- It has been reported that several noncoding RNAs may regulate animals ’ response to heat stress which can be easily induced by hyperthermia in summer. - However, the expression and functions of circRNAs in the pituitary of sows and whether they participate in heat stress adaption are still unclear.. - Results: In this study, we found that high temperature over the thermoneutral zone of sows during the summer increased the serum heat shock protein 70 (HSP70) level, decreased the superoxide dismutase (SOD) vitality and prolactin (PRL) concentration, and induced heat stress in sows. - Then, we explored circRNA in the pituitary of heat- stressed and normal sows using RNA sequencing and bioinformatics analysis. - Furthermore, the predicted circRNA – miRNA – mRNA interactions showed that some circRNAs might sponge miRNAs to regulate pituitary-specific genes and heat shock protein family members, indicating circRNA ’ s roles in pituitary hormone secretion and heat stress response.. - Conclusions: Our results provided a meaningful reference to understand the functions of circRNA in the porcine pituitary and the mechanisms by which circRNA may participate in animals ’ response to heat stress.. - brains [10], and other common tissues [11], the information about circRNAs in the pig pituitary is little known.. - Circular RNA in the pituitary of sheep have been analyzed and indicated to have functions in embryo pituitary development and endocrine regula- tion [16]. - Espe- cially in the south of China, high temperatures last half a year, which easily cause heat stress and negatively influ- ences sow health and performance. - Recent studies have found the expression alteration of ncRNAs in the mam- malian heat stress response and indicated that ncRNAs may be novel regulators during heat stress [18]. - In homeothermic animals, such as rat and mice, heat stress altered miRNAs expression patterns in the rat small intestine [19] and increased the expression of miR-1, miR-21, and miR-24 in the mice heart to generate a car- dioprotective phenotype resistant to I/R injury [20]. - Heat stress may also manipulate miRNA levels by affecting the enzymes that involved in miRNAs’ biogenesis [21].. - Besides, heat stress could alter lncRNAs expression.. - Two lncRNA species including B2 RNA in mice and Alu RNA in humans accumulate at relatively low levels during normal cellular growth, however, their abundance transiently increases by as much as 40-fold under certain conditions of heat stress [22, 23]. - Since circRNAs are also non-coding RNAs and could func- tion as a type of ceRNA capable of sequestering miRNA activity, it is reasonable to speculate that cir- cRNAs may vary their expression and play a role in response to heat stress. - In addition, it has reported that heat stress could affect circRNAs expression and biogenesis in plants [24, 25]. - However, little attention has been focused on circRNAs ’ alteration and func- tion in animals during heat stress. - The results of our study may help to better understand the roles of circRNAs in the regulation of the pituitary function and the response to heat stress.. - Another 6 sows were obtained during the summer months of 2017, with a high average temperature C), and desig- nated the heat stress (HS) group. - The body weight (in the range of 250 ± 10 kg) and parity (between 5 and 7) were balanced in the 2 test groups. - First, differentially expressed circRNAs in heat stress and all pig miRNAs and mRNAs candidates were chose. - The temperature and humidity of the swine pen in the winter and summer months were recorded every day in this study. - The SOD vitality of serum in the HS group was lower comparing with the TN group (Fig. - At the same time, the level of the heat stress–sensitive protein HSP70 increased significantly in the HS group (Fig. - 1d) in the HS group.. - The results above indicated that sows obtained in sum- mer suffered from heat stress.. - Abundance and characteristics of circular RNA in the sow pituitary. - Identification of circular RNA in the sow pituitary. - 1 The influence of heat stress on hormones and the antioxidative index. - a The superoxide dismutase (SOD) vitality of sow serum in the thermoneutral (TN) and heat stress (HS) groups and (b – d) the level of heat shock protein 70 (HSP70), cortisol, and prolactin (PRL) of sow serum in the TN and HS groups. - Thus, we identified 59 dif- ferentially expressed circRNAs in the sow pituitary be- tween the TN and HS groups (Additional file 3: Table S3). - 0.05), except circRNA6541 and cir- cRNA2600, which were highly expressed in the HS. - In the constructed po- tential circRNA–miRNA–mRNA associations, there were 110 miRNAs interacted with 10,598 mRNA transcripts and 51 differentially expressed circRNAs (Additional file 4: Table S4). - In order to learn the potential functions of the differentially expressed cir- cRNAs in the pituitary, we used GO and KEGG. - a The number of circRNA and back-spliced reads identified in the pituitaries of the thermoneutral (TN) and heat stress (HS) groups. - d Distribution of the exon number of circRNA in the 6 pituitaries of the sows. - functional genes as well as the heat stress regulated genes, screened effective information from the previous circRNA-miRNA-mRNA network (Additional file 4:. - Table S7-A) and heat stress (Fig. - pituitary under heat stress, we found that 42 ceRNA pairs (Additional file 8: Table S8-A) may regulate pituitary- specific genes and 154 ceRNA pairs (Additional file 8: Table S8-B) may be involved in heat stress.. - Finally, we looked for the conserved ceRNA pairs related to pituitary spe- cific genes or heat stress genes by checking for the conser- vation of miRNA-target interactions (Additional file 10:. - Whereas there were no conserved ceRNA interactions related to pituitary specific genes and heat stress among pigs and mice.. - During the summer, high temperature exceeds the upper limit of the thermoneutral zone will induce heat stress in the sows. - HS_P1 = heat stress_pituitary 1. - HS_P2 = heat stress_pituitary 2. - HS_P3 = heat stress_pituitary 3. - Therefore, the sows suf- fered from heat stress most of the time during our ex- perimental period. - Also, the THI is an index that can be used for evaluating if the animals are under heat stress, and a THI range of 72 to 89 is considered to be moder- ate heat stress [44]. - In our study, the calculated THI above 72 during summer months could also be an indi- cator of heat stress. - It has been reported that heat stress reduced animal antioxidant activity [45] and altered the. - The SOD vitality was reduced in the HS group, indicating the sows’ impaired antioxidant activity under heat stress. - HSP70 is a univer- sal marker of heat stress, and its expression rapidly in- creased when animals were subjected to heat stress [47, 48]. - 0.05) in the categories of biological process, molecular function, and cellular components. - However, there was a decline in the serum cortisol level in our study (Fig. - It has been reported that cortisol levels increased in the early stage of heat stress and then. - decreased as the heat stress continued [51]. - The different alteration of cortisol concentrations may be depended on whether the heat stress is acute or chronic [52]. - In this study, sows were in chronic heat stress during the summer months. - As cortisol is the hormone that regu- late thermogenesis, the decreased cortisol level in heat stress is benefit for reducing the metabolic heat produc- tion [50].. - But there is no information about circRNAs in the pig pituitary. - So our study was the first to explore the circRNA expression profile of the pig pituitary, and we identified 12,035 cir- cRNAs in the pig pituitary using high-throughput se- quencing. - Therefore, the abun- dance of circRNA in pig pituitary may also indicate that circRNAs have crucial functions in the pig pituitary.. - Studies have reported that cir- cRNAs have functions during heat stress in plants [24].. - Therefore, we analyzed the circRNA expres- sion changes in the sow pituitary between normal and heat stress conditions. - It has been reported that heat stress alters the circRNA size and the exons numbers in Arabidopsis species [24]. - So, we speculated that circRNAs may be the new regulators of the heat stress response.. - There were 32 pituitary-specific genes that had been reported in the Tissue Specific Genes Database (TiSGeD) database [70]. - The results indicated that these circRNAs may play important roles in secre- tion of hormones, including FSHB, GH1, and PRL, which could be a possible regulatory mechanism of the endocrine status alteration during heat stress. - Since many investiga- tions pointed out that the HSP and HSF are the key players in animals’ heat stress response [71]. - So the circRNA- mediated network may contribute to the response and adaptation to heat stress. - Whereas there were no conserved ceRNA interactions related to pituitary specific genes or heat stress among pigs and mice.. - In addition, the metabolic pathways were enriched in the KEGG pathway analysis. - In sum- mary, the results of our analysis suggested that the cir- cRNA may participate in the metabolic regulation of pigs under heat stress.. - Our study is the first to analyze the circRNA expression profile in sow pituitary and identify the differentially expressed circRNAs in pituitary of sows under heat stress. - These results suggested the sow pituitary has abundant circRNAs and the differentially expressed cir- cRNAs may paly functions on hormone synthesis and stress response by regulating the specific pituitary genes and heat stress–related genes. - Therefore, our study pro- vided a reference for investigating the functions of circRNAs in pituitary regulation and heat stress response.. - 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Effects of heat stress on carbohydrate and lipid metabolism in growing pigs
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