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| Status |
Public on Aug 02, 2024 |
| Title |
Essential functions of RNA helicase Vasa in Drosophila spermatogenesis, from maintenance of germline stem cells to passage through meiosis between Drosophila melanogaster and Drosophila simulans [RNA-seq] |
| Organism |
Drosophila melanogaster |
| Experiment type |
Expression profiling by high throughput sequencing
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| Summary |
DEAD-box RNA helicase Vasa is required for gonad development and fertility in multiple animals. In Drosophila, Vasa performs essential functions in oogenesis, including the maintenance of germline stem cells (GSCs), piRNA silencing of mobile elements, translation regulation, and primordial germ cell specification. Despite its evident significance, the mechanistic basis of Vasa action and its precise role in spermatogenesis become incomprehensible. Several papers affirm the fertility of males carrying vasa mutations. However, it is also shown that Vasa is essential for piRNA-mediated repression of Stellate genes needed for the maintenance of male fertility.Here we found that loss-of-function vasa mutations led to a rapid decline in GSC maintenance in the testes, a severe loss of total germ cell content, and a strong decrease in male fertility over time. With the aid of analysis of small RNA libraries, we revealed that collapse of piRNA biogenesis in the absence of vasa expression. Despite that, we did not reveal increasing cell death events in the early germ cells of vasa mutant testes. The introduction of the transgene rhino copy, encoding a nuclear component of the piRNA pathway, in vasa mutant background allowed us to rescue premeiotic stages of spermatogenesis, including GSC maintenance and the development of spermatogonia and spermatocytes. However, the progression of spermatocytes through meiosis and the fertility of the rhino transgene-rescued males were disrupted by strong Stellate gene derepression owing to the absence of corresponding piRNAs. We have shown that Vasa functions in spermatogenesis are essential at two separate developmental stages: in GSCs for their maintenance and in spermatocytes for the repression of Stellate genes.
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| Overall design |
To study the contribution of Vasa in Stellate silencing in more detail, we prepared and sequenced on the Illumina platform whole-transcriptome libraries and 18–29 nt small RNAs isolated from the testes of vasa mutants, control heterozygous males, and males with the transgenic construct rhi-GFP in the background of vasa mutations.
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| Contributor(s) |
Kotov AA |
| Citation(s) |
39184915 |
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| Submission date |
Jun 16, 2024 |
| Last update date |
Sep 27, 2024 |
| Contact name |
Alexei A. Kotov |
| E-mail(s) |
kotov_alexei@mail.ru
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| Organization name |
Koltzov Institute of Developmental Biology of the Russian Academy of Sciences
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| Street address |
26 Vavilov Street
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| City |
Moscow |
| ZIP/Postal code |
119334 |
| Country |
Russia |
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| Platforms (1) |
| GPL25244 |
Illumina NovaSeq 6000 (Drosophila melanogaster) |
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| Samples (3) |
| GSM8332113 |
D. melanogaster vigEP812 +/-, testis |
| GSM8332114 |
D. melanogaster vigEP812 -/-, testis |
| GSM8332115 |
D. melanogaster vigEP812 -/- RhiGFP, testis |
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| Relations |
| BioProject |
PRJNA1124564 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE269988_vasa_Ste_SuSte_RNAseq.xlsx |
5.3 Kb |
(ftp)(http) |
XLSX |
SRA Run Selector |
| Raw data are available in SRA |
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