run_metadata: 76361
This data as json
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| 76361 | SRR24844105 | SRX20608296 | SRS17908189 | SRP441411 | PRJNA980839 | Adad1 knockout and misense mutant RNAseq | PRJNA980839 | Other | The double stranded RNA binding protein Adad1 adenosine deaminase domain containing 1 is a member of the adenosine deaminase acting on RNAs Adar protein family with germ cell specific expression. In mice Adad1 is necessary for sperm differentiation however its function outside of mammals has not been investigated. Here through an N ethyl N nitrosourea ENU based forward genetic screen we identified an adad1 mutant zebrafish line that develop as sterile males. Further histological examination revealed complete lack of germ cells in adult mutant fish however germ cells populated the gonad proliferated and entered meiosis in larval and juvenile fish. Although meiosis was initiated in adad1 mutant testes the spermatocytes failed to progress beyond the zygotene stage. Thus Adad1 is essential for meiosis and germline maintenance in zebrafish. We tested if spermatogonial stem cells were affected using nanos2 RNA FISH and a label retaining cell LRC assay and found that the mutant testes had fewer LRCs and nanos2 expressing cells compared to wild type siblings suggesting that failure to maintain the spermatogonial stem cells resulted in germ cell loss by maturity. To identify potential molecular processes regulated by Adad1 we sequenced bulk mRNA from mutants and wild type testes and found mis regulation of genes involved in RNA stability and modification pointing to a potential broader role in post transcriptional regulation. Our findings suggest that the RNA regulatory protein Adad1 is required for fertility through regulation of spermatogonial stem cell maintenance in zebrafish. | adad1 wildtype sibling control A | adad1 WT A | strain:Tuebingen|age:45dpf|dev stage:Juvenile|collection date:2021 03|geo loc name:USA:Boston|sex:male|tissue:Testis|genotype:WildtypeA|BioSampleModel:Model organism or animal | RNAseq of Danio rerio adad1 wildtype sibling: male testes | adad1 WT A | adad1 WT A | Total RNA was isolated from 45 dpf adad1Y67X Tg [ziwi:eGFP]uc1 and adad1M392K Tg [ddx4:eGFP] testes using the RNeasy micro kit Qiagen. Since the testes were very small at this age 5 testes were pooled together for each biological replicate 4 biological replicates for each genotype. Isolated RNA was sent to Genewiz at Azenta Life Sciences NJ for library preparation with poly A selection method and 150 bp paired end sequencing on an Illumina HiSeq platform. | RNA-Seq | TRANSCRIPTOMIC | Oligo-dT | PAIRED | ILLUMINA | Illumina HiSeq 2500 | SRP441411 | adad1-WT-A_R1_001.fastq.gz adad1-WT-A_R2_001.fastq.gz | fastq fastq | 8011954200.0 | 26706514.0 | adad1 WT A R1 001.fastq.gz | 0:150 1:150 | A:2090202005;C:1898897650;G:2026356221;T:1996371474;N:126850 | 150 | 150 | 2090202005 | 1898897650 | 2026356221 | 1996371474 | 126850 | SRX20608296 | SRS17908189 | SRA1650047 | University of Massachusetts Boston|Biology | University of Massachusetts Boston | 2 | 0.91189 | 0.91021 | 0.06102 | 0.06139 | 0.68525 | 0.68718 | 0.50676 | 0.50889 | 150 | 150 | B | B | biological fallback assumption | illumina | hiseq_era | unknown | poly_a | unknown | bulk | bulk | bulk | United States | 2023-06-07 | Juvenile | Juvenile | Gonad | Reproductive System |