Involvement of Membrane Progestin Receptor Beta (mPRβ/Paqr8) in Sex Pheromone Progestin-Induced Expression of Luteinizing Hormone in the Pituitary of Male Chinese Black Sleeper (Bostrychus Sinensis)

Abstract

Our previous studies showed that 17α, 20β-dihydroxy-4-pregnen-3-one (DHP) acted as a sex pheromone to induce reproductive success in Chinese black sleeper (Bostrychus sinensis), but its functional mechanism remains unclear. In the present study, we cloned the cDNAs of the gonadotropin subunits (cgα, fshβ, and lhβ), and found that, in exposure to 5 nM DHP, transcript levels of lhβ significantly increased in the pituitary at 6 h post exposure; plasma 11-KT levels increased at 24 h post exposure in mature male fish. In contrast, DHP exposure failed to increase the transcript levels of lhβ in the pituitary of immature male fish, suggesting that the responsiveness to DHP depends on reproductive status. Interestingly, expression of progestin and adipoQ receptor 8 (paqr8, also known as mPRβ) and progesterone receptor membrane component 2 significantly increased in the olfactory rosette of male fish at late meiosis stage following a co-injection of human chorionic gonadotropin (HCG) and luteinizing hormone releasing hormone-A3 (LHRH-A3), while no increases of other progestin receptors were observed. Moreover, Paqr8 protein was localized in the dendritic knobs of the olfactory sensory neurons, which were activated following the in vivo exposure to DHP. The DHP-induced expression of lhβ in pituitary was not inhibited by RU486, an antagonist of nuclear progesterone receptor. Taken together, our results suggested that sex pheromone DHP increased the expression of lhβ transcript in the pituitary and plasma 11-KT levels of mature male, important for reproduction; and Paqr8 might be involved in responding to sex pheromone DHP in the olfactory rosette of male B. sinensis.

Nuclear Progestin Receptor Mediated Linkage of Blood Coagulation and Ovulation

“…4E, Supplemental Table 5) (28) was conducted using a Q5 site-directed mutagenesis kit (NEB, Ipswich, MA, USA) according to the manufacturer’s protocol. Human embryonic kidney cells (HEK 293T) were used for the transactivation assays (34,35). Briefly, cells were maintained at 37 °C with 5 % CO 2 in phenol red-free Dulbecco’s modified eagle’s medium (Hyclone, USA) containing penicillin/streptomycin (Meilunbio, Dalian, China) and supplemented with 10 % fetal bovine serum (FBS, Gibco, Brazil).…”

“…4E, Supplemental Table 6) was conducted using a Q5 site-directed mutagenesis kit (NEB, Ipswich, MA, USA) according to the manufacturer's protocol. Human embryonic kidney cells (HEK 293T) were used for the transactivation assays (Chen et al, 2010;Zhang et al, 2018). Briefly, cells were maintained at 37 °C with 5 % CO 2 in phenol red-free Dulbecco's modified eagle's medium (Hyclone, USA) containing penicillin/streptomycin (Meilunbio, Dalian, China) and supplemented with 10 % fetal bovine serum (FBS, Gibco, Brazil).…”

Nuclear Progestin Receptor–mediated Linkage of Blood Coagulation and Ovulation

“…5) (28) was conducted using a Q5 site-directed mutagenesis kit (NEB, Ipswich, MA, USA) according to the manufacturer's protocol. Human embryonic kidney cells (HEK 293T) were used for the transactivation assays (34,35). Briefly, cells were maintained at 37 °C with 5 % CO2 in phenol red-free Dulbecco's modified eagle's medium (Hyclone, USA) containing penicillin/streptomycin (Meilunbio, Dalian, China) and supplemented with 10 % fetal bovine serum (FBS, Gibco, Brazil).…”