To determine whether conditional depletion of Progesterone Receptor Membrane Component (PGRMC) 1 and PGRMC2 affected ovarian follicle development, follicle distribution was assessed in ovaries of young (≈ 3 month-old) and middle-aged (≈6 month-old) control (Pgrmc1/2fl/fl) and double conditional PGRMC1/2 knockout (Pgrmc1/2d/d) mice. This study revealed that the distribution of primary, preantral and antral follicles was not altered in Pgrmc1/2d/d mice, regardless of the age. Although the number of primordial follicles was similar at ≈ 3 months of age, their numbers were reduced by ≈ 80% in 6-month old Pgrmc1/2d/d mice compared to age-matched Pgrmc1/2fl/fl mice. The Pgrmc1/2d/d mice were generated using Pgr-cre mice, so ablation of Pgrmc1 and Pgrmc2 in the ovary was restricted to peri-ovulatory follicles and subsequent corpora lutea (CL). In addition, the vascularization of CL was attenuated in Pgrmc1/2d/d mice, although mRNA levels of Vascular Endothelial Growth Factor A (Vegfa) were elevated. Moreover, depletion of Pgrmc1 and Pgrmc2 altered the gene expression profile in the non-luteal component of the ovary such that Vegfa expression, a stimulator of primordial follicle growth, was elevated; Kit Ligand expression, another stimulator of primordial follicle growth, was suppressed and Anti-Mullerian Hormone, an inhibitor of primordial follicle growth, was enhanced compared to Pgrmc1/2fl/fl mice. These data reveal that luteal cell depletion of Pgrmc1 and 2 alters the expression of growth factors within the non-luteal component of the ovary which could account for the premature demise of the adult population of primordial follicles.
John J Peluso, Xiufang Liu, Tracy Uliasz, Cindy A. Pru, Nicole C. Kelp and James Pru
Melissa Lynn McCallum, Cindy A. Pru, Andrea R Smith, Nicole C Kelp, Marc Foretz, Benoit Viollet, Min Du and James Pru
Adenosine monophosphate-activated protein kinase (AMPK) is a highly conserved heterotrimeric complex that acts as an intracellular energy sensor. Based on recent observations of AMPK expression in all structures of the female reproductive system, we hypothesized that AMPK is functionally required for maintaining fertility in the female. This hypothesis was tested by conditionally ablating the two catalytic alpha subunits of AMPK, Prkaa1 and Prkaa2, using Pgr-cre mice. After confirming the presence of PRKAA1, PRKAA2 and the active phospho-PRKAA1/2 in the gravid uterus by immunohistochemistry, control (Prkaa1/2fl/fl) and double conditional knockout mice (Prkaa1/2d/d) were placed into a six-month breeding trial. While the first litter size was comparable between Prkaa1/2fl/fl and Prkaa1/2d/d female mice (p=0.8619), the size of all subsequent litters was dramatically reduced in Prkaa1/2d/d female mice (p=0.0015). All Prkaa1/2d/d female mice experienced premature reproductive senescence or dystocia by the fourth parity. This phenotype manifested despite no difference in estrous cycle length, ovarian histology in young and old nulliparous or multiparous animals, mid-gestation serum progesterone levels, or uterine expression of Esr1 or Pgr between Prkaa1/2fl/fl and Prkaa1/2d/d female mice suggesting that the hypothalamic-pituitary-ovary axis remained unaffected by PRKAA1/2 deficiency. However, an evaluation of uterine histology from multiparous animals identified extensive endometrial fibrosis and disorganized stromal-glandular architecture indicative of endometritis, a condition that causes subfertility or infertility in most mammals. Interestingly, Prkaa1/2d/d female mice failed to undergo artificial decidualization. Collectively, these findings suggest that AMPK plays an essential role in endometrial regeneration following parturition and tissue remodeling that accompanies decidualization.