A Testis-Specific Long Noncoding RNA, Start, Is a Regulator of Steroidogenesis in Mouse Leydig Cells
The testis expresses many long noncoding RNAs (lncRNAs), but their functions and overview of lncRNA variety are not well understood. The mouse Prss/Tessp locus contains six serine protease genes and two lncRNAs that have been suggested to play important roles in spermatogenesis. Here, we found a nov...
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doaj-aedf6664851947cb9938f3656dbc04fa2021-04-08T13:38:31ZengFrontiers Media S.A.Frontiers in Endocrinology1664-23922021-04-011210.3389/fendo.2021.665874665874A Testis-Specific Long Noncoding RNA, Start, Is a Regulator of Steroidogenesis in Mouse Leydig CellsKai Otsuka0Shin Matsubara1Akira Shiraishi2Natsumi Takei3Yui Satoh4Miho Terao5Shuji Takada6Shuji Takada7Tomoya Kotani8Tomoya Kotani9Honoo Satake10Atsushi P. Kimura11Atsushi P. Kimura12Graduate School of Life Science, Hokkaido University, Sapporo, JapanBioorganic Research Institute, Suntory Foundation for Life Sciences, Kyoto, JapanBioorganic Research Institute, Suntory Foundation for Life Sciences, Kyoto, JapanGraduate School of Life Science, Hokkaido University, Sapporo, JapanGraduate School of Life Science, Hokkaido University, Sapporo, JapanDepartment of Systems BioMedicine, National Research Institute for Child Health and Development, Tokyo, JapanDepartment of Systems BioMedicine, National Research Institute for Child Health and Development, Tokyo, JapanDepartment of NCCHD Child Health and Development, Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University, Tokyo, JapanGraduate School of Life Science, Hokkaido University, Sapporo, JapanDepartment of Biological Sciences, Faculty of Science, Hokkaido University, Sapporo, JapanBioorganic Research Institute, Suntory Foundation for Life Sciences, Kyoto, JapanGraduate School of Life Science, Hokkaido University, Sapporo, JapanDepartment of Biological Sciences, Faculty of Science, Hokkaido University, Sapporo, JapanThe testis expresses many long noncoding RNAs (lncRNAs), but their functions and overview of lncRNA variety are not well understood. The mouse Prss/Tessp locus contains six serine protease genes and two lncRNAs that have been suggested to play important roles in spermatogenesis. Here, we found a novel testis-specific lncRNA, Start (Steroidogenesis activating lncRNA in testis), in this locus. Start is 1822 nucleotides in length and was found to be localized mostly in the cytosol of germ cells and Leydig cells, although nuclear localization was also observed. Start-knockout (KO) mice generated by the CRISPR/Cas9 system were fertile and showed no morphological abnormality in adults. However, in adult Start-KO testes, RNA-seq and qRT-PCR analyses revealed an increase in the expression of steroidogenic genes such as Star and Hsd3b1, while ELISA analysis revealed that the testosterone levels in serum and testis were significantly low. Interestingly, at 8 days postpartum, both steroidogenic gene expression and testosterone level were decreased in Start-KO mice. Since overexpression of Start in two Leydig-derived cell lines resulted in elevation of the expression of steroidogenic genes including Star and Hsd3b1, Start is likely to be involved in their upregulation. The increase in expression of steroidogenic genes in adult Start-KO testes might be caused by a secondary effect via the androgen receptor autocrine pathway or the hypothalamus-pituitary-gonadal axis. Additionally, we observed a reduced number of Leydig cells at 8 days postpartum. Collectively, our results strongly suggest that Start is a regulator of steroidogenesis in Leydig cells. The current study provides an insight into the overall picture of the function of testis lncRNAs.https://www.frontiersin.org/articles/10.3389/fendo.2021.665874/fulllong noncoding RNAspermatogenesistestissteroidogenesisleydig cellknockout mouse |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Kai Otsuka Shin Matsubara Akira Shiraishi Natsumi Takei Yui Satoh Miho Terao Shuji Takada Shuji Takada Tomoya Kotani Tomoya Kotani Honoo Satake Atsushi P. Kimura Atsushi P. Kimura |
spellingShingle |
Kai Otsuka Shin Matsubara Akira Shiraishi Natsumi Takei Yui Satoh Miho Terao Shuji Takada Shuji Takada Tomoya Kotani Tomoya Kotani Honoo Satake Atsushi P. Kimura Atsushi P. Kimura A Testis-Specific Long Noncoding RNA, Start, Is a Regulator of Steroidogenesis in Mouse Leydig Cells Frontiers in Endocrinology long noncoding RNA spermatogenesis testis steroidogenesis leydig cell knockout mouse |
author_facet |
Kai Otsuka Shin Matsubara Akira Shiraishi Natsumi Takei Yui Satoh Miho Terao Shuji Takada Shuji Takada Tomoya Kotani Tomoya Kotani Honoo Satake Atsushi P. Kimura Atsushi P. Kimura |
author_sort |
Kai Otsuka |
title |
A Testis-Specific Long Noncoding RNA, Start, Is a Regulator of Steroidogenesis in Mouse Leydig Cells |
title_short |
A Testis-Specific Long Noncoding RNA, Start, Is a Regulator of Steroidogenesis in Mouse Leydig Cells |
title_full |
A Testis-Specific Long Noncoding RNA, Start, Is a Regulator of Steroidogenesis in Mouse Leydig Cells |
title_fullStr |
A Testis-Specific Long Noncoding RNA, Start, Is a Regulator of Steroidogenesis in Mouse Leydig Cells |
title_full_unstemmed |
A Testis-Specific Long Noncoding RNA, Start, Is a Regulator of Steroidogenesis in Mouse Leydig Cells |
title_sort |
testis-specific long noncoding rna, start, is a regulator of steroidogenesis in mouse leydig cells |
publisher |
Frontiers Media S.A. |
series |
Frontiers in Endocrinology |
issn |
1664-2392 |
publishDate |
2021-04-01 |
description |
The testis expresses many long noncoding RNAs (lncRNAs), but their functions and overview of lncRNA variety are not well understood. The mouse Prss/Tessp locus contains six serine protease genes and two lncRNAs that have been suggested to play important roles in spermatogenesis. Here, we found a novel testis-specific lncRNA, Start (Steroidogenesis activating lncRNA in testis), in this locus. Start is 1822 nucleotides in length and was found to be localized mostly in the cytosol of germ cells and Leydig cells, although nuclear localization was also observed. Start-knockout (KO) mice generated by the CRISPR/Cas9 system were fertile and showed no morphological abnormality in adults. However, in adult Start-KO testes, RNA-seq and qRT-PCR analyses revealed an increase in the expression of steroidogenic genes such as Star and Hsd3b1, while ELISA analysis revealed that the testosterone levels in serum and testis were significantly low. Interestingly, at 8 days postpartum, both steroidogenic gene expression and testosterone level were decreased in Start-KO mice. Since overexpression of Start in two Leydig-derived cell lines resulted in elevation of the expression of steroidogenic genes including Star and Hsd3b1, Start is likely to be involved in their upregulation. The increase in expression of steroidogenic genes in adult Start-KO testes might be caused by a secondary effect via the androgen receptor autocrine pathway or the hypothalamus-pituitary-gonadal axis. Additionally, we observed a reduced number of Leydig cells at 8 days postpartum. Collectively, our results strongly suggest that Start is a regulator of steroidogenesis in Leydig cells. The current study provides an insight into the overall picture of the function of testis lncRNAs. |
topic |
long noncoding RNA spermatogenesis testis steroidogenesis leydig cell knockout mouse |
url |
https://www.frontiersin.org/articles/10.3389/fendo.2021.665874/full |
work_keys_str_mv |
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