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Series GSE69417 Query DataSets for GSE69417
Status Public on Aug 10, 2016
Title Developmental regulation of piRNAs during spermatogenesis in Drosophila melanogaster
Organism Drosophila melanogaster
Experiment type Non-coding RNA profiling by high throughput sequencing
Summary The piRNA pathway is studied in great detail in Drosophila female germline. In this study we show that unlike the female germline where all Piwi proteins are expressed throughout oogenesis, Ago3 - a Piwi family protein shows a spatial expression male germline. To understand dynamics of piRNA pathway during spermatogonia and primary spermatocyte stages of male germline development, we used arrest mutants. The bag of marbles (bam) and benign gonial cell neoplasm (bgcn) mutants have only early mitotic dividing germline cells in the testes due to failure to progress to primary spermatocyte stage, the cannonball (can) and spermatocyte arrest (sa) mutant germline cells cannot progress beyond primary spermatocyte stage. To investigate the dynamics of the piRNA pathway during spermatogenesis in spermatogonia and primary spermatocyte stages, we used testicular tissues from these stage-specific arrested mutants. While we used entire bam and bgcn mutant testes for spermatogonia purification, we while we manually removed the apical regions of can and sa mutant testes to exclude mitotically dividing undifferentiated germline cells for primary spermatocytes purification. Our results show that piRNAs mapping to transposons are more abundant in spermatogonia, whereas those mapping to Suppressor of Stellate [Su(Ste)] and AT-chX are mostly expressed in primary spermatocytes. Furthermore we observed that transposon-mapping piRNAs with ping-pong signature are more abundant in spermatogonia albeit still detectable in primary spermatocytes where Ago3 is not expressed. These results suggest that robust piRNA production via ping-pong cycle takes place in spermatogonia, and to a lesser extent in primary spermatocytes even in the absence of Ago3. Consistently, piRNAs from ago3 mutant testes also exhibit the ping-pong signature, confirming that a non-canonical ping-pong cycle is acting during spermatogenesis. Our study provides a developmental dimension to the piRNA pathway and uncovers a new mechanism used in the male germline to silence transposons.
Overall design The difference in piRNA from spermatogonia and primary spermatocyte stages was studied by comparing small RNAs from bam and bgcn mutant testis, which represent spermatogonia stages with the small RNAs from apex removed can and sa testis, representing primary spermatocyte stages. In the study we also studied effect of loss of Piwi family proteins Aub and Ago3, which have different spatial expression during male germline development.
Contributor(s) Anand A, Kai T
Citation(s) 27208314
Submission date Jun 01, 2015
Last update date May 15, 2019
Contact name Toshie Kai
Organization name Temasek Life Sciences Laboratory
Street address 1 Research Link, National University of Singapore
City Singapore
ZIP/Postal code 117604
Country Singapore
Platforms (1)
GPL13304 Illumina HiSeq 2000 (Drosophila melanogaster)
Samples (9)
GSM1700916 bam mutant small RNA-Seq
GSM1700917 bgcn mutant small RNA-Seq
GSM1700918 can mutant small RNA-Seq
BioProject PRJNA285464
SRA SRP058915

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Supplementary file Size Download File type/resource
GSE69417_RAW.tar 500.0 Kb (http)(custom) TAR (of XLSX)
GSE69417_aub-ago3-transposon-mapping-small-RNAs.xlsx 63.3 Kb (ftp)(http) XLSX
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Raw data are available in SRA
Processed data provided as supplementary file
Processed data are available on Series record

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