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| Status |
Public on Aug 04, 2016 |
| Title |
17beta-HSD type 12-like is responsible for maturation-inducing hormone synthesis during oocyte maturation in masu salmon |
| Organism |
Oncorhynchus masou masou |
| Experiment type |
Expression profiling by high throughput sequencing
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| Summary |
The maturation-inducing hormone 17alpha,20beta-dihydroxy-4-pregnen-3-one (DHP) was first identified in the amago salmon. However, although carbonyl reductase-like 20beta-hydroxysteroid dehydrogenase (CR/20beta-HSD) was reported to convert 17alpha-hydroxyprogesterone (17alpha-P) to DHP in rainbow trout, we previously found that CR/20beta-HSD mRNA was not up-regulated in stimulated granulosa cells from masu salmon, which suggests that DHP is synthesized by a different enzyme. Accordingly, the present study aimed to identify the specific 20beta-hydroxysteroid dehydrogenase (20beta-HSD) responsible for DHP production by granulosa cells during final oocyte maturation in masu salmon. Granulosa layers were isolated from ovarian follicles at one month before ovulation and incubated with or without forskolin, which was used to mimic luteinizing hormone. Subsequent RNA-sequencing yielded ~12 million reads, with an average length of 51 bp, and 71,062 contigs of >100 bp were constructed. Of the 953 contigs that were exclusively constructed from the reads of forskolin-induced granulosa layers, tBlastx analysis identified one contig (#f103496) that matched 17beta-hydroxysteroid dehydrogenase type 12. We found that mammalian cells transfected with full-length omhsd17beta12l exhibited considerable 20beta-HSD activity, as indicated by efficient conversion of exogenous 17alpha-P to DHP. In addition, we found that omhsd17beta12l mRNA levels were consistently low in follicles during vitellogenic growth; however, the levels increased significantly during final oocyte maturation. The levels of omhsd17beta12l mRNA were also considerably increased in granulosa layers in which 20beta-HSD activity was induced by salmon pituitary extract. Therefore, we suggest that omhsd17beta12l, not CR/20beta-HSD, is the 20beta-HSD responsible for DHP production by granulosa cells in masu salmon during final oocyte maturation.
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| Overall design |
Comparison of mRNA levels between control and forskolin-incubated sample.
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| Contributor(s) |
Ijiri S, Hagihara S |
| Citation(s) |
27967235 |
| BioProject |
PRJDB4997 |
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| Submission date |
Aug 03, 2016 |
| Last update date |
May 15, 2019 |
| Contact name |
Shigeho Ijiri |
| E-mail(s) |
[email protected]
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| Phone |
+81 138 40 5617
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| Organization name |
Graduate School of Fisheries Sciences, Hokkaido University
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| Department |
Division of Marine Life Sciences
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| Street address |
Minato-cho 3-1-1
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| City |
Hakodate |
| State/province |
Hokkaido |
| ZIP/Postal code |
041-8611 |
| Country |
Japan |
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| Platforms (1) |
| GPL22274 |
Illumina Genome Analyzer IIx (Oncorhynchus masou masou) |
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| Samples (2) |
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| Relations |
| SRA |
DRP003193 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE85134_RAW.tar |
1.1 Mb |
(http)(custom) |
TAR (of TXT) |
SRA Run Selector |
| Raw data are available in SRA |
| Processed data provided as supplementary file |
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