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Results: 4

Fig. 3

Fig. 3. From: MAPK3/1 (ERK1/2) in Ovarian Granulosa Cells Are Essential for Female Fertility.

ERK1/2 extensively regulate the expression of FSH/LH-target genes during ovulation. (A) Summary of gene expression profiling data. Genes regulated more than fourfold between 0 and 4 hours after hCG treatment were defined as LH-target genes; those that failed to respond to hCG in the Erk1/2gc−/− cells were defined as ERK1/2-dependent genes. (B to D) qRT-PCR shows the expression of selected FSH/eCG and LH/hCG target genes in GCs of WT and Erk1/2gc−/− mice after hCG treatment. Error bars denote SD.

Heng-Yu Fan, et al. Science. ;324(5929):938-941.
Fig. 2

Fig. 2. From: MAPK3/1 (ERK1/2) in Ovarian Granulosa Cells Are Essential for Female Fertility.

ERK1/2 are required for the terminal differentiation of GCs during ovulation. (A and B) BrdU staining of WT and Erk1/2gc−/− ovary sections at 8 hours after hCG treatment. Scale bars, 62.5 µm. (C to F) In situ hybridization shows the expression of Cyp11a1 mRNA in ovaries of WT and Erk1/2gc−/− mice at 48 hours after hCG treatment. Histology of the ovaries is shown by hematoxylin staining (C and D); localization of Cyp11a1 mRNA is shown by dark-field images (E and F). Scale bars, 250 µm.

Heng-Yu Fan, et al. Science. ;324(5929):938-941.
Fig. 1

Fig. 1. From: MAPK3/1 (ERK1/2) in Ovarian Granulosa Cells Are Essential for Female Fertility.

ERK1/2 mediate LH-induced oocyte maturation, ovulation, and luteinization. (A) Breeding assays, superovulation assays, and oocyte maturation (germinal vesicle breakdown; GVBD) rate in the different mouse genotypes. An asterisk (*) indicates that the data point is zero. Error bars denote SD. (B to E) Hematoxylin and eosin (H&E) staining of ovaries from immature WT and Erk1/2gc−/− mice treated with hCG for 8 hours (B and C) or 48 hours (D and E). CL: corpus luteum. In (B) and (C), arrows indicate the nuclear configuration of the oocyte nucleus. Scale bars, 31.25 µm. In (D) and (E), arrows indicate nonovulated COCs in preovulatory follicles. Scale bars, 250 µm. (F) Changes in estradiol and progesterone concentrations in serum after hCG treatment of eCG-primed 3-week-old WT and Erk1/2gc−/− females. D23: postnatal day 23; eCG: 23-day-old female mice treated with eCG for 48 hours; hCG D1 and hCG D2: 23-day-old females treated with eCG for 48 hours followed by hCG for 24 hours (D1) or 48 hours (D2).

Heng-Yu Fan, et al. Science. ;324(5929):938-941.
Fig. 4

Fig. 4. From: MAPK3/1 (ERK1/2) in Ovarian Granulosa Cells Are Essential for Female Fertility.

C/EBPβ is a key mediator of ERK1/2 activity in preovulatory GCs. (A) Immature WT or Erk1/2gc−/− mice were primed with eCG for 24 hours. GCs were collected and cultured overnight, and some Erk1/2gc−/− GCs were transfected with an ERK2 expression plasmid. The cells were then infected with an adenoviral vector encoding C/EBPβ for 4 hours and further treated with AREG with (+) or without (−) U0126 for another 4 hours. Total RNA or protein was extracted. LH, hCG, and AREG downstream genes were quantified by qRT-PCR. Western blots document AREG-induced phosphorylation of ERK1/2 and C/EBPβ and overexpression of C/EBPβ. Error bars denote SD. (B) H&E staining shows the absence of CLs in the ovary of adult Cebpbgc−/− mice. Follicles are indicated by arrows. Scale bar, 250 µm. (C) The number of ovulated COCs decreased at 16 hours after hCG injection in Cebpbgc−/− mice. (D) Summary. LH, cAMP, and protein kinase A (PKA) induce AREG and EREG, which activate RAS and ERK1/2. Activated ERK1/2 is required to (i) induce and activate CEBPβ and genes essential for oocyte maturation, ovulation, and luteinization; (ii) maintain AREG, EREG, and BTC by inducing Ptgs2/PGE and activation of EP2, cAMP, and PKA; and (iii) silence the FSH-regulated program. LH may also transactivate RAS and ERK1/2 directly and regulate C/EBPβ expression by other pathways. FSH and LH pathways are shown in green and red, respectively.

Heng-Yu Fan, et al. Science. ;324(5929):938-941.

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