Exploitation of genetic and physiological determinants of embryonic resistance to elevated temperature to improve embryonic survival in dairy cattle during heat stress

Heat stress causes large reductions in fertility in lactating dairy cows. The magnitude and geographical extent of this problem is increasing because improvements in milk yield have made it more difficult for cows to regulate body temperature during warm weather. There have been efforts to improve fertility during heat stress by exploiting determinants of oocyte and embryonic responses to elevated temperature. Among these determinants are genotype, stage of development, and presence of cytoprotective molecules in the reproductive tract. One effective strategy for increasing pregnancy rate during heat stress is to use embryo transfer to bypass effects of elevated temperature on the oocyte and early embryo. Pregnancy success to embryo transfer in the summer can be further improved by exposure of embryos to insulin-like growth factor-I during culture before transfer. Among the cytoprotective molecules that have been examined for enhancing fertility during heat stress are bovine somatotropin and various antioxidants. To date, an effective method for delivery of these molecules to increase fertility during heat stress has not been identified. Genes in cattle exist for regulation of body temperature and for cellular resistance to elevated temperature. Although largely unidentified, the existence of these genes offers the possibility for their incorporation into dairy breeds through crossbreeding or on an individual-gene basis. In summary, physiological or genetic manipulation of the cow to improve embryonic resistance to elevated temperature is a promising approach for enhancing fertility of lactating dairy cows.     

Cellular and functional adaptation to thermal stress in ovarian granulosa cells in mammals

Climate change represents a significant environmental challenge to human welfare. One of many negative impacts may be on animal reproduction. Elevated ambient temperature unfavourably influences reproductive processes in mammals. High temperature can affect reproductive processes such as follicle development and may alter follicular fluid concentrations of amino acids, fatty acids, minerals, enzymes, antioxidants defence and growth factors. These impacts may lead to inferior oocyte competence and abnormal granulosa cell (GCs) function. Mammalian oocytes are enclosed by GCs that secret hormones and signalling molecules to promote oocyte competence. GCs are essential for proper follicular development, oocyte maturation, ovulation, and luteinization. Many environmental stressors, including thermal stress, affect GC function and alter oocyte development and growth. Several studies documented a link between elevated ambient temperature and increased generation of cellular reactive oxygen species (ROS). ROS can damage DNA, reduce cell proliferation, and induce apoptosis in GCs, thus altering oocyte development. Additionally, thermal stress induces upregulation of thermal shock proteins, such as HSP70 and HSP90. This review provides an update on the influence of thermal stress on GCs of mammals. Discussions include impacts to steroidogenesis (estradiol and progesterone), proliferation and cell cycle transition, apoptosis, oxidative stress (ROS), antioxidants related genes, heat shock proteins (HSPs) and endoplasmic reticulum responses.     

Susceptibility of bovine germinal vesicle-stage oocytes from antral follicles to direct effects of heat stress in vitro

Delineation of maternal versus direct effects of heat stress in reducing development at the germinal vesicle (GV) stage is challenging, because oocytes spontaneously resume meiosis after removal from antral follicles. The use of S-roscovitine (inhibitor of p34(cdc2)/cyclin B kinase) to hold bovine oocytes at the GV stage without compromising early embryo development was previously validated in our laboratory. The objective of the present study was to assess the direct effects of an elevated temperature commonly seen in heat-stressed dairy cows on cumulus-oocyte complexes (COCs) held at the GV stage using 50 microM S-roscovitine. During roscovitine culture, GV-stage COCs (antral follicle diameter, 3-8 mm) were cultured at 38.5 or 41 degrees C. Thereafter, oocytes were removed from roscovitine medium and allowed to undergo in vitro maturation, fertilization, and culture. Zona pellucida hardening (solubility to 0.5% pronase), nuclear stage (Hoechst 33342), cortical granule type (lens culinaris agglutinin-fluorescein isothiocyanate [FITC]), and early embryo development were evaluated. Culture of GV-stage COCs at 41 degrees C increased the proportion that had type III cortical granules and reduced the proportion that progressed to metaphase II after in vitro maturation. Effects of 41 degrees C on zona pellucida hardening, fertilization (penetration, sperm per oocyte, pronuclear formation, and monospermic and putative embryos), and cleavage of putative zygotes were not noted. However, culture of GV-stage COCs at 41 degrees C for 6 h decreased the proportion of 8- to 16-cell embryos, whereas 41 degrees C for 12 h reduced blastocyst development. In summary, antral follicle COCs are susceptible to direct effects of elevated body temperature, which may account in part for reduced fertility in heat-stressed cows.     

The balance between stress resilience and vulnerability is regulated by corticotropin‐releasing hormone during the critical postnatal period for sensory development

Determining whether a stressful event will lead to stress‐resilience or vulnerability depends probably on an adjustable stress response set point, which is most likely effective during postnatal sensory development and involves the regulation of corticotrophin‐releasing hormone (CRH) expression. During the critical period of thermal‐control establishment in 3‐day‐old chicks, heat stress was found to render resilient or sensitized response, depending on the ambient temperature. These two different responses were correlated with the amount of activation of the hypothalamic–pituitary–adrenal (HPA) axis. The expression of CRH mRNA in the hypothalamic paraventricular nucleus was augmented during heat challenge a week after heat conditioning in chicks which were trained to be vulnerable to heat, while it declined in chicks that were trained to be resilient. To study the role of CRH in HPA‐axis plasticity, CRH or Crh‐antisense were intracranially injected into the third ventricle. CRH caused an elevation of both body temperature and plasma corticosterone level, while Crh‐antisense caused an opposite response. Moreover, these effects had long term implications by reversing a week later, heat resilience into vulnerability and vice versa. Chicks that had been injected with CRH followed by exposure to mild heat stress, normally inducing resilience, demonstrated, a week later, an elevation in body temperature, and Crh mRNA level similar to heat vulnerability, while Crh‐antisense injected chicks, which were exposed to harsh temperature, responded in heat resilience. These results demonstrate a potential role for CRH in determining the stress resilience/vulnerability balance. © 2014 Wiley Periodicals, Inc. Develop Neurobiol 75: 842–853, 2015     

Mouse liver is more resistant than skeletal muscle to heat-induced apoptosis

During passive heat stress, shifting of blood flow from the hepato-splanchnic to peripheral regions produces less favorable physiological conditions in the liver than in the skeletal muscle. We were wondering if the two organs differ in susceptibility to heat injury and thus examined the effects of heat shock exposure on apoptotic and heat stress-related markers in the gastrocnemius muscle and liver of mice. During heat exposure, mice had a peak core body temperature of 41.1 ± 0.7 °C. Heat-exposed mice showed higher levels of reactive oxygen species (ROS), cleaved caspases, fragmented DNA, and Drp1 protein expression in the gastrocnemius muscles than control mice. These changes were not observed in the livers of heat-exposed mice. Furthermore, the levels of glucocorticoid receptor, HSP70, and HSF1 proteins were significantly elevated in the gastrocnemius muscles of heat-exposed mice compared with that of control mice. The livers of heat-exposed mice also revealed increased expression of HSP70 but no changes in the other proteins. These results demonstrate that heat exposure induces significantly lower levels of the stress response and apoptosis in the liver than in the skeletal muscle of mice. The liver tissue resistance against heat stress is associated with low levels of heat-induced ROS production and mitochondrial fission protein expression.     

Exposure to exogenous egg cortisol does not rescue juvenile Chinook salmon body size,condition, or survival from the effects of elevated water temperatures

Climate change is leading to altered temperature regimes which are impacting aquatic life, particularly for ectothermic fish. The impacts of environmental stress can be translated across generations through maternally derived glucocorticoids, leading to altered offspring phenotypes. Although these maternal stress effects are often considered negative, recent studies suggest this maternal stress signal may prepare offspring for a similarly stressful environment (environmental match). We applied the environmental match hypothesis to examine whether a prenatal stress signal can dampen the effects of elevated water temperatures on body size, condition, and survival during early development in Chinook salmon Oncorhynchus tshawytscha from Lake Ontario, Canada. We exposed fertilized eggs to prenatal exogenous egg cortisol (1,000 ng/ml cortisol or 0 ng/ml control) and then reared these dosed groups at temperatures indicative of current (+0°C) and future (+3°C) temperature conditions. Offspring reared in elevated temperatures were smaller and had a lower survival at the hatchling developmental stage. Overall, we found that our exogenous cortisol dose did not dampen effects of elevated rearing temperatures (environmental match) on body size or early survival. Instead, our eyed stage survival indicates that our prenatal cortisol dose may be detrimental, as cortisol‐dosed offspring raised in elevated temperatures had lower survival than cortisol‐dosed and control reared in current temperatures. Our results suggest that a maternal stress signal may not be able to ameliorate the effects of thermal stress during early development. However, we highlight the importance of interpreting the fitness impacts of maternal stress within an environmentally relevant context.     

Effects of l-leucine in ovo feeding on thermotolerance,growth and amino acid metabolism under heat stress in broilers

Recently, we found that in ovo feeding of l-leucine (l-Leu) afforded thermotolerance, stimulated lipid metabolism and modified amino acid metabolism in male broiler chicks. However, the effects of in ovo feeding of l-Leu on thermoregulation and growth performance until marketing age of broilers are still unknown. In this study, we investigated the effects of in ovo feeding of l-Leu on body weight (BW) gain under control thermoneutral temperature or chronic heat stress. We measured changes of body temperature and food intake, organ weight, as well as amino acid metabolism and plasma metabolites under acute and chronic heat stress in broilers. A total of 168 fertilized Chunky broiler eggs were randomly divided into 2 treatment groups in experiments. The eggs were in ovo fed with l-Leu (34.5 µmol/500 µl per egg) or sterile water (500 µl/egg) during incubation. After hatching, male broilers were selected and assigned seven to nine replicates (one bird/replicate) in each group for heat challenge experiments. Broilers (29- or 30-day-old) were exposed to acute heat stress (30 ± 1°C) for 120 min or a chronic heat cyclic and continued heat stress (over 30 ± 1°C; ages, 15 to 44 days). In ovo feeding of l-Leu caused a significant suppression of enhanced body temperature without affecting food intake, plasma triacylglycerol, non-esterified fatty acids, ketone bodies, glucose, lactic acid or thyroid hormones under acute heat stress. Daily body temperature was significantly increased by l-Leu in ovo feeding under chronic heat stress. Interestingly, in ovo feeding of l-Leu caused a significantly higher daily BW gain compared with that of the control group under chronic heat stress. Moreover, some essential amino acids, including Leu and isoleucine, were significantly increased in the liver and decreased in the plasma by l-Leu in ovo feeding under acute heat stress. These results suggested that l-Leu in ovo feeding afforded thermotolerance to broilers under acute heat stress mainly through changing amino acid metabolism until marketing age.     

Adverse impact of heat stress on embryo production: causes and strategies for mitigation

The production of embryos by superovulation is often reduced in periods of heat stress. The associated reduction in the number of transferable embryos is due to reduced superovulatory response, lower fertilization rate, and reduced embryo quality. There are also reports that success of in vitro fertilization procedures is reduced during warm periods of the year. Heat stress can compromise the reproductive events required for embryo production by decreasing expression of estrus behavior, altering follicular development, compromising oocyte competence, and inhibiting embryonic development. While preventing effects of heat stress can be difficult, several strategies exist to improve embryo production during heat stress. Among these strategies are changing animal housing to reduce the magnitude of heat stress, utilization of cows with increased resistance to heat stress (i.e., cows with lower milk yield or from thermally-adapted breeds), and manipulation of physiological and cellular function to overcome deleterious consequences of heat stress. Effects of heat stress on estrus behavior can be mitigated by use of estrus detection aids or utilization of ovulation synchronization treatments to allow timed embryo transfer. There is some evidence that embryonic survival can be improved by antioxidant administration and that pharmacological treatments can be developed that reduce the degree of hyperthermia experienced by cows exposed to heat stress.     

Interactive effects of high temperature and elevated carbon dioxide concentration on cowpea [Vigna unguiculata (L.) Walp.]

Limitations in carbohydrate supplies have been implicated as a factor responsible for reproductive failure under heat stress. Heat stress affects two stages of reproductive development in cowpea [Vigna unguiculata (L.) Walp.], and genotypes are available with tolerance and sensitivity to heat during these different stages. The objectives of this study were to determine the responses of these cowpea lines to ambient and elevated [CO₂], under heat stress and optimal temperature, and test whether differences in carbohydrate supplies due to genotypes, CO₂ enrichment and heat stress are associated with differences in sensitivity to heat during reproductive development. Plants were grown in reach-in growth chambers and subjected to day/night temperatures of either 33/20 or 33/30°C, and [CO₂] levels of either 350 or 700 μmol mol^-1. Under intermediate night temperature (33/20°C), all lines set substantial numbers of pods. Under high night temperature (33/30°C) with either ambient or elevated [CO₂], one heat-sensitive line produced no flowers and the other set no pods, whereas the heat-tolerant line abundantly set pods. High night temperature reduced the overall carbohydrate content of the plants, especially peduncle sugars, and caused decreases in photosynthetic rates. The high pod set of the heat-tolerant line, under high night temperature, was associated with higher levels of sugars in peduncles compared with the heat-sensitive lines. The heat-tolerant line accumulated substantial shoot biomass, exhibited less accumulation of starch in leaves, and possibly had less down-regulation of photosynthesis in response to CO₂ enrichment and heat stress than the heat-sensitive lines. Elevated [CO₂] resulted in higher overall carbohydrate levels in heat-sensitive lines (starch in leaves, stems and peduncles), but it did not increase their heat tolerance with respect to flower production or pod set. Heat-induced damage to floral buds and anthers in the sensitive lines was associated with low sugars levels in peduncles, indicating that heat had greater effects on assimilate demand than on leaf assimilate supply. The heat-tolerant line was the most responsive genotype to elevated [CO₂] with respect to pod production under either high or intermediate temperatures.     

Hyperthermia modifies muscle metaboreceptor and baroreceptor modulation of heat loss in humans

The relative influence of muscle metabo- and baroreflex activity on heat loss responses during post-isometric handgrip (IHG) exercise ischemia remains unknown, particularly under heat stress. Therefore, we examined the separate and integrated influences of metabo- and baroreceptor-mediated reflex activity on sweat rate and cutaneous vascular conductance (CVC) under increasing levels of hyperthermia. Twelve men performed 1 min of IHG exercise at 60% of maximal voluntary contraction followed by 2 min of ischemia with simultaneous application of lower body positive pressure (LBPP, +40 mmHg), lower body negative pressure (LBNP, -20 mmHg), or no pressure (control) under no heat stress. On separate days, trials were repeated under heat stress conditions of 0.6°C (moderate heat stress) and 1.4°C (high heat stress) increase in esophageal temperature. For all conditions, mean arterial pressure was greater with LBPP and lower with LBNP than control during ischemia (all P ≤ 0.05). No differences in sweat rate were observed between pressure conditions, regardless of the level of hyperthermia (P > 0.05). Under moderate heat stress, no differences in CVC were observed between pressure conditions. However, under high heat stress, LBNP significantly reduced CVC by 21 ± 4% (P ≤ 0.05) and LBPP significantly elevated CVC by 14 ± 5% (P ≤ 0.05) relative to control. These results show that sweating during post-IHG exercise ischemia is activated by metaboreflex stimulation, and not by baroreflexes. In contrast, our results suggest that baroreflexes can influence the metaboreflex modulation of CVC, but only at greater levels of hyperthermia.     

To be or not to be--determinants of embryonic survival following heat shock

Elevated temperature can reduce developmental competence of the preimplantation embryo. Whether an embryo survives elevated temperature depends on its genotype, stage of development, exposure to regulatory molecules and redox status. Following fertilization, the embryo is very sensitive to heat shock. By Days 4-5 after insemination, however, the embryo has acquired increased resistance to elevated temperature. One system that may potentiate embryonic survival at later stages of embryonic development is the apoptosis response-inhibition of apoptosis responses at Day 4 exacerbated effects of heat shock on development. Embryo responses to heat shock at Days 4-5 also depend upon genotype because Bos indicus embryos are more resistant than embryos from non-adapted B. taurus. Some experiments (although not all) indicate that survival following heat shock can be increased by reducing oxygen tension, suggesting involvement of reactive oxygen species or hypoxia-induced factors. Embryonic responses to heat shock are also affected by regulatory molecules that act to modify cellular physiology and improve cell survival. The best characterized of these is insulin-like growth factor-1 (IGF-1). Actions of IGF-1 to allow development following heat shock are independent of its anti-apoptotic actions because inhibition of the phosphatidylinositol-3 kinase pathway through which IGF-1 blocks apoptosis does not prevent thermoprotective effects of IGF-1 on development. Identification of specific determinants of embryonic survival creates the opportunity for new strategies to improve pregnancy rates in animals exposed to heat stress. Many environmental perturbations activate similar cellular responses. Therefore, molecular and cellular systems that improve embryonic survival to heat shock may confer protection from other embryotoxic conditions.     

Mitigation of heat stress-related complications by a yeast fermentate product

Heat stress results in a multitude of biological and physiological responses which can become lethal if not properly managed. It has been shown that heat stress causes significant adverse effects in both human and animals. Different approaches have been proposed to mitigate the adverse effects caused by heat stress, among which are special diet and probiotics. We characterized the effect of the yeast fermentate EpiCor (EH) on the prevention of heat stress-related complications in rats. We found that increasing the body temperature of animals from 37.1±0.2 to 40.6±0.2 °C by exposure to heat (45 °C for 25 min) resulted in significant morphological changes in the intestine. Villi height and total mucosal thickness decreased in heat-stressed rats pre-treated with PBS in comparison with control animals not exposed to the heat. Oral treatment of rats with EH before heat stress prevented the traumatic effects of heat on the intestine. Changes in intestinal morphology of heat-stressed rats, pre-treated with PBS resulted in significant elevation of lipopolysaccharides (LPS) level in the serum of these animals. Pre-treatment with EH was effective in the prevention of LPS release into the bloodstream of heat-stressed rats. Our study revealed that elevation of body temperature also resulted in a significant increase of the concentration of vesicles released by erythrocytes in rats, pre-treated with PBS. This is an indication of a pathological impact of heat on the erythrocyte structure. Treatment of rats with EH completely protected their erythrocytes from this heat-induced pathology. Finally, exposure to heat stress conditions resulted in a significant increase of white blood cells in rats. In the group of animals pre-treated with EH before heat stress, the white blood cell count remained the same as in non-heated controls. These results showed the protective effect of the EH product in the prevention of complications, caused by heat stress.     

Hemodynamic responses to heat stress in the resting and exercising human leg: insight into the effect of temperature on skeletal muscle blood flow

Heat stress increases limb blood flow and cardiac output (Q) in humans, presumably in sole response to an augmented thermoregulatory demand of the skin circulation. Here we tested the hypothesis that local hyperthermia also increases skeletal muscle blood flow at rest and during exercise. Hemodynamics, blood and tissue oxygenation, and muscle, skin, and core temperatures were measured at rest and during exercise in 11 males across four conditions of progressive whole body heat stress and at rest during isolated leg heat stress. During whole body heat stress, leg blood flow (LBF), Q, and leg (LVC) and systemic vascular conductance increased gradually with elevations in muscle temperature both at rest and during exercise (r(2) = 0.86-0.99; P < 0.05). Enhanced LBF and LVC were accompanied by reductions in leg arteriovenous oxygen (a-vO(2)) difference and increases in deep femoral venous O(2) content and quadriceps tissue oxygenation, reflecting elevations in muscle and skin perfusion. The increase in LVC occurred despite an augmented plasma norepinephrine (P < 0.05) and was associated with elevations in muscle temperature (r(2) = 0.85; P = 0.001) and arterial plasma ATP (r(2) = 0.87; P < 0.001). Isolated leg heat stress accounted for one-half of the increase in LBF with severe whole body heat stress. Our findings suggest that local hyperthermia also induces vasodilatation of the skeletal muscle microvasculature, thereby contributing to heat stress and exercise hyperemia. The increased limb muscle vasodilatation in these conditions of elevated muscle sympathetic vasoconstrictor activity is closely related to the rise in arterial plasma ATP and local tissue temperature.     

Heat stress increases apical glucose transport in the chicken jejunum

In chickens, elevated environmental temperature reduces food intake. We have previously reported that, during heat stress, the intestinal mucosa has an increased capacity to take up sugars. To investigate whether the effects of warm environment on sugar uptake are an intestinal adaptation to lower energy intake or a response attributable to heat stress, we examined the glucose transport kinetics of apical and basolateral membranes of the jejunum and the mucosal morphology of broiler chickens maintained in climatic chambers for 2 wk. Experimental groups were 1) control ad libitum (CAL), fed ad libitum and in thermoneutral conditions (20 degrees C); 2) heat stress ad libitum (HSAL), fed ad libitum and kept in a heated environment (30 degrees C); and 3) control pair-fed (CPF), maintained in thermoneutral conditions and fed the same amount of food as that consumed by the HSAL group. Both the CPF and the HSAL groups showed reduced body weight gain, but only the HSAL chickens had lower plasma thyroid hormones and higher corticosterone than CAL and CPF groups. The fresh weight and length of the jejunum were only reduced in the HSAL group. The activity and expression of apical sodium-dependent glucose transporter 1 (SGLT-1) were increased by approximately 50% in the HSAL chickens, without effects in the CPF group. No changes in K(d) or in SGLT-1 and glucose transporter-2 K(m) were observed in the pair-fed and heated birds. These results support the view that increased intestinal hexose transport capacity is entirely dependent on adaptations of apical SGLT-1 expression to heat stress and is not due to reduced food intake.     

Environmental temperature and human growth in early life

A link between adult human body size and environmental temperature, evolved through adaptation to heat stress, was first recognized a century ago and is now well accepted in human biology. Increasing heat stress favours smaller body size and an increased ratio of surface area to mass. However, possible effects of temperature on growth in early life have not been considered. Many developing country populations inhabit relatively hot environments compared to industrialized populations, but growth faltering in developing countries is invariably attributed to the combination of poor nutrition and infection. The aim of this study was to model the theoretical effect of growth faltering on cooling capacity in early life. Data on weight, length and total energy expenditure were selected from the literature. Three types of growth (normal growth, stunting and wasting) were considered, and the relationship between body size and heat production investigated. Relative to normal growth, stunting causes little increase in the area-to-mass ratio until 2 years of age, and results in little increase in cooling capacity during infancy. Wasting increases the area-to-mass ratio by > 15% in early infancy, and by 10% thereafter, equivalent to a similar increase in cooling capacity. These findings demonstrate theoretically that growth faltering can relieve heat stress in both infancy and childhood. The hypothesis that heat stress plays a role in human growth faltering in hot environments therefore merits empirical investigation.     

Supplemental ascorbic acid does not affect inferred heat loss in broiler chickens exposed to elevated temperature

The provision of supplemental ascorbic acid has been reported to lower the body temperature of chickens maintained at elevated environmental temperatures. Since body temperature is the net effect of heat production and heat loss, it is not known if the reductions in body temperature were due to a lower heat production or an increase in heat loss. The purpose of this work was to determine if supplemental ascorbic acid facilitates heat loss in chickens exposed to an elevated temperature. On day 12 post-hatch broiler chickens were implanted intra-abdominally with a thermo-sensitive radio transmitter. The following day, birds were placed inside an indirect calorimeter maintained at 34 C for 24 h and provided water containing 0 or 400 ppm ascorbic acid. Oxygen consumption, carbon dioxide production, heat production, respiratory exchange ratio, and body core temperature were measured for 3 h; beginning 21 h after the birds were placed inside the calorimeter. No differences were observed in heat production or body core temperature between birds provided or not and 400 ppm ascorbic acid. This suggests that ascorbic acid has no effect on heat loss. Birds provided ascorbic acid did exhibit a significantly lower respiratory exchange ratio suggesting a greater utilization of lipid for energy production. Although lipid has a lower heat increment compared with protein and carbohydrate, the significance of this finding to birds exposed to elevated temperature is not known. In conclusion, under the conditions of this study the provision of supplemental ascorbic acid to broiler chickens maintained at an elevated temperature did not affect heat loss as inferred from measured heat production and body core temperature.     

Interactions between oxygen tension and glucose concentration that modulate actions of heat shock on bovine oocytes during in vitro maturation

Exposure of oocytes to elevated temperature (i.e. heat shock) during maturation can reduce fertilization rate and development of the resultant embryos. Given the possible role of free radicals in actions of heat shock on cellular function, we tested the hypothesis that a high oxygen environment exacerbates the magnitude of deleterious effects of heat shock on in vitro maturation of bovine oocytes. A preliminary experiment was performed to establish conditions for oocyte maturation that would be independent of oxygen concentration. Oocytes were matured in a modified tissue culture medium-199 (mTCM-199) or modified synthetic oviduct fluid (mSOF) containing 5.6 or 20 mM glucose and under either high (atmospheric oxygen, approximately 21%, v/v) or low oxygen (5%, a value approximating oxygen content of the follicle). For oocytes matured in mTCM-199, development was greater in high oxygen than in low oxygen, whereas development was unaffected by oxygen using mSOF (mediumxoxygen, P<0.05). Accordingly, mSOF was used as the maturation medium in a second study to test the effect of oxygen concentration on the magnitude of actions of heat shock during maturation. Maturation was at 38.5 degrees C for 22 h (control) or 41 degrees C for 12h and 38.5 degrees C for 10h (heat shock). Heat shock slightly decreased cleavage rate, regardless of the maturation conditions, and decreased blastocyst development under all maturation conditions except for the group matured under high oxygen and high glucose (temperature x glucose for oocytes under low oxygen, P<0.05). The percentage of oocytes becoming blastocysts for control and heat shocked oocytes was 25.9% versus 22.5% (low oxygen -- 5.6 mM glucose), 41.6% versus 34.9% (low -- 20 mM), 41.7% versus 35.0% (high -- 5.6 mM), and 37.6% versus 37.5% (high -- 20 mM). In conclusion, under an oxygen tension that approached physiological conditions, heat shock during in vitro maturation reduced oocyte competence for fertilization and subsequent development.     

Alcohol stress,membranes, and chaperones

Ethanol, which affects all body organs, exerts a number of cytotoxic effects, most of them independent of cell type. Ethanol treatment leads to increased membrane fluidity and to changes in membrane protein composition. It can also interact directly with membrane proteins, causing conformational changes and thereby influencing their function. The cytotoxic action may include an increased level of oxidative stress. Heat shock protein molecular chaperones are ubiquitously expressed evolutionarily conserved proteins which serve as critical regulators of cellular homeostasis. Heat shock proteins can be induced by various forms of stresses such as elevated temperature, alcohol treatment, or ischemia, and they are also upregulated in certain pathological conditions. As heat shock and ethanol stress provoke similar responses, it is likely that heat shock protein activation also has a role in the protection of membranes and other cellular components during alcohol stress.     

Yield response of field‐grown soybean exposed to heat waves under current and elevated [CO2]

Elevated atmospheric CO₂ concentration ([CO₂]) generally enhances C₃ plant productivity, whereas acute heat stress, which occurs during heat waves, generally elicits the opposite response. However, little is known about the interaction of these two variables, especially during key reproductive phases in important temperate food crops, such as soybean (Glycine max). Here, we grew soybean under elevated [CO₂] and imposed high‐ (+9°C) and low‐ (+5°C) intensity heat waves during key temperature‐sensitive reproductive stages (R1, flowering; R5, pod‐filling) to determine how elevated [CO₂] will interact with heat waves to influence soybean yield. High‐intensity heat waves, which resulted in canopy temperatures that exceeded optimal growth temperatures for soybean, reduced yield compared to ambient conditions even under elevated [CO₂]. This was largely due to heat stress on reproductive processes, especially during R5. Low‐intensity heat waves did not affect yields when applied during R1 but increased yields when applied during R5 likely due to relatively lower canopy temperatures and higher soil moisture, which uncoupled the negative effects of heating on cellular‐ and leaf‐level processes from plant‐level carbon assimilation. Modeling soybean yields based on carbon assimilation alone underestimated yield loss with high‐intensity heat waves and overestimated yield loss with low‐intensity heat waves, thus supporting the influence of direct heat stress on reproductive processes in determining yield. These results have implications for rain‐fed cropping systems and point toward a climatic tipping point for soybean yield when future heat waves exceed optimum temperature.