PMID- 38381411 OWN - NLM STAT- MEDLINE DCOM- 20240222 LR - 20240227 IS - 1552-5783 (Electronic) IS - 0146-0404 (Print) IS - 0146-0404 (Linking) VI - 65 IP - 2 DP - 2024 Feb 1 TI - Deficiency of Acetyltransferase nat10 in Zebrafish Causes Developmental Defects in the Visual Function. PG - 31 LID - 10.1167/iovs.65.2.31 [doi] LID - 31 AB - PURPOSE: N4-acetylcytidine (ac4C) is a post-transcriptional RNA modification catalyzed by N-acetyltransferase 10 (NAT10), a critical factor known to influence mRNA stability. However, the role of ac4C in visual development remains unexplored. METHODS: Analysis of public datasets and immunohistochemical staining were conducted to assess the expression pattern of nat10 in zebrafish. We used CRISPR/Cas9 and RNAi technologies to knockout (KO) and knockdown (KD) nat10, the zebrafish ortholog of human NAT10, and evaluated its effects on early development. To assess the impact of nat10 knockdown on visual function, we performed comprehensive histological evaluations and behavioral analyses. Transcriptome profiling and real-time (RT)-PCR were utilized to detect alterations in gene expression resulting from the nat10 knockdown. Dot-blot and RNA immunoprecipitation (RIP)-PCR analyses were conducted to verify changes in ac4C levels in both total RNA and opsin mRNA specifically. Additionally, we used the actinomycin D assay to examine the stability of opsin mRNA following the nat10 KD. RESULTS: Our study found that the zebrafish NAT10 protein shares similar structural properties with its human counterpart. We observed that the nat10 gene was prominently expressed in the visual system during early zebrafish development. A deficiency of nat10 in zebrafish embryos resulted in increased mortality and developmental abnormalities. Behavioral and histological assessments indicated significant vision impairment in nat10 KD zebrafish. Transcriptomic analysis and RT-PCR identified substantial downregulation of retinal transcripts related to phototransduction, light response, photoreceptors, and visual perception in the nat10 KD group. Dot-blot and RIP-PCR analyses confirmed a pronounced reduction in ac4C levels in both total RNA and specifically in opsin messenger RNA (mRNA). Additionally, by evaluating mRNA decay in zebrafish treated with actinomycin D, we observed a significant decrease in the stability of opsin mRNA in the nat10 KD group. CONCLUSIONS: The ac4C-mediated mRNA modification plays an essential role in maintaining visual development and retinal function. The loss of NAT10-mediated ac4C modification results in significant disruptions to these processes, underlining the importance of this RNA modification in ocular development. FAU - Yang, Hou-Zhi AU - Yang HZ AD - Tianjin Medical University, Tianjin, China. FAU - Zhuo, Donghai AU - Zhuo D AD - School of Medicine, Nankai University, Tianjin, China. FAU - Huang, Zongyu AU - Huang Z AD - Tianjin Medical University, Tianjin, China. FAU - Luo, Gan AU - Luo G AD - Tianjin Medical University, Tianjin, China. AD - Department of Spinal Surgery, Tianjin Union Medical Center, Tianjin, China. FAU - Liang, Shuang AU - Liang S AD - Tianjin Central Hospital of Gynecology and Obstetrics, Tianjin, China. FAU - Fan, Yonggang AU - Fan Y AD - School of Medicine, Nankai University, Tianjin, China. FAU - Zhao, Ying AU - Zhao Y AD - School of Medicine, Nankai University, Tianjin, China. FAU - Lv, Xinxin AU - Lv X AD - School of Medicine, Nankai University, Tianjin, China. FAU - Qiu, Caizhen AU - Qiu C AD - School of Medicine, Nankai University, Tianjin, China. FAU - Zhang, Lingzhu AU - Zhang L AD - School of Medicine, Nankai University, Tianjin, China. FAU - Liu, Yang AU - Liu Y AD - Department of Spinal Surgery, Tianjin Union Medical Center, Tianjin, China. FAU - Sun, Tianwei AU - Sun T AD - Tianjin Medical University, Tianjin, China. AD - Department of Spinal Surgery, Tianjin Union Medical Center, Tianjin, China. FAU - Chen, Xu AU - Chen X AD - Tianjin Central Hospital of Gynecology and Obstetrics, Tianjin, China. AD - Tianjin Key Laboratory of Human Development and Reproductive Regulation, Tianjin, China. FAU - Li, Shan-Shan AU - Li SS AD - School of Medicine, Nankai University, Tianjin, China. FAU - Jin, Xin AU - Jin X AD - School of Medicine, Nankai University, Tianjin, China. AD - Tianjin Central Hospital of Gynecology and Obstetrics, Tianjin, China. AD - Tianjin Key Laboratory of Human Development and Reproductive Regulation, Tianjin, China. LA - eng PT - Journal Article PL - United States TA - Invest Ophthalmol Vis Sci JT - Investigative ophthalmology & visual science JID - 7703701 RN - EC 2.3.1.- (Acetyltransferases) RN - 1CC1JFE158 (Dactinomycin) RN - 0 (Opsins) RN - 0 (Rod Opsins) RN - 63231-63-0 (RNA) RN - 0 (RNA, Messenger) SB - IM MH - Humans MH - Animals MH - *Acetyltransferases MH - *Zebrafish/genetics MH - Dactinomycin MH - Opsins MH - Rod Opsins MH - RNA/genetics MH - RNA, Messenger/genetics PMC - PMC10893899 COIS- Disclosure: H.-Z. Yang, None; D. Zhuo, None; Z. Huang, None; G. Luo, None; S. Liang, None; Y. Fan, None; Y. Zhao, None; X. Lv, None; C. Qiu, None; L. Zhang, None; Y. Liu, None; T. Sun, None; X. Chen, None; S.-S. Li, None; X. Jin, None EDAT- 2024/02/21 12:42 MHDA- 2024/02/22 06:43 PMCR- 2024/02/21 CRDT- 2024/02/21 11:33 PHST- 2024/02/22 06:43 [medline] PHST- 2024/02/21 12:42 [pubmed] PHST- 2024/02/21 11:33 [entrez] PHST- 2024/02/21 00:00 [pmc-release] AID - 2793385 [pii] AID - IOVS-23-38079 [pii] AID - 10.1167/iovs.65.2.31 [doi] PST - ppublish SO - Invest Ophthalmol Vis Sci. 2024 Feb 1;65(2):31. doi: 10.1167/iovs.65.2.31.