한국가금학회지 (Korean Journal of Poultry Science)

  • 제44권1호
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  • Pages.1-9
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  • 2017
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  • 1225-6625(pISSN)
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  • 2287-5387(eISSN)
한국가금학회 (The Korean Society of Poultry Science)
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DOI QR Code

Lipopolysaccride 감염처리가 닭의 품종간 스트레스연관 유전자 발현에 미치는 영향

Effects of Lipopolysaccride-induced Stressor on the Expression of Stress-related Genes in Two Breeds of Chickens

  • 장인석 (경남과학기술대학교 동물생명과학과) ;
  • 손시환 (경남과학기술대학교 동물생명과학과) ;
  • 문양수 (경남과학기술대학교 동물생명과학과)
  • Jang, In Surk (Department of Animal Science and Biotechnology, Gyeongnam National University of Science and Technology) ;
  • Sohn, Sea Hwan (Department of Animal Science and Biotechnology, Gyeongnam National University of Science and Technology) ;
  • Moon, Yang Soo (Department of Animal Science and Biotechnology, Gyeongnam National University of Science and Technology)
  • 투고 : 2016.11.29
  • 심사 : 2017.02.13
  • 발행 : 2017.03.31
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초록

본 연구는 한국재래계(KNC)와 백색레그혼(WLH)에서 lipopolysaccharide(LPS)감염 스트레스가 닭의 품종간 스트레스 연관 유전자들의 발현에 미치는 영향을 비교 분석하고자 실시되었다. 공시계를 대상으로 생리식염수(대조구)와 LPS(처리구)를 복강에 투여한 후, 시간(0, 48 hr) 및 처리별 각 개체로부터 간 조직을 취하고, microarray 및 quantitative RT-PCR(qRT-PCR) 분석을 하였다. 처리에 따른 유전자 발현차이를 보면, KNC(대조구)와 KNC에 LPS를 처리한 경우(KNC-LPS)를 비교한 결과, 대조구 대비 2배 이상 유전자의 발현이 증가한 유전자의 수는 1,044개, 발현이 감소한 유전자의 수는 1,000개였다. WLH(대조구)를 WLH-LPS와 비교한 경우, 유전자의 발현이 증가한 유전자의 수는 1,193개, 발현이 감소한 유전자의 수는 1,072개였다. LPS 처리에 따른 스트레스 연관 유전자들의 microarray 발현에서 스트레스연관 유전자들의 발현은 두 품종 모두에서 감소하였으며, 품종 간 차이는 없는 것으로 나타났다. Microarray의 결과를 바탕으로 HSP90, HMGCR, ATF4, SREBP1, XBP1 등의 유전자 발현을 qRT-PCR을 이용하여 검증한 결과, 대조구와 LPS 감염구 간에 유의적 차이를 나타내었다(P<0.05). 세포 수준의 스트레스(ER 스트레스)에서 ATF4, XBP1, SREBP1은 화이트레그혼에서 microarray와 qRT-PCR에서와 같이 이들 유전자들의 발현이 억제되는 것을 보여주었다. 그러나 한국 재래계에서는 ATF4를 제외한 유전자들은 LPS에 의해 영향을 받지 않거나(XBP1), 오히려 증가(SREBP)하는 양상을 보였다. ER-stress 연관 유전자들의 발현 양상으로 볼 때, KNC이 WLH에 비하여 LPS 감염에 더 민감하게 반응하는 것으로 보인다. HMGCR은 두 품종간에 LPS에 의한 상호작용이 없는 것으로 보아, HMGCR 발현에 의한 감염 차이점을 찾을 수 없었다. 한국재래계에서 HSP70은 LPS 처리 후에 대조구에 비하여 약 2.5배 이상 높은 발현을 보였으나, 백색 레그혼에서는 낮은 발현 양상을 나타내었다. 스트레스 지표 유전자들의 종류뿐만 아니라, 스트레스 종류(예: 환경스트레스, 감염스트레스)에 따라 유전자들의 발현 반응에 차이가 있음을 보여주었다. LPS 감염스트레스에 따른 스트레스연관 유전자 발현연구는 닭의 품종별 질병 저항성 및 동물복지 관련 지표의 탐색에 기여할 것으로 사료된다.

The objective of the present study was to determine the expression of genes associated with lipopolysaccharide (LPS)-induced stressor in two breeds of chickens: the Korean native chicken (KNC) and the White Leghorn chicken (WLH). Forty chickens per breed, aged 40 weeks, were randomly allotted to the control (CON, administered the saline vehicle) and LPS-injected stress groups. Samples were collected at 0 and 48 h post-LPS injection, and total RNA was extracted from the chicken livers for RNA microarray and quantitative real-time polymerase chain reaction (qRT-PCR) analyses. In response to LPS, 1,044 and 1,193 genes were upregulated, and 1,000 and 1,072 genes were downregulated in the KNC and WLH, respectively, using a ${\geq}2$-fold cutoff change. A functional network analysis revealed that stress-related genes were downregulated in both KNC and WLH after LPS infection. The results obtained from the qRT-PCR analysis of mRNA expression of heat shock 90 (HSP90), 3-hydroxy-3-methylglutaryl-CoA reductase (HMGCR), activating transcription factor 4 (ATF4), sterol regulatory element-binding protein 1 (SREBP1), and X-box binding protein 1 (XBP1) were confirmed by the results of the microarray analysis. There was a significant difference in the expression of stress-associated genes between the control and LPS-injected KNC and WLH groups. The qRT-PCR analysis revealed that the stress-related $HSP90{\alpha}$ and HMGCR genes were downregulated in both LPS-injected KNC and WLH groups. However, the HSP70 and $HSP90{\beta}$ genes were upregulated only in the LPS-injected KNC group. The results suggest that the mRNA expression of stress-related genes is differentially affected by LPS stimulation, and some of the responses varied with the chicken breed. A better understanding of the LPS-induced infective stressors in chicken using the qRT-PCR and RNA microarray analyses may contribute to improving animal welfare and husbandry practices.

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