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Abstract
Oxytocin (OT) and vasopressin (VP) superfamily neuropeptides are distributed in not only vertebrates but also diverse invertebrates. However, no VPergic innervation of invertebrates has ever been documented. In the ascidian, Ciona intestinalis Type A (Ciona robusta), an OT/VP superfamily peptide was identified, and the Ciona vasopressin (CiVP) induces oocyte maturation and ovulation. In the present study, we characterize the innervation and phenotypes of genetically modified Ciona: CiVP promoter-Venus transgenic and CiVP mutants. CiVP promoter-Venus transgenic Ciona demonstrated that CiVP gene was highly expressed in the cerebral ganglion and several nerves. Fluorescence was also detected in the ovary of young CiVP promoter-Venus transgenic ascidians, suggesting that the CiVP gene is also expressed temporarily in the ovary of young ascidians. Furthermore, a marked decrease of post-vitellogenic (stage III) follicles was observed in the ovary of CiVP mutants, whereas pre-vitellogenic (stage I) and vitellogenic (stage II) follicles were increased in the mutant ovary, compared with that of wildtype Ciona. Gene expression profiles showed that the expression of various genes, including genes related to ovarian follicle growth, was altered in the ovary of CiVP mutants. Altogether, these results indicated that CiVP, mainly as a neuropeptide, plays pivotal roles in diverse biological functions, including growth of early-stage ovarian follicles via regulation of the expression of a wide variety of genes. This is the first report describing a VP gene promoter-transgenic and VP gene-edited invertebrate and also on its gene expression profiles and phenotypes.
Highlights
Oxytocin (OT) and vasopressin (VP) superfamily peptides are one of the most widely distributed neuropeptides and/or neurohypophysial hormones in both vertebrates and invertebrates [1,2,3,4,5,6,7,8,9,10]
To examine Ciona vasopressin (CiVP) gene expression in detail, we constructed transgenic ascidians introduced by the 5-kb CiVP promoter region-conjugated with the ORF of a fluorescent protein, Venus
This fluorescent signal was in good agreement with the localization of the CiVP mRNA in wildtype Ciona larva but not in any earlier developmental stages as observed in our previous study [36], suggesting that the fluorescence derived from CiVP promoter-Venus construct indicates the expression of the CiVP gene in the transgenic Ciona
Summary
Oxytocin (OT) and vasopressin (VP) superfamily peptides are one of the most widely distributed neuropeptides and/or neurohypophysial hormones in both vertebrates and invertebrates [1,2,3,4,5,6,7,8,9,10] These peptides are typically comprised of 9 to 14 amino acids, and share Cys and Cys, which form an intramolecular disulfide bridge essential for biological activities. The vertebrate OT family peptides share a neutral aliphatic amino acid (Leu, Ile, Val or Thr), whereas the vertebrate VP family peptides contain a basic one (Lys or Arg) at position 8 These superfamily peptides play pleiotropic roles in reproductive behaviors, osmoregulation, food intake, learning, and social behavior and are implicated with pathological processes such as anxiety and autism [1, 2, 11,12,13]. We present the expression profiles of CiVP promoter-Venus-Ciona transgenic larvae, young adults, and adults, and comparative transcriptomes between the wildtype and the genome-edited CiVP-knockout Ciona
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