Arabidopsis

In our previous study, 25 Hsf gene homologues were identified and analyzed in maize. However, the exact functions of these gene family members are unclear at present. Here, we report the isolation and characterization of a strongly heat-induced HsfA2 gene from maize, ZmHsf04, and show its function in the heat/salt-stress response. Subcellular location assays showed that the ZmHsf04 protein is localized to the nucleus. Transcription activation data demonstrate that an AHA2 domain protein is necessary for the transcriptional activity of ZmHsf04. Furthermore, the function of ZmHsf04 was characterized in detail in transgenic plants of Arabidopsis thaliana Col-0. Overexpression of ZmHsf04 in Arabidopsis enhanced thermotolerance and positively regulated short-term-acquired thermotolerance. In addition, ZmHsf04 significantly improved salt tolerance and increased the sensitivity to abscisic acid in transgenic Arabidopsis. Overexpression of ZmHsf04 in Col-0 plants significantly up-regulated the expression of heat-specific HSP genes (AtHsp25.3-P, AtHsp18.2-CI, and AtHsp70B) and stress-related genes (AtAPX2 and AtGolS1) compared to the wild type. In summary, we have characterized a transcriptional regulator, ZmHsf04, which has important functions in enhancing heat and salt tolerance in plants.

Overexpression of Three Heat Shock Proteins Protects Monochamus alternatus (Coleoptera: Cerambycidae) From Thermal Stress

Ambient temperature is an important factor limiting the abundance and distribution of insects, and heat shock protein (Hsp) gene expression is sensitive to extremes of cold and heat. In order to explore the role of Hsps during thermal stress and development in Monochamus alternatus Hope (Coleoptera: Cerambycidae), we cloned and characterized full-length Hsp genes, including MaHsp60, MaHsp70, and MaHsp90. M. alternatus were exposed to different temperatures (−15, −5, 5, 15, 25, 35, and 40℃) for 1 h and was allowed to recover at 25℃ for 1 h. Following the treatments, we investigated the expression of the Hsps by quantitative real-time polymerase chain reaction. In third instar larvae, MaHsp60, MaHsp70, and MaHsp90 expression was upregulated in response to cold and heat, but the three Hsps were especially sensitive to heat, specifically at 35℃ and 40℃. After heating M. alternatus to 35℃, the expression of MaHsp60, MaHsp70, and MaHsp90 was higher than at 5℃ and 25℃ in nearly all developmental stages. MaHsp60, MaHsp70, and MaHsp90 expression was highest in later pupal, early adult, and early adult stages, respectively. These results suggest that compared with normal ambient temperatures, thermal stress could induce high expression of the three Hsps.

Heat stress induces expression of HSP genes in genetically divergent chickens

BackgroundChickens are animals that are sensitive to thermal stress, which may decrease their production level in terms that it affects feed intake and thus, decreasing body weight gain. The Heat Shock Factors (HSF) and Heat Shock Proteins (HSP) genes are involved in the key cellular defense mechanisms during exposure in hot environments. Aimed with this study to analyze the expression of HSF1, HSF3, HSP70 and HSP90 genes in two local breeds (Peloco and Caneluda) and a commercial broiler line (Cobb 500®) to verify differences in resistance of these chicken to Heat stress treatment. Chicken were submitted to heat stress under an average temperature of 39°C ± 1.ResultsUnder stress environment, the HSP70 and HSP90 genes were more expressed in backyard chickens than in broiler. There was a difference in HSP70 and HSP90 expression between Caneluda and Cobb and between Peloco and Cobb under stress and comfort environment respectively. HSP70 expression is higher in local breeds during heat stress than in a commercial broiler line. No significant differences were observed in the expression of HSF1 and HSF3 genes between breeds or environments.ConclusionsHSP70 and HSP90 genes are highly expressed, HSF1 and HSF3 genes did not have high expression in all genetic groups. HSP70 and HSP90 are highly expressed in Peloco and Caneluda within heat stress, these breeds proved to be very resistant to high temperature.

Effect of mannan-oligosaccharides (MOS) supplementation on performance, immunity and HSP70 gene expression in broiler chicken during hot-dry summer

An experiment was conducted on broiler chicks (n=120), reared in cages on a standard diet up to 14th day of age and thereafter up to 42nd day (14-42d) on test diets with or without MOS. The 14th day old chicks were randomly distributed into three dietary treatment groups viz., T1 (control diet), T2 (control diet with MOS @ 0.3%) and T3 (control diet with MOS @ 0.5%). Each dietary treatment comprised of four replicates of eight birds each. Experiment was carried out during hot-dry summer. It was found that production indices improved (P&gt;0.001) significantly during all phases. Cellular and humoral immunity at 28th day of age improved significantly (P&gt;0.05) due to MOS supplementation. The percentage of hemoglobin, protein, AST and ALT increased significantly (P&gt;0.001) in MOS supplemented group at 28th as well as 42nd day of age. While H:L ratio, serum corticosterone and cholesterol levels decreased significantly (P&gt;0.001) in MOS supplemented groups. Supplementation of MOS at the both levels caused significant (P&gt;0.001) down regulation of HSP70 expression in jejunum tissues during 28th and 42nd day. Based on this study it was concluded that supplementation of 0.3% MOS in broiler diets significantly improved their performance and welfare during heat stressed conditions.

Evaluation of adaptability to different seasons in goat breeds of semi-arid region in India through differential expression pattern of heat shock protein genes

The present study was conducted to examine differential expression pattern of HSP genes and adaptability in Indian goat breeds of semi-arid region. The study was conducted in five animals from each breed viz. Barbari, Sirohi, and Jhakrana during winter, thermo-neutral and summer seasons. The respiratory rate (RR) and rectal temperature (RT) of the goats were recorded at 09:00 h during the study period. The blood samples were collected for RNA isolation, cDNA synthesis, and quantitative analysis of HSP genes expression by quantitative RT-PCR. The RR increased significantly (p < 0.01) during summer as compared to winter and thermo-neutral season however, RT did not change (p > 0.05) during different seasons. The expression of HSP genes was significantly (p < 0.01) increased during summer (high THI) as compared to thermo-neutral season in all the goat breeds. Among HSPs, only HSP90 was upregulated (p < 0.01) in Jhakrana goats during winter as compared to thermo-neutral season. The deviation in expression of HSP genes during summer and winter with respect to thermo-neutral season was minimum in Barbari goats. Therefore, it can be concluded that Barbari goats possessed better adaptability during summer and winter as compared to Sirohi and Jhakrana goats in semi-arid climatic conditions of India.

Expression of Hsp70, Igf1, and Three Oxidative Stress Biomarkers in Response to Handling and Salt Treatment at Different Water Temperatures in Yellow Perch, Perca flavescens.

Stress is a major factor that causes diseases and mortality in the aquaculture industry. The goal was to analyze the expression of stress-related biomarkers in response to different stressors in yellow perch, which is an important aquaculture candidate in North America and highly sensitive to handling in captivity. Three fish groups were established, each having four replicates, and subjected to water temperatures of 14, 20, and 26°C and acute handling stress was performed followed by a salt treatment for 144h at a salinity of 5 ppt. Serum and hepatic mRNA levels of heat shock protein (hsp70), insulin-like growth factor 1 (Igf1), glutathione peroxidase (Gpx), superoxide dismutase 1 (Sod1), and glutathione reductase (Gsr) were quantified at seven times interval over 144 h using ELISA and RT-qPCR. Handling stress caused a significant down-regulation in Hsp70, Gpx, Sod1, and Gsr at a water temperature of 20°C compared to 14 and 26°C. Igf1 was significantly upregulated at 20°C and down-regulated at 14 and 26°C. Salt treatment had a transient reverse effect on the targeted biomarkers in all groups at 72 h, then caused an upregulation after 144 h, compared to the control groups. The data showed a negative strong regulatory linear relationship between igf1 with hsp70 and anti-oxidative gene expressions. These findings could provide valuable new insights into the stress responses that affect fish health and could be used to monitor the stress.