Heat shock impairs DNA synthesis and down-regulates gene expression for leptin and Ob-Rb receptor in concanavalin A-stimulated bovine peripheral blood mononuclear cells

This study verified whether leptin or its long isoform receptor (Ob-Rb) genes are expressed in proliferating lymphocytes from bovine species, and whether their expression changes with increased temperatures. Peripheral blood mononuclear cells (PBMC) from five Holstein cows were incubated in the presence of concanavalin A, and alternatively subjected for 65 h to each of the following treatments (T): 39 degrees C continuously (T39) or three 13-h cycles at 40 (T40), 41 (T41) or 42 degrees C (T42), respectively, which were alternated with two 13-h cycles at 39 degrees C. T39 mimicked normothermia; T40, 41 and 42 mimicked conditions of hyperthermia alternated with normothermia. PBMC proliferation declined under T42. Compared with T39, levels of mRNA for leptin was lower under T42, whereas mRNA for Ob-Rb was lower in lymphocytes cultured both under T41 and T42. DNA synthesis was positively correlated with leptin mRNA. This study supports the concept that severe heat stress impairs proliferation of bovine PBMC, confirms that bovine lymphocytes express Ob-Rb gene, and provides the first experimental evidence that bovine lymphocytes express gene for leptin, and that increased temperatures are associated with altered gene expression for leptin and Ob-Rb.     

热应激对牛奶中乳蛋白含量和组分的影响及作用机制

热应激不仅影响奶牛健康,而且易导致奶牛"热应激乳蛋白降低征"的发生,从而影响牛奶品质。本文综述了近些年来热应激影响牛奶中乳蛋白含量及酪蛋白和乳清蛋白组分的研究进展,探讨了热应激诱导的"热应激乳蛋白降低征"发生机理,以期为缓解奶牛热应激和提高牛奶品质提供参考。     

Effect of Heat Stress on Milk Production,Rectal Temperature,Respiratory Rate and Blood Chemistry in Holstein,Jersey and Australian Milking Zebu Cows

The effect of heat stress on changes in milk production, rectal temperature, respiratory rate and blood chemistry was evaluated in three groups of six mature Holstein, Jersey and Australian Milking Zebu (AMZ) dairy cows. These animals were subjected to a cool environment when the mean temperature-humidity index (THI) was 72+/-1.4 (dry bulb temperature of 22.2-24.4 degrees C and relative humidity of 100-60%) during the month of December. This experiment was repeated during the hotter month of July of the following year, when the mean THI was 93+/-3.1 (dry bulb temperature of 35.6-43.9 degrees C and relative humidity 95-35%). Holstein cows produced more (p <0.01) milk than AMZ and Jersey cows during the cooler months of the year and all the cows were dry during the hotter months from June until September. Heat stress increased (p<0.01) rectal temperature and respiratory rate in all three breeds. Heat stress had no effect on blood pH in Holstein and AMZ cows but lowered (p <0.01) blood pH from 7.42 to 7.34 in Jersey cows. In addition, heat stress lowered (p <0.01) blood pCO2 (kPa), bicarbonate (HCO3, mmol/L), base excess (BE, mmol/L) and plasma chloride (Cl-, mmol/L) in all three breeds. The total haemoglobin (THb, g/dl) was elevated (p <0.01) in all three breeds when they were subjected to heat stress. Heat stress increased (p<0.01) oxygen saturation (O2SAT, %) in Jersey and AMZ cows but lowered it (p <0.01) in Holstein cows. On the other hand, heat stress increased (p <0.01)pO2 (kPa) in Holstein and Jersey cows but lowered it (p <0.01) in AMZ cows. Heat stress increased (p <0.01) plasma potassium (K, mmol/L) and calcium (Ca, mmol/L) only in Holstein and Jersey cows but lowered them (p<0.01) in AMZ cows. The plasma glucose (GLU, mmol/L) increased (p<0.01) with heat stress in Holstein and AMZ cows but decreased (p <0.01) in Jersey cows. Heat stress increased (p<0.01) plasma creatinine (CR, (mol/L) but lowered (p<0.01) plasma creatinine phosphokinase (CPK, IU/L), aspartate aminotransferase (AST, IU/L) and blood urea nitrogen (BUN, mmol/L) in all three breeds. These results indicate that heat-stressed Holstein and AMZ cows were able to maintain their acid-base balance with a marginal change in their pH of 0.02 when their rectal temperatures increased by 0.47 and 0.38 degrees C, respectively. When heat stress increased the rectal temperature in Jersey cows by 0.70 degrees C, the pH decreased (p<0.01) from 7.42 to 7.34. However, even with this decrease 0.08 the pH is still within the lower physiological limit of 7.31.     

Spontaneous and mitogen-induced RNA synthesis by blood lymphocytes from bovine leukemia virus-infected and normal cows

Peripheral blood lymphocytes (PBL) from normal and bovine leukemia virus (BLV)-infected cattle were prepared by density gradient technique and incubated with and without phytohaemagglutinin (PHA) and pokeweed mitogen (PWM). RNA synthesis was determined at different periods of incubation by 3H-uridine incorporation. PBL from BLV-infected cows with persistent lymphocytosis (PL) showed the highest spontaneous RNA synthesis. PBL from BLV-infected cows with normal lymphocyte counts synthesized more RNA than cells from normal animals. Decreased mitogen responses were observed in PBL from infected cows with PL in comparison to normal and BLV-infected cattle without PL. PHA and PWM did not show significant differences in their degree of stimulation of RNA synthesis.     

Influence of induced heat stress on HSP70 in buffalo lymphocytes

Heat stress in farm animals, such as cattle and buffalo during summer and post-summer seasons is a problem for livestock producers. The effect of heat stress becomes pronounced when heat stress is accompanied with ambient humidity impairing the immune status, growth, production and reproductive performance of animals. Increase in HSP70 levels from cell cultures in presence of different stressors often does not reflect the physiological adaptability of animals governing thermal regulation. In this study we directly compared the effect of different heat stress conditions with the immune status and HSP70 expression patterns from buffalo lymphocytes both in vivo and in vitro. Murrah buffalo calves were exposed to induced heat stress with two experimental treatments: hot-dry (42 °C with existing relative humidity) or hot humid (35 °C with 70% relative humidity) condition in psychometric chamber, 4 h daily for 12 days and compared with control animals maintained in an experimental shed under natural conditions. There was >200-fold increase in serum-HSP70 levels in both heat stress conditions compared with control. Furthermore, the immune status of the calves failed to activate the level of HSP70 expression in serum lymphocytes. Lymphocytes cultured in vitro at higher temperature exert 2.5-fold increase in HSP70 concentration. This study is the first of its kind to demonstrate more complex expression pattern of buffalo serum-HSP70 level as a thermo adaptive response compared with in vitro treated cells. Results from this study indicate that serum-HSP70 levels could be used as a sensitive biomarker for heat stress management in large farm animals.     

In vitro migration responses of neutrophils from cows and calves

The directional (chemotactic) and random migration activities of neutrophils from cows and newborn and 2-week-old calves were determined by use of the chemotaxis-under-agarose assay. Blood samples were stored for 2, 24, or 48 hours and at 4 or 25 C before testing. During the assay, cells were incubated at 17, 27, or 37 C. The assay was found suitable for testing the directional and random migration activities of neutrophils from cattle. Directional migration of neutrophils was diminished (P less than or equal to 0.05) when cells were incubated at 17 or 27 C, compared with data from incubation at 37 C. Random migration of neutrophils was unaffected by test incubation temperature. Significant (P less than or equal to 0.05) differences were found between cows and calves regarding the percentage number and viability and the directional and random migration activities of neutrophils. Neutrophils from cows were adversely affected to a greater extent by prolonged sample storage times or low storage temperature than were neutrophils from calves. Results indicate that a sample storage time of up to 24 hours, a sample storage temperature of 25 C, and a test incubation temperature of 37 C provided optimal conditions for testing the migratory activities of neutrophils from cattle.     

Effect of xylazine in heifers under thermoneutral or heat stress conditions

A study was performed to assess the effect of xylazine HCl (0.1 mg/kg of body weight, IV) in heifers maintained at thermoneutrality (18 C, 42% humidity) or under heat stress (33 C, 63% humidity) conditions. Xylazine caused 50 and 70% decreases in serum insulin concentrations in the thermoneutral and heat-stressed heifers, respectively. Xylazine-induced hypoinsulinemia was associated with hyperglycemia. In the thermoneutral group, serum glucose concentrations increased from a basal concentration of 75 mg/dl to 150 mg/dl after 15 minutes. In the heat stress group, the serum glucose concentration increased from 65 mg/dl to 105 mg/dl. Hyperglycemia peaked at 2 hours and remained high for 6 hours after xylazine administration. Heat-stressed heifers took a longer time (107 minutes) to stand than did heifers under thermoneutral conditions (41 minutes). The time to regain sensation to pain was significantly prolonged in heat-stressed heifers. Xylazine had no effect on body temperature and respiration rate in heifers under the thermoneutral condition, whereas it markedly induced hyperthermia and suppressed respiration rate in the heat-stressed heifers. Furthermore, the pulse rate was slightly decreased in thermoneutral heifers and was markedly decreased in the heat-stressed heifers.     

Bovine leukosis virus in sheep,lymphocyte modification and surface-immunoglobulin-bearing cell numbers

Lymphocytes from sheep experimentally infected with bovine leukosis virus (BLV) and from non-infected normal sheep were examined for the presence of surface Ig by an immunofluorescence test. Surface Ig-bearing lymphocytes in blood from BLV-infected sheep increased when lymphocyte counts of blood were elevated in comparison with normal animals. The mitogen stimulation of cultured lymphocytes from BLV-infected sheep and from non-infected normal sheep was determined by measuring ³H-thymidine incorporation. Peripheral blood lymphocytes (PBL) from BLV-infected leukemic sheep with elevated PBL counts responded poorly to phytohemagglutinin M and concanavalin A but responded well to lipopolysaccharide compared with lymphocytes from uninfected animals. In BLV-infected preleukemic sheep with low PBL counts, stimulation indices of mitogen responses of lymphocytes with phytohemagglutinin M, concanavalin A, and pokeweed mitogen were low compared with those of lymphocytes from uninfected animals. The results indicated that B cells were affected by BLV infection in sheep as suggested by the increased number of surface Ig-bearing lymphocytes and that significant alteration of mitogen stimulation of lymphocytes occured in sheep with BLV infection.     

Effect of prolonged heat stress in single-comb white leghorn hens on progeny resistance to Salmonella enteritidis organ invasion.

In our laboratory, we have often had difficulty infecting neonatal chickens with invasive salmonellae when ambient temperatures exceed 30 C. We hypothesized that this increased resistance in chicks during warmer months may be associated with heat stress-associated maternal factors. Presently, single-comb white leghorn hens were separated into a non-heat-stressed group, reared under temperatures from approximately 10 to 24 C, and a heat-stressed group, in which environmental temperature was incrementally elevated to near 37 C and maintained for the duration of the 13-wk study. For Expt. 1, eggs from heat-stressed or control hens, collected on days 8-14 of the study, were pooled respective to treatment and incubated. At the time of egg collection, mean hen-day egg production was 51.83% or 65% for heat-stressed or control hens, respectively. On day of hatch, progeny from hens in each group were orally challenged with 0.9 x 10(4) colony-forming units (CFU) Salmonella enteritidis (SE). Rates of SE organ invasion of 97.3% or 94.4% were obtained in progeny from heat-stressed or control hens, respectively. In Expt. 2, eggs from heat-stressed or control hens from days 30-42 of the study were collected and pooled by treatment for incubation. Mean hen-day egg production was 46.5% or 72.85% for heat-stressed or control hens, respectively. On day of hatch, progeny were orally challenged with either 2.2 x 10(3) or 2.2 x 10(4) CFU SE. A 100% incidence in SE organ invasion was observed in all groups. In Expt. 3, eggs were collected from days 43 through 56 of the study. Mean hen-day egg production was 19.8% or 76.8% for heat-stressed or control hens, respectively. On day of hatch, progeny were orally challenged with 2 x 10(3) CFU SE. Rates of SE organ invasion of 95.8% or 95.6% were obtained in progeny from heat-stressed or control hens, respectively. These data suggest that factors other than elevated temperature may be responsible for seasonal resistance to invasive salmonellae infection in neonatal chickens observed in our laboratory during warmer months in Texas.     

Comparative studies on temperature threshold for heat shock protein 70 induction in young and adult Murrah buffaloes

To know the temperature threshold for heat shock protein 70 (HSP70) induction in lymphocytes and to assess physiological changes, if any, in relation to HSP70 induction in young and adult Murrah buffaloes, this study was divided into two parts: I. In vivo study: where assay of HSP70 was performed in blood samples collected from acutely exposed young and adult Murrah buffaloes (n = 6) inside a climatic chamber at 40, 42 and 45 °C for 4 h and thermoneutral temperature (22 °C). Physiological parameters viz., rectal temperature, respiratory rate, pulse rate and skin temperature of different body parts were monitored to assess magnitude of stress in the animals owing to thermal exposure II. For in vitro study, equal numbers of lymphocyte cells were separated from blood collected from young and adult buffaloes and were subjected to four temperature treatments (38, 40, 42 and 45 °C) for 4 h. A significant increase (p < 0.05) in all the physiological parameters in both young and adult buffaloes was observed after exposure to 40, 42 and 45 °C for 4 h as compared to 38 °C. The average plasma HSP70 concentrations (ng/ml) were significantly higher (p < 0.05) at 40, 42 and 45 °C as compared to 38 °C in both young and adult and were higher in young than adult buffaloes at 38 and 45 °C. Heat shock protein 70 level in lymphocyte lysate showed highest concentration after 3-h exposure to all temperatures (40, 42 and 45 °C) in both young and adult buffaloes. The intensity of changes of all physiological parameters was more in young animals than in the adults indicating the greater susceptibility of younger animals to heat stress and was found to be changed at around 40 °C when animals were exposed to different temperatures, indicating the possibility that HSP70 production may be initiated at this temperature which is 2 or 3 °C higher than core body temperature.     

Correlation of hematological changes and serum and monocyte inhibition with the early suppression of phytohemagglutinin stimulation of lymphocytes in experimental infectious bursal disease.

Several experiments were conducted to study the mechanism of infectious bursal disease virus induced suppression of phytohemagglutinin stimulation of peripheral blood lymphocytes. Infectious bursal disease virus inoculation of one week old chicks resulted in significant suppression of phytohemagglutinin stimulation during the first three days after inoculation as demonstrated by a whole blood assay. Mild thymic necrosis was seen on day 3. Hematological changes during this time consisted of increased numbers of circulating lymphocytes and monocytes in infected chickens. Absolute monocyte counts remained elevated even after phytohemagglutinin stimulation had returned to normal. Furthermore, even after a 72.3% reduction in the monocyte population in leukocyte preparations, there was still marked viral induced suppression of phytohemagglutinin stimulation. An elevation in the absolute number of circulating large immature lymphocytes correlated with suppression of phytohemagglutinin stimulation. Sera from infected and control chickens depressed phytohemagglutinin stimulation of lymphocytes from control chickens at the 5 and 10% concentration. At the 1% concentration, inhibiton by control sera was considerably less than the inhibition by infected sera. The relationship between these findings and the mechanism of viral induced suppression of T-lymphocyte function is discussed.     

自然生产条件下热应激周期变化揭示泌乳中期奶牛出现“热应激乳蛋白降低征”

本试验在连续3年时间里测定了上海地区热应激周期变化对泌乳中期奶牛生产性能和牛奶品质的影响。通过实地测定并计算分析,绘制了上海地区热应激周期变化图谱,揭示了整个热应激周期中不同热应激程度的分布状况。研究对比了自然生产环境下无热应激与中度热应激对奶牛生产性能和牛奶品质的影响,发现中度热应激极显著降低了奶牛采食量、产奶量、乳脂校正乳产量、能量校正乳产量、乳脂率、乳蛋白含量、总固体含量(P<0.01),而且显著增加了乳中尿素氮含量(P<0.05)。在热应激周期变化研究中发现,中度热应激显著升高泌乳奶牛的直肠温度和呼吸频率(P<0.05),而且呼吸频率比直肠温度对热应激变化的反应更快、更敏感。热应激周期变化对奶牛干物质采食量、产奶量的影响取决于热应激程度,2012年整个热应激周期的热应激程度比较低,热应激周期变化对奶牛干物质采食量、产奶量无显著影响(P>0.05),但是2013年热应激程度更加严重,热应激周期变化对奶牛干物质采食量、产奶量产生了极显著影响(P<0.01)。在牛奶品质中,受热应激影响最大的是乳蛋白合成量(P<0.01)。2012年和2013年2个热应激周期变化对其他乳成分含量没有显著影响(P>0.05),但是两年的热应激周期变化都导致乳蛋白含量显著下降(P<0.05)和乳中尿素氮含量显著升高(P<0.05)。尤其值得注意的是,2012年热应激周期变化并没有导致奶牛采食量下降(P>0.05),而且产奶量也没有显著性差异(P>0.05),但是仍然出现了乳蛋白含量下降和乳中尿素氮含量升高(P<0.05)。这表明热应激周期变化改变了泌乳中期奶牛氮代谢的途径,发生了氮营养重分配(repartitioning)现象,而且这种现象不依赖于采食量和产奶量,可以称之为“热应激乳蛋白降低征”(heat-stressed milk protein decrease syndrome,HS-MPD)。     

Effect of heat stress on early embryonic development in the beef cow

Hereford and Hereford X Angus cows (n = 31) were utilized to determine the effects of heat stress on early embryonic development and survival. After acclimation to handling, cows were cannulated via the jugular vein on d 7 and assigned to either a control (C) chamber environment of 22 C, 35% relative humidity (RH) or one of two heat stress treatments. Ambient temperature was maintained at 37 C for 12 h followed by a decrease to 33 C for the remainder of the day in both treatment groups. Relative humidity was maintained at 27% in treatment 1 (TRT 1) and 38% in treatment 2 (TRT 2). On d 8 to 16, daily measurements of respiration rate (RES), rectal temperature (REC) and water intake were taken along with samples of blood, which were analyzed for hematocrit (HEM) and plasma concentration of protein (PP), progesterone (P4), estradiol-17 beta (E2), thyroxine (T4) and glucose (GLU). The uterus was recovered and flushed with saline on d 17 to recover the conceptus and uterine contents. Conceptus (if present) and corpus luteum (CL) wet weight were determined. Cows subjected to TRT 2 had increased RES and REC (P less than .01), while HEM was decreased (P less than .05) compared with C cows. Plasma T4 concentration was decreased (P less than .10) in TRT 2 compared with TRT 1 and cows, while P4 concentration were not significantly different. Corpora lutea wet weights were reduced (P less than .10) in heat-stressed cows vs C cows.(ABSTRACT TRUNCATED AT 250 WORDS)     

奶牛在适应热应激过程中机体产生的代谢变化

当外界环境的温度超过奶牛机体的热适温区时,细胞和组织及新陈代谢都会发生变化,机体对热应激反应的防御系统被激活,表现为开始储存热量、迅速增加蒸发散热。如果持续高温,内分泌系统将会做出一定的调节,促使动物适应这种变化。作者主要阐述了在热应激产生的过程中,奶牛细胞内热应激蛋白,内分泌及能量代谢发生的一些变化,并简述了奶牛对热应激的适应。     

热应激对奶牛生理和免疫功能的影响及其机理

奶牛热应激综合征是指奶牛受到超过本身体温调节能力的高温刺激而产生的非特异性防御反应。奶牛发生热应激后会激活体内的下丘脑-垂体-肾上腺轴,改变机体的神经内分泌调节网络,引起奶牛体内皮质醇激素和多种激素水平变化,协同作用于机体以抵抗热应激对自身的影响;此外,热应激奶牛采食量和消化率普遍降低,营养物质摄入不足,机体处于能量负平衡状态。在这种状态下,奶牛会通过增加体内糖、脂肪和蛋白质的代谢来为机体提供能量从而缓解热应激。然而,严重热应激会导致奶牛代谢功能紊乱及免疫系统损伤,最终导致奶牛消化率、产奶量、繁殖率下降,而疾病易感风险性增加,从而影响奶牛生产的经济效益,给畜牧业带来巨大的经济损失。目前,关于奶牛热应激的研究较多,且多集中于产奶性能和繁殖性能等方面,而有关生理和免疫机能的研究报道较少。作者阐述了奶牛发生热应激时皮质醇激素的变化和调节、三大代谢过程的改变及免疫细胞和相关细胞因子的表达分泌等过程,旨在更加深入地了解热应激对奶牛生理状态及免疫功能的影响,从而为奶牛热应激综合征的防控、诊断和治疗提供理论依据。     

In vitro and in vivo effects of corticosteroids on peripheral blood lymphocytes from ponies.

The in vitro and in vivo effects of corticosteroids on peripheral blood lymphocytes (PBL) from ponies were studied. Prednisolone inhibited lymphocyte stimulation by phytohemagglutin (PHA) in a dose-dependent manner, without inducing lysis even at large doses. The PBL from horses heterozygous for the combined immunodeficiency trait responded to corticosteroid treatment the same as did PBL from normal ponies. Removal of the corticosteroid after incubation with PBL from normal ponies partially restored responsiveness of these cells to PHA. Chronic in vivo treatment of ponies with corticosteroids caused a marked decrease in the absolute numbers of circulating lymphocytes. Most remaining lymphocytes had detectable surface immunoglobulin and C3 receptors, suggesting a greater decrease in the T-lymphocyte population. In spite of this, there was little change in the in vitro PHA- or keyhole limpet hemocyanin-sensitized ponies. In general, the corticosteroid effects of lysis, as well as the mitogenic and antigenic responses of PBL from ponies, were similar to those previously reported for human lymphocytes.     

N-acetyl-beta-D-glucosaminidase activities, milk somatic cell counts, and blood leukocyte and erythrocyte counts in cows after heat-induced stress or after intravenous administration of adrenocorticotropic hormone

Six midlactation, nonpregnant cows were subjected to heat-induced stress or to repeated injections of adrenocorticotropic hormone (ACTH). The heat stress (experiment 1) consisted of subjecting the cows to 7 days of fluctuating temperatures (21 C during the day, 32 C during the night). The ACTH-induced stress (experiment 2) was accomplished by giving each of the cows 100 IU of ACTH twice daily for 4 consecutive days. The cows' milk somatic cell counts (MSCC), milk N-acetyl-beta-D-glucosaminidase (NAGase) activity, quarter milk production (ie, milk production per mammary quarter), blood leukocyte counts, and plasma concentrations of NAGase were monitored in both experiments. Although heat stress did not result in significant differences in NAGase, heat stress did significantly (P less than 0.05) decrease milk production. The ACTH administrations resulted in increased milk NAGase activity, MSCC, and blood leukocyte counts, and in decreased plasma NAGase activity and quarter milk production. Therefore, milk NAGase activity and MSCC were not affected by short-term heat stress, but were increased by ACTH-induced stress. Blood erythrocyte concentrations were not affected by heat stress or by ACTH-induced stress.     

Effects of recombinant bovine granulocyte-macrophage colony-stimulating factor on bovine peripheral blood neutrophil functions in vitro and in vivo

Effects of recombinant bovine granulocyte-macrophage colony-stimulating factor (rboGM-CSF) on bactericidal activity of bovine peripheral blood neutrophils in vitro and in vivo were studied. In in vitro experiment, bovine blood neutrophils were cultured for 9 hr in media containing 0.005, 0.05 or 0.5 microg/ml of rboGM-CSF. Neutrophils treated with rboGM-CSF showed significantly higher luminol-dependent chemiluminescence (LDCL) than control cells. In in vivo experiment, neutrophils isolated from cows injected 5.0 microg/kg of rboGM-CSF showed significantly higher Nitrobluetetrazolium (NBT) reduction value than that from control cows 24 hr post injection. Total leukocyte counts of cows injected rboGM-CSF sharply decreased 6 hr post injection and recovered to normal level 2 days post injection. Body temperature of these cows rose 6 hr post injection and back to normal level at 24 hr post injection. It was suggested that rboGM-CSF enhanced bactericidal activity of bovine neutrophils both in vitro and in vivo.     

Elevated temperature (heat stress) in vitro reduces androstenedione and estradiol and increases progesterone secretion by follicular cells from bovine dominant follicles

Dairy cattle are susceptible to heat stress-induced reductions in fertility; however, direct effects of hyperthermia on specific reproductive functions are difficult to determine in vivo. The objective of this experiment was to examine the effect of elevated temperature in vitro on follicular steroidogenesis, to gain insight into specific follicular responses associated with heat stress. Dominant follicles were obtained from Holstein heifers on day 6 post-estrus (luteal phase; n = 4) or day 8, 36 h after an injection with 25 mg PGF(2alpha) to induce regression of the corpus luteum (follicular phase; n = 4). Pieces of follicle wall were isolated from dominant follicles and cultured for 96 h with 0, 2 or 100 ng/ml LH or FSH at 37, 39 or 41 degrees C. Concentrations of androstenedione, estradiol and progesterone were determined in culture media collected every 24h. During the last 48 h of culture, basal secretion of androstenedione and estradiol by pieces of follicle wall was lower at 41 degrees C than at 37 or 39 degrees C (P < 0.05). In contrast, cumulative secretion of progesterone by pieces of follicle wall in medium alone was higher at 41 degrees C than at 37 or 39 degrees C (P < 0.05). Pieces of follicle wall responded to treatment with both low (2 ng/ml) and high (100 ng/ml) doses of gonadotropins at all temperatures. However, gonadotropin-induced secretion of androstenedione and estradiol was generally lower, whereas gonadotropin-induced secretion of progesterone was higher at 41 degrees C and sometimes at 39 than at 37 degrees C. The changes in basal steroidogenesis and in responses to gonadotropins suggest that follicular cells begin to luteinize at elevated temperatures in vitro. Premature luteinization of follicular cells in vivo has been associated with reduced fertility in cattle with persistent follicles, suggesting that the premature differentiation of follicular cells observed in the current study may be responsible, in part, for the reduced fertility of dairy cattle under heat-stressed conditions.     

HSP90 gene expression induced by aspirin is associated with damage remission in a chicken myocardial cell culture exposed to heat stress

1. To understand the potential protection of heat shock protein 90 (HSP90) induced by aspirin against heat stress damage in chicken myocardial cells, enzyme activities related to stress damage, cytopathological changes, the expression and distribution of HSP90, and HSP90 mRNA levels in the myocardial cells exposed to heat stress (42°C) for different durations with or without aspirin administration (1 mg/ml, 2 h prior) in vitro were investigated.

2. Significant increase of enzyme levels in the supernatant of heat-stressed myocardial cells and cellular lesions characterised by acute degeneration, karyopyknosis and karyorrhexis were observed, compared to non-treated cells. However, the lesions of cells treated with aspirin were milder, characterised by earlier recovery of enzyme levels to the control levels and no obvious heat stress-related cellular necrosis.

3. Stronger positive signals in the cytoplasm and longer retention of HSP90 signal in nuclei were observed in aspirin-treated myocardial cells than those of only heat-stressed cells. HSP90 level in the aspirin-treated myocardial cells was 11.1-fold higher than that in non-treated cells, and remained at a high level at the early stage of heat stress, whereas it was just 4.1-fold higher in only heat-stressed cells and returned rapidly to a low level.

4. Overexpression of HSP90 mRNA in aspirin-treated cells was observed throughout the experiment, whereas HSP90 mRNA decreased significantly only in heat-stressed cells.

5. The early higher HSP90 expression induced by aspirin during heat stress was accompanied by decreased heat stress damage, suggesting that aspirin might play an important role in preventing myocardial cells from heat stress damage in vitro.