{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,19]],"date-time":"2026-09-19T06:56:44Z","timestamp":1789801004503,"version":"4.0.1"},"reference-count":65,"publisher":"MDPI AG","issue":"20","license":[{"start":{"date-parts":[[2023,10,13]],"date-time":"2023-10-13T00:00:00Z","timestamp":1697155200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["32002355"],"award-info":[{"award-number":["32002355"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["BK20200933"],"award-info":[{"award-number":["BK20200933"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["JSSCTD202224"],"award-info":[{"award-number":["JSSCTD202224"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Jiangsu Provincial Natural Science Foundation of China","award":["32002355"],"award-info":[{"award-number":["32002355"]}]},{"name":"Jiangsu Provincial Natural Science Foundation of China","award":["BK20200933"],"award-info":[{"award-number":["BK20200933"]}]},{"name":"Jiangsu Provincial Natural Science Foundation of China","award":["JSSCTD202224"],"award-info":[{"award-number":["JSSCTD202224"]}]},{"name":"Jiangsu Shuangchuang Innovation Group Project","award":["32002355"],"award-info":[{"award-number":["32002355"]}]},{"name":"Jiangsu Shuangchuang Innovation Group Project","award":["BK20200933"],"award-info":[{"award-number":["BK20200933"]}]},{"name":"Jiangsu Shuangchuang Innovation Group Project","award":["JSSCTD202224"],"award-info":[{"award-number":["JSSCTD202224"]}]},{"name":"A Project Funded by the Priority Academic Program Development of Jiangsu Higher Education Institutions (PAPD)","award":["32002355"],"award-info":[{"award-number":["32002355"]}]},{"name":"A Project Funded by the Priority Academic Program Development of Jiangsu Higher Education Institutions (PAPD)","award":["BK20200933"],"award-info":[{"award-number":["BK20200933"]}]},{"name":"A Project Funded by the Priority Academic Program Development of Jiangsu Higher Education Institutions (PAPD)","award":["JSSCTD202224"],"award-info":[{"award-number":["JSSCTD202224"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Animals"],"abstract":"<jats:p>Semen delivery practice is crucial to the efficiency of artificial insemination using high-quality boar sperm. The present study aimed to evaluate the effect of a common semen delivery method, a Styrofoam box, under elevated temperatures on boar sperm quality and functionality and to investigate the underlying molecular responses of sperm to the temperature rise. Three pooled semen samples from 10 Duroc boars (3 ejaculates per boar) were used in this study. Each pooled semen sample was divided into two aliquots. One aliquot was stored at a constant 17 \u00b0C as the control group. Another one was packaged in a well-sealed Styrofoam box and placed in an incubator at 37 \u00b0C for 24 h to simulate semen delivery on hot summer days and subsequently transferred to a refrigerator at 17 \u00b0C for 3 days. The semen temperature was continuously monitored. The semen temperature was 17 \u00b0C at 0 h of storage and reached 20 \u00b0C at 5 h, 30 \u00b0C at 14 h, and 37 \u00b0C at 24 h. For each time point, sperm quality and functionality, apoptotic changes, expression levels of phosphorylated AMPK, and heat shock proteins HSP70 and HSP90 were determined by CASA, flow cytometry, and Western blotting. The results showed that elevated temperature during delivery significantly deteriorated boar sperm quality and functionality after 14 h of delivery. Storage back to 17 \u00b0C did not recover sperm motility. An increased temperature during delivery apparently promoted the conversion of sperm early apoptosis to late apoptosis, showing a significant increase in the expression levels of Bax and Caspase 3. The levels of phosphorylated AMPK were greatly induced by the temperature rise to 20 \u00b0C during delivery but reduced thereafter. With the temperature elevation, expression levels of HSP70 and HSP90 were notably increased. Our results indicate that a temperature increase during semen delivery greatly damages sperm quality and functionality by promoting sperm apoptosis. HSP70 and HSP90 could participate in boar sperm resistance to temperature changes by being associated with AMPK activation and anti-apoptotic processes.<\/jats:p>","DOI":"10.3390\/ani13203203","type":"journal-article","created":{"date-parts":[[2023,10,13]],"date-time":"2023-10-13T10:04:39Z","timestamp":1697191479000},"page":"3203","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":20,"title":["Temperature Elevation during Semen Delivery Deteriorates Boar Sperm Quality by Promoting Apoptosis"],"prefix":"10.3390","volume":"13","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-5189-0299","authenticated-orcid":false,"given":"Junwei","family":"Li","sequence":"first","affiliation":[{"name":"College of Veterinary Medicine, Yangzhou University, Yangzhou 225009, China"},{"name":"Jiangsu Co-Innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou University, Yangzhou 225009, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wenming","family":"Zhao","sequence":"additional","affiliation":[{"name":"College of Veterinary Medicine, Yangzhou University, Yangzhou 225009, China"},{"name":"Jiangsu Co-Innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou University, Yangzhou 225009, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jiaqiao","family":"Zhu","sequence":"additional","affiliation":[{"name":"College of Veterinary Medicine, Yangzhou University, Yangzhou 225009, China"},{"name":"Jiangsu Co-Innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou University, Yangzhou 225009, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shuaibiao","family":"Wang","sequence":"additional","affiliation":[{"name":"DanAg Agritech Consulting (Zhengzhou) Co., Ltd., Zhengzhou 450000, China"},{"name":"Royal Veterinary College, London NW1 0TU, UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Huiming","family":"Ju","sequence":"additional","affiliation":[{"name":"College of Veterinary Medicine, Yangzhou University, Yangzhou 225009, China"},{"name":"Jiangsu Co-Innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou University, Yangzhou 225009, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shufang","family":"Chen","sequence":"additional","affiliation":[{"name":"Ningbo Academy of Agricultural Science, Ningbo 315040, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4752-6654","authenticated-orcid":false,"given":"Athina","family":"Basioura","sequence":"additional","affiliation":[{"name":"Department of Agriculture, School of Agricultural Sciences, University of Western Macedonia, 53100 Florina, Greece"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0622-6513","authenticated-orcid":false,"given":"Gra\u00e7a","family":"Ferreira-Dias","sequence":"additional","affiliation":[{"name":"CIISA\u2014Centre for Interdisciplinary Research in Animal Health, Faculty of Veterinary Medicine, University of Lisbon, 1300-477 Lisbon, Portugal"},{"name":"Associate Laboratory for Animal and Veterinary Sciences (AL4AnimalS), 1300-477 Lisbon, Portugal"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9071-8363","authenticated-orcid":false,"given":"Zongping","family":"Liu","sequence":"additional","affiliation":[{"name":"College of Veterinary Medicine, Yangzhou University, Yangzhou 225009, China"},{"name":"Jiangsu Co-Innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou University, Yangzhou 225009, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2023,10,13]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"15","DOI":"10.1186\/s40813-017-0062-5","article-title":"Boar management and semen handling factors affect the quality of boar extended semen","volume":"3","author":"Arsenakis","year":"2017","journal-title":"Porc. Health Manag."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"10367","DOI":"10.3168\/jds.2017-13676","article-title":"A 100-Year Review: Stress physiology including heat stress","volume":"100","author":"Collier","year":"2017","journal-title":"J. Dairy Sci."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"1363","DOI":"10.1111\/rda.14214","article-title":"Effect of season, genetic line and temperature during transport on sperm motility of commercial insemination doses of pooled boar semen: A retrospective study","volume":"57","author":"Porflidt","year":"2022","journal-title":"Reprod. Domest. Anim."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"54","DOI":"10.1016\/j.theriogenology.2021.08.033","article-title":"Factors influencing the response of spermatozoa to agitation stress: Implications for transport of extended boar semen","volume":"175","author":"Paschoal","year":"2021","journal-title":"Theriogenology"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"1715","DOI":"10.1111\/jeb.13710","article-title":"Combined effects of rearing and testing temperatures on sperm traits","volume":"33","author":"Harrison","year":"2020","journal-title":"J. Evol. Biol."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"236","DOI":"10.1016\/j.jtherbio.2019.07.008","article-title":"Mitochondrial OXPHOS is involved in the protective effects of L-arginine against heat-induced low sperm motility of boar","volume":"84","author":"Li","year":"2019","journal-title":"J. Therm. Biol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"97","DOI":"10.1007\/s11250-020-02516-y","article-title":"Susceptibility of boar spermatozoa to heat stress using in vivo and in vitro experimental models","volume":"53","author":"Stone","year":"2021","journal-title":"Trop. Anim. Health Prod."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"2623","DOI":"10.1007\/s00018-012-1165-4","article-title":"Heat stress response of male germ cells","volume":"70","author":"Kim","year":"2013","journal-title":"Cell Mol. Life Sci."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1093\/molehr\/gaz063","article-title":"Stress response pathways in the male germ cells and gametes","volume":"26","author":"Santiago","year":"2020","journal-title":"Mol. Hum. Reprod."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"727","DOI":"10.1016\/j.bbagen.2016.01.017","article-title":"Resveratrol and the mitochondria: From triggering the intrinsic apoptotic pathway to inducing mitochondrial biogenesis, a mechanistic view","volume":"1860","author":"Nabavi","year":"2016","journal-title":"Biochim. Biophys. Acta"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"505","DOI":"10.3892\/mmr.2013.1506","article-title":"Berberine induces selective apoptosis through the AMPK-mediated mitochondrial\/caspase pathway in hepatocellular carcinoma","volume":"8","author":"Yang","year":"2013","journal-title":"Mol. Med. Rep."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"2143","DOI":"10.1016\/j.bbamem.2013.05.026","article-title":"AMP-activated kinase, AMPK, is involved in the maintenance of plasma membrane organization in boar spermatozoa","volume":"1828","author":"Gil","year":"2013","journal-title":"Biochim. Biophys. Acta"},{"key":"ref_13","doi-asserted-by":"crossref","unstructured":"Martin-Hidalgo, D., Hurtado de Llera, A., Calle-Guisado, V., Gonzalez-Fernandez, L., Garcia-Marin, L., and Bragado, M.J. (2018). AMPK Function in Mammalian Spermatozoa. Int. J. Mol. Sci., 19.","DOI":"10.3390\/ijms19113293"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"55","DOI":"10.1016\/j.abb.2016.08.016","article-title":"Inhibition of autophagy enhances heat-induced apoptosis in human non-small cell lung cancer cells through ER stress pathways","volume":"607","author":"Xie","year":"2016","journal-title":"Arch. Biochem. Biophys."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"35","DOI":"10.1007\/s11010-013-1568-z","article-title":"TAK1 activates AMPK-dependent cell death pathway in hydrogen peroxide-treated cardiomyocytes, inhibited by heat shock protein-70","volume":"377","author":"Chen","year":"2013","journal-title":"Mol. Cell. Biochem."},{"key":"ref_16","doi-asserted-by":"crossref","unstructured":"Wang, T., Yu, Q., Chen, J., Deng, B., Qian, L., and Le, Y. (2010). PP2A mediated AMPK inhibition promotes HSP70 expression in heat shock response. PLoS ONE, 5.","DOI":"10.1371\/journal.pone.0013096"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"849","DOI":"10.1042\/bj20021813","article-title":"Heat-shock protein 90 and Cdc37 interact with LKB1 and regulate its stability","volume":"370","author":"Boudeau","year":"2003","journal-title":"Biochem. J."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"123","DOI":"10.5534\/wjmh.2014.32.3.123","article-title":"Understanding the role of heat shock protein isoforms in male fertility, aging and apoptosis","volume":"32","author":"Purandhar","year":"2014","journal-title":"World J. Mens. Health"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"233","DOI":"10.1007\/s11259-017-9689-6","article-title":"Relative abundance of heat shock proteins and clusterin transcripts in spermatozoa collected from boar routinely utilised in an artificial insemination centre: Preliminary results","volume":"41","author":"Zannoni","year":"2017","journal-title":"Vet. Res. Commun."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"275","DOI":"10.1016\/j.canlet.2010.10.014","article-title":"Targeting heat shock proteins in cancer","volume":"332","author":"Jego","year":"2013","journal-title":"Cancer Lett."},{"key":"ref_21","doi-asserted-by":"crossref","unstructured":"Li, J., Zhao, W., Zhu, J., Ju, H., Liang, M., Wang, S., Chen, S., Ferreira-Dias, G., and Liu, Z. (2023). Antioxidants and Oxidants in Boar Spermatozoa and Their Surrounding Environment Are Associated with AMPK Activation during Liquid Storage. Vet. Sci., 10.","DOI":"10.3390\/vetsci10030214"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"2089","DOI":"10.2527\/jas.2005-766","article-title":"Determination of intracellular reactive oxygen species and high mitochondrial membrane potential in Percoll-treated viable boar sperm using fluorescence-activated flow cytometry","volume":"84","author":"Guthrie","year":"2006","journal-title":"J. Anim. Sci."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"677","DOI":"10.1016\/S0093-691X(03)00081-5","article-title":"Assessment of fresh and frozen-thawed boar semen using an Annexin-V assay: A new method of evaluating sperm membrane integrity","volume":"60","author":"Pena","year":"2003","journal-title":"Theriogenology"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"1743","DOI":"10.1242\/bio.017954","article-title":"Energy metabolic state in hypothermically stored boar spermatozoa using a revised protocol for efficient ATP extraction","volume":"5","author":"Nguyen","year":"2016","journal-title":"Biol. Open"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"601","DOI":"10.1002\/mrd.23643","article-title":"Update on artificial insemination: Semen, techniques, and sow fertility","volume":"90","author":"Mellagi","year":"2022","journal-title":"Mol. Reprod. Dev."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"328","DOI":"10.1016\/j.anireprosci.2018.03.035","article-title":"Effect of vibration emissions during shipping of artificial insemination doses on boar semen quality","volume":"192","author":"Schulze","year":"2018","journal-title":"Anim. Reprod. Sci."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"62","DOI":"10.1016\/j.theriogenology.2020.05.014","article-title":"Heat stress and bull fertility","volume":"153","author":"Morrell","year":"2020","journal-title":"Theriogenology"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"375","DOI":"10.1016\/j.theriogenology.2020.10.002","article-title":"Acute mild heat stress alters gene expression in testes and reduces sperm quality in mice","volume":"158","author":"Rizzoto","year":"2020","journal-title":"Theriogenology"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"131","DOI":"10.1016\/j.theriogenology.2023.01.023","article-title":"Effects of heat stress on sperm quality of French Bulldogs","volume":"199","author":"Lyrio","year":"2023","journal-title":"Theriogenology"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"846981","DOI":"10.3389\/fvets.2022.846981","article-title":"The Mechanism of Heat Stress Resistance During Spermatogenesis in Turpan Black Sheep","volume":"9","author":"Song","year":"2022","journal-title":"Front. Vet. Sci."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"48","DOI":"10.1186\/1751-0147-50-48","article-title":"Stress and its influence on reproduction in pigs: A review","volume":"50","author":"Einarsson","year":"2008","journal-title":"Acta Vet. Scand."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"99","DOI":"10.1016\/S0378-4320(99)00035-4","article-title":"Viability, susceptibility to peroxidation and fatty acid composition of boar semen during liquid storage","volume":"58","author":"Cerolini","year":"2000","journal-title":"Anim. Reprod. Sci."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"69","DOI":"10.1016\/j.cryobiol.2012.11.006","article-title":"The protective effect of a 17 \u00b0C holding time on boar sperm plasma membrane fluidity after exposure to 5 \u00b0C","volume":"66","author":"Casas","year":"2013","journal-title":"Cryobiology"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"533","DOI":"10.1530\/jrf.0.0850533","article-title":"Effect of temperature on the fluidity of boar sperm membranes","volume":"85","author":"Canvin","year":"1989","journal-title":"J. Reprod. Fertil."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"1477","DOI":"10.1016\/S0093-691X(00)00290-9","article-title":"Evaluation on sperm quality of freshly ejaculated boar semen during in vitro storage under different temperatures","volume":"53","author":"Zou","year":"2000","journal-title":"Theriogenology"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"953021","DOI":"10.3389\/fvets.2022.953021","article-title":"Temperature limits for storage of extended boar semen from the perspective of the sperm\u2019s energy status","volume":"9","author":"Henning","year":"2022","journal-title":"Front. Vet. Sci."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"155","DOI":"10.1530\/REP-21-0507","article-title":"Cellular responses and microRNA profiling in bovine spermatozoa under heat shock","volume":"164","author":"Rodrigues","year":"2022","journal-title":"Reproduction"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"1039","DOI":"10.1002\/mrd.22871","article-title":"Reactive oxygen species as mediators of sperm capacitation and pathological damage","volume":"84","author":"Aitken","year":"2017","journal-title":"Mol. Reprod. Dev."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"14","DOI":"10.1016\/j.rbmo.2014.09.018","article-title":"Causes, effects and molecular mechanisms of testicular heat stress","volume":"30","author":"Durairajanayagam","year":"2015","journal-title":"Reprod. Biomed. Online"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"1096","DOI":"10.2164\/jandrol.112.016535","article-title":"Reactive oxygen species and sperm function--in sickness and in health","volume":"33","author":"Aitken","year":"2012","journal-title":"J. Androl."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"502","DOI":"10.1093\/molehr\/gav014","article-title":"Oxidative stress and human spermatozoa: Diagnostic and functional significance of aldehydes generated as a result of lipid peroxidation","volume":"21","author":"Moazamian","year":"2015","journal-title":"Mol. Hum. Reprod."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"198","DOI":"10.1016\/j.acthis.2016.09.003","article-title":"Germ cell apoptosis and expression of Bcl-2 and Bax in porcine testis under normal and heat stress conditions","volume":"119","author":"Fan","year":"2017","journal-title":"Acta Histochem."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"969","DOI":"10.1002\/mrd.22841","article-title":"Scrotal insulation and sperm production in the boar","volume":"84","author":"Parrish","year":"2017","journal-title":"Mol. Reprod. Dev."},{"key":"ref_44","doi-asserted-by":"crossref","unstructured":"Capela, L., Leites, I., Rom\u00e3o, R., Lopes-da-Costa, L., and Pereira, R.M.L.N. (2022). Impact of Heat Stress on Bovine Sperm Quality and Competence. Animals, 12.","DOI":"10.3390\/ani12080975"},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"516","DOI":"10.1095\/biolreprod.104.034520","article-title":"Functional role of caspases in heat-induced testicular germ cell apoptosis","volume":"72","author":"Vera","year":"2005","journal-title":"Biol. Reprod."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"843","DOI":"10.1016\/j.bbrc.2003.10.191","article-title":"Suppression of cytochrome c release and apoptosis in testes with heat stress by minocycline","volume":"312","author":"Matsuki","year":"2003","journal-title":"Biochem. Biophys. Res. Commun."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"555","DOI":"10.1042\/CS20110625","article-title":"AMP-activated protein kinase, stress responses and cardiovascular diseases","volume":"122","author":"Wang","year":"2012","journal-title":"Clin. Sci."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"311","DOI":"10.1007\/s12013-011-9153-0","article-title":"Activation of AMP-activated protein kinase contributes to doxorubicin-induced cell death and apoptosis in cultured myocardial H9c2 cells","volume":"60","author":"Chen","year":"2011","journal-title":"Cell Biochem. Biophys."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"762","DOI":"10.1038\/labinvest.2010.36","article-title":"Hydrogen peroxide inhibits mTOR signaling by activation of AMPKalpha leading to apoptosis of neuronal cells","volume":"90","author":"Chen","year":"2010","journal-title":"Lab. Investig."},{"key":"ref_50","doi-asserted-by":"crossref","unstructured":"Hurtado de Llera, A., Martin-Hidalgo, D., Gil, M.C., Garcia-Marin, L.J., and Bragado, M.J. (2012). AMP-activated kinase AMPK is expressed in boar spermatozoa and regulates motility. PLoS ONE, 7.","DOI":"10.1371\/journal.pone.0038840"},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"541","DOI":"10.1631\/jzus.B1400341","article-title":"Effects of heat stress on serum insulin, adipokines, AMP-activated protein kinase, and heat shock signal molecules in dairy cows","volume":"16","author":"Min","year":"2015","journal-title":"J. Zhejiang Univ. Sci. B"},{"key":"ref_52","doi-asserted-by":"crossref","unstructured":"Adesina, P.A., Saeki, I., Yamamoto, Y., and Suzuki, T. (2023). N-butyrate increases heat shock protein 70 through heat shock factor 1 and AMP-activated protein kinase pathways in human intestinal Caco-2 cells. Arch. Biochem. Biophys., 736.","DOI":"10.1016\/j.abb.2023.109525"},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"1889","DOI":"10.1111\/jcmm.12881","article-title":"Heat shock protein 70 and AMP-activated protein kinase contribute to 17-DMAG-dependent protection against heat stroke","volume":"20","author":"Tsai","year":"2016","journal-title":"J. Cell. Mol. Med."},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"1248","DOI":"10.1016\/j.juro.2009.11.009","article-title":"Heat shock protein and heat shock factor expression in sperm: Relation to oligozoospermia and varicocele","volume":"183","author":"Ferlin","year":"2010","journal-title":"J. Urol."},{"key":"ref_55","doi-asserted-by":"crossref","first-page":"630","DOI":"10.1038\/nrm3658","article-title":"Chaperone machines for protein folding, unfolding and disaggregation","volume":"14","author":"Saibil","year":"2013","journal-title":"Nat. Rev. Mol. Cell Biol."},{"key":"ref_56","doi-asserted-by":"crossref","unstructured":"Lymbery, R.A., Evans, J.P., and Kennington, W.J. (2020). Post-ejaculation thermal stress causes changes to the RNA profile of sperm in an external fertilizer. Proc. Biol. Sci., 287.","DOI":"10.1098\/rspb.2020.2147"},{"key":"ref_57","first-page":"261","article-title":"Seasonal variation in heat shock proteins (hsp70 and hsp90) and their association with frozen semen quality and fertility in buffaloes","volume":"42","author":"Parmar","year":"2021","journal-title":"Cryo Lett."},{"key":"ref_58","doi-asserted-by":"crossref","unstructured":"Ikwegbue, P.C., Masamba, P., Oyinloye, B.E., and Kappo, A.P. (2017). Roles of Heat Shock Proteins in Apoptosis, Oxidative Stress, Human Inflammatory Diseases, and Cancer. Pharmaceuticals, 11.","DOI":"10.3390\/ph11010002"},{"key":"ref_59","doi-asserted-by":"crossref","first-page":"534","DOI":"10.1002\/mrd.20367","article-title":"Immunolocalization of heat shock protein 70 (Hsp 70) in boar spermatozoa and its role during fertilization","volume":"72","author":"Spinaci","year":"2005","journal-title":"Mol. Reprod. Dev."},{"key":"ref_60","doi-asserted-by":"crossref","first-page":"828","DOI":"10.1016\/j.theriogenology.2015.10.029","article-title":"Loss of heat shock protein 70 from apical region of buffalo (Bubalus bubalis) sperm head after freezing and thawing","volume":"85","author":"Varghese","year":"2016","journal-title":"Theriogenology"},{"key":"ref_61","doi-asserted-by":"crossref","first-page":"385","DOI":"10.1111\/j.1439-0531.2007.00918.x","article-title":"Comparative immunolocalization of heat shock proteins (Hsp)-60, -70, -90 in boar, stallion, dog and cat spermatozoa","volume":"43","author":"Volpe","year":"2008","journal-title":"Reprod. Domest. Anim."},{"key":"ref_62","doi-asserted-by":"crossref","first-page":"13","DOI":"10.1016\/j.anireprosci.2017.09.009","article-title":"HSP90 maintains boar spermatozoa motility and mitochondrial membrane potential during heat stress","volume":"187","author":"Bragado","year":"2017","journal-title":"Anim. Reprod. Sci."},{"key":"ref_63","doi-asserted-by":"crossref","first-page":"105","DOI":"10.1007\/s12192-008-0068-7","article-title":"Guidelines for the nomenclature of the human heat shock proteins","volume":"14","author":"Kampinga","year":"2009","journal-title":"Cell Stress Chaperones"},{"key":"ref_64","doi-asserted-by":"crossref","first-page":"2592","DOI":"10.4161\/cc.5.22.3448","article-title":"Heat shock proteins 27 and 70: Anti-apoptotic proteins with tumorigenic properties","volume":"5","author":"Garrido","year":"2006","journal-title":"Cell Cycle"},{"key":"ref_65","doi-asserted-by":"crossref","first-page":"106227","DOI":"10.1016\/j.anireprosci.2019.106227","article-title":"Effect of boar semen supplementation with recombinant heat shock proteins during summer","volume":"211","author":"Bragado","year":"2019","journal-title":"Anim. Reprod. 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