Transcriptome Functional Analysis of Mammary Gland of Cows in Heat Stress and Thermoneutral Condition

Heat stress (HS) exerts significant effects on the production of dairy animals through impairing health and biological functions. However, the molecular mechanisms related to the effect of HS on dairy cow milk production are still largely unknown. The present study employed an RNA-sequencing approach to explore the molecular mechanisms associated with a decline in milk production by the functional analysis of differentially expressed genes (DEGs) in mammary glands of cows exposed to HS and non-heat-stressed cows. The results of the current study reveal that HS increases the rectal temperature and respiratory rate. Cows under HS result in decreased bodyweight, dry matter intake (DMI), and milk yield. In the current study, a total of 213 genes in experimental cow mammary glands was identified as being differentially expressed by DEGs analysis. Among identified genes, 89 were upregulated, and 124 were downregulated. Gene Ontology functional analysis found that biological processes, such as immune response, chaperone-dependent refolding of protein, and heat shock protein binding activity, were notably affected by HS. The Kyoto Encyclopedia of Genes and Genomes enrichment analysis found that almost all of the top-affected pathways were related to immune response. Under HS, the expression of heat shock protein 90 kDa beta I (HSP90B1) and heat shock 70 kDa protein 1A was upregulated, while the expression of bovine lymphocyte antigen (BoLA) and histocompatibility complex, class II, DRB3 (BoLA-DRB3) was downregulated. We further explored the effects of HS on lactation-related genes and pathways and found that HS significantly downregulated the casein genes. Furthermore, HS increased the expression of phosphorylation of mammalian target of rapamycin, cytosolic arginine sensor for mTORC1 subunit 2 (CASTOR2), and cytosolic arginine sensor for mTORC1 subunit 1 (CASTOR1), but decreased the phosphorylation of Janus kinase-2, a signal transducer and activator of transcription factor-5. Based on the findings of DMI, milk yield, casein gene expression, and the genes and pathways identified by functional annotation analysis, it is concluded that HS adversely affects the immune function of dairy cows. These results will be beneficial to understand the underlying mechanism of reduced milk yield in HS cows.

Download Full-text

Genomewide analysis of circular RNA in pituitaries of normal and heat-stressed sows

BMC Genomics ◽

10.1186/s12864-019-6377-7 ◽

2019 ◽

Vol 20 (1) ◽

Author(s):

Haojie Zhang ◽

Baoyu Hu ◽

Jiali Xiong ◽

Ting Chen ◽

Qianyun Xi ◽

...

Keyword(s):

Heat Stress ◽

Heat Shock ◽

Heat Shock Protein ◽

Noncoding Rna ◽

Target Genes ◽

Hormone Secretion ◽

Circular Rna ◽

Differentially Expressed ◽

Pcr Analysis ◽

Pituitary Hormone

Abstract Background As a newly characterized type of noncoding RNA, circular RNA (circRNA) has been shown to have functions in diverse biological processes of animals. 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. In total, 12,035 circRNAs were detected, with 59 circRNAs differentially expressed, including 42 up-regulated and 17 down-regulated circRNAs in pituitaries of the heat-stressed sows. Six randomly selected circRNAs were identified through reverse transcription PCR followed by DNA sequencing and other 7 randomly selected differentially expressed circRNAs were verified by quantitative real-time PCR analysis. The predicted target genes regulated by circRNAs through sponging microRNAs (miRNAs) were enriched in metabolic pathway. 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.

Download Full-text

Proteomic Analysis Identifies Differences in Multiple Myeloma (MM) Cells Sensitive and Resistant to AKT Inhibition.

Blood ◽

10.1182/blood.v106.11.3402.3402 ◽

2005 ◽

Vol 106 (11) ◽

pp. 3402-3402

Author(s):

Alissa Huston ◽

Alexey Leontovich ◽

Michael Timm ◽

Yazan Alsayed ◽

Ujjal Singha ◽

...

Keyword(s):

Heat Shock ◽

Heat Shock Protein ◽

Cell Lines ◽

Proteomic Analysis ◽

Molecular Mechanisms ◽

Treatment Time ◽

Control Sample ◽

Differentially Expressed ◽

Akt Inhibitor ◽

The Mean

Abstract Prior studies have demonstrated that MM cells with PTEN mutation and high AKT activity are more sensitive to inhibitors of the PI3K/Akt/mTOR pathway. However, the molecular mechanisms that regulate the differential response to these agents are not well characterized. The objective of this study was to determine proteins that are different between MM cell lines with higher AKT activity (OPM2) and lower AKT activity (MM.1S) in response to the AKT inhibitor, perifosine. Methods: MM cell lines (MM.1S and OPM2) were treated with serial concentration of perifosine (KRX-0401, Keryx, NY, NY, provided by the NCI). Proteomic analysis using the nanoscale BD Clontech antibody-based protein microarray technique was performed using cells treated with perifosine (10uM for 16 hrs) or vehicle (sterile water) as control. Apoptosis was determined using Annexin V/PI FACS analysis at 24 and 48 hrs. The treatment time and concentration were chosen so that it did not induce more than 25% apoptosis to ensure adequate analysis of changes in signaling pathways. The antibody microarray is a technique that detects differences in protein abundance between the treated and control sample with each experiment by hybridizing fluorescently labeled (Cy3 and Cy5) protein mixtures onto slides spotted with 512 human monoclonal antibodies. Two microarray slides were used for each experiment. The slides were scanned using the Axon GenePix 4000B scanner. Two ratios were generated from the spot images for each protein target. The mean of the ratios of Cy5/Cy3 of both slides were analyzed using Clontech software and used to calculate an Internally Normalized Ratio (INR = (Ratio1/Ratio2, ratios 1 and 2 correspond to slides 1 and 2) for each spot on the array. The INR values were input into GeneSpring 6.0 software (Silicon Genetics, CA). The data was normalized to the mean INR of the two cell lines. Proteins whose expression level changed relative to control greater than 1.3 fold were determined. Unsupervised clustering demonstrated a different protein signature between MM.1S and OPM2 in response to perifosine. There were 144 proteins differentially expressed by 1.3 fold between MM.1S and OPM2. Proteins that were downregulated in OPM2 as compared to MM.1S included those in the PI3K pathway and cell cycle regulation such as PTEN, p70S6Kinase, the AKT substrate GSK-3, eEF-2 kinase, eIF-4g, Ku-80, cyclin A, E2F-2, CDK2, CDK7, and c-myc; proteins involved in apoptosis such as p21WAF, caspase 4 and 8, FADD, and PARP; kinases such as PKAc, PKA RI, ERK2, and JNK1; and other proteins regulating apoptosis and proliferation including p53, the NF-kB inhibitor IkB, and the heat shock protein HSP70. The nanoscale protein array is a useful and rapid technique that may be used to identify differences between resistant and sensitive cells to novel therapeutic agents. We identified proteins that are differentially expressed between MM cells sensitive and relatively resistant to the AKT inhibitor perifosine. Further analysis of the role of these proteins in the mechanism of resistance/sensitivity to perifosine is being performed. Future use of inhibitors of NF-kB (bortezomib) or heat shock protein inhibitors in conjunction with perifosine may overcome resistance induced by these proteins in MM cells with low AKT activity. Supported in part by an ASH scholar award and an MMRF grant.

Download Full-text

Integrated analysis of co-expression, conserved genes and gene families reveal core regulatory network of heat stress response in Cleistogenes songorica, a xerophyte perennial desert plant

BMC Genomics ◽

10.1186/s12864-020-07122-8 ◽

2020 ◽

Vol 21 (1) ◽

Author(s):

Qi Yan ◽

Xifang Zong ◽

Fan Wu ◽

Jie Li ◽

Tiantian Ma ◽

...

Keyword(s):

Heat Stress ◽

Comparative Analysis ◽

Heat Shock ◽

Heat Shock Protein ◽

Expression Analysis ◽

Regulatory Network ◽

Transcriptome Profiling ◽

Gene Families ◽

Differentially Expressed ◽

Conserved Genes

Abstract Background As global warming continues, heat stress (HS) is becoming an increasingly significant factor limiting plant growth and reproduction, especially for cool-season grass species. The objective of this study was to determine the transcriptional regulatory network of Cleistogenes songorica under HS via transcriptome profiling, identify of gene families and comparative analysis across major Poaceae species. Results Physiological analysis revealed significantly decreased leaf relative water content (RWC) but increased proline (Pro) content in C. songorica under 24 h of HS. Transcriptome profiling indicated that 16,028 and 14,645 genes were differentially expressed in the shoots and roots of C. songorica under HS, respectively. Two subgenomes of C. songorica provide equal contribution under HS on the basis of the distribution and expression of differentially expressed genes (DEGs). Furthermore, 216 DEGs were identified as key evolutionarily conserved genes involved in the response to HS in C. songorica via comparative analysis with genes of four Poaceae species; these genes were involved in the ‘response to heat’ and ‘heat acclimation’. Notably, most of the conserved DEGs belonged to the heat-shock protein (HSP) superfamily. Similar results were also obtained from co-expression analysis. Interestingly, hub-genes of co-expression analysis were found to overlap with conserved genes, especially heat-shock protein (HSP). In C. songorica, 84 HSP and 32 heat-shock transcription factor (HSF) genes were identified in the allotetraploid C. songorica genome, and might have undergone purifying selection during evolutionary history based on syntenic and phylogenetic analysis. By analysing the expression patterns of the CsHSPs and CsHSFs, we found that the transcript abundance of 72.7% of the CsHSP genes and of 62.5% of the CsHSF genes changed under heat stress in both the shoots and roots. Finally, a core regulatory network of HS was constructed on the basis of the CsHSP, CsHSF and other responsive genes in C. songorica. Conclusions Regulatory network and key genes were comprehensively analysed and identified in C. songorica under HS. This study improves our knowledge of thermotolerance mechanisms in native grasses, and also provides candidate genes for potential applications in the genetic improvement of grasses.

Download Full-text

Expression Profiling of Heat Shock Protein Genes as Putative Early Heat-Responsive Members in Lettuce

Horticulturae ◽

10.3390/horticulturae7090312 ◽

2021 ◽

Vol 7 (9) ◽

pp. 312

Author(s):

Yeeun Kang ◽

Suk-Woo Jang ◽

Hee Ju Lee ◽

Derek W. Barchenger ◽

Seonghoe Jang

Keyword(s):

Heat Treatment ◽

Heat Stress ◽

Heat Shock ◽

Heat Shock Protein ◽

Differentially Expressed Genes ◽

Expression Patterns ◽

Differentially Expressed ◽

Cool Season ◽

Potential Link ◽

Hsp Genes

High temperatures due to global warming can cause harmful effects on the productivity of lettuce, a cool-season crop. To identify lettuce heat shock protein (HSP) genes that could be involved in early responses to heat stress in plants, we compared RNA transcriptomes between lettuce plants with and without heat treatment of 37 °C for 1 h. Using transcriptome sequencing analyses, a total of 7986 differentially expressed genes (DEGs) were identified including the top five, LsHSP70A, LsHSP70B, LsHSP17.3A, LsHSP17.9A and LsHSP17.9B, which were the most highly differentially expressed genes. In order to investigate the temporal expression patterns of 24 lettuce HSP genes with a fold-change greater than 100 under heat stress, the expression levels of the genes were measured by qRT-PCR at 0, 1, 4, 8, 14, and 24 h time points after heat treatment. The 24 LsHSP genes were classified into three groups based on the phylogenetic analysis and/or major domains available in each protein, and we provided a potential link between the phylogenetic relationships and expression patterns of the LsHSP genes. Our results showed putative early heat-responsive lettuce HSP genes that could be possible candidates as breeding guides for the development of heat-tolerant lettuce cultivars.

Download Full-text

INCREASING THE ACTIVITY OF ANTIOXIDANT SYSTEM IN BROILER CHICKS DURING SUMMER SEASON: 1-EFFECT OF SOME ANTI-HEAT STRESS AGENTS ON BROILER PERFORMANCE, IMMUNE RESPONSE AND HEAT SHOCK PROTEIN 70

Egyptian Poultry Science Journal ◽

10.21608/epsj.2016.33380 ◽

2016 ◽

Vol 36 (1) ◽

pp. 355-374

Author(s):

S. Youssef ◽

Nessrin Selim ◽

M. Galal

Keyword(s):

Immune Response ◽

Heat Stress ◽

Heat Shock ◽

Heat Shock Protein ◽

Antioxidant System ◽

Heat Shock Protein 70 ◽

Summer Season ◽

Broiler Chicks ◽

Broiler Performance

Download Full-text

The effect of heat stress on bull sperm quality and related HSPs expression

Animal Biology ◽

10.1163/15707563-00002507 ◽

2016 ◽

Vol 66 (3-4) ◽

pp. 321-333 ◽

Cited By ~ 4

Author(s):

Yunyun Cheng ◽

Songcai Liu ◽

Ying Zhang ◽

Dan Su ◽

Gang Wang ◽

...

Keyword(s):

Heat Stress ◽

Heat Shock ◽

Heat Shock Proteins ◽

Heat Shock Protein ◽

Protein Expression ◽

Molecular Mechanisms ◽

Sperm Quality ◽

Heat Shock Protein Expression ◽

Shock Proteins

Heat stress dramatically decreases bull sperm quality and has recently received more attention due to the warmer global climate and more intensive production. However, no data exist regarding sperm quality or the related molecular mechanisms under heat stress. Recent studies showed that inducible heat shock proteins (HSPs) play an important role in the dairy heat stress regulation. In this article, to investigate the impacts of heat stress on sperm quality and the associated molecular mechanisms, sperm quality and enzyme activities concerning acrosome reaction were assessed in Simmental, Limousin and Yanbian bulls under heat stress. Subsequently, changes in heat shock protein expression profiles of Simmental bulls were observed, because we observed that sperm quality of these bulls was most sensitive to heat stress. Finally, the relationship between sperm quality and heat shock protein expression under heat stress was analyzed. The results show that summer heat stress decreased the sperm quality of the three bull breeds significantly. Moreover, different levels of heat stimulation induced various enzyme activity changes, among which the activity change in acrosomal enzyme was the most remarkable. Furthermore, the expression of heat shock proteins in the sperm was influenced by the imposed heat stress, among which the expression levels of HSP60 and HSP70 were increased while HSP90 decreased. In summary, our data show that heat stress seriously affects sperm quality and that HSP90 was most sensitive, although it should be noted that seasonal effects may confound these results. This change in heat shock protein expression may be the major factor that affected the sperm quality of the bulls. The findings may provide a new hypothesis for how heat stress impacts reproduction mechanistically.

Download Full-text

Differential Morpho-Physiological and Transcriptomic Responses to Heat Stress in Two Blueberry Species

International Journal of Molecular Sciences ◽

10.3390/ijms22052481 ◽

2021 ◽

Vol 22 (5) ◽

pp. 2481

Author(s):

Jodi Callwood ◽

Kalpalatha Melmaiee ◽

Krishnanand P. Kulkarni ◽

Amaranatha R. Vennapusa ◽

Diarra Aicha ◽

...

Keyword(s):

Heat Stress ◽

Molecular Mechanisms ◽

Stomatal Closure ◽

Enrichment Analysis ◽

Leaf Size ◽

Vaccinium Corymbosum ◽

Climatic Conditions ◽

Differentially Expressed ◽

Transcriptomic Changes ◽

Go Terms

Blueberries (Vaccinium spp.) are highly vulnerable to changing climatic conditions, especially increasing temperatures. To gain insight into mechanisms underpinning the response to heat stress, two blueberry species were subjected to heat stress for 6 and 9 h at 45 °C, and leaf samples were used to study the morpho-physiological and transcriptomic changes. As compared with Vaccinium corymbosum, Vaccinium darrowii exhibited thermal stress adaptation features such as small leaf size, parallel leaf orientation, waxy leaf coating, increased stomatal surface area, and stomatal closure. RNAseq analysis yielded ~135 million reads and identified 8305 differentially expressed genes (DEGs) during heat stress against the control samples. In V. corymbosum, 2861 and 4565 genes were differentially expressed at 6 and 9 h of heat stress, whereas in V. darrowii, 2516 and 3072 DEGs were differentially expressed at 6 and 9 h, respectively. Among the pathways, the protein processing in the endoplasmic reticulum (ER) was the highly enriched pathway in both the species: however, certain metabolic, fatty acid, photosynthesis-related, peroxisomal, and circadian rhythm pathways were enriched differently among the species. KEGG enrichment analysis of the DEGs revealed important biosynthesis and metabolic pathways crucial in response to heat stress. The GO terms enriched in both the species under heat stress were similar, but more DEGs were enriched for GO terms in V. darrowii than the V. corymbosum. Together, these results elucidate the differential response of morpho-physiological and molecular mechanisms used by both the blueberry species under heat stress, and help in understanding the complex mechanisms involved in heat stress tolerance.

Download Full-text

Integrated Insight into the Molecular Mechanisms of Spontaneous Abortion during Early Pregnancy in Pigs

International Journal of Molecular Sciences ◽

10.3390/ijms22126644 ◽

2021 ◽

Vol 22 (12) ◽

pp. 6644

Author(s):

Xupeng Zang ◽

Ting Gu ◽

Wenjing Wang ◽

Chen Zhou ◽

Yue Ding ◽

...

Keyword(s):

Immune Response ◽

Spontaneous Abortion ◽

Molecular Mechanisms ◽

Gene Set Enrichment Analysis ◽

Differentially Expressed ◽

Gene Set ◽

Cerna Network ◽

Major Interest ◽

End Stage ◽

Insight Into

Due to the high rate of spontaneous abortion (SAB) in porcine pregnancy, there is a major interest and concern on commercial pig farming worldwide. Whereas the perturbed immune response at the maternal–fetal interface is an important mechanism associated with the spontaneous embryo loss in the early stages of implantation in porcine, data on the specific regulatory mechanism of the SAB at the end stage of the implantation remains scant. Therefore, we used high-throughput sequencing and bioinformatics tools to analyze the healthy and arresting endometrium on day 28 of pregnancy. We identified 639 differentially expressed lncRNAs (DELs) and 2357 differentially expressed genes (DEGs) at the end stage of implantation, and qRT-PCR was used to verify the sequencing data. Gene set variation analysis (GSVA), gene set enrichment analysis (GSEA), and immunohistochemistry analysis demonstrated weaker immune response activities in the arresting endometrium compared to the healthy one. Using the lasso regression analysis, we screened the DELs and constructed an immunological competitive endogenous RNA (ceRNA) network related to SAB, including 4 lncRNAs, 11 miRNAs, and 13 genes. In addition, Blast analysis showed the applicability of the constructed ceRNA network in different species, and subsequently determined HOXA-AS2 in pigs. Our study, for the first time, demonstrated that the SAB events at the end stages of implantation is associated with the regulation of immunobiological processes, and a specific molecular regulatory network was obtained. These novel findings may provide new insight into the possibility of increasing the litter size of sows, making pig breeding better and thus improving the efficiency of animal husbandry production.

Download Full-text