In silico predicted transcriptional regulatory control of steroidogenesis in spawning female fathead minnows (Pimephales promelas)

Abstract

Oocyte development and maturation (or oogenesis) in spawning female fish is mediated by interrelated transcriptional regulatory and steroidogenesis networks. This study integrates a transcriptional regulatory network (TRN) model of steroidogenic enzyme gene expressions with a flux balance analysis (FBA) model of steroidogenesis. The two models were functionally related. Output from the TRN model (as magnitude gene expression simulated using extreme pathway (ExPa) analysis) was used to re-constrain linear inequality bounds for reactions in the FBA model. This allowed TRN model predictions to impact the steroidogenesis FBA model. These two interrelated models were tested as follows: First, in silico targeted steroidogenic enzyme gene activations in the TRN model showed high co-regulation (67–83%) for genes involved with oocyte growth and development (cyp11a1, cyp17-17,20-lyase, 3β-HSD and cyp19a1a). Whereas, no or low co-regulation corresponded with genes concertedly involved with oocyte final maturation prior to spawning (cyp17-17α-hydroxylase (0%) and 20β-HSD (33%)). Analysis (using FBA) of accompanying steroidogenesis fluxes showed high overlap for enzymes involved with oocyte growth and development versus those involved with final maturation and spawning. Second, the TRN model was parameterized with in vivo changes in the presence/absence of transcription factors (TFs) during oogenesis in female fathead minnows (Pimephales promelas). Oogenesis stages studied included: PreVitellogenic-Vitellogenic, Vitellogenic-Mature, Mature-Ovulated and Ovulated-Atretic stages. Predictions of TRN genes active during oogenesis showed overall elevated expressions for most genes during early oocyte development (PreVitellogenic-Vitellogenic, Vitellogenic-Mature) and post-ovulation (Ovulated-Atretic). Whereas ovulation (Mature-Ovulated) showed highest expression for cyp17-17α-hydroxylase only. FBA showed steroid hormone productions to also follow trends concomitant with steroidogenic enzyme gene expressions. General trends predicted by in silico modeling were similar to those observed in vivo. The integrated computational framework presented was capable of mechanistically representing aspects of reproductive function in fish. This approach can be extended to study reproductive effects under exposure to adverse environmental or anthropogenic stressors.