Cold Stress Group Research Articles

Gene Expression Profile and Co-Expression Network of Pearl Gentian Grouper under Cold Stress by Integrating Illumina and PacBio Sequences.

Simple SummaryIn this study, we investigated the liver transcriptomic responses of pearl gentian grouper towards cold stress. Some cold-related key genes and biological pathways were screened, of which energy-related metabolic pathways and genes had higher expression levels under cold stress. This suggested that energy homeostasis plays a crucial role in the physiological adjustments of pearl gentian grouper when exposed to the cold stress environment. Our results will expedite the understanding of different fishes adaptive mechanisms to profound environmental temperature changes and provide insights into the molecular breeding of cold-tolerant pearl gentian grouper varieties.Pearl gentian grouper (Epinephelus fuscoguttatus ♀ × Epinephelus lanceolatus ♂) is a fish of high commercial value in the aquaculture industry in Asia. However, this hybrid fish is not cold-tolerant, and its molecular regulation mechanism underlying cold stress remains largely elusive. This study thus investigated the liver transcriptomic responses of pearl gentian grouper by comparing the gene expression of cold stress groups (20, 15, 12, and 12 °C for 6 h) with that of control group (25 °C) using PacBio SMRT-Seq and Illumina RNA-Seq technologies. In SMRT-Seq analysis, a total of 11,033 full-length transcripts were generated and used as reference sequences for further RNA-Seq analysis. In RNA-Seq analysis, 3271 differentially expressed genes (DEGs), two low-temperature specific modules (tan and blue modules), and two significantly expressed gene sets (profiles 0 and 19) were screened by differential expression analysis, weighted gene co-expression networks analysis (WGCNA), and short time-series expression miner (STEM), respectively. The intersection of the above analyses further revealed some key genes, such as PCK, ALDOB, FBP, G6pC, CPT1A, PPARα, SOCS3, PPP1CC, CYP2J, HMGCR, CDKN1B, and GADD45Bc. These genes were significantly enriched in carbohydrate metabolism, lipid metabolism, signal transduction, and endocrine system pathways. All these pathways were linked to biological functions relevant to cold adaptation, such as energy metabolism, stress-induced cell membrane changes, and transduction of stress signals. Taken together, our study explores an overall and complex regulation network of the functional genes in the liver of pearl gentian grouper, which could benefit the species in preventing damage caused by cold stress.

Short-term cold stress can reduce the abundance of antibiotic resistance genes in the cecum and feces in a pig model

Comprehensive studies on the effects of cold stress on antibiotic resistance genes (ARGs) in the intestines and feces remain scarce. In this study, pigs were selected as the animal model and divided into a normal temperature group and a 48-h short-term cold stress group. The ARG profiles in fecal, cecal content and cecal mucosa samples were analyzed. The results showed that the normalized abundance of ARGs in the cecal mucosa samples in the cold stress group was significantly higher than that in the normal temperature group, while the normalized ARG abundances in the fecal and cecal content samples were significantly lower than those in the normal temperature group (P < 0.05). The bacterial community composition (especially Firmicutes) was the major driver impacting the ARG profile and accounted for 32.2% of the variation in the ARG profile, followed by metabolites (especially creatinine and oxypurinol) and mobile genetic elements (MGEs) (especially plasmids and insertion elements). And it was found that creatinine and oxypurinol can reduce the abundance of ARGs and Firmicutes in the in vitro experiment. The results indicate that short-term cold stress can reduce the abundance of ARGs in the cecum and feces of pigs, providing reference data for environmental safety.

The Mediation of miR-34a/miR-449c for Immune Cytokines in Acute Cold/Heat-Stressed Broiler Chicken.

Simple SummaryIn the intensive and scale poultry industry, the level of heat stress (HS) directly affects the growth, development, and production performance of poultry. To alleviate the adverse effects of stress in broilers, microRNA (miRNA) was regarded as a potential regulator of immune cytokines. In this study, through the sequencing analysis of spleens after cold/heat stress, we found that 33 and 37 miRNA were differentially expressed in the heat stress group compared with the normal (NS) group and cold stress (CS) group, respectively. The differential miRNA were mainly involved in biological processes such as the cytokine–cytokine receptor interaction. To further understand the miRNA-mediated effect of heat stress on the immune level of chickens, we selected miR-34a and miR-449c as the research objects, predicted and verified that interleukin 2 (IL-2) and interleukin 12α (IL-12α) were the target genes of miR-34a and miR-449c. Coupled with the analysis of the expression of other cytokines, we found that miRNA could change the expression of immune cytokines directly or indirectly. This discovery provides a new insight into the mediation of miRNA for immune cytokines in acute cold/heat stressed broiler chicken.An increasing amount of evidence has revealed that microRNAs (miRNAs) participated in immune regulation and reaction to acute cold and heat stresses. As a new type of post-transcriptional regulatory factor, miRNA has received widespread attention; However, the specific mechanism used for this regulation still needs to be determined. In this study, thirty broilers at the same growth period were divided into three groups and treated with different temperature and humidity of CS (10–15 °C and 90% Relative Humidity (RH)), HS (39 °C and 90% RH), and NS (26 °C and 50–60% RH) respectively. After 6 h, splenic tissues were collected from all study groups. miRNA sequencing was performed to identify the differentially expressed miRNAs (DEMs) between HS, CS, and NS. We found 33, 37, and 7 DEMs in the HS-NS, HS-CS, CS-NS group. Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis showed that DEMs were significantly enriched in cytokine–cytokine receptor interaction and functioned as the cellular responders to stress. We chose two miRNA, miR-34a and miR-449c, from the same family and differential expressed in HS-CS and HS-NS group, as the research objects to predict and verify the target genes. The dual-luciferase reporter assay and quantitative real-time PCR (qRT-PCR) confirmed that two cytokines, IL-2 and IL-12α, were the direct target genes of miR-34a and miR-449c. To further understand the mediation mechanism of miRNAs in acute cold/heat-stressed broiler chicken, a splenic cytokines profile was constructed. The results showed that IL-1β was strongly related to acute heat stress in broiler chicken, and from this we predicted that the increased expression of IL-1β might promote the expression of miR-34a, inducing the upregulation of interferon-γ (INF-γ) and IL-17. Our finds have laid a theoretical foundation for the breeding of poultry resistance and alleviation of the adverse effects of stress.

Transcriptome analysis reveals molecular mechanisms responsive to acute cold stress in the tropical stenothermal fish tiger barb (Puntius tetrazona)

BackgroundTropical stenothermal fish exhibit special tolerance and response to cold stress. However current knowledge of the molecular mechanisms response to cold stress in aquatic ectotherms is largely drawn from eurythermal or extreme stenothermal species. The tiger barb Puntius tetrazona is a tropical stenothermal fish, with great popularity in aquarium trade and research.ResultsTo investigate the response mechanism of P. tetrazona to low temperature, fish were exposed to increasing levels of acute cold stress. Histopathological analysis showed that the brain, gill, liver and muscle tissues appeared serious damage after cold stress (13 °C). Brain, gill, liver and muscle tissues from control (CTRL) groups (27 °C) and COLD stress groups (13 °C) of eight-month fish (gender-neutral) were sampled and assessed for transcriptomic profiling by high-throughput sequencing. 83.0 Gb of raw data were generated, filtered and assembled for de novo transcriptome assembly. According to the transcriptome reference, we obtained 392,878 transcripts and 238,878 unigenes, of which 89.29% of the latter were annotated. There were 23,743 differently expressed genes (DEGs) been filtered from four pairs of tissues (brain, gill, liver and muscle) between these cold stress and control groups. These DEGs were mainly involved in circadian entrainment, circadian rhythm, biosynthesis of steroid and fatty acid. There were 64 shared DEGs between the four pairs of groups, and five were related to ubiquitylation/deubiquitylation. Our results suggested that ubiquitin-mediated protein degradation might be necessary for tropical stenothermal fish coping with acute cold stress. Also, the significant cold-induced expression of heat shock 70 kDa protein (HSP70) and cold-induced RNA-binding protein (CIRBP) was verified. These results suggested that the expression of the molecular chaperones HSP70 and CIRBP in P. tetrazona might play a critical role in coping with acute cold stress.ConclusionsThis is the first transcriptome analysis of P. tetrazona using RNA-Seq technology. Novel findings about tropical stenothermal fish under cold stress (such as HSP70 and CIRBP genes) are presented here. This study contributes new insights into the molecular mechanisms of tropical stenothermal species response to acute cold stress.

Characterization of Gut Microbiota in Prenatal Cold Stress Offspring Rats by 16S rRNA Sequencing.

Simple SummaryPrenatal stress, including prenatal cold stress has long-term effects on offspring’s physical and mental health. Our previous study showed a reduction of anxiety-like behavior in offspring rats suffered from prenatal cold stress. It is well-known that gut microbiota was involved in a variety of physiological activities, such as emotion, cognition, and behavior. However, information on the comparison between prenatal cold stress and gut microbiota in offspring is limited. The current study compared the gut microbiota composition of the prenatal cold stress and non-stress offspring rats. Cold stressed during gestation period showed to change the offspring gut microbiota composition, and Bacteroides and Lactobacillus were significantly increased in prenatal cold stress offspring rat guts. With the hope, cold stress-induced negative effects of animals can be prevented by microbiological interventions.Our previous study showed a reduction of anxiety-like behavior in offspring rats suffered from prenatal cold stress; whether this was related to changes in the offspring gut microbiota is unclear. To obtain the evidence for the role of the gut microbiota in prenatal cold stress offspring, 16S rRNA sequencing technology was used. Male and female offspring rat feces were collected from a room temperature group and a prenatal cold stress group (n ≥ 8) for microbial DNA extraction, followed by 16S rRNA sequencing. The results indicated that prenatal cold stress could change the offspring’s gut microbiota composition. Prenatal cold stress significantly upregulates Lactobacillus, Lactobacillus_gasseri, Bacteroides, and Bacteroides-acidifaciens in female offspring, whereas prenatal cold stress significantly reduced Lachnospiraceae and Prevotellaceae in male offspring. These data showed the characterization of gut microbiota in prenatal cold stress offspring rats, and these data suggest that microbiological intervention in the future can potentially prevent the negative effects caused by cold stress to animals.

Differential expression of small heat shock proteins in the brain of broiler embryo; the effects of embryonic thermal manipulation.

Broilers are more vulnerable to high temperatures than mammals due to the feather cover, lack of sweat glands, fast growth and intensive breeding in commercial systems. Thermal stresses affect the function of various organs and change the expression profiles of hundreds of genes in the different tissues of broilers. Thermal manipulation (TM) during embryogenesis can increase heat tolerance in growing broilers. Small heat shock proteins (SHSPs) are a group of HSPs which participate in many cellular functions like response to different stressors. However, their role in the thermotolerance has not been fully elucidated. Ninety fertilized eggs were randomly divided into three groups (30 eggs/group; 10 eggs/replicate). Normal control (NC) eggs were incubated at 37.5°C throughout the incubation period whereas heat stress (HS) and cold stress (CS) groups were kept at 41°C and 33°C from 15 to 17th day of incubation for 3h each day, respectively. On day 20, samples from the cerebrums were harvested for histopathology and mRNA expression analyses of HSPB1, HSPB5, HSPB8, and HSPB9. There were no significant differences in survivability, defected embryos, hatchability, and body weight among treatments. TM had no major deleterious effects on the cerebral tissue except for mild degeneration in the HS group. HSPB1, HSPB5, HSPB8, and HSPB9 were expressed in the presence and absence of TM. All SHSP genes tested were downregulated in response to TM except for HSPB9 which was upregulated in the HS group. The highest change in gene expression due to TM observed for HSPB1. This study presents a broader understanding of mechanisms underlying response to TM in broilers. The results suggest that HSPB1, HSPB5, HSPB8, and HSPB9 are involved in thermotolerance in broilers and SHSPs could be involved in the gene expression profiling of TM. It may propose the use of nutritional supplements in the poultry industry to modulate SHSPs.

Exposure to suboptimal ambient temperature during specific gestational periods and adverse outcomes in mice.

Exposure to suboptimal ambient temperature during pregnancy has been reported as a potential teratogen of fetal development. However, limited animal evidence is available regarding the impact of extreme temperatures on maternal pregnancy and the subsequent adverse pregnancy outcomes. Our objective in this study is to investigate the relationship between temperature and maternal stress during pregnancy in mice. This study used the Naval Medical Research Institute (NMRI) mice during the second and third pregnant weeks with the gestational day (GD) (GD 6.5-14.5 and GD 14.5-17.5). Mice were exposed to suboptimal ambient temperature (1 °C, 5 °C, 10 °C, 15 °C, 40 °C, 42 °C, 44 °C, 46 °C, and 48 °C for the experimental group and 23 °C for the control group) 1 h per day, 7 days a weekin each trimester. Measurements of placental development (placental weight [PW] and placental diameter [PD]) and fetal growth (fetal weight [FW] and crown-to-rump length [CRL]) between experimental and control groups were compared using analysis of variance (ANOVA). Data on the occurrence of preterm birth (PTB) and abnormalities were also collected. The results showed that exposure to both cold and heat stress in the second and third weeks of pregnancy caused significant decreases in measurements of placental development (PW and PD) and fetal growth (FW and CRL). For all temperature exposures, 15 °C was identified as the optimal temperature in the development of the embryo. Most PTB occurrences were observed in high-temperature stress groups, with the highest PTB number seen in the exposure group at 48 °C, whereas PTB occurred only at 1 °C among cold stress groups. In the selected exposure experiments, an approximate U-shaped relation was observed between temperature and number of abnormality occurrence. The highest percentage of these anomalies occurred at temperatures of 1 °C and 48 °C, while no abnormalities were observed at 15 °C and in the control group. Our findings strengthened the evidence that exposure to suboptimal ambient temperatures may trigger adverse pregnancy outcomes and worsen embryo and fetal development in mice.

Kortikosteroid ve Antihistaminik Enjeksiyonların Sıçan Mesane Dokusunda Soğuk Kaynaklı Stres Üzerine Etkileri: Deneysel Bir Çalışma

Objective: Cold is a stress-inducing factor that can cause changes in bladder function in various ways. The present study is intended to investigate the effects of corticosteroid and antihistaminic treatment on acutely and chronically induced cold stress.Material and Methods: Forty-two female rats were randomly divided into seven groups as follows: Control group; Acute cold-stress (ACS) group; ACS+ corticosteroid (CORT) group; ACS+CORT + Antihistaminic group; Cronic cold-stres (CCS) group; CSS+CORT group; CSS+CORT+Antihistaminic group. On the 15th day after these treatments, bladders of the rats were harvested for histopathological examinations under general anesthesia and fixed with 10% neutral buffered formaline. Hematoxylin and eosin and toluidine blue stainings were performed.Results: The comparison based on mast cell count yielded the highest value in the CCS group in comparison to the control group. The lowest value was harvested in the CCS + CORT group. The comparison between the CCS groups revealed the highest polymorphonuclear leucocyte (PNL) values in the lamina propria in the CCS group, whereas the CORT and CORT + Antihistaminic treatments were found to have significantly decreased the PNL values in the lamina propria. While the PNL counts in the epithelium were high in the ACS and CCS groups, the results in the ACS and CCS groups that were treated with CORT and/or antihistaminic were revealed to be similar with those in the control group. It was discovered that antihistaminic injection in addition to CORT decreased the lymphocyte counts in epithelium in CCS more efficiently than CORT alone did.Conclusion: The present research revealed that corticosteroid treatment reduced inflammatory cell infiltration and decreased mast cell count. A more evident amelioration was observed particularly in chronic cold stress.

Identification, functional prediction, and key lncRNA verification of cold stress-related lncRNAs in rats liver

Cold stimulation reduces the quality of animal products and increases animal mortality, causing huge losses to the livestock industry in cold regions. Long non-coding RNAs (lncRNAs) take part in many biological processes through transcriptional regulation, intracellular material transport, and chromosome remodeling. Although cold stress-related lncRNAs have been reported in plants, no research is available on the characteristic and functional analysis of lncRNAs after cold stress in rats. Here, we built a cold stress animal model firstly. Six SPF male Wistar rats were randomly divided to the acute cold stress group (4 °C, 12 h) and the normal group (24 °C, 12 h). lncRNA libraries were constructed by high-throughput sequencing (HTS) using rat livers. 2,120 new lncRNAs and 273 differentially expressed (DE) lncRNAs were identified in low temperature environments. The target genes of DElncRNA were predicted by cis and trans, and then functional and pathway analysis were performed to them. GO and KEGG analysis revealed that lncRNA targets were mainly participated in the regulation of nucleic acid binding, cold stimulation reaction, metabolic process, immune system processes, PI3K-Akt signaling pathway and pathways in cancer. Next, a interaction network between lncRNA and its targets was constructed. To further reveal the mechanism of cold stress, DElncRNA and DEmRNA were extracted to reconstruct a co-expression sub-network. We found the key lncRNA MSTRG.80946.2 in sub-network. Functional analysis of key lncRNA targets showed that targets were significantly enriched in fatty acid metabolism, the PI3K-Akt signaling pathway and pathways in cancer under cold stress. qRT-PCR confirmed the sequencing results. Finally, hub lncRNA MSTRG.80946.2 was characterized, and verified its relationship with related mRNAs by antisense oligonucleotide (ASO) interference and qRT-PCR. Results confirmed the accuracy of our analysis. To sum up, our work was the first to perform detailed characterization and functional analysis of cold stress-related lncRNAs in rats liver. lncRNAs played crucial roles in energy metabolism, growth and development, immunity and reproductive performance in cold stressed rats. The MSTRG.80946.2 was verified by network and experiments to be a key functional lncRNA under cold stress, regulating ACP1, TSPY1 and Tsn.

Embryonic Thermal Manipulation Affects the Antioxidant Response to Post-Hatch Thermal Exposure in Broiler Chickens.

Simple SummaryThe broiler chicken is one of the most important livestock species in the world, as it occupies a major role in the modern human diet. Due to uneven artificial selection pressures, the broiler has increased in size over the past few decades at the expense of its ability to withstand oxidative damage, the latter of which is often a byproduct of thermal stress. In order to attenuate the effects of heat stress, thermal manipulation (TM), which involves changes in incubation temperature at certain points of embryonic development, is increasingly being presented as a way in which to improve broiler thermotolerance. Therefore, the objective of this study was to investigate how TM might affect broiler response to post-hatch thermal stress in the context of the genes that help combat oxidative damage, namely the catalase, NADPH oxidase 4 (NOX4), and superoxide dismutase 2 (SOD2) genes. Expression of all three aforementioned genes differed significantly between TM and control chickens after exposure to cold and heat stress. Conclusively, TM may act as a viable mode of preventative treatment for broilers at risk of thermally induced oxidative stress.Thermal stress is a major source of oxidative damage in the broiler chicken (Gallus gallus domesticus) due to the latter’s impaired metabolic function. While heat stress has been extensively studied in broilers, the effects of cold stress on broiler physiologic and oxidative function are still relatively unknown. The present study aimed to understand how thermal manipulation (TM) might affect a broiler’s oxidative response to post-hatch thermal stress in terms of the mRNA expression of the catalase, NADPH oxidase 4 (NOX4), and superoxide dismutase 2 (SOD2) genes. During embryonic days 10 to 18, TM was carried out by raising the temperature to 39 °C at 65% relative humidity for 18 h/day. To induce heat stress, room temperature was raised from 21 to 35 °C during post-hatch days (PD) 28 to 35, while cold stress was induced during PD 32 to 37 by lowering the room temperature from 21 to 16 °C. At the end of the thermal stress periods, a number of chickens were euthanized to extract hepatic and splenic tissue from the heat-stressed group and cardiac, hepatic, muscular, and splenic tissue from the cold-stressed group. Catalase, NOX4, and SOD2 expression in the heart, liver, and spleen were decreased in TM chickens compared to controls after both cold and heat stress. In contrast, the expression levels of these genes in the breast muscles of the TM group were increased or not affected. Moreover, TM chicks possessed an increased body weight (BW) and decreased cloacal temperature (TC) compared to controls on PD 37. In addition, TM led to increased BW and lower TC after both cold and heat stress. Conclusively, our findings suggest that TM has a significant effect on the oxidative function of thermally stressed broilers.

Heat shock protein 70 protects the quail cecum against oxidant stress, inflammatory injury, and microbiota imbalance induced by cold stress

The intent of this study was to investigate the effects of cold stress on oxidative indexes, inflammatory factors, and microbiota in the quail cecum. A total of 192 male quails (15-day-old) were randomly divided into 12 groups (16 in each group) and were exposed to acute (up to 12h) and chronic (up to 20 D) cold stress at 12 ± 1°C. After cold stress treatment, we examined morphological damage, oxidative stress indexes, inflammatory factors, and intestinal microbiota. Results of morphological examination showed that both acute and chronic cold stress can lead to cecal tissue injury. In addition, both acute and chronic cold stress, especially chronic cold stress can influence the activity of oxidative stress mediators. Glutathione (GSH) and glutathione peroxidase (GSH-Px) activities decreased significantly (p < 0.05), while the nitric oxide (NO) content and inducible nitric oxide synthase (iNOS) activity increased significantly (p < 0.05). Moreover, mRNA levels of inflammatory factors cyclooxygenase-2 (COX-2), prostaglandin E synthase (PTGES), and heat shock protein 70 (Hsp70) were higher in both acute and chronic cold stress groups when compared with the control group (p < 0.05). Furthermore, the intestinal microbiota was changed in both the acute and chronic cold stress groups. These results suggested that cold stress caused oxidative stress and inflammatory injury in cecal tissues, influenced cecal microbiota, and increased expression of Hsp70, which may contribute in protecting the cecum against cold stress in quails.

Effects and transcriptional responses in the hepatopancreas of red claw crayfish Cherax quadricarinatus under cold stress.

The red claw crayfish, Cherax quadricarinatus, is an economically important freshwater crustacean that cannot tolerate low temperature, which diminishes survival via unknown mechanisms. Herein, physiological regulation of C. quadricarinatus was investigated following exposure to low temperature stress at 9 ± 2 °C for 4 weeks. Hepatopancreas tissue was tested for nonspecific enzyme activity, histological structure, and transcriptome sequencing analyses. The results showed that the activities of nonspecific enzymes were inhibited following low temperature stress. Ultrastructural observation revealed that the hepatopancreas structure was oxidatively damaged at low temperature, with numerous autophagic vesicles visible. Apoptosis in the hepatopancreas was significantly increased in the cold stress group, indicating diminished function. Transcriptome sequencing identified 2615 differentially expressed genes (DEGs) following low temperature stress, of which 1147 and 1468 were up- and down-regulated, respectively. Functional analysis of DEGs indicated involvement in substance metabolism, antioxidant defences, signal transduction, and immune responses. Therefore, chronic cold stress can suppress metabolism and cause oxidative damage and immune deficiency in crayfish. The findings provide fundamental molecular information for further study of the regulatory mechanisms of cold tolerance in red claw crayfish.

Effects of prenatal maternal cold stress on spontaneous, exploratory and anxious behaviors in offspring

To study the effects of prenatal cold stress on the behavior and mood of offspring in pregnant rats. Six SPF-class Wister pregnant rats were randomly divided into normal temperature control group and cold stress group with 3 rats in each group. The pregnant female rats in the normal temperature control group were kept in the environment of (22 ±2)℃, and the pregnant female rats in the cold stress group were placed in the artificial intelligence climate chamber at(4 ±0.1)℃ for 7 days before the birth, and the young rats were divided into normal temperature after the young rats were born. After the young rats were born, they were divided into normal temperature control group of male rats (MR, 22), normal temperature control group of mother rats (FR, 15), cold stress group of male rats (MC, 15), and cold stress group of female rats (FC, 15) .In the fourth generation of the offspring, the open field experiment and the elevated cross maze test were carried out. In the open field experiment, there was no significant difference in spontaneous activity and exploration behavior between the normal temperature control group and the cold stress group (P＞0.05). In the elevated plus maze experiment, the retention time of the open arms, the number of open arms and the distance of the male and female rats in the cold stress group were significantly higher than those in the normal temperature control group (P＜0.05). Prenatal maternal cold stress has no significant effect on spontaneous activity, exploration behavior and activity level of offspring, but the offspring have obvious abnormal behaviors with reduced anxiety behavior.

Activation of the MAPK signaling pathway induces upregulation of pro-apoptotic proteins in the hippocampi of cold stressed adolescent mice

Stress induces many non-specific responses in the hippocampus, especially during adolescence. Low environmental temperature is known to induce stress, but its influence on the hippocampus, especially in adolescent mice is not clear. We compared apoptotic-related protein levels and MAPK signaling pathway activation in hippocampal neurons of adolescent mice under low temperature conditions (4 °C for 12 h) with western blotting and immunohistochemistry. Western bolt results demonstrated that the levels of phospho-JNK, phospho-p38, and cleaved-caspase 3 significantly increased, while the ratio of Bcl-XL/Bax decreased, in the cold stress group. The results of immunohistochemistry (IHC) and Nissl staining demonstrated that the protein optical density of caspase 3 increased and Nissl bodies decreased in the cold stress group compared with controls. Thus, we conclude that cold exposure initiates activation of the MAPK signaling pathway and subsequently induces the upregulation of pro-apoptotic proteins in the hippocampi of adolescent mice. Overall our study reveals the relationship between cold stress and apoptosis in adolescent mice.

The Study of Cold Stress - Induced Changes in Rat Mesovarium's Mast Cell Size and Structure in Relation with Estrous Cycle

The climate of Mongolia is dry and extreme, with cold seasons during the 5-7 months of year, which is one of the causes of cold-borne disorders occurring in people. In our country, there are cases of women with inflammatory disease of reproductive organ too. Although the researches on ovarian mast cells have been common so far, the researches on mesovarium mast cells, which have good blood and nerve supply, are pretty uncommon. The researches on cold stress induced changes in mesovarium mast cells’ number, size and structure in relation with menstrual cycle hasn’t been done yet in our country. In our study, we have chosen 16 rats in control group and 48 rats in cold stress group, a total of 64 rats. The cold stress group was divided again into 3 groups (16:16:16) and underwent daily cold stress (refrigerator-15°C) from 8:00 am-11:00 am for 7, 14, 21 days in order to generate cold stress model. The number, shape, structural changes, size, perimeter, length and width of the mesovarium mast cell were determined. The determination of estrous cycles was carried out at 8 am on test day by taking a smear from the vagina using Papanicolaou test and the phases of estrous cycle was determined by histological test.

COPD rat model is more susceptible to cold stress and PM2.5 exposure and the underlying mechanism

The purpose of this study is to verify the hypothesis that chronic obstructive pulmonary disease (COPD) model rat is more susceptible to cold stress and fine particulate matter (PM2.5) exposure than the healthy rat, and explore the related mechanism. COPD rat model, established with cigarette smoke and lipopolysaccharide intratracheal instillation, were exposed to cold stress (0 °C) and PM2.5 (0, 3.2, 12.8 mg/ml). After that, the levels of superoxide dismutase, inducible nitric oxide synthase (iNOS), tumor necrosis factor alpha (TNF-α), monocyte chemotactic protein 1 (MCP-1) and angiotensin Ⅱ (Ang-Ⅱ) in lung were measured, as well as the expression levels of lung 8-hydroxy-2-deoxyguanosine (8-OHdG), nuclear factor kappa B (NF-κB), heme-oxygenase-1 (HO-1) and nuclear factor erythroid-2-related factor 2 (Nrf2). There were significant positive relationships between PM2.5 and lung level of iNOS, TNF-α, MCP-1 and Ang-Ⅱ, lung function and pathologic damage in COPD rats. The HO-1, NF-κB and 8-OHdG were found highly expressed in COPD rat lung, particularly at the higher PM2.5 dose of cold stress groups, while Nrf2 was found declined. Thus, COPD rats may be more susceptible to cold stress and PM2.5 exposure. Cold stress may aggravate PM2.5-induced toxic effects in the lung of COPD rats through increasing Ang-Ⅱ/NF-κB signaling pathway and suppressing Nrf2 signaling pathway.

The role of heat shock protein 70 in oxidant stress and inflammatory injury in quail spleen induced by cold stress.

The aim of this study was to investigate the role of heat shock protein 70 (Hsp70) in oxidative stress and inflammatory damage in the spleen of quails which were induced by cold stress. One hundred ninety-two 15-day-old male quails were randomly divided into 12 groups and kept at 12 ± 1°C to examine acute and chronic cold stress. We first detected the changes in activities of antioxidant enzymes in the spleen tissue under acute and chronic cold stress. The activities of glutathione peroxidase (GSH-Px) fluctuated in acute cold stress groups, while they were significantly decreased (p < 0.05) after chronic cold stress. The activities of superoxide dismutase (SOD), inducible nitric oxide synthase (iNOS), and nitric oxide (NO) content were decreased significantly (p < 0.05) in both of the acute and chronic cold stress groups. Malondialdehyde (MDA) content was significantly increased (p < 0.05) under cold stress except the 0.5h group of acute cold stress. Besides, histopathological analysis showed that quail's spleen tissue was inflammatory injured seriously in both the acute and chronic cold stress groups. Additionally, the inflammatory factors (cyclooxygenase-2 (COX-2), prostaglandin E synthase (PTGES), iNOS, nuclear factor-kappa B (NF-κB), and tumor necrosis factor-a (TNF-α)) and Hsp70 mRNA levels were increased in both of the acute and chronic cold stress groups compared with the control groups. These results suggest that oxidative stress and inflammatory injury could be induced by cold stress in spleen tissues of quails. Furthermore, the increased expression of Hsp70 may play a role in protecting the spleen against oxidative stress and inflammatory damage caused by cold stress.

Effects of both cold and heat stress on the liver of the giant spiny frog (Quasipaa spinosa): stress response and histological changes.

Ambient temperature-associated stress can affect normal physiological functions in ectotherms. To assess the effects of cold or heat stress on amphibians, giant spiny frogs (Quasipaa spinosa) were acclimated at 22°C followed by exposure to 5°C or 30°C for 0, 3, 6, 12, 24 and 48 h, respectively. Histological alterations, apoptotic index, generation of mitochondrial reactive oxygen species (ROS), antioxidant activity indices and stress-response gene expression in frog livers were subsequently determined. Results showed that many fat droplets appeared after 12 h of heat stress and the percentage of melanomacrophage centres significantly changed after 48 h at both stress conditions. Furthermore, the mitochondrial ROS levels were elevated in a time-dependent manner up to 6 h and 12 h in the cold and heat stress groups, respectively. The activities of superoxide dismutase, glutathione peroxidase and catalase were successively increased with increasing periods of cold or heat exposure, and their gene expression levels showed similar changes in both stress conditions. Most tested heat shock protein (HSP) genes were sensitive to temperature exposure, and the expression profiles of most apoptosis-related genes was significantly upregulated at 3 and 48 h under cold and heat stress, respectively. Apoptotic index at 48 h under cold stress was significantly higher than that under heat stress. Notably, lipid droplets, HSP30, HSP70 and HSP110 might be suitable bioindicators of heat stress. The results of these alterations at physiological, biochemical and molecular levels might contribute to a better understanding of the stress response of Q. spinosa, and perhaps amphibians more generally, under thermal stress.

Effect of Mahuang Gancao Ganjiang Decoction on Fusion and Fission of Mitochondria and Apoptosis of Lymphocytes in Mice under Cold Stress.

Mahuang Gancao Ganjiang Decoction (MGGD) can effectively alleviate the symptoms of the patients suffering from exogenous cold stress. However, the curative mechanism has not been fully clarified. This study was designed to investigate the effect of MGGD on the apoptosis of lymphocytes induced by cold stress in mice. The model mice were randomly divided into four groups: the normal control group (no handling mice), cold stress group, MGGD + cold stress group, and MGGD group. Lymphocytes of the mice were isolated from the peripheral blood. Electron microscopy analysis revealed cold stress resulted in mitochondrial fragmentation. Accompanied with the change of morphology of mitochondria, ATP production and the activity of respiratory chain complex decreased in these cells. Western blot analysis showed that these cells expressed decreased fusion-related proteins Mitofusin 1 (Mfn1), Mitofusin 2 (Mfn2), and optic atrophy protein-1 (Opa-1) and increased fission-related proteins dynamin-related protein 1 (Drp1) and fission 1 (Fis-1); our results also show that decreased mitochondrial fusion induces cell apoptosis during cold stress. Meanwhile, we found MGGD can inhibit cell apoptosis induced by cold stress through regulating expression level of Mfn1, Mfn2, Drp1, Fis-1, and Opa-1. These findings are very significant for understanding how MGGD regulates cold-stress-induced cell apoptosis.

Transcriptome analysis of the Antarctic psychrotrophic bacterium Psychrobacter sp. G in response to temperature stress

The key functional genes involved in temperature adaption of the Antarctic psychrotrophic bacterium Psychrobacter sp. G. were identified by transcriptomic sequencing. We analyzed the global transcriptional profile of Psychrobacter sp. G under cold stress (0°C) and heat stress (30°C), with the optimal growth temperature 20°C as the control. There were large alterations of the transcriptome profile, including significant upregulation of 11 and 12 transcripts as well as significant downregulation of 47 and 42 transcripts in the cold and heat stress groups, respectively, compared to the control. The expression of various genes encoding enzymes and transcriptional regulators, including PfpI and TetR family transcriptional regulators under heat stress, as well as the expression of DEAD/DEAH box helicase and the IclR family of transcriptional regulators under cold stress, were upregulated significantly. The expression of several genes, most affiliated with TonB-dependent receptor and siderophore receptor, was downregulated significantly under both heat and cold stress. Many of the genes associated with the metabolism of fatty acid and ABC transporters were regulated differentially under different temperature stress. The results of this survey of transcriptome and temperature stress-relevant genes contribute to our understanding of the stress-resistant mechanism in Antarctic bacteria.