Lipka Aleksandra, Paukszto Lukasz, Majewska Marta, Jastrzebski Jan Pawel, Panasiewicz Grzegorz, Szafranska Bozena
Department of Gynecology and Obstetrics, School of Medicine, Collegium Medicum, University of Warmia and Mazury in Olsztyn, Niepodległości Str 44, 10-045, Olsztyn, Poland.
Department of Plant Physiology, Genetics and Biotechnology, Faculty of Biology and Biotechnology, University of Warmia and Mazury in Olsztyn, Oczapowskiego Str 1A, 10-719, Olsztyn, Poland.
Funct Integr Genomics. 2019 May;19(3):421-435. doi: 10.1007/s10142-019-00663-6. Epub 2019 Feb 18.
Our pioneering data provide the first comprehensive view of placental transcriptome of the beaver during single and multiple gestation. RNA-Seq and a de novo approach allowed global pattern identification of C. fiber placental transcriptome. Non-redundant beaver transcriptome comprised 211,802,336 nt of placental transcripts, grouped into 128,459 contigs and clustered into 83,951 unigenes. An Ensembl database search revealed 14,487, 14,994, 15,004, 15,267 and 15,892 non-redundant homologs for Ictidomys tridecemlineatus, Rattus norvegicus, Mus musculus, Homo sapiens and Castor canadensis, respectively. Due to expression levels, the identified transcripts were divided into two sets: non-redundant and highly expressed (FPKM > 2 in at least three examined samples), analysed simultaneously. Among 17,009 highly expressed transcripts, 12,147 had BLASTx hits. GO annotations (175,882) were found for 4301 transcripts that were assigned to biological process (16,386), cellular component (9149) and molecular function (8338) categories; 666 unigenes were also classified into 122 KEGG pathways. Comprehensive analyses were performed for 411 and 3078 highly expressed transcripts annotated with a list of processes linked to 'placenta' (31 GO terms) or 'embryo' (324 GO terms), respectively. Among transcripts with entire CDS annotation, 281 (placenta) and 34 (embryo) alternative splicing events were identified. A total of 8499 putative SNVs (~ 6.2 SNV/transcript and 1.7 SNV/1 kb) were predicted with 0.1 minimum frequency and maximum variant quality (p value 10e). Our results provide a broad-based characterization of the global expression pattern of the beaver placental transcriptome. Enhancement of transcriptomic resources for C. fiber should improve understanding of crucial pathways relevant to proper placenta development and successful reproduction.
我们的开创性数据首次全面展现了河狸单胎和多胎妊娠期间胎盘转录组的情况。RNA测序和从头测序方法使得能够对河狸胎盘转录组的全局模式进行识别。非冗余的河狸转录组包含211,802,336个核苷酸的胎盘转录本,被组装成128,459个重叠群,并聚类为83,951个单基因。通过Ensembl数据库搜索,分别发现了14,487、14,994、15,004、15,267和15,892个与三线松鼠、褐家鼠、小家鼠、智人和加拿大河狸的非冗余同源物。根据表达水平将鉴定出的转录本分为两组:非冗余和高表达(在至少三个检测样本中FPKM>2),同时进行分析。在17,009个高表达转录本中,有12,147个有BLASTx比对结果。对4301个转录本进行了GO注释(175,882条),这些转录本被分配到生物过程(16,386条)、细胞成分(9149条)和分子功能(8338条)类别;666个单基因也被归类到122条KEGG通路中。分别对411个和3078个高表达转录本进行了综合分析,这些转录本分别用与“胎盘”(31个GO术语)或“胚胎”(324个GO术语)相关的过程列表进行注释。在具有完整CDS注释的转录本中,鉴定出281个(胎盘)和34个(胚胎)可变剪接事件。共预测了8499个推定的单核苷酸变异(~6.2个单核苷酸变异/转录本和1.7个单核苷酸变异/1kb),最小频率为0.1,最大变异质量(p值10e)。我们的结果为河狸胎盘转录组的全局表达模式提供了广泛的特征描述。增强河狸的转录组资源应有助于更好地理解与胎盘正常发育和成功繁殖相关的关键途径。