Tanaka Yasuko, Morishita Yoshiyuki, Ishibashi Kenichi
Department of Medical Physiology, School of Pharmacy, Meiji Pharmaceutical University, 2-522-1 Noshio, Kiyose, Tokyo 204-8588, Japan.
Division of Nephrology, Department of Medicine, Jichi Medical University, 3311-1, Yakushiji, Shimotsuke-city, Tochigi 329-0498, Japan.
Biochem Biophys Rep. 2015 Mar 28;1:16-21. doi: 10.1016/j.bbrep.2015.03.009. eCollection 2015 May.
Although AQP10 is mainly expressed in the human GI tract, its physiological role is unclear. In fact, we previously reported that mouse AQP10 is a pseudogene. It is possible that AQP10 is also a pseudogene in other animals.
Genome databases were searched for AQP10 orthologs and the genomic DNA of each candidate pseudogene was sequenced to confirm its mutations. The expression of the AQP10 mRNA was examined by RT-PCR in the small intestine where human AQP10 is highly expressed.
The genomic database of some mammals had insertions and deletions in the exons of the AQP10 gene, including cattle (), sheep () and goats (). In the bovine AQP10 gene, exon 1 and 5 had deletions resulting in a frame-shift or a premature termination, respectively, which were confirmed by the direct exon sequencing of the genomic DNA. In the RT-PCR experiments, the PCR primer sets for exon 1/2 and exon 4/5 failed to detect the bands for AQP10 mRNA in the duodenum and jejunum. Similar AQP10 gene mutations were also confirmed in the genomic DNA from sheep and goats. Although these animals were derived from porcine ancestors, the exons of the swine () AQP10 gene were complete without mutations. Therefore, AQP10 gene might have turned to a pseudogene around 65 million years before when cattle evolved from porcine ancestors.
AQP10 of ruminantia which regurgitate and rechew their food may have lost its role possibly due to the redundant expression of other aquaglyceroporins.
尽管水通道蛋白10(AQP10)主要在人类胃肠道中表达,但其生理作用尚不清楚。事实上,我们之前报道过小鼠AQP10是一个假基因。在其他动物中,AQP10也有可能是假基因。
在基因组数据库中搜索AQP10直系同源基因,并对每个候选假基因的基因组DNA进行测序以确认其突变。通过逆转录聚合酶链反应(RT-PCR)检测人AQP10高表达的小肠中AQP10 mRNA的表达。
一些哺乳动物的基因组数据库显示,AQP10基因的外显子存在插入和缺失,包括牛()、绵羊()和山羊()。在牛AQP10基因中,外显子1和5有缺失,分别导致移码或提前终止,这通过基因组DNA的直接外显子测序得到证实。在RT-PCR实验中,外显子1/2和外显子4/5的PCR引物组未能在十二指肠和空肠中检测到AQP10 mRNA的条带。在绵羊和山羊的基因组DNA中也证实了类似的AQP10基因突变。尽管这些动物起源于猪的祖先,但猪()AQP10基因的外显子是完整的,没有突变。因此,在大约6500万年前牛从猪的祖先进化而来时,AQP10基因可能已变成假基因。
反刍和重新咀嚼食物的反刍动物的AQP10可能由于其他水甘油通道蛋白的冗余表达而失去了其作用。