Molecular signals governing cremaster muscle development: clues for cryptorchidism

Cryptorchidism affects 2-4% of newborn boys. Testicular descent requires the gubernaculum to differentiate into cremaster muscle (CM) during androgen-mediated inguino-scrotal descent, but the cellular mechanisms regulating this remodeling remain elusive. β-Catenin, a marker of canonical Wnt signaling, promotes myogenic genes and cellular adhesion. We aimed to determine if androgen receptor (AR) blockade altered β-catenin and its downstream myogenic proteins within the CM.

Gubernacula from male rats (n=12) and rats treated with anti-androgen, flutamide (n=12) at E19, D0, D2 were processed for immunohistochemistry. Antibodies against β-catenin, embryonic myosin, and myogenin were visualized by confocal microscopy.

At E19, β-catenin immuno-reactivity (IR) localized to the CM membrane. By D2, cytoplasmic β-catenin-IR was noted with overall β-catenin-IR decreasing. Myogenic proteins resided primarily in cells containing β-catenin on their plasma membrane. Embryonic myosin-IR was high at E19 and then decreased by D2, while myogenin-IR increased. AR blockade increased cytoplasmic β-catenin at D2 and reduced levels of both myogenic proteins.

Myogenic proteins are present in CM cells containing β-catenin. AR blockade did not alter cellular adhesion via β-catenin. In contrast, blocking AR prevented β-catenin entering the nucleus and impaired CM myogenesis. Mutations in this pathway may result in idiopathic cryptorchidism.