DIFFERENTIAL EXPRESSION OF DISTINCT MESSENGER-RNAS FOR OVINE TROPHOBLAST PROTEIN-1 AND RELATED SHEEP TYPE-I INTERFERONS
DIFFERENTIAL EXPRESSION OF DISTINCT MESSENGER-RNAS FOR OVINE TROPHOBLAST PROTEIN-1 AND RELATED SHEEP TYPE-I INTERFERONS
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DOI：
10.1095/biolreprod48.4.768
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发表时间：
1993-04-01
影响因子：
3.6
通讯作者：
IMAKAWA, K
中科院分区：
文献类型：
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作者：
NEPHEW, KP;WHALEY, AE;IMAKAWA, K
绵羊滋养层蛋白-1（oTP-1）是一种新型的I型干扰素（IFN），是由绵羊孕体分泌的抗黄体溶解物质，主要负责母体识别妊娠。本研究的目的是：1）研究绵羊基因组中是否有多个不同的基因编码oTP-1和其他I型IFN; 2）检测在孕体发育过程中oTP-1和其他IFN mRNA的表达。从第25天（第0天=发情期）绵羊孕体高分子量DNA构建的亚基因组文库中分离I型IFN的基因。分离六个克隆并从-1000至+900（相对于帽位点的碱基）进行核苷酸测序。氨基酸序列分析表明，其中4个克隆为不同的oTP-1基因，另外2个克隆为o 9和o 12，与oTP-1的氨基酸同源性分别为71%和54%。通过逆转录-聚合酶链反应（RT-PCR）分析从第13-45天孕体提取的总细胞RNA（tcRNA），定量检测oTP-1和I型IFN基因编码的mRNA的存在。从第75天胎盘和成人淋巴细胞获得的总细胞RNA也通过RT-PCR分析，再加上PCR反应产物与特异性DNA探针的Southern印迹杂交。对PCR产物进行测序以确认引物特异性，并鉴定了对应于四个oTP-1基因中的两个和两个相关IFN克隆（o 9和o 12）的mRNA。此外，PCR产物的定量显示，在检测的两个oTP-1基因中，一个在第13-20天高度表达，而转录物在第30天和第45天可微弱检测到。相比之下，检测的其他oTP-1基因仅在第13-20天弱表达。杂交信号的光密度分析显示，IFN α 9 mRNA在第75天胎盘中检测到，但在孕体（第13-45天）和成人淋巴细胞中仅微弱检测到。IFN o 12 mRNA在淋巴细胞中相对于检查的其他组织是丰富的。总的来说，这些结果表明存在不同的oTP-1和相关的I型IFN基因。这些数据表明，这些基因在怀孕期间显示差异，组织特异性表达和发育调节。
An antiluteolytic substance secreted by the ovine conceptus and primarily responsible for maternal recognition of pregnancy is ovine trophoblast protein-1 (oTP-1), a new type I interferon (IFN). The objectives of this research were 1 ) to investigate whether multiple, distinct genes encode oTP-1 and other type I IFNs in the ovine genome and 2) to examine expression of oTP-1 and other IFN mRNAs during conceptus development. Genes for type I IFNs were isolated from a subgenomic library constructed from Day 25 (Day 0 = estrus) ovine conceptus high-molecular-weight DNA. Six clones were isolated and nucleotide-sequenced from -1000 to +900 (bases relative to cap site). Comparisons of inferred amino acid sequences demonstrated that four clones were distinct oTP-1 genes and that two clones, defined as o9 and o12, were related type I IFNs (deduced aa homology of o9 and o12 to oTP-1 was 71% and 54%, respectively). The presence of mRNAs encoded by oTP-1 and type I IFN genes was examined quantitatively via reverse transcription-polymerase chain-reaction (RT-PCR) analysis of total cellular RNA (tcRNA) extracted from Day 13-45 concepti. Total cellular RNA obtained from Day 75 placenta and adult lymphocytes was also analyzed by RT-PCR, coupled with Southern blot hybridization of the PCR reaction products with specific DNA probes. PCR products were sequenced in order to confirm primer specificity, and mRNAs corresponding to two of the four oTP-1 genes and to both related IFN clones (o9 and o12) were identified. Furthermore, quantitation of the PCR products revealed that of the two oTP-1 genes examined, one was highly expressed on Days 13-20 and transcripts were weakly detectable on Days 30 and 45. In contrast, the other oTP-1 gene examined was weakly expressed on Days 13-20 only. Densitometric analysis of hybridization signals revealed that IFN o9 mRNA was detected in Day 75 placenta but only weakly detected in conceptus (Days 13-45) and adult lymphocytes. IFN o12 mRNA was abundant in lymphocytes relative to the other tissues examined. Collectively, these results demonstrate the existence of distinct oTP-1 and related type I IFN genes- The data suggest that these genes display differential, tissue-specific expression and developmental regulation during pregnancy.