Brugaletta Giorgio, Greene Elizabeth, Ramser Alison, Maynard Craig W, Tabler Travis W, Sirri Federico, Anthony Nicholas B, Orlowski Sara, Dridi Sami
Department of Poultry Science, Center of Excellence for Poultry Science, University of Arkansas, Fayetteville, AR, United States.
Department of Agricultural and Food Sciences, Alma Mater Studiorum - University of Bologna, Bologna, Italy.
Front Vet Sci. 2022 May 25;9:905225. doi: 10.3389/fvets.2022.905225. eCollection 2022.
Heat stress (HS) is devastating to poultry production sustainability due its detrimental effects on performance, welfare, meat quality, and profitability. One of the most known negative effects of HS is feed intake depression, which is more pronounced in modern high-performing broilers compared to their ancestor unselected birds, yet the underlying molecular mechanisms are not fully defined. The present study aimed, therefore, to determine the hypothalamic expression of a newly involved pathway, hypoxia/oxygen homeostasis, in heat-stressed broiler-based research lines and jungle fowl. Three populations of broilers (slow growing ACRB developed in 1956, moderate growing 95RB from broilers available in 1995, and modern fast growing MRB from 2015) and unselected Jungle fowl birds were exposed to cyclic heat stress (36°C, 9 h/day for 4 weeks) in a 2 × 4 factorial experimental design. Total RNAs and proteins were extracted from the hypothalamic tissues and the expression of target genes and proteins was determined by real-time quantitative PCR and Western blot, respectively. It has been previously shown that HS increased core body temperature and decreased feed intake in 95RB and MRB, but not in ACRB or JF. HS exposure did not affect the hypothalamic expression of HIF complex, however there was a line effect for HIF-1α ( = 0.02) with higher expression in JF under heat stress. HS significantly up regulated the hypothalamic expression of hemoglobin subunits (HBA1, HBBR, HBE, HBZ), and HJV in ACRB, HBA1 and HJV in 95RB and MRB, and HJV in JF, but it down regulated FPN1 in JF. Additionally, HS altered the hypothalamic expression of oxygen homeostasis- up and down-stream signaling cascades. Phospho-AMPK was activated by HS in JF hypothalamus, but it decreased in that of the broiler-based research lines. Under thermoneutral conditions, p-AMPK was higher in broiler-based research lines compared to JF. Ribosomal protein S6K1, however, was significantly upregulated in 95RB and MRB under both environmental conditions. HS significantly upregulated the hypothalamic expression of NF-κB2 in MRB, RelB, and TNFα in ACRB, abut it down regulated RelA in 95RB. The regulation of HSPs by HS seems to be family- and line-dependent. HS upregulated the hypothalamic expression of HSP60 in ACRB and 95RB, down regulated HSP90 in JF only, and decreased HSP70 in all studied lines. Taken together, this is the first report showing that HS modulated the hypothalamic expression of hypoxia- and oxygen homeostasis-associated genes as well as their up- and down-stream mediators in chickens, and suggests that hypoxia, thermotolerance, and feed intake are interconnected, which merit further in-depth investigations.
热应激(HS)对家禽生产的可持续性具有破坏性,因为它会对生产性能、福利、肉质和盈利能力产生不利影响。HS最广为人知的负面影响之一是采食量下降,与未经选育的祖先禽类相比,现代高性能肉鸡的这种情况更为明显,但其潜在的分子机制尚未完全明确。因此,本研究旨在基于热应激的肉鸡品系和原鸡,确定新涉及的缺氧/氧稳态途径在下丘脑的表达。三个肉鸡群体(1956年培育的生长缓慢的ACRB、1995年可得肉鸡中生长中等的95RB以及2015年的现代快速生长的MRB)和未经选育的原鸡,按照2×4析因实验设计,暴露于周期性热应激(36°C,每天9小时,持续4周)。从下丘脑组织中提取总RNA和蛋白质,分别通过实时定量PCR和蛋白质免疫印迹法测定靶基因和蛋白质的表达。先前的研究表明,热应激会使95RB和MRB的核心体温升高、采食量下降,但ACRB或原鸡则不会。热应激暴露并未影响HIF复合物在下丘脑的表达,然而,HIF-1α存在品系效应(P = 0.02),热应激下原鸡中的表达较高。热应激显著上调了ACRB中血红蛋白亚基(HBA1、HBBR、HBE、HBZ)和HJV、95RB和MRB中HBA1和HJV以及原鸡中HJV在下丘脑的表达,但下调了原鸡中FPN1的表达。此外,热应激改变了氧稳态上下游信号级联在下丘脑的表达。热应激使原鸡下丘脑的磷酸化AMPK活化,但在基于肉鸡的品系中则降低。在热中性条件下,基于肉鸡的品系中磷酸化AMPK高于原鸡。然而,在两种环境条件下,核糖体蛋白S6K1在95RB和MRB中均显著上调。热应激显著上调了MRB中NF-κB2、ACRB中RelB和TNFα在下丘脑的表达,但下调了95RB中RelA的表达。热应激对热休克蛋白(HSPs)的调节似乎具有家族和品系依赖性。热应激上调了ACRB和95RB中HSP60在下丘脑的表达,仅下调了原鸡中HSP90的表达,并降低了所有研究品系中HSP70的表达。综上所述,这是第一份表明热应激调节鸡下丘脑缺氧和氧稳态相关基因及其上下游介质表达的报告,并表明缺氧、耐热性和采食量相互关联,值得进一步深入研究。