Hsps Genes Research Articles

Glycyrrhiza glabra and Saussurea costus extracts mitigate arsenic-induced nephrotoxicity via orchestrating Nrf2/HO-1 and NF-kB crosstalk and repressing the expression of HSPs genes, and apoptosis in broiler chickens

This research aimed to evaluate the role of Glycyrrhiza glabra (GGE) and Saussurea costus (Costus) extracts in alleviating arsenic (As2O3)-evoked nephrotoxicity in chickens. Seven groups (36 chicks/group) were treated daily for 42 days as follows: control group; GGE group, obtained GGE (500 mg/kg feed); Costus group was given Costus orally (300 mg/kg bodyweight); As group, received As2O3 (30 mg/kg diet); As + GGE; As + Costus; As + GGE + Costus. The administration of GGE and/or Costus in As-intoxicated birds significantly decreased (p < 0.001) creatinine level relative to As group. Additionally, both extracts markedly reduced (p < 0.001) the renal malondialdehyde content and enhanced the antioxidant enzymes performance, which function via Nrf2/HO-1 signaling. Moreover, They downregulated the renal inflammatory cascade by suppressing the expression of TNF-α and NF-κB genes compared to As group. Furthermore, a significant decline in the levels of HSPs mRNA was recorded following treatment with GGE and/or Costus. Conclusively, GGE and Costus can be accounted as prospective antidotes in controlling As poisoning.

Protective Effects of 17-βE2 on the Primary Hepatocytes of Rainbow Trout (Oncorhynchus mykiss) Under Acute Heat Stress.

The rainbow trout (Oncorhynchus mykiss) is a typical cold-water species. However, due to global warming, it has experienced prolonged high-temperature stress. Research indicates that thermotolerance in rainbow trout varies by sex at multiple physiological levels. Specifically, females exhibit higher thermotolerance, which may be attributed to estrogen-mediated signal transduction pathways. This study involved culturing primary hepatocytes from rainbow trout and exposing them to estradiol and estrogen receptor antagonists to assess estradiol's protective effects. The analysis focused on expression of ER, HSPs genes, hepatocyte viability, and antioxidant indices. Four experimental groups were treated with 17-βE2 at concentrations of 0, 0.1, 1, and 10 μM/mL for durations of 4, 8, 12, 24, and 48 h at 18 °C. 17-βE2 treatment led to increased hepatocyte viability and enhanced SOD, GSH-Px, and CAT levels but decreased MDA levels. hsp70a, hsp90β, era1, and erβ1 levels were notably higher, with the optimal 17-βE2 concentration being 1.0 μM/mL. Following heat stress (24 °C), the addition of 1.0 μM/mL 17-βE2 improved hepatocyte viability and increased SOD, GSH-Px, and CAT levels, while MDA content initially decreased before rising. The gene expression of hsp70a, hsp90β, era1, and erβ1 was significantly elevated compared to controls. Flow cytometry analysis showed increased apoptosis after heat exposure; however, 17-βE2 treatment significantly reduced the heat stress-induced effects (p < 0.05). In conclusion, 17-βE2 and mild heat stress collaboratively enhanced the expression of HSPs and estrogen receptors, thereby providing protection to hepatocytes from heat stress damage, indicating a beneficial protective role of estradiol in rainbow trout hepatocytes.

Acclimation to higher temperature and antioxidant supplemented diets improved rainbow trout (Oncorhynchus mykiss) resilience to heatwaves

Coldwater species are challenged with increasing water temperatures and fluctuations over their upper thermal limits. This study evaluated the potential of acclimation to higher temperature and dietary antioxidants capacity to mitigate the adverse effects of heat shocks in rainbow trout. To this end, rainbow trout fingerlings were acclimated at optimal (14 °C) and high (20 °C) temperatures and fed on selenium (5 mg/kg) and polyphenol (2 g/kg) supplemented diets for 60 days and then were exposed to heat shocks by increasing water temperature up to 30 °C. Growth performance, survival rate, haemato-immunological parameters, and expression of HSP70α, HSP70β, HSP90β, and IL-1β genes were measured to evaluate the hypothesises. The rainbow trout acclimated to 20 °C and fed on antioxidants supplemented diets showed a significantly higher aftershock survival rate. Moreover, fish acclimated to higher temperature showed higher red blood cell counts as well as serum total protein and albumin during the acclimation trial and heat shocks phase. Acclimation to higher temperature and feeding on antioxidants remarkably enhanced fish immune and antioxidant capacity in comparison to fish adapted to cold water and fed on the basal diet measured by improved respiratory burst and lysozyme activities and upregulation of IL-1β expression during exposure of fish to heat shocks. Furthermore, fish acclimated to higher temperature, especially those fed on antioxidant supplemented diets, showed lower expression levels of HSPs genes during the heat shock phase, indicating that high heat shocks were less stressful for these fish in comparison to cold water acclimated fish. This finding was also supported by lower cortisol levels during heat shocks in fish acclimated to higher temperature. In conclusion, the results of this study indicated that acclimation to higher temperature and/or fed on diets supplemented by selenium and polyphenol, can help to mitigate the adverse effects of the heat shock in rainbow trout.

The Role of Heat Shock Proteins in Plant Protection from Oxidative Stress

The review considers the recent progress on the role of heat shock proteins (HSPs), as well as transcription factors of heat shock proteins (HSFs) in protecting plants from oxidative stress induced by various types of abiotic and biotic stresses. HSPs are pleiotropic proteins involved in various intracellular processes and performing many important functions. In particular, HSPs increase plant resistance to stress by protecting the structure and activity of proteins of the antioxidant system. Overexpression of Hsps genes under stressful conditions, leading to an increased content of HSP, can be used as a marker of oxidative stress. Plant HSFs are encoded by large gene families with variable sequences, expression and function. Plant HSFs regulate transcription of a wide range of stress-induced genes, including HSPs and other chaperones, reactive oxygen species scavengers, enzymes involved in protective metabolic reactions and osmolytic biosynthesis, or other transcriptional factors. Genome-wide analysis of Arabidodpsis, rice, poplar, lettuce and wheat revealed a complex network of interaction between Hsps and Hsfs gene families that form plant protection against oxidative stress. Plant protection systems are discussed, with special emphasis on the role of HSPs and HSFs in plant response to stress, which will be useful for the development of technologies to increase productivity and stress resistance of plant crops.

The ameliorative effect of bergamot oil nano-emulsion in stressed rabbit bucks: Influence on blood biochemical parameters, redox status, immunity indices, inflammation markers, semen quality, testicular changes and the expression of HSPs genes

This work explored the activities of bergamot oil nano-emulsion (NBG) in modulating blood biochemical parameters, redox status, immunity indices, inflammation markers, semen quality, testicular changes and the expression of HSPs genes in stressed rabbit bucks. Twenty-four mature rabbit bucks (5 months) were randomly divided into three groups; control group (NBG0) received 1 ml of distilled water, while the other two groups received NBG orally at doses of 50 and 100 mg/kg (bw) twice a week. The present study's findings revealed that treated groups had lower values of total and direct bilirubin, triglyceride, lactate dehydrogenase, and creatinine compared with NBG0 group (p < 0.05). NBG100 group recorded the greatest of total protein, albumin, GPx, T3 and T4 values as well as the lowest values of uric acid, MDA, and indirect bilirubin. Both treated groups showed significantly reduced 8-OhDG, Amyloid A, TLR 4, while significantly increased nitric oxide, IgA, IgM, TAC, and SOD levels. Semen characteristics such as volume, sperm count, sperm motility, normal sperm, and vitality were significantly higher in the NBG100 group compared to the NBG50 and NBG0 groups, whereas sperm abnormalities and dead sperm were significantly reduced. HSP70, HSP72, and HSPA9 gene overexpression showed that testicular integrity was maintained after buck received oral doses of 50 or 100 mg/kg of NBG. Existing findings indicate that oral administration of NBG improves heat tolerance in rabbit bucks primarily as e result of its antioxidant and anti-inflammatory effects.

ZNF330/NOA36 interacts with HSPA1 and HSPA8 and modulates cell cycle and proliferation in response to heat shock in HEK293 cells

BackgroundThe human genome contains nearly 20.000 protein-coding genes, but there are still more than 6,000 proteins poorly characterized. Among them, ZNF330/NOA36 stand out because it is a highly evolutionarily conserved nucleolar zinc-finger protein found in the genome of ancient animal phyla like sponges or cnidarians, up to humans. Firstly described as a human autoantigen, NOA36 is expressed in all tissues and human cell lines, and it has been related to apoptosis in human cells as well as in muscle morphogenesis and hematopoiesis in Drosophila. Nevertheless, further research is required to better understand the roles of this highly conserved protein.ResultsHere, we have investigated possible interactors of human ZNF330/NOA36 through affinity-purification mass spectrometry (AP-MS). Among them, NOA36 interaction with HSPA1 and HSPA8 heat shock proteins was disclosed and further validated by co-immunoprecipitation. Also, “Enhancer of Rudimentary Homolog” (ERH), a protein involved in cell cycle regulation, was detected in the AP-MS approach. Furthermore, we developed a NOA36 knockout cell line using CRISPR/Cas9n in HEK293, and we found that the cell cycle profile was modified, and proliferation decreased after heat shock in the knocked-out cells. These differences were not due to a different expression of the HSPs genes detected in the AP-MS after inducing stress.ConclusionsOur results indicate that NOA36 is necessary for proliferation recovery in response to thermal stress to achieve a regular cell cycle profile, likely by interaction with HSPA1 and HSPA8. Further studies would be required to disclose the relevance of NOA36-EHR interaction in this context.

Identification of global transcriptional variations in human PBMCs two months post heat injury helps categorization heat tolerant or heat intolerant phenotypes.

Thermal intolerance may limit activity in hostile environments. Following heat illness two physiologically distinct phenotypes evolve: Heat Tolerant (HT) and Heat Intolerant (HI). The recognition that heat illness alters gene expression justified revisiting the established physiological concept of HI. We used a DNA microarray to examine the global transcriptional response in Peripheral Blood Mononuclear Cells (PMBCs) from HI and HT phenotypes, categorized 2-mo post heat injury using a functional physiological Heat Tolerance Test (HTT, 40°C)-Recovery (R, 24°C) protocol. The impact of recurrent heat stress was studied in vitro using PBMCs from controls (participants with no history of heat injury), HI and HT (categorized by functional HTT) with a customized NanoString array. There were significant differences under basal conditions between the HI and HT. HI were more immunological alerted. Almost no shared genes were found between end-HTT and recovery phases, suggesting vast cellular plasticity. In HI, mitochondrial function was dysregulated, canonical pathways associated with exercise endurance-NRF2 and insulin were downregulated, while AMPK and PPAR were upregulated. HT exhibited reciprocal responses, suggesting that energy dysregulation found in HI interfered with performance in the heat. The endoplasmic-reticulum stress response was also suppressed in HI. In vitro HTT (43°C) abolished differences between HI and HT PBMCs including the HSPs genes, while controls showed profound HSPs upregulation.

Phenotypic and genotypic traits governing adaptive capacity of indigenous Salem Black goats subjected to protein energy alterations in the diet

A study was conducted to assess the different proportions of protein energy ratio on the adaptive capabilities of Salem Black goats based on changes in both phenotypic and selective genotypic traits. Forty adult Salem Black does (average body weight (BW) 16.0 kg) were divided into five groups, GI (n = 8; ICAR recommended; crude protein (CP) 80 g and total digestible nutrients (TDN) 358 g); GII (n = 8; 50 % Low Protein and Normal Energy; CP 40 g and TDN- 358 g); GIII (n = 8; Normal Protein and 50 % Low Energy; CP 80 g and TDN 179 g), GIV (n = 8; 50 % Low Protein and 50 % Low Energy; CP 40 g and TDN 179 g) and GV (n = 8; 70 % Low Protein and 70 % Low Energy; CP 24 g and TDN 107 g). The results indicated that the alteration in protein energy composition of the diet did not influence any of the physiological and endocrine variables. However, the alteration in the feed composition changed the molecular adaptation process by inducing changes (P < 0.05) in the expression pattern of selective (heat shock protein) HSPs genes. Further, it was observed that the gene expression patterns were sensitive to energy component of the diet rather than protein content. This was evident from the fact that when energy content was not compromised the Salem black goat was able to mount appropriate stress response. Thus, the study signified the importance of ensuring optimum nutrient requirement to cope with the existing environmental condition. In addition, the study also identified heat shock factor 1 (HSF1), heat shock factor binding protein 1 (HSBP1), HSP27, HSP60, HSP70, HSP90 and HSP110 could serve as reliable biomarkers for nutritional stress in Salem Black goat.

Analysis of APXs and HSPs genes responsible to respond to heat stress in tomato plants cultivated in Central Sulawesi

Analysis of APXs and HSPs genes responsible to respond to heat stress in tomato plants cultivated in Central Sulawesi

Betalain health-promoting effects after ingestion in Caenorhabditis elegans are mediated by DAF-16/FOXO and SKN-1/Nrf2 transcription factors.

Betalain-rich extracts have been used for many years by their nutraceutical potential. However, the study of their bioactivities has always been hampered by their difficult obtention. To explain their mode of action, seventeen pure betalains were tested in vivo using the animal model C. elegans. Four betalains, named indicaxanthin, indoline carboxylic acid-betacyanin, phenylalanine-betaxanthin, and dopaxanthin, behaved as extraordinary in vivo antioxidants and anti-aging compounds, by increasing the lifespan of C. elegans up to 16.82%, 16.65%, 16.53%, and 12.93%, respectively. The first microarrays performed with betalains and biological confirmation with different mutant strains showed that this life extension is due to a reduction of oxidative stress and the activation of the transcription factors DAF-16/FOXO and SKN-1/Nrf2. They are involved in longevity and oxidative stress resistance pathways and lead to overexpression of HSPs genes, involved in resistance to cancer and Alzheimer's, opening novel research lines in the search for effective plant-based treatments.

Comparative transcriptome analysis of genes involved in response to thermal stress and leaf colour change of Acer palmatum

Exterior leaf colour is an important trait for ornamental foliage plants. Accumulating evidence indicates that thermal stress (TS) has a significant effect on the colour of plant leaves, but the molecular mechanism underlying TS-induced change in leaf colour between red and green of Acer palmatum is poorly understood. In this study, a comparative transcriptome analysis was performed using RNA-seq, in which A. palmatum was exposed to 23 °C for 4 d (A1_23) and 8 d (A2_23) and 28 °C for 4 d (A1_28) and 8 d (A2_28). There were 20,146 (12,304 up and 7842 down) and 9570 (3700 up and 5870 down) differentially expressed genes (DEGs) in the A1_28 and A2_28 samples compared to the A1_23 and A2_23 samples, respectively. Three hundred forty-six DEGs were co-upregulated, and 640 DEGs were co-downregulated in A1_28 and A2_28. Bioinformatics analyses revealed that A. palmatum cell resistance to TS occurs through upregulated interacting Hsp90-Hsp83-Hsp20 and GST-GSH-HSPs or through small Hsps genes and sterol biosynthesis genes that regulate membrane fluidity. In addition, our results demonstrated that A. palmatum leaves reduce the expression of the negative regulator PP2C and interleukin receptor kinase to activate PP2C substrates, such as MAPK, to regulate TS signalling pathways. Moreover, our results showed that the anthocyanin and carotenoids biosynthetic related genes significantly upregulated, and the chlorophyll biosynthetic and chloroplast constitutive related genes significantly downregulated, which were the direct molecular mechanisms of TS changing leaf colour from red return to green in A. palmatum. Our findings provide key genetic resources for further investigation of the molecular mechanisms of TS changing leaf colour in A. palmatum and other related species.

Identification and expression profiling of all Hsp family member genes under salinity stress in different poplar clones

Heat shock proteins (Hsps) play a key role for regulation of the changes during different stress conditions including salinity, drought, heavy metal and extreme temperature. Molecular based studies on the response mechanisms of forest trees to abiotic stresses started in 2006 when Populus trichocarpa genome sequence was completed as a model tree species. In recent years, bioinformatic analyzes have been carried out to determine functional gene regions of tree species. In this study, sHsp, Hsp40, Hsp60, Hsp90 and Hsp100 gene family members were identified in poplar genome. Some bioinformatics analyses were conducted, such as: identification of DNA/protein sequences, chromosomal localization, gene structure, calculation of genomic duplications, determination of phylogenetic groups, examination of protected motif regions, identification of gene ontology categories, modeling of protein 3D structure, determination of miRNA targeting genes, examination of sHsp, Hsp40, Hsp60, Hsp90 and Hsp100 gene family members in transcriptome data during salinity stress. As a result of bioinformatic analyzes made on P. trichocarpa genome; 60, 145, 49, 34, 12 and 90 genes belonging to members of sHsp, Hsp40, Hsp60, Hsp70, Hsp90 and Hsp100 protein families were firstly defined within the scope of this study. A total of 390 genes belonging to all Hsps gene families were characterized using different bioinformatics tools. In addition, salinity stress was applied to Populus tremula L. (Samsun) naturally grown in Turkey, Hybrid poplar species I-214 (Populus euramericana Dode. Guinier) and Black Poplar species (Populus nigra L.), Geyve and N.03.368.A clones. The expression levels of the selected Hsps genes were determined by the qRT-PCR method. After salt stress application in various poplar clones, expression levels of genes including PtsHsp-11, PtsHsp-21, PtsHsp-36, PtHsp40-113, PtHsp40-117, PtHsp60-31, PtHsp60-33, PtHsp60-38, PtHsp60-49, PtHsp70-09, PtHsp70-12, 33, PtHsp90-09, PtHsp90-12, PtHsp100-21, and PtHsp100-75 were increased. The role of the Hsps genes during salt stress has been revealed. Together with detailed bioinformatics analyses, gene expression analysis greatly contributes to understand functions of these gene family members. This research serves as a blueprint for future studies and offers a significant clue for the further study of the functions of this important gene family. Moreover, determined genes in this study can also be used for cloning studies in agricultural practices.

Expression of heat-shock protein genes in Apis mellifera meda (Hymenoptera: Apidae) after exposure to monoterpenoids and infestation by Varroa destructor mites (Acari: Varroidae)

Heat shock proteins (hsps) protect proteins in eukaryotic cells from damage. Expression of hsps in insects subject to different environmental stimuli is poorly characterized. Here, levels of expression of the hsps genes (hsp40, hsp70, and hsp90) were recorded in Apis mellifera Linnaeus (Hymenoptera: Apidae) workers after exposure to sublethal concentrations of thymol, eucalyptol, α-pinene, trans-anethole, diallyl disulfide and infestation with Varroa mites. Our results show a dose-dependent up-regulation in the levels of all the hsps tested after the bees were treated with thymol, eucalyptol and α-pinene. Although these up-regulated expressions were statistically significant for hsp70 and hsp90 when the bees were treated with thymol and eucalyptol, they were not significant when treated with α-pinene. In addition, significant down-regulated expressions of the hsp genes were recorded in the diallyl disulfide treatment. The transcriptions of all the hsps tested were significantly down-regulated when pupae were infested with different numbers (0-5) of Varroa mites. Thus, it is likely that hsps can be used as biomarkers of survival when honey bees are under toxic and pathogenic stress, but this needs to be confirmed.

De novo Transcriptome Analysis of Chinese Citrus Fly, Bactrocera minax (Diptera: Tephritidae), by High-Throughput Illumina Sequencing.

The Chinese citrus fly, Bactrocera minax (Enderlein), is one of the most devastating pests of citrus in the temperate areas of Asia. So far, studies involving molecular biology and physiology of B. minax are still scarce, partly because of the lack of genomic information and inability to rear this insect in laboratory. In this study, de novo assembly of a transcriptome was performed using Illumina sequencing technology. A total of 20,928,907 clean reads were obtained and assembled into 33,324 unigenes, with an average length of 908.44 bp. Unigenes were annotated by alignment against NCBI non-redundant protein (Nr), Swiss-Prot, Clusters of Orthologous Groups (COG), Gene Ontology (GO), and Kyoto Encyclopedia of Genes and Genomes Pathway (KEGG) database. Genes potentially involved in stress tolerance, including 20 heat shock protein (Hsps) genes, 26 glutathione S-transferases (GSTs) genes, and 2 ferritin subunit genes, were identified. These genes may play roles in stress tolerance in B. minax diapause stage. It has previously been found that 20E application on B. minax pupae could avert diapause, but the underlying mechanisms remain unknown. Thus, genes encoding enzymes in 20E biosynthesis pathway, including Neverland, Spook, Phantom, Disembodied, Shadow, Shade, and Cyp18a1, and genes encoding 20E receptor proteins, ecdysone receptor (EcR) and ultraspiracle (USP), were identified. The expression patterns of 20E-related genes among developmental stages and between 20E-treated and untreated pupae demonstrated their roles in diapause program. In addition, 1,909 simple sequence repeats (SSRs) were detected, which will contribute to molecular marker development. The findings in this study greatly improve our genetic understanding of B. minax, and lay the foundation for future studies on this species.

Transcriptional profiling induced by pesticides employed in organic agriculture in a wild population of Chironomus riparius under laboratory conditions

Copper (Cu) and azadirachtin (AZA-A+B) are pesticides allowed in organic agriculture whose environmental risk and toxicity for aquatic wildlife is only partially known. Reverse Transcription Polymerase Chain Reaction was used to assess the molecular effect of acute and short-term exposure (3, 24h) of Cu (0.01, 0.05, 1, 10, 25mgl−1) and AZA-A+B (0.2, 0.3, 0.4, 0.5, 1mgl−1) on the expression of five candidate genes (hsp70, hsc70, hsp40, hsp10 and cyP450) in a non-target species, Chironomus riparius. Fourth-instar larvae were collected from a mountain stream polluted by agricultural land run-off. All genes were responsive to both pesticides but each gene had a specific response to the different experimental concentrations and exposure times. A few similarities in transcriptional profiling were observed, such as a linear concentration-dependent response of hsp70 after 24h of exposure (at ≥1mgl−1 of Cu and ≥0.2mgl−1 of AZA-A+B) and an up-regulation regardless of the concentration of hsc70 after 24h of exposure (at ≥0mgl−1 of Cu and ≥0.2mgl−1 of AZA-A+B and the up-regulation of hsp70 after 3h of exposure at ~LC50 (Cu-LC50=26.1±2.5mgl−1, AZA-A+B-LC50=1.1±0.2mgl−1). According to the results, hsp40, hsp10 and cyP450 may be defined as pesticide-dependent (i.e., hsp40 and hsp10 seemed to responded mainly to AZA-A+B and cyP450 to Cu), while hsc70 as time-dependent regardless of the pesticide (i.e., hsc70 responded only after 24h of treatment with Cu and AZA-A+B). This study gives new insights on the potential role of the C. riparius's hsps and cyP450 genes as sensitive biomarkers for freshwater monitoring.

Transcriptome profiling of two Iberian freshwater fish exposed to thermal stress

The congeneric freshwater fish Squalius carolitertii and S. torgalensis inhabit different Iberian regions with distinct climates; Atlantic in the North and Mediterranean in the South, respectively. While northern regions present mild temperatures, fish in southern regions often experience harsh temperatures and droughts. Previous work with two hsp70 genes suggested that S. torgalensis is better adapted to harsher thermal conditions than S. carolitertii as a result of the different environmental conditions. We present a transcriptomic characterisation of these species' thermal stress responses. Through differential gene expression analysis of the recently available transcriptomes of these two endemic fish species, comprising 12 RNA-seq libraries from three tissues (skeletal muscle, liver and fins) of fish exposed to control (18°C) and test (30°C) conditions, we intend to lay the foundations for further studies on the effects of temperature given predicted climate changes.Results showed that S. carolitertii had more upregulated genes, many of which are involved in transcription regulation, whereas S. torgalensis had more downregulated genes, particularly those responsible for cell division and growth. However, both species displayed increased gene expression of many hsps genes, suggesting that they are able to deal with protein damage caused by heat, though with a greater response in S. torgalensis. Together, our results suggest that S. torgalensis may have an energy saving strategy during short periods of high temperatures, re-allocating resources from growth to stress response mechanisms. In contrast, S. carolitertii regulates its metabolism by increasing the expression of genes involved in transcription and promoting the stress response, probably to maintain homoeostasis. Additionally, we indicate a set of potential target genes for further studies that may be particularly suited to monitoring the responses of Cyprinidae to changing temperatures, particularly for species living in similar conditions in the Mediterranean Peninsulas.

Heat shock proteins expression during thermal risk exposure in the temperate xerothermic ant Formica cinerea

The abiotic conditions of the desert habitat fluctuate in a circadian rhythm of hot days and cold nights. Species living in desert habitats evolved many adaptations to increase their chances of survival. However, abiotic conditions in xerothermic habitats of a temperate climate are much different. Diurnal fluctuations are not as strong, but animals have to cope with seasonal changes and hibernate during the winter, which may potentially influence their adaptations to critical temperature conditions. We attempted to assess heat resistance adaptations using the example of a widely distributed xerothermic ant Formica cinerea. Using Real-Time PCR, we measured the expression of three heat shock protein genes (Hsp60, Hsp75, Hsp90) and assessed the adaptations of F. cinerea to enable foraging in risk prone conditions. The analysis of gene expression using the Generalized Linear Model surprisingly indicated that there was no significant effect of temperature when comparing workers from the control (23ºC) with workers foraging on the surface of hot sand (47-54ºC). As a next step we tried to estimate the threshold of a thermal resistance with the use of thermal chambers. Expression of all Hsps genes increase compare to the control group, expression of Hsp60 and Hsp90 continued up to 45ºC.

Molecular Cloning and mRNA Expression of Heat Shock Protein Genes and Their Response to Cadmium Stress in the Grasshopper Oxya chinensis

Heat shock proteins (Hsps) are highly conserved molecular chaperones that are synthesized in response to stress. In this study, we cloned the full-length sequences of the Grp78 (glucose-regulated protein 78), Hsp70, Hsp90, and Hsp40 genes from the Chinese rice grasshopper Oxya chinensis. The full-length cDNA sequences of OcGrp78, OcHsp70, OcHsp90, and OcHsp40 contain open reading frames of 1947, 1920, 2172, and 1042 bp that encode proteins of 649, 640, 724, and 347 amino acids, respectively. Fluorescent real-time quantitative PCR (RT-qPCR) was performed to quantify the relative transcript levels of these Hsp genes in different tissues and developmental stages. The mRNAs encoding these four Hsp genes were present at all developmental stages and in all tissues examined but were expressed at varying levels. Additionally, we investigated the mRNA expression profiles of these four Hsps in O. chinensis subjected to Cadmium (Cd) stress. OcGrp78, OcHsp70, OcHsp90, and OcHsp40 mRNA expression was induced under acute Cd stress; the levels reached a maximum within a short time (6 h), were reduced significantly at 12 h, and were lowered to or below control levels by 48 h. Regarding induction efficiency, OcHsp70 was the most sensitive gene to acute Cd stress. Chronic Cd exposure showed that dietary Cd treatment induced increased OcGrp78, OcHsp90, and OcHsp40 expression. However, dietary Cd induced a significant reduction of OcHsp70 expression. In the period tested, no significant difference in the mortality of the grasshoppers was observed. Our results suggest that these four Hsps genes, especially OcHsp70, are sensitive to acute Cd stress and could be used as molecular markers for toxicology studies. However, our results also indicate that OcHsp70 is not suitable for use as a molecular marker of chronic Cd contamination.

The Role of Inducible Hsp70, and Other Heat Shock Proteins, in Adaptive Complex of Cold Tolerance of the Fruit Fly (Drosophila melanogaster)

BackgroundThe ubiquitous occurrence of inducible Heat Shock Proteins (Hsps) up-regulation in response to cold-acclimation and/or to cold shock, including massive increase of Hsp70 mRNA levels, often led to hasty interpretations of its role in the repair of cold injury expressed as protein denaturation or misfolding. So far, direct functional analyses in Drosophila melanogaster and other insects brought either limited or no support for such interpretations. In this paper, we analyze the cold tolerance and the expression levels of 24 different mRNA transcripts of the Hsps complex and related genes in response to cold in two strains of D. melanogaster: the wild-type and the Hsp70- null mutant lacking all six copies of Hsp70 gene.Principal FindingsWe found that larvae of both strains show similar patterns of Hsps complex gene expression in response to long-term cold-acclimation and during recovery from chronic cold exposures or acute cold shocks. No transcriptional compensation for missing Hsp70 gene was seen in Hsp70- strain. The cold-induced Hsps gene expression is most probably regulated by alternative splice variants C and D of the Heat Shock Factor. The cold tolerance in Hsp70- null mutants was clearly impaired only when the larvae were exposed to severe acute cold shock. No differences in mortality were found between two strains when the larvae were exposed to relatively mild doses of cold, either chronic exposures to 0°C or acute cold shocks at temperatures down to -4°C.ConclusionsThe up-regulated expression of a complex of inducible Hsps genes, and Hsp70 mRNA in particular, is tightly associated with cold-acclimation and cold exposure in D. melanogaster. Genetic elimination of Hsp70 up-regulation response has no effect on survival of chronic exposures to 0°C or mild acute cold shocks, while it negatively affects survival after severe acute cold shocks at temperaures below -8°C.

Peripheral blood mononuclear cells: a potential cellular system to understand differential heat shock response across native cattle (Bos indicus), exotic cattle (Bos taurus), and riverine buffaloes (Bubalus bubalis) of India

Circulating leukocytes can be used as an effective model to understand the heat stress response of different cattle types and buffaloes. This investigation aimed to determine the temporal profile of HSPs (HSP40, HSP60, HSP70, and HSP90) expression in circulating peripheral blood mononuclear cells (PBMCs) of Murrah buffaloes, Holstein-Friesian (HF), and Sahiwal cows in response to sublethal heat shock at 42 °C. The viability data indicated HF PBMCs to be the most affected to the heat shock, whereas Sahiwal PBMCs were least affected, indicating its better survivability during the heat stress condition. The qRT-PCR expression data showed significant increase in mRNA expression of the analyzed HSPs genes after heat stimuli to the PBMCs under in vitro condition. In each case, the HSPs were most upregulated at 2 h after the heat stress. Among the HSPs, HSP70 was relatively more expressed followed by HSP60 indicating the action of molecular chaperones to stabilize the native conformation of proteins. However, PBMCs from different cattle types and buffaloes showed difference in the extent of transcriptional response. The level of expression of HSPs throughout the time period of heat stress was highest in buffaloes, followed by HF and Sahiwal cows. The higher abundance of HSP70 mRNA at each time point after heat stress showed prolonged effect of heat stress in HF PBMCs. The data presented here provided initial evidence of transcriptional differences in PBMCs of different cattle types and buffaloes and warrant further research.