GEOapplication/xmlftp://ftp.ncbi.nlm.nih.gov/geo/series/GSE299nnn/GSE299271/primaryOK200TranscriptomicsDichotomyctere nigroviridisExpression profiling by high throughput sequencinghttps://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE299271GEOGSEfalseNew insights into SREB orphan receptor roles in the ovary, using comparative transcriptomics across 3 fishesGreen spotted pufferfish ovary in vitro assays were completed using methods modified from Rajeswari et al., 2020. Briefly, female D. nigroviridis (n = 14) were euthanized in 300 mg/L buffered MS-222 (Syncaine®, tricaine methanesulfonate, Syndel, Ferndale, WA, USA), weighed, and measured (total length (mm), TL). Green spotted pufferfish ovaries from each fish were partitioned into 5 sections of approximately equal size. Each of four of these sections were immersed into an assigned well within a 24-well plate containing 990 μl of culture media (90% Leibovitz’s L-15 medium [Sigma Cat# L1518], 1% Penicillin-Streptomycin [Gibco™ Cat# 15070-063], 0.5% bovine serum albumin [Sigma Cat# 81053N]), with either a DMSO control or one of 3 potential agonist treatments, 8535 (5 μM), PT-91 (5 μM), or PNX-20 (100 nM). Agonists were dissolved in DMSO and added to their respective wells in 10μl aliquots to achieve desired final concentrations in a final volume of 1ml in each well. The fifth section of ovary was photographed on a Sedgewick rafter cell with a Jenoptik ProgRes® C5 microscope camera to characterize egg stages for each fish. Four plates containing n=4-5 replicates per treatment were incubated for 6 hours at 28 ℃, each plate completed on different days to replicate the assay temporally. After incubation, ovarian tissues were snap frozen in RNAlater™ (Invitrogen Cat# AM7020) for transcriptome analysis and culture media were stored in glass tubes at -80°C for further steroid quantification analysis via liquid chromatography tandem mass spectrometry (LC-MS/MS).2026/08/17GSE299271GSM9036476GSM9036477GSM9036478GSM9036479GSM9036483GSM9036472GSM9036484GSM9036473GSM9036485GSM9036474GSM9036475GSM9036480GSM9036481GSM9036470GSM9036471GSM903648230566299271Dichotomyctere nigroviridis[42563402]