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Establishment of an organ culture system to induce Sertoli cell differentiation from undifferentiated mouse gonads
Organ culture systems are useful for elucidating the process of testicular differentiation from mammalian undifferentiated genetically male gonads, as they permit various experiments, including experiments involving the control of gene expression. However, without addition of testicular differentiat...
Autores principales: | , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
The Japanese Society of Veterinary Science
2020
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7192728/ https://www.ncbi.nlm.nih.gov/pubmed/32092744 http://dx.doi.org/10.1292/jvms.20-0036 |
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author | HASEGAWA, Chinatsu YOKOYAMA, Toshifumi UMEMURA, Yuria KAWANISHI, Kohei MIURA, Yuuka TAKADA, Nanako OHNO, Shuji ONARU, Kanoko OMOTEHARA, Takuya HIRANO, Tetsushi MANTANI, Yohei Miki, Takanori HOSHI, Nobuhiko |
author_facet | HASEGAWA, Chinatsu YOKOYAMA, Toshifumi UMEMURA, Yuria KAWANISHI, Kohei MIURA, Yuuka TAKADA, Nanako OHNO, Shuji ONARU, Kanoko OMOTEHARA, Takuya HIRANO, Tetsushi MANTANI, Yohei Miki, Takanori HOSHI, Nobuhiko |
author_sort | HASEGAWA, Chinatsu |
collection | PubMed |
description | Organ culture systems are useful for elucidating the process of testicular differentiation from mammalian undifferentiated genetically male gonads, as they permit various experiments, including experiments involving the control of gene expression. However, without addition of testicular differentiation-related factors, it is difficult to induce the formation of testis cord from immature gonads by a time point earlier 12 tail somites (ts) that corresponding to 11.0 days post coitum (dpc). In this study, we attempted to establish an organ culture system that induces testis formation from immature gonads (around 8 ts: 10.5 dpc) just before Sry (sex-determining region of the Y chromosome) expression. A paired gonad-mesonephros complex of around 8 ts was placed in the groove of an agarose gel block and put the semi-cylindrical piece of agarose gel to maintain the gonad morphology. The gonads were cultured in the gas phase for 96 hr. As a result, testis cord-like structures appeared in many genetically male gonads. Cells expressing the Sertoli cell markers Sox9 (SRY-box 9) and Amh (anti-Müllerian hormone) were observed, while granulosa cell marker Foxl2 (forkhead box L2) was not detected. In addition, Sox9- and Amh-expressing cells were observed throughout the entire gonad in many individuals. Amh mRNA expression was also upregulated. Surprisingly, formation of a partial testicular structure was observed from more immature gonads (6 ts). These results show that our gonadal organ culture system is useful for elucidating the regulation mechanism of Sry expression in undifferentiated bipotential gonads. |
format | Online Article Text |
id | pubmed-7192728 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | The Japanese Society of Veterinary Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-71927282020-05-06 Establishment of an organ culture system to induce Sertoli cell differentiation from undifferentiated mouse gonads HASEGAWA, Chinatsu YOKOYAMA, Toshifumi UMEMURA, Yuria KAWANISHI, Kohei MIURA, Yuuka TAKADA, Nanako OHNO, Shuji ONARU, Kanoko OMOTEHARA, Takuya HIRANO, Tetsushi MANTANI, Yohei Miki, Takanori HOSHI, Nobuhiko J Vet Med Sci Anatomy Organ culture systems are useful for elucidating the process of testicular differentiation from mammalian undifferentiated genetically male gonads, as they permit various experiments, including experiments involving the control of gene expression. However, without addition of testicular differentiation-related factors, it is difficult to induce the formation of testis cord from immature gonads by a time point earlier 12 tail somites (ts) that corresponding to 11.0 days post coitum (dpc). In this study, we attempted to establish an organ culture system that induces testis formation from immature gonads (around 8 ts: 10.5 dpc) just before Sry (sex-determining region of the Y chromosome) expression. A paired gonad-mesonephros complex of around 8 ts was placed in the groove of an agarose gel block and put the semi-cylindrical piece of agarose gel to maintain the gonad morphology. The gonads were cultured in the gas phase for 96 hr. As a result, testis cord-like structures appeared in many genetically male gonads. Cells expressing the Sertoli cell markers Sox9 (SRY-box 9) and Amh (anti-Müllerian hormone) were observed, while granulosa cell marker Foxl2 (forkhead box L2) was not detected. In addition, Sox9- and Amh-expressing cells were observed throughout the entire gonad in many individuals. Amh mRNA expression was also upregulated. Surprisingly, formation of a partial testicular structure was observed from more immature gonads (6 ts). These results show that our gonadal organ culture system is useful for elucidating the regulation mechanism of Sry expression in undifferentiated bipotential gonads. The Japanese Society of Veterinary Science 2020-02-21 2020-04 /pmc/articles/PMC7192728/ /pubmed/32092744 http://dx.doi.org/10.1292/jvms.20-0036 Text en ©2020 The Japanese Society of Veterinary Science This is an open-access article distributed under the terms of the Creative Commons Attribution Non-Commercial No Derivatives (by-nc-nd) License. (CC-BY-NC-ND 4.0: https://creativecommons.org/licenses/by-nc-nd/4.0/) |
spellingShingle | Anatomy HASEGAWA, Chinatsu YOKOYAMA, Toshifumi UMEMURA, Yuria KAWANISHI, Kohei MIURA, Yuuka TAKADA, Nanako OHNO, Shuji ONARU, Kanoko OMOTEHARA, Takuya HIRANO, Tetsushi MANTANI, Yohei Miki, Takanori HOSHI, Nobuhiko Establishment of an organ culture system to induce Sertoli cell differentiation from undifferentiated mouse gonads |
title | Establishment of an organ culture system to induce Sertoli cell
differentiation from undifferentiated mouse gonads |
title_full | Establishment of an organ culture system to induce Sertoli cell
differentiation from undifferentiated mouse gonads |
title_fullStr | Establishment of an organ culture system to induce Sertoli cell
differentiation from undifferentiated mouse gonads |
title_full_unstemmed | Establishment of an organ culture system to induce Sertoli cell
differentiation from undifferentiated mouse gonads |
title_short | Establishment of an organ culture system to induce Sertoli cell
differentiation from undifferentiated mouse gonads |
title_sort | establishment of an organ culture system to induce sertoli cell
differentiation from undifferentiated mouse gonads |
topic | Anatomy |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7192728/ https://www.ncbi.nlm.nih.gov/pubmed/32092744 http://dx.doi.org/10.1292/jvms.20-0036 |
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