Activational and organizational actions of gonadal hormones on the sexual dimorphism of the α6-subunit containing GABAA receptor in Wistar rats with neuropathic pain

Sex differences in pain perception and response to analgesics are well documented, yet the underlying causes remain poorly understood. Here we investigate the sexual dimorphism in the function of α₆GABA_A receptors in neuropathic pain, focusing on activational and organizational actions of gonadal hormones. Using the nerve ligation model in rats, we found that the positive allosteric modulator, PZ-ll-029 (30 nmol, it), produced a robust antiallodynic effect in females but not in males. Ovariectomy abolished this effect, while a single dose of estradiol (20 μg/kg sc, −24 h), that returned to physiological serum levels, partially restored it, indicating that the activational effect of estradiol is crucial for α₆GABA_A receptor-mediated antiallodynia in females. Interestingly, adult or neonatal (at postnatal day 3) orchidectomy did not alter the male's insensitivity to PZ-ll-029, even after estradiol treatment. However, neonatal female's virilization (with testosterone propionate 120 μg/rat at postnatal day 3) induced a male-like insensitivity to PZ-ll-029, that was partial when the ovaries were present and complete after adult ovariectomy. These findings reveal that the neonatal organizational effects of testosterone determine the sex-specific insensitivity of α₆GABA_A receptors to modulate neuropathic pain, while the activational effects of estradiol can partly maintain the female-typical response, despite early androgen exposure. Our results provide new insights into hormonal regulation of pain modulation and suggest that both developmental exposure and adult status should be considered in basic research and preclinical studies investigating sex-based dimorphisms.