Biology Department, Woods Hole Oceanographic Institution, Woods Hole, MA 02543, USA.
Fish Shellfish Immunol. 2010 Dec;29(6):1003-9. doi: 10.1016/j.fsi.2010.08.008. Epub 2010 Aug 20.
Epizootic shell disease is a poorly understood condition that has significantly affected the American lobster fishery in New England (northeastern US) since the 1990s. Here we present the results of a study to identify changes in gene expression in lobsters exhibiting symptoms of epizootic shell disease. Suppressive subtractive hybridization (SSH) was used to compare gene expression between cDNA pools from diseased (symptomatic) and apparently healthy (asymptomatic) lobsters. Subsequently, quantitative real-time polymerase chain reaction (qPCR) was used to measure expression of nine genes that were differentially-expressed in the SSH analysis, in seven tissues (muscle, gill, heart, hepatopancreas, brain, branchiostegite, gonad) dissected from individual symptomatic and asymptomatic lobsters. Expression of arginine kinase (involved in cellular energetics) was significantly decreased in muscle of symptomatic lobsters. Expression of hemocyanin (a respiratory hemolymph protein involved in oxygen transport) was highest in hepatopancreas and showed highly variable expression with a trend toward higher expression in asymptomatic individuals. Alpha-2 macroglobulin (involved in the innate immune system) was most highly expressed in the ovary, particularly of symptomatic lobsters. The ESTs produced through this study add to the fledgling field of crustacean genomics and revealed three genes that could be further evaluated in lobsters of varying shell disease severity, molt stage, and reproductive condition, for possible implication in epizootic shell disease.
流行病壳病是一种尚未被充分了解的疾病,自 20 世纪 90 年代以来,它已对新英格兰(美国东北部)的美洲龙虾渔业造成了重大影响。在这里,我们呈现了一项研究的结果,该研究旨在确定表现出流行病壳病症状的龙虾中基因表达的变化。抑制性消减杂交(SSH)被用于比较来自患病(有症状)和明显健康(无症状)龙虾的 cDNA 池之间的基因表达。随后,定量实时聚合酶链反应(qPCR)用于测量在 SSH 分析中差异表达的九个基因在七个组织(肌肉、鳃、心脏、肝胰腺、脑、鳃盖骨、性腺)中的表达,这些组织是从单个有症状和无症状龙虾中解剖出来的。有症状龙虾肌肉中的精氨酸激酶(参与细胞能量学)表达显著降低。血蓝蛋白(一种参与氧气运输的呼吸血液蛋白)在肝胰腺中的表达最高,且在无症状个体中表现出高度可变的表达趋势,表达水平较高。α-2 巨球蛋白(参与先天免疫系统)在卵巢中表达最高,尤其是在有症状的龙虾中。通过这项研究产生的 EST 丰富了甲壳类动物基因组学的新兴领域,并揭示了三个基因,这些基因可以在不同壳病严重程度、蜕皮阶段和生殖状况的龙虾中进一步评估,以确定其在流行病壳病中的可能影响。