title: Suppression of Neu1 sialidase delays the absorption of yolk sac in medaka (Oryzias latipes) accompanied with the accumulation of α2-3 sialo-glycoproteins.
abstract: Sialidase catalyzes the removal of sialic acids from glycoconjugates. Recently, medaka sialidase Neu1 has been cloned and its enzymatic properties were investigated. Although enzymatic properties of this sialidase, such as optimal pH and substrate specificity, exhibits high similarity with human NEU1, Neu1 physiological functions in fish are still unclear. Here, to understand Neu1 significance in medaka embryogenesis, sialidase translation knockdown was carried out with one-cell stage fertilized egg using morpholino oligo injection. Neu1 exhibited desialylation of α2-3 sialic acid linkage in vitro and lysosomal localization in medaka caudal fin primary cells. Chloroquine treatment, inhibitor of lysosomal enzymes, caused an accumulation of α2-3 sialo-glycoproteins in the primary cells. During the embryogenesis neu1 mRNA level was elevated until 3.5 day post fertilization (dpf) while an initial decrease of α2-3 sialo-glycoprotein was observed around the same developmental stage. Neu1 knockdown by morpholino oligo induced some abnormal phenotypes such as delay of yolk sac absorption and small embryos. Sialidase-knockdown embryos also showed increase of heart rate in 5.5 and 6.5 dpf. Furthermore, about 37% decrease of hatching rate was observed in Neu1-MO treated embryos compared with control MO. Embryos showing severe phenotypes stopped embryogenesis at the late stage of development. Alteration of embryonic sialo-glycoproteins induced by morpholino injection was examined by lectin blotting to clarify the mechanism of abnormal development. As a result, degradation of several α2-3 sialo-glycoproteins was suppressed in Neu1-MO embryo, possibly induced by the interruption of lysosomal desialylation toward yolk glycoprotein. Our results suggest that medaka Neu1 could be crucial for embryonic development through the degradation of yolk sac nutrition.
authors.affiliation: Faculty of Fisheries, Kagoshima University, Kagoshima, Japan.
authors.forename: Sena
authors.initials: S
authors.lastname: Ryuzono
authors.affiliation: Faculty of Fisheries, Kagoshima University, Kagoshima, Japan.
authors.forename: Ryo
authors.initials: R
authors.lastname: Takase
authors.affiliation: Faculty of Fisheries, Kagoshima University, Kagoshima, Japan.
authors.forename: Yuko
authors.initials: Y
authors.lastname: Kamada
authors.affiliation: Faculty of Science, Kagoshima University, Kagoshima, Japan.
authors.forename: Takanori
authors.initials: T
authors.lastname: Ikenaga
authors.affiliation: Faculty of Fisheries, Kagoshima University, Kagoshima, Japan; The United Graduate School of Agricultural Sciences, Kagoshima University, Kagoshima, Japan.
authors.forename: Petros Kingstone
authors.initials: PK
authors.lastname: Chigwechokha
authors.affiliation: Faculty of Fisheries, Kagoshima University, Kagoshima, Japan; The United Graduate School of Agricultural Sciences, Kagoshima University, Kagoshima, Japan.
authors.forename: Masaharu
authors.initials: M
authors.lastname: Komatsu
authors.affiliation: Faculty of Fisheries, Kagoshima University, Kagoshima, Japan; The United Graduate School of Agricultural Sciences, Kagoshima University, Kagoshima, Japan. Electronic address: shiozaki@fish.kagoshima-u.ac.jp.
authors.forename: Kazuhiro
authors.initials: K
authors.lastname: Shiozaki
mesh_terms: D000818:Animals; D006023:Glycoproteins / Q000235:genetics / Q000378:metabolism; D009439:Neuraminidase / Q000235:genetics / Q000378:metabolism*; D009990:Oryzias / Q000196:embryology* / Q000378:metabolism*; D012333:RNA, Messenger / Q000378:metabolism; D015017:Yolk Sac / Q000378:metabolism
chemical_list: D006023:Glycoproteins; D012333:RNA, Messenger; D009439:Neuraminidase
keywords: Development; Glycoprotein; Lysosome; Medaka; Sialidase