Background <p>Follicle-stimulating hormone (FSH) is a glycoprotein involved in oogenesis and subsequent maturation. An inadequate level of this hormone critically disrupts pre-vitellogenic oocyte progression. This disruption is a major bottleneck in the captive maturation of Asian catfish, <i>Clarias magur</i>.</p> Materials and methods <p>The current study investigates the potential biological molecular interaction of recombinant human follicle-stimulating hormone (r-hFSH) and FSHR of <i>Clarias magur</i> through comprehensive in silico analysis. Subsequent in vivo research was conducted on the progression of pre-vitellogenic oocytes in <i>Clarias magur</i> by r-hFSH induction and shower simulation consisting of three treatments and one control, viz. C: Control, S: Shower, H: Hormone and SH: Shower and hormone.</p> Results <p>The in silico evaluation revealed that r-hFSH had a better interaction with the <i>C. magur</i> FSH receptor (FSHR) than native FSHβ. These predicted affinity values confirm the trend observed in our docking scores. They provide quantitative support for the conclusion that the recombinant human ligand forms a highly stable complex with the <i>C. magur</i> receptor. In a subsequent 60-day in vivo experiment, weekly r-hFSH administration significantly elevated serum biomarker. Hormone stimulation, with and without shower, effectively advanced oocytes from the pre-vitellogenic stage to spawning. It showed superior fecundity, fertilization, and hatching rates. This demonstrates its potential as a reliable strategy to induce oocyte development and improve captive breeding success in <i>C. magur</i>.</p> Conclusion <p>The current finding emphasizes a novel opportunity for regulating oogenesis and the advancement of pre-vitellogenic oocyte development to maturation in captive farmed <i>C. magur</i>, utilizing solely r-hFSH.</p>