2021 Fiscal Year Research-status Report
A study of newly-identified testis-specific long non-coding RNAs in mouse spermatogenesis
| Project/Area Number |
20K18103
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| Research Institution | International University of Health and Welfare |
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Principal Investigator |
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| Project Period (FY) |
2020-04-01 – 2023-03-31
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| Keywords | testis-specific lncRNAs / spermatogenesis / arsenic |
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| Outline of Annual Research Achievements |
Testis-specific long non-protein coding RNAs (lncRNAs) are greatly focused as important regulators for mammalian spermatogenesis. Using IHC analysis, we found new testis-specific lncRNAs (1700101O22Rik and 1700108J01Rik), which are exclusively expressed in the mouse spermatogenetic cells. For functional study, we tried in vivo knockdown of these lncRNA using microinjection of retrovirus into the seminiferous tubules of mouse. We also tried in vitro functional analysis using mouse spermatogenetic cells (GC-1 cells). In vitro study, arsenic exposure of GC-1 cells showed down-regulation of 1700108J01Rik. Therefore, 1700108J01Rik may be an important target gene of arsenic and the downregulated expression of 1700108J01Rik may be closely related to male reproductive toxicity induced by arsenic.
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| Current Status of Research Progress |
Current Status of Research Progress
3: Progress in research has been slightly delayed.
Reason
Because of the pandemic outbreak of coronavirus, the performing of experiments were delayed than expected.
In vivo functional analysis, our target cells (spermatogonia and spermatocytes) were not successfully transfected by retrovirus. The transfection mainly expressed in the sertoli cells, in which the target testis-specific lncRNAs were not positive.
Currently, I am planning to perform RNAseq analysis in order to know the possible mechanisms or functions related to the target lncRNAs.
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| Strategy for Future Research Activity |
The main purpose of this study is to investigate the function of newly-identified testis-specific lncRNAs. From the in situ hybridization analysis, the target lncRNAs showed the stage-specific and spermatogenesis-related cells suggesting that these lncRNAs may involved in the mouse spermatogenesis.
As a supplemental part, I also tried arsenic exposure to mouse spermatogenetic cell lines.
As a future plan, I want to perform the RNA-sequence analysis of target gene knockdown and arsenic-exposed spermatogenetic cells to study the spermatogenetic related pathways and mechanisms.
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| Causes of Carryover |
I would like to perform RNA-seq analysis and investigate the possible target regulators for the target testis-specific lncRNAs in mouse spermatogenesis.
As a future plan, I would like to continue the pathway analysis or new regulators related to male reproductive functions and infertility.
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