Milani Diogo, Palacios-Gimenez Octavio M, Cabral-de-Mello Diogo C
Departamento de Biologia, Instituto de Biociências/IB, Universidade Estadual Paulista (UNESP), Rio Claro, Brazil.
Cytogenet Genome Res. 2017;151(1):36-40. doi: 10.1159/000458468. Epub 2017 Mar 2.
In this study, we describe a strategy to determine the presence of B chromosomes in the living grasshopper Abracris flavolineata by FISH using U2 snDNA as a probe in interphase hemolymph nuclei. In individuals without B chromosomes, (0B) 2 dot signals were noticed, corresponding to A complement U2 snDNA clusters. In +1B and +2B individuals, 4 or 8 additional signals were noticed, respectively. In all cases, the absence or presence of 1 or 2 B chromosomes correlated in hemolymph and in somatic or germline tissues, validating the efficiency of the marker. Our data suggest that the B chromosome of A. flavolineata is present in all somatic tissues. B-carrying individuals showed the same number of B chromosomes in germ and somatic cells, suggesting that the B is mitotically stable. The marker was used to compare B chromosome frequency in the analyzed population with a sample collected previously, in order to test for B frequency changes and differences of B chromosome prevalence among sexes, but no statistically significant differences were noticed. The identification of living animals harboring B chromosomes will be very useful in future studies of B chromosome transmission, as well as in functional studies involving RNA analysis, thus contributing to the understanding of evolutionary history and the possible role of the B chromosome in A. flavolineata.
在本研究中,我们描述了一种策略,通过在间期血淋巴细胞核中使用U2 snDNA作为探针进行荧光原位杂交(FISH),来确定活的黄缘阿布拉蝗(Abracris flavolineata)中B染色体的存在情况。在没有B染色体的个体(0B)中,观察到2个点状信号,对应于A染色体组的U2 snDNA簇。在+1B和+2B个体中,分别观察到4个或8个额外的信号。在所有情况下,血淋巴细胞以及体细胞或生殖细胞组织中1条或2条B染色体的缺失或存在情况都是相关的,验证了该标记的有效性。我们的数据表明,黄缘阿布拉蝗的B染色体存在于所有体细胞组织中。携带B染色体的个体在生殖细胞和体细胞中显示相同数量的B染色体,表明B染色体在有丝分裂过程中是稳定的。该标记用于比较分析群体与先前收集样本中B染色体的频率,以测试B染色体频率的变化以及不同性别之间B染色体流行率的差异,但未观察到统计学上的显著差异。鉴定携带B染色体的活体动物在未来B染色体传递研究以及涉及RNA分析的功能研究中将非常有用,从而有助于理解进化历史以及B染色体在黄缘阿布拉蝗中的可能作用。