Gene duplication of C-type natriuretic peptide-4 (CNP4) in teleost lineage elicits subfunctionalization of ancestral CNP
Gene duplication of C-type natriuretic peptide-4 (CNP4) in teleost lineage elicits subfunctionalization of ancestral CNP
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DOI：
10.1007/s00441-022-03596-y
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发表时间：
2022-02-16
影响因子：
3.6
通讯作者：
Tsukada, Takehiro
中科院分区：
文献类型：
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作者：
Katayama, Yukitoshi;Saito, Ami;Tsukada, Takehiro
在鳗鱼中发现了多种类型的利钠肽（NP）家族，包括4种CNP（CNP 1 -4）、ANP、BNP和VNP。在这里，我们成功地从鳗鱼（一种基底硬骨鱼）和斑马鱼（一种后来的分支硬骨鱼）的大脑中克隆了额外的cnp基因。这些基因被鉴定为硬骨鱼谱系中第三轮全基因组复制（3R）产生的cnp 4的旁系同源物，因此分别被命名为鳗鱼cnp 4 b和斑马鱼cnp 4-like。为了研究其表达的组织学模式，我们采用了一种新开发的原位杂交（ISH）链反应，除了传统的ISH，使用短发夹DNA。鳗鱼cnp 4 b在延髓中表达，而鳗鱼cnp 4a（前cnp 4）的mRNA定位于视前区。在斑马鱼脑中，cnp 4样mRNA是检测不到的，而已知的cnp 4在视前区和延髓中表达。结合RT-PCR和ISH检测到的cnp 4a和cnp 4 b在鳗鱼外周组织中的不同mRNA分布，我们认为祖先硬骨鱼中cnp 4s的亚功能化仅在基础硬骨鱼中保留。有趣的是，cnp 4 b表达神经元的舌咽-迷走神经运动复合体的延髓共定位与胆碱乙酰转移酶，这表明Cnp 4 b参与吞咽和呼吸功能，由迷走神经调制。由于硬骨鱼Cnp 4是哺乳动物CNP的直系同源物，硬骨鱼Cnp 4的确定定位将有助于未来的研究，旨在破译CNP的生理功能。
The diversified natriuretic peptide (NP) family, consisting of four CNPs (CNP1-4), ANP, BNP, and VNP, has been identified in the eel. Here, we successfully cloned additional cnp genes from the brain of eel (a basal teleost) and zebrafish (a later branching teleost). The genes were identified as paralogues of cnp4 generated by the third round of whole genome duplication (3R) in the teleost lineage, thereby being named eel cnp4b and zebrafish cnp4-like, respectively. To examine the histological patterns of their expressions, we employed a newly developed in situ hybridization (ISH) chain reaction using short hairpin DNAs, in addition to conventional ISH. Eel cnp4b was expressed in the medulla oblongata, while mRNAs of eel cnp4a (former cnp4) were localized in the preoptic area. In the zebrafish brain, cnp4-like mRNA was undetectable, while the known cnp4 was expressed in both the preoptic area and medulla oblongata. Together with the different mRNA distribution of cnp4a and cnp4b in eel peripheral tissues determined by RT-PCR and ISH, it is suggested that subfunctionalization by duplicated cnp4s in ancestral teleosts has been retained only in basal teleosts. Intriguingly, cnp4b-expressing neurons in the glossopharyngeal-vagal motor complex of the medulla oblongata were co-localized with choline acetyltransferase, suggesting an involvement of Cnp4b in swallowing and respiration functions that are modulated by the vagus. Since teleost Cnp4 is an ortholog of mammalian CNP, the identified localization of teleost Cnp4 will contribute to future studies aimed at deciphering the physiological functions of CNP.