Changes in fatty acid profiles of thermo-intolerant and thermo-tolerant marine diatoms during temperature stress

Fatty acid composition and degree of fatty acid saturation during temperature stress in thermo-intolerant (Phaeodactylum tricornutum) and thermo-tolerant (Chaetoceros muelleri) marine diatoms were investigated. A greater number of fatty acids were observed in C. muelleri than in P. tricornutum regardless of treatment. The major fatty acids detected were 14:0, 16:0, 16:1, 16:2, 16:3, 18:0, 18:1(n-9)c, 18:2(n-6) and 20:5(n-3) with additional fatty acids 18:1(n-9)t and 20:4(n-6) detected in C. muelleri. Short duration (2 h) temperature increase above optimal growth temperature had a greater effect on fatty acid composition in C. muelleri than in P. tricornutum and the degree of fatty acid saturation was affected more by temperature in C. muelleri than in P. tricornutum during both short and long duration (24 h) treatments. Total protein assay results suggest that P. tricornutum, but not C. muelleri, was undergoing stress under our growing conditions although lipids in both diatoms were affected by increased temperature. Immunodetection of proteins with anti-rubisco indicates that the rubisco large subunit was undergoing greater turnover in C. muelleri than in P. tricornutum. However, the integrity of rubisco as a suitable indicator of lipid status needs further study. This work supports the hypothesis that a particular temperature, and not treatment duration, has the greater effect on changes in fatty acid composition. Furthermore, changes in fatty acid composition and degree of fatty acid saturation occurred more quickly in the diatoms in response to increased temperature than previously observed in nutrient starvation studies. Since diatom lipids represent an important resource for growth and reproduction of marine animals, the rapid alteration of their lipid composition under temperatures normally encountered in marine environments warrants further study.     

Functional analysis of sigH expression in Corynebacterium glutamicum

The sigH gene of Corynebacterium glutamicum encodes ECF sigma factor sigmaH. The gene apparently plays an important role in other stress responses as well as heat stress response. In this study, we found that deleting the sigH gene made C. glutamicum cells sensitive to the thiol-specific oxidant diamide. In the sigH mutant strain, the activity of thioredoxin reductase markedly decreased, suggesting that the trxB gene encoding thioredoxin reductase is probably under the control of sigmaH. The expression of sigH was stimulated in the stationary growth phase and modulated by diamide. In addition, the SigH protein was required for the expression of its own gene. These data indicate that the sigH gene of C. glutamicum stimulates and regulates its own expression in the stationary growth phase in response to environmental stimuli, and participates in the expression of other genes which are important for survival following heat and oxidative stress response.     

C-fos expression in rat brain during heat stress

Rats, under urethane anesthesia, 0, 20, 40 or 80 min after the start of heat stress (42°C) were sacrificed for determination of c-fos expression in different brain regions. In situ hybridization and immunocytochemistry methods were used, respectively, for determination of c-fos mRNA and protein, respectively. In general, either colon temperature (T_CO), mean arterial pressure (MAP), local cerebral blood flow (CBF) or c-fos expression in different brain regions (including the preoptic area, supraoptic nuclei, paraventricular nuclei, thalamus, amygdala, nucleus tract solitarii, area postrema and ventrolateral medulla) increased at 20–40 min after the start of heat exposure. However, the heatstroke, which appears as profound decreases in both MAP and local CBF and increases in T_CO, was produced 80 min after heat stress. The c-fos expression was heavily induced in all these brain regions after the onset of heatstroke. The data suggest that c-fos expression in rat brain during heatstroke is associated with hyperthermia, arterial hypotension or cerebral ischemia.     

Effects of juvenile hormone and 20-hydroxyecdysone on alkaline phosphatase activity in Drosophila under normal and heat stress conditions

The effect of 20-hydroxyecdysone (20E) and the juvenile hormone (JH) on the activity of the alkaline phosphatase (ALP) has been studied in young females of wild-type Drosophila virilis and Drosophila melanogaster under normal conditions and under heat stress (38 degrees C). Both 20E feeding of the flies and JH application led to a substantial rise in ALP activity. ALP activity was also measured in young females of a JH-deficient strain of D. melanogaster, apterous(56f). A decrease in the enzyme activity was observed in the mutant females as compared to wild type. A rise in JH and 20E levels was found not to prevent the response of ALP to heat stress, but to change its stress-reactivity. Mechanisms of regulation of dopamine (DA) level by gonadotropins in Drosophila are discussed.     

Capacity for protein synthesis following heat stimulus of Drosophila associates with heat tolerance but does not underlie the latitudinal tolerance cline

Across populations of Drosophila melanogaster along the Australian eastern coastline latitudinal clines occur in both heat-knockdown tolerance and hardened heat-knockdown tolerance – low latitude tropical populations being more tolerant. A latitudinal cline also occurs for rates of total protein synthesis following a mild heat stress, with tropical populations having higher rates. Since the control of protein synthesis following heat stress is an important component of the cellular heat-shock response, we hypothesised that the higher rates of synthesis that follow a heat stimulus lead to higher knockdown tolerance and underpins the cline. However, levels of heat-stimulated total protein synthesis have been negatively related to heat-hardening capacity, a somewhat conflicting result. Here we examine the relationship between these physiological and adaptive traits in a set of 40 family lines derived from a hybrid laboratory population established by crossing populations from either end of the latitudinal transect. Among these lines high levels of heat-stimulated total protein synthesis were associated with both low basal and low heat-hardened adult knockdown time, confirming the importance of a negative relationship between protein synthesis and thermal tolerance. This result, when considered along with the directions of the latitudinal clines in protein synthesis and tolerance, suggests that variation in rates of heat-stimulated total protein synthesis is not a factor contributing to the latitudinal cline in heat tolerance. Given the robustness of this negative relationship we discuss possible explanations and future experiments to elucidate how the cellular heat stress response might facilitate increased knockdown tolerance.     

Effect of feeding Saccharomyces Cerevisiae on performance of dairy cows during summer heat stress

Effects of feeding a culture of Saccharomyces cerevisiae to lactating cows on their lactational performance during heat stress were determined. Multiparous Holstein cows (n = 723) calving during the summer months from two dairy farms were randomly assigned to a diet containing no yeast culture (control; n = 361) or 30 g/d of a S. cerevisiae yeast culture (YC; n = 362) fed from 20 to 140 d in milk (DIM). Cows were milked twice daily and the production of milk and milk components was measured every 2 weeks. Dry matter (DM) intakes from 6 pens were measured daily and pen temperature and humidity were evaluated hourly from June to November. Rectal temperature was measured in 88 cows (22/treatment/farm), once weekly, and blood was sampled from a subset of 120 cows at 58 and 100 DIM for measurements of plasma glucose, nonesterified fatty acids, 3-OH-butyrate, insulin, and urea N concentrations. Daily temperature, humidity and the temperature-humidity index in the study pens did not differ between treatments, and rectal temperature of cows in the control and YC treatments differed with days postpartum. Intake of DM was similar between diets, but cows fed YC produced 1.2 kg/d more milk, more milk true protein, solids-not-fat and lactose than that produced by control cows. However, energy-corrected milk yield, and concentrations of true protein, solids-not-fat and lactose did not differ between treatments. Feeding YC did not influence plasma metabolites, insulin, or body condition score of cows, but urea N concentrations were reduced. Feeding a yeast culture of S. cerevisiae improved yields of milk and milk components in heat-stressed multiparous Holstein cows.     

Effects of exposure to short-term heat stress on male reproductive fitness in a soil arthropod

Ambient temperature is a key environmental factor influencing a variety of aspects of the ecology and evolution of ectotherms. Reproductive traits have been suggested to be more sensitive to thermal stress than other life history traits. This study investigated the direct and indirect effects of heat shock on male reproductive success in the widespread springtail Orchesella cincta. Male springtails were exposed to four temperature treatments: heat hardening (35.2 °C for 1 h), heat shock (37.2 °C for 1 h), heat hardening + heat shock (35.2 °C for 1 h, followed 15 h later by 37.2 °C for 1 h), and control (20 °C). The heat shock gene Hsp70 showed high expression in all the heat treatments, indicating that the treatments indeed induced thermal stress. Significant mortality was only found in the treatment with heat shock, both with and without heat hardening. A direct effect of heat treatment was found on time to first reproduction, which was significantly longer after heat shock (with or without heat hardening) than in the control treatment. There was no difference among treatments in the number of spermatophores produced in the first reproductive instar. Heat treatment also had indirect effects on male reproductive success. Females chose significantly more spermatophores from control males than from males that received heat shock, heat hardening or both. A high percentage of spermatophores produced by heat shocked males caused reproductive failure in females, but no significant differences among treatments were found.Our results suggest that not all traits were equally affected by the heat stress. Heat hardening did not protect reproductive traits against the negative effects of heat shock. The indirect effects of heat shock on reproduction may be equally important as the direct effects.     

Cold stenothermal cave-dwelling beetles do have an HSP70 heat shock response

The response to high temperatures in adults of two cold stenothermal cave-dwelling leptodirins, Neobathyscia mancinii and Neobathyscia pasai (Coleoptera, Cholevidae) was evaluated by determinating levels of gene expression of two members of the family of heat shock proteins 70 kDa by qPCR. In both species, hsc70 mRNA level was constant with increasing temperature, whereas a significant increase in the inducible member (hsp70) mRNA was observed, higher in N. pasai. This difference could be due to their in-cave distribution: N. pasai colonizes the cave entrance where the temperature is more variable than the internal part where N. mancinii is confined. These results demonstrated for the first time the occurrence of a heat shock response in troglobite insects and suggest the correlation between the intensity of this response and the adaptation to the cave environment.     

Cell damage and recovery in cryopreserved microphytobenthic diatoms

Two pennate microphytobenthic diatoms, Amphora coffeaeformis (Agardh) Kutzing and Navicula transitans var. derasa f. delicatula Heimdal, were cryopreserved and monitored on thawing to track the mechanical injuries and their post-preservation recovery. Cells were subjected to (1) direct freezing in liquid nitrogen and (2) two-step cooling with and without the cryoprotectant, dimethyl sulfoxide (Me(2)SO). Mechanical injury due to exposure to low temperature differed between the two species. While A. coffeaeformis cells were intact and could survive even direct freezing without a cryoprotectant, N. delicatula cell chloroplasts were damaged. However, the two-step cooling along with a cryoprotectant minimized the mechanical injury to cells of both species thereby enhancing the post-thaw viability.     

OGT functions as a catalytic chaperone under heat stress response: a unique defense role of OGT in hyperthermia

Protein O-GlcNAcylation is proceeded by O-linked GlcNAc transferase (OGT) in nucleocytoplasm and is involved in many biological processes although its physiological role is not clearly defined. To identify the functional significance of O-GlcNAcylation, we investigated heat stress effects on protein O-GlcNAcylation. Here, we found that protein O-GlcNAcylation was significantly increased in vivo during acute heat stress in mammalian cells and simultaneously, the enhanced protein O-GlcNAcylation was closely associated with cell survival in hyperthermia. Our results demonstrate that hyperthermal cytotoxicity may considerably be facilitated under the condition of insufficient level of protein O-GlcNAcylation inside cells. Furthermore, OGT reaction might be crucial for triggering thermotolerance to recover hyperthermal sensitivity without particular induction of heat shock proteins (hsps). Thus, we propose that OGT can respond rapidly to heat stress through the enhancement of nucleocytoplasmic protein O-GlcNAcylation for a rescue from the early phase of hyperthermal cytotoxicity.     

The combined effect of salt stress and heat shock on proteome profiling in Suaeda salsa

Under natural conditions or in the field, plants are often subjected to a combination of different stresses such as salt stress and heat shock. Although salt stress and heat shock have been extensively studied, little is known about how their combination affects plants. We used proteomics, coupled with physiological measurements, to investigate the effect of salt stress, heat shock, and their combination on Suaeda salsa plants. A combination of salt stress and heat shock resulted in suppression of CO₂ assimilation and the photosystem II efficiency. Approximately 440 protein spots changed their expression levels upon salt stress, heat shock and their combination, and 57 proteins were identified by MS. These proteins were classified into several categories including disease/defense, photosynthesis, energy production, material transport, and signal transduction. Some proteins induced during salt stress, e.g. choline monooxygenase, chloroplastic ATP synthase subunit beta, and V-type proton ATPase catalytic subunit A, and some proteins induced during heat shock, e.g. heat shock 70 kDa protein, probable ion channel DMI1, and two component sensor histidine kinase, were either unchanged or suppressed during a combination of salt stress and heat shock. In contrast, the expression of some proteins, including nucleoside diphosphate kinase 1, chlorophyll a/b binding protein, and ABC transporter I family member 1, was specifically induced during a combination of salt stress and heat shock. The potential roles of the stress-responsive proteins are discussed.     

The capacity of Drosophila to heat harden associates with low rates of heat-shocked protein synthesis

The thermotolerance of a species or of an ecotype is important for determining its habitat range and vigour, and considerable research has focused on identifying underlying physiological, biochemical and genetic bases of thermotolerance traits. Rates of protein synthesis in tissues when organisms experience a sudden heat stress as occurs on rare hot days may be important to avoid heat-induced paralysis and to survive. While natural variation in Drosophila melanogaster thermotolerance has been associated with heat-shock gene expression, little attention has been given to examining the thermo-protective role of protein synthesis generally. Using two independently derived sets of single-pair mating lines, we characterised variation in rates of protein synthesis in dissected ovarian tissues, both before and after a heat shock applied at different severities in the two sets. In both sets of lines heat-shocked protein synthesis rates were negatively associated with the increase in heat knockdown tolerance after hardening. These associations occurred in a different sex in each set. Variation in rates of Hsp70 synthesis failed to associate with levels of heat tolerance or general protein synthesis. Our results suggest heritable variation in the rate of protein synthesis following heat stress, independently of Hsp70 variation, contributes to heat tolerance variation in this species.     

Serotonin synthesis inhibition by pre-treatment of p-CPA alters sleep-electrophysiology in an animal model of acute and chronic heat stress

The effects of pre-treatment of para-chlorophenylalanine (p-CPA) on sleep–wake electroencephalograms (EEG) have been demonstrated in three age groups of rats subjected to heat stress. Each age group for both p-CPA pre-treated and untreated subjects was sub-divided into three groups: (i) acute heat stress—subjected to a single heat exposure for 4 h at 38 °C; (ii) chronic heat stress—exposed for 21 days daily for 1 h in the incubator at 38 °C; and (iii) handling control groups. Digital polygraphic sleep recordings were performed just after the heat exposure from acute stressed rats and on the 22nd day from chronic stressed rats. The analyses of results demonstrated that many changes associated with sleep-EEG (either in sleep–wake parameter or in EEG frequencies) due to acute and chronic heat stress were reversed (changes were analyzed; P<0.05 or better) in p-CPA pre-treated groups of rats. However, differential observations between acute and chronic heat stress groups of subjects were recorded, which are thought to have happened due to acclimatization of subjects to the hot environment. The results of present study supported the previous hypothesis about the significant involvement of serotonin in sleep–wake parameters and also demonstrated its participation in brain electrophysiological alterations in stressed conditions.