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Abnormal development of zebrafish after knockout and knockdown of ribosomal protein L10a.
- Source :
-
Scientific reports [Sci Rep] 2019 Dec 02; Vol. 9 (1), pp. 18130. Date of Electronic Publication: 2019 Dec 02. - Publication Year :
- 2019
-
Abstract
- 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.
- Subjects :
- Animals
CRISPR-Cas Systems
DEAD-box RNA Helicases genetics
Erythropoiesis genetics
GATA1 Transcription Factor genetics
Gene Knockdown Techniques
Gene Knockout Techniques
Germ Cells physiology
Oligonucleotides, Antisense
RNA-Binding Proteins metabolism
Tumor Suppressor Protein p53 genetics
Zebrafish embryology
Zebrafish Proteins metabolism
Embryo, Nonmammalian abnormalities
Gene Expression Regulation, Developmental
Hemoglobins genetics
RNA-Binding Proteins genetics
Zebrafish genetics
Zebrafish Proteins genetics
Subjects
Details
- Language :
- English
- ISSN :
- 2045-2322
- Volume :
- 9
- Issue :
- 1
- Database :
- MEDLINE
- Journal :
- Scientific reports
- Publication Type :
- Academic Journal
- Accession number :
- 31792295
- Full Text :
- https://doi.org/10.1038/s41598-019-54544-w