Genomic abnormalities affecting mussels (Mytilus edulis-galloprovincialis) in France are related to ongoing neoplastic processes, evidenced by dual flow cytometry and cell monolayer analyses
Genomic abnormalities affecting mussels (Mytilus edulis-galloprovincialis) in France are related to ongoing neoplastic processes, evidenced by dual flow cytometry and cell monolayer analyses
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DOI：
10.1016/j.jip.2018.08.003
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发表时间：
2018-09-01
影响因子：
3.4
通讯作者：
Benjamin, Morga
中科院分区：
文献类型：
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作者：
Abdellah, Benadelmouna;Alice, Saunier;Benjamin, Morga
在法国自2014年以来贻贝大量异常死亡的背景下，2015年和2016年使用流式细胞仪（FCM）研究了从2000只贻贝采集的半血循环细胞的DNA含量和细胞周期特征。这些贻贝是从分布在法国大西洋沿岸从布列塔尼南部到佩尔图伊夏朗泰地区的 12 个野生和养殖蓝贻贝种群中取样的。在这些调查中，经常检测到各种遗传异常，倍性特征揭示了与各自不同的卫生状况相对应的对比图谱，即具有高细胞遗传学质量（HCQ）的健康贻贝与具有低细胞遗传学质量（LCQ）的患病贻贝。在目前的工作中，FCM 和血细胞单层技术相结合，以确定观察到的与死亡率水平显着相关的遗传异常的推定原因。 FCM 和细胞单层方法允许定义新的阈值，将 HCQ 贻贝与 LCQ 贻贝区分开来。 HCQ 组贻贝的 FCM 直方图显示一个单一或主要占优势的二倍体 (2n) 细胞核群体和大部分正常血细胞。血淋巴细胞单层分析显示主要为嗜酸性粒细胞，其特征在于正常大小的细胞核和具有大量颗粒的大细胞质。相反，LCQ 组的 FCM 直方图显示，除了正常的二倍体 (2n) 细胞核外，细胞核群体在广泛的倍性范围内显示出非整倍体模式，包括二倍体-三倍体 (2-3n)、四倍体-五倍体 (4-5n) 和七倍体-八倍体水平 (7-8n)。相应的血淋巴细胞单层显示出播散性肿瘤疾病的细胞特征，具有频繁的异常间变性细胞，表现出明显数量的有丝分裂图，具有正常和异常的染色体分离模式。这些肿瘤细胞呈圆形，具有减少的无颗粒细胞质和大（11-12μm）到非常大（高达21μm）的圆形或卵圆形细胞核，对应于先前通过FCM分析检测到的4-5n和7-8n细胞核。这些特征表明，通过 FCM 检测到的遗传异常与正在影响法国蓝贻贝的持续肿瘤过程有关，至少自 2014 年严重影响法国蓝贻贝种群的死亡率以来是这样。
In the context of the abnormal mass mortality of mussels in France since 2014, Flow CytoMetry (FCM) was used in 2015 and 2016 to study the DNA content and cell cycle characteristics of hemic circulating cells collected from 2000 mussels. The mussels were sampled from 12 wild and cultivated blue mussels stocks distributed along the French Atlantic coast from the south Brittany to Pertuis Charentais areas. During these surveys, various genetic abnormalities were frequently detected, and ploidy characteristics revealed contrasting profiles that corresponded to respective contrasting sanitary status, i.e. healthy mussels with high cytogenetic quality (HCQ) versus diseased mussels with low cytogenetic quality (LCQ). In the present work, FCM and hemocytology cell mono layer techniques were combined in order to determine the putative causes of the observed genetic abnormalities that were significantly associated with mortality levels. FCM and cell monolayer approaches permitted the definition of new threshold values delimiting HCQ mussels from LCQ ones. FCM histograms of mussels from the HCQ group showed one single or a largely dominant population of diploid (2n) nuclei and a large majority of normal hemocytes. Hemolymph cell-monolayer analyses showed predominantly acidophil granulocytes characterized by nuclei of normal size and a large cytoplasm with numerous granulations. In contrast, FCM histograms for the LCQ group showed, in addition to the normal diploid (2n) nuclei, populations of nuclei that displayed aneuploidy patterns in a broad ploidy range, including diploid-triploid (2-3n), tetraploid-pentaploid (4-5n) and heptaploid-octaploid levels (7-8n). The corresponding hemolymph cell-monolayer showed cellular features characteristic of disseminated neoplasia disease with frequent abnormal anaplastic cells that exhibited noticeable numbers of mitotic figures with both normal and aberrant chromosomes segregation patterns. These neoplastic cells were a rounded shape with a reduced, granulation-free cytoplasm and large (11-12 mu m) to very large (up to 21 mu m) round or ovoid nuclei that correspond to the 4-5n and 7-8n nuclei previously detected by FCM analyses. These characteristics suggest that the genetic abnormalities detected by means of FCM were related to an ongoing neoplastic process that is affecting blue mussels in France, at least since the onset in 2014 of the mortality that heavily impacted French blue mussels stocks.