Martos-Sitcha Juan Antonio, Campinho Marco Antonio, Mancera Juan Miguel, Martínez-Rodríguez Gonzalo, Fuentes Juan
Centre of Marine Sciences (CCMar), Universidade do Algarve, Campus de Gambelas, Faro 8005-139, Portugal Departamento de Biología, Facultad de Ciencias del Mar y Ambientales, Universidad de Cádiz, Puerto Real, Cádiz E-11510, Spain Instituto de Ciencias Marinas de Andalucía, Consejo Superior Investigaciones Científicas (ICMAN-CSIC), Puerto Real, Cádiz E-11510, Spain.
Centre of Marine Sciences (CCMar), Universidade do Algarve, Campus de Gambelas, Faro 8005-139, Portugal.
J Exp Biol. 2015 Mar;218(Pt 5):684-93. doi: 10.1242/jeb.114546. Epub 2015 Jan 8.
Aquaporins (AQPs) are specific transmembrane water channels with an important function in water homeostasis. In terrestrial vertebrates, AQP2 function is regulated by vasopressin (AVP) to accomplish key functions in osmoregulation. The endocrine control of aquaporin function in teleosts remains little studied. Therefore, in this study we investigated the regulatory role of vasotocin (AVTR) and isotocin (ITR) receptors in Aqp1 paralog gene function in the teleost gilthead sea bream (Sparus aurata). The complete coding regions of Aqp1a, Aqp1b, AVTR V1a2-type, AVTR V2-type and ITR from sea bream were isolated. A Xenopus oocyte-swelling assay was used to functionally characterize AQP1 function and regulation by AVT and IT through their cognate receptors. Microinjection of oocytes with Aqp1b mRNA revealed regulation of water transport via PKA (IBMX+forskolin sensitive), whereas Aqp1a mRNA injection had the same effect via PKC signaling (PDBU sensitive). In the absence of expressed receptors, AVT and IT (10(-8) mol l(-1)) were unable to modify AQP1 function. AVT regulated AQP1a and AQP1b function only when the AVTR V2-type was co-expressed. IT regulated AQP1a function, but not AQP1b, only when ITR was present. Considering that Aqp1a and Aqp1b gene expression in the sea bream intestine is highly salinity dependent in vivo, our results in ovo demonstrate a regulatory role for AVT and IT in AQP1 function in the sea bream in the processing of intestinal fluid to achieve osmoregulation.
水通道蛋白(AQPs)是特定的跨膜水通道,在水稳态中具有重要功能。在陆生脊椎动物中,AQP2的功能受抗利尿激素(AVP)调节,以完成渗透调节中的关键功能。硬骨鱼中水通道蛋白功能的内分泌控制仍鲜为人知。因此,在本研究中,我们调查了硬骨鱼金头鲷(Sparus aurata)中血管紧张素(AVTR)和异速加压素(ITR)受体在Aqp1旁系同源基因功能中的调节作用。分离了金头鲷的Aqp1a、Aqp1b、AVTR V1a2型、AVTR V2型和ITR的完整编码区。采用非洲爪蟾卵母细胞肿胀试验从功能上表征AQP1的功能以及抗利尿激素(AVT)和异速加压素(IT)通过其同源受体对AQP1功能的调节。向卵母细胞显微注射Aqp1b mRNA显示水运输受蛋白激酶A调节(对异丁基甲基黄嘌呤+福斯高林敏感),而注射Aqp1a mRNA通过蛋白激酶C信号传导产生相同效果(对佛波酯敏感)。在未表达受体的情况下,AVT和IT(10⁻⁸ mol·l⁻¹)无法改变AQP1的功能。只有当共表达AVTR V2型时,AVT才调节AQP1a和AQP1b的功能。只有当存在ITR时,IT才调节AQP1a的功能,而不调节AQP1b的功能。鉴于金头鲷肠道中Aqp1a和Aqp1b基因表达在体内高度依赖盐度,我们在卵内的研究结果表明,AVT和IT在金头鲷处理肠液以实现渗透调节的过程中对AQP1功能具有调节作用。