{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,12]],"date-time":"2025-10-12T03:19:47Z","timestamp":1760239187730,"version":"build-2065373602"},"reference-count":18,"publisher":"MDPI AG","issue":"10","license":[{"start":{"date-parts":[[2020,10,21]],"date-time":"2020-10-21T00:00:00Z","timestamp":1603238400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Diagnostics"],"abstract":"<jats:p>Mucopolysaccharidosis type II (MPS II) is an X-linked disorder resulting from a deficiency in iduronate 2-sulfatase (IDS), which is reported to be caused by gene mutations in the iduronate 2-sulfatase (IDS) gene. Many IDS mutation sites have not yet had their causal relationship with MPS II characterized. We employed a gain-of-function strategy whereby we microinjected different mutated zebrafish ids (z-ids) mRNAs corresponded to human IDS gene into zebrafish embryos, and then measured their total IDS enzymatic activity and observed the occurrence of defective phenotypes during embryonic development. We examined three known mutation sites for human IDS genes (h-IDS) associated with MPS II symptoms, including h-IDS-P86L, -S333L and -R468W, which corresponded to z-ids-P80L, -S327L and -R454W. When these three mutated z-ids mRNAs were overexpressed in zebrafish embryos, the IDS enzymatic activity of the total proteins extracted from the injected embryos was not increased compared with the endogenous IDS of the untreated embryos, which suggests that the IDS enzymatic activity of these three mutated z-ids was totally lost, as expected. Additionally, we observed defective phenotypes in these injected embryos, resulting from the failed IDS enzyme breakdown, which, in turn, has a dominant negative effect on the endogenous wild-type IDS function. These phenotypes were similar to the clinical symptoms observed in MPS II pathogenesis. We further studied six uncharacterized IDS mutation sites as identified by the Taiwanese MPS newborn screening programs. We propose a novel IDS enzyme activity assay combined with phenotypic observation in zebrafish embryos, as an alternative platform for quickly providing a valuable index for preliminarily assessment of any identified IDS point mutation gene that has not yet been characterized, in the context of its role in MPS II development.<\/jats:p>","DOI":"10.3390\/diagnostics10100854","type":"journal-article","created":{"date-parts":[[2020,10,21]],"date-time":"2020-10-21T10:14:22Z","timestamp":1603275262000},"page":"854","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["Effect of Mutated ids Overexpression on IDS Enzyme Activity and Developmental Phenotypes in Zebrafish Embryos: A Valuable Index for Assessing Critical Point-Mutations Associated with Mucopolysaccharidosis Type II Occurrence in Humans"],"prefix":"10.3390","volume":"10","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-9570-0218","authenticated-orcid":false,"given":"Cheng-Yung","family":"Lin","sequence":"first","affiliation":[{"name":"Institute of Biomedical Sciences, MacKay Medical College, New Taipei City 25245, Taiwan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9619-0990","authenticated-orcid":false,"given":"Hsiang-Yu","family":"Lin","sequence":"additional","affiliation":[{"name":"Institute of Biomedical Sciences, MacKay Medical College, New Taipei City 25245, Taiwan"},{"name":"Department of Medicine, MacKay Medical College, New Taipei City 25245, Taiwan"},{"name":"Department of Pediatrics, MacKay Memorial Hospital, Taipei 10449, Taiwan"},{"name":"Department of Medical Research, MacKay Memorial Hospital, New Taipei City 25160, Taiwan"},{"name":"MacKay Junior College of Medicine, Nursing and Management, Taipei 11260, Taiwan"},{"name":"Department of Medical Research, China Medical University Hospital, China Medical University, Taichung 40402, Taiwan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9776-2092","authenticated-orcid":false,"given":"Chih-Kuang","family":"Chuang","sequence":"additional","affiliation":[{"name":"Department of Medical Research, MacKay Memorial Hospital, New Taipei City 25160, Taiwan"},{"name":"College of Medicine, Fu-Jen Catholic University, Taipei 24205, Taiwan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Po-Hsiang","family":"Zhang","sequence":"additional","affiliation":[{"name":"Institute of Biomedical Sciences, MacKay Medical College, New Taipei City 25245, Taiwan"},{"name":"Department of Medical Research, MacKay Memorial Hospital, New Taipei City 25160, Taiwan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ru-Yi","family":"Tu","sequence":"additional","affiliation":[{"name":"Department of Medical Research, MacKay Memorial Hospital, New Taipei City 25160, Taiwan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Shuan-Pei","family":"Lin","sequence":"additional","affiliation":[{"name":"Department of Medicine, MacKay Medical College, New Taipei City 25245, Taiwan"},{"name":"Department of Pediatrics, MacKay Memorial Hospital, Taipei 10449, Taiwan"},{"name":"Department of Medical Research, MacKay Memorial Hospital, New Taipei City 25160, Taiwan"},{"name":"Department of Infant and Child Care, National Taipei University of Nursing and Health Sciences, Taipei 11219, Taiwan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Huai-Jen","family":"Tsai","sequence":"additional","affiliation":[{"name":"Institute of Biomedical Sciences, MacKay Medical College, New Taipei City 25245, Taiwan"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2020,10,21]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"D\u2019Avanzo, F., Rigon, L., Zanetti, A., and Tomanin, R. (2020). Mucopolysaccharidosis Type II: One Hundred Years of Research, Diagnosis, and Treatment. Int. J. Mol. Sci., 21.","DOI":"10.3390\/ijms21041258"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"960","DOI":"10.1002\/ajmg.a.32781","article-title":"Incidence of the mucopolysaccharidoses in Taiwan, 1984\u20132004","volume":"149","author":"Lin","year":"2009","journal-title":"Am. J. Med. Genet. Part A"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"227","DOI":"10.1016\/j.ymgme.2017.05.016","article-title":"Epidemiology of mucopolysaccharidoses","volume":"121","author":"Khan","year":"2017","journal-title":"Mol. Genet. Metab."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"449","DOI":"10.1002\/humu.9128","article-title":"Mutational spectrum of the iduronate 2 sulfatase gene in 25 unrelated Korean Hunter syndrome patients: Identification of 13 novel mutations","volume":"21","author":"Kim","year":"2003","journal-title":"Hum. Mutat."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"395","DOI":"10.1007\/s10038-005-0266-4","article-title":"Mutational and structural analysis of Japanese patients with mucopolysaccharidosis type II","volume":"50","author":"Kato","year":"2005","journal-title":"J. Hum. Genet."},{"key":"ref_6","first-page":"3421","article-title":"The mucopolysaccharidoses","volume":"Volume 136","author":"Scriver","year":"2001","journal-title":"The Metabolic and Molecular Bases of Inherited Disease"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"84","DOI":"10.1186\/s13023-018-0816-4","article-title":"Status of newborn screening and follow up investigations for Mucopolysaccharidoses I and II in Taiwan","volume":"13","author":"Chuang","year":"2018","journal-title":"Orphanet J. Rare Dis."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"176","DOI":"10.1016\/j.jpeds.2018.09.063","article-title":"Taiwan National Newborn Screening Program by Tandem Mass Spectrometry for Mucopolysaccharidoses Types I, II, and VI","volume":"205","author":"Chan","year":"2019","journal-title":"J. Pediatr."},{"key":"ref_9","doi-asserted-by":"crossref","unstructured":"Lin, H.-Y., Tu, R.-Y., Chern, S.-R., Lo, Y.-T., Fran, S., Wei, F.-J., Huang, S.-F., Tsai, S.-Y., Chang, C.-Y., and Lee, C.-L. (2020). Identification and Functional Characterization of IDS Gene Mutations Underlying Taiwanese Hunter Syndrome (Mucopolysaccharidosis Type II). Int. J. Mol. Sci., 21.","DOI":"10.3390\/ijms21010114"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"43","DOI":"10.1016\/j.matbio.2009.09.001","article-title":"A novel functional role of iduronate-2-sulfatase in zebrafish early development","volume":"29","author":"Moro","year":"2010","journal-title":"Matrix Biol."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"1643","DOI":"10.1093\/hmg\/ddx069","article-title":"Perturbations in cell signaling elicit early cardiac defects in mucopolysaccharidosis type II","volume":"26","author":"Costa","year":"2017","journal-title":"Hum. Mol. Genet."},{"key":"ref_12","doi-asserted-by":"crossref","unstructured":"Stainier, D.Y.R., Raz, E., Lawson, N.D., Ekker, S.C., Burdine, R.D., Eisen, J.S., Ingham, P.W., Schulte-Merker, S., Yelon, D., and Weinstein, B.M. (2017). Guidelines for morpholino use in zebrafish. PLoS Genet., 13.","DOI":"10.1371\/journal.pgen.1007000"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"2262","DOI":"10.1093\/hmg\/ddy131","article-title":"FGF signaling deregulation is associated with early developmental skeletal defects in animal models for mucopolysaccharidosis type II (MPS II)","volume":"27","author":"Bellesso","year":"2018","journal-title":"Hum. Mol. Genet."},{"key":"ref_14","unstructured":"Westerfield, M. (1995). The Zebrafish Book, University of Oregon Press. [3rd ed.]."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"486","DOI":"10.1089\/zeb.2013.0903","article-title":"Normal Function of Myf5 During Gastrulation Is Required for Pharyngeal Arch Cartilage Development in Zebrafish Embryos","volume":"10","author":"Lin","year":"2013","journal-title":"Zebrafish"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"e49175","DOI":"10.7554\/eLife.49175","article-title":"Extracellular Pgk1 enhances neurite outgrowth of motoneurons through Nogo66\/NgR-independent targeting of NogoA","volume":"8","author":"Lin","year":"2019","journal-title":"eLife"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"237","DOI":"10.1136\/adc.79.3.237","article-title":"Mutation analysis in 57 unrelated patients with MPS II (Hunter\u2019s disease)","volume":"79","author":"Vafiadaki","year":"1998","journal-title":"Arch. Dis. Child."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"29","DOI":"10.1016\/j.cca.2006.01.001","article-title":"Detection of hunter syndrome (mucopolysaccharidosis type II) in Taiwanese: Biochemical and linkage studies of the iduronate-2-sulfatase gene defects in MPS II patients and carriers","volume":"369","author":"Lin","year":"2006","journal-title":"Clin. Chim. 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