GEOapplication/xmlftp://ftp.ncbi.nlm.nih.gov/geo/series/GSE313nnn/GSE313040/primaryOK200TranscriptomicsFundulus heteroclitusExpression profiling by high throughput sequencinghttps://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE313040GEOGSEfalseNew insights into SREB orphan receptor roles in the ovary, using comparative transcriptomics across 3 fishesMummichog (Fundulus heteroclitus) ovary in vitro assays were completed using methods modified from Rajeswari et al., 2020. Briefly, female F. heteroclitus (n = 20) were euthanized in 300 mg/L buffered MS-222 (Syncaine®, tricaine methanesulfonate, Syndel, Ferndale, WA, USA), weighed, and measured (total length (mm), TL). 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 dimethyl sulfoxide (DMSO) control or one of three 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=4-6 replicates per treatment were incubated for 6 hours at 28 ℃ and each plate represented a temporal replicate. 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).2026/08/17GSE313040GSM9359623GSM9359624GSM9359613GSM9359625GSM9359614GSM9359615GSM9359626GSM9359627GSM9359616GSM9359628GSM9359617GSM9359618GSM9359619GSM9359620GSM9359621GSM935962234850313040Fundulus heteroclitus[42563402]