Abnormal development of zebrafish after knockout and knockdown of ribosomal protein L10a

• Published in: Scientific reports Vol. 9; no. 1; pp. 18130 - 11
• Main Authors: Palasin, Kunwadee, Uechi, Tamayo, Yoshihama, Maki, Srisowanna, Naparee, Choijookhuu, Narantsog, Hishikawa, Yoshitaka, Kenmochi, Naoya, Chotigeat, Wilaiwan
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 02-12-2019; Nature Publishing Group
• Subjects: 38; 38/23; 38/70; 45; 45/29; 45/44; 45/77; 45/90; 631/136/2086; 631/208/199; 64; 64/116; Animals; Antisense oligonucleotides; Apoptosis; Clonal deletion; CRISPR; CRISPR-Cas Systems; Cytology; Danio rerio; DEAD-box RNA Helicases - genetics; Edema; Embryo, Nonmammalian - abnormalities; Embryogenesis; Embryonic growth stage; Embryos; Erythrocytes; Erythropoiesis - genetics; Fertilization; GATA-1 protein; GATA1 Transcription Factor - genetics; Gene deletion; Gene Expression Regulation, Developmental; Gene Knockdown Techniques; Gene Knockout Techniques; Genomes; Germ Cells - physiology; Hemoglobins - genetics; Humanities and Social Sciences; Morphology; mRNA; multidisciplinary; Oligonucleotides, Antisense; p53 Protein; Ribosomal protein L10a; RNA-Binding Proteins - genetics; RNA-Binding Proteins - metabolism; Science; Science (multidisciplinary); Tumor Suppressor Protein p53 - genetics; Yolk; Yolk sac; Zebrafish; Zebrafish - embryology; Zebrafish - genetics; Zebrafish Proteins - genetics; Zebrafish Proteins - metabolism
• ISSN: 2045-2322; 2045-2322
• DOI: 10.1038/s41598-019-54544-w

In this study, to investigate the secondary function of Rpl10a in zebrafish development, morpholino antisense oligonucleotides (MOs) were used to knock down the zebrafish ribosomal protein L10a (
rpl10a
). At 25 hpf (hours post-fertilization), embryos injected with the
rpl10a
MO showed an abnormal morphology, including short bodies, curved tails, and small yolk sac extensions. We observed pigment reductions, edema, larger yolk sacs, smaller eyes and smaller yolk sac extensions at 50 hpf. In addition, reductions in the expression of primordial germ cell (PGC) marker genes (
nanos1
and
vasa
) were observed in
rpl10a
knockdown embryos. A rescue experiment using a
rpl10a
mRNA co-injection showed the recovery of the morphology and red blood cell production similar to wild-type. Moreover, the CRISPR-Cas9 system was used to edit the sequence of
rpl10a
exon 5, resulting in a homozygous 5-bp deletion in the zebrafish genome. The mutant embryos displayed a morphology similar to that of the knockdown animals. Furthermore, the loss of
rpl10a
function led to reduced expression of
gata1
,
hbae3
, and
hbbe1
(erythroid synthesis) and increased
tp53
expression. Overall, the results suggested that Rpl10a deficiency caused delays in embryonic development, as well as apoptosis and anemia, in zebrafish.