A CONSTRAINT-BASED STOICHIOMETRIC MODEL OF THE STEROIDOGENIC NETWORK OF ZEBRAFISH (DANIO RERIO)

Abstract

The metabolic process of steroidogenesis exhibits a complex biochemical network topology as the activity of various steroidogenic enzymes control cholesterol metabolism to steroid hormone derivatives. In this paper, a stoichiometric reconstruction of a sub-set of 65 reactions from the zebrafish (Danio rerio) steroidogenic network is presented and simulated using uniform reaction constraints. The reconstruction defined a set of 65 enzyme catalyzed reactions and 37 exchange or transport reactions for steroid metabolites. The reconstructed reactions were inclusive of cholesterol and androgen/estrogen metabolism. Biased (statement of network objective function) and un-biased (no statement of objective function) analyses were applied to identify network properties dependent on reaction stoichiometry. Random sampling of flux distributions through the network identified highly-correlated reaction sets that corresponded to the catalysis of steroid metabolites of physiological relevance. Subsequently, optimal flux distributions through network pathways were determined for the production of the three steroidogenic metabolites of: 11-deoxycorticosterone, testosterone and 17β-estradiol. Furthermore, flux variability analyses revealed and confirmed optimal network fluxes through physiologically feasible pathways. The stoichiometric dependence of reactions was also confirmed by conducting deletions of reactions utilized for the optimal production of 17β-estradiol. This paper demonstrates the potential application of constraint-based reconstruction and simulation techniques in enabling the construction of deterministic and predictive physiological models. This acknowledgement is poignant considering the susceptibility of the steroidogenic network to environmental and anthropogenic stressors.