New insights into SREB orphan receptor roles in the ovary, using comparative transcriptomics across 3 fishes

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.