Overexpression of Cyclin D3 Improves Decidualization Defects in Hoxa-10−/− Mice

Abstract

Uterine decidualization, a crucial process for implantation, is a tightly regulated process encompassing proliferation, differentiation, and polyploidization of uterine stromal cells. Hoxa (Homeobox A)-10, a homeobox transcription factor, is highly expressed in decidualizing stromal cells. Targeted gene deletion experiments have demonstrated marked infertility resulting from severely compromised decidualization in Hoxa-10(-/-) mice. However, the underlying mechanism by which Hoxa-10 regulates stromal cell differentiation remains poorly understood. Cyclin D3, a G(1) phase cell-cycle regulatory protein involved in stromal cell proliferation and decidualization, is significantly reduced in Hoxa-10(-/-) mice. The expression of cyclin D3 in the pregnant mouse uterus parallels stromal cell decidualization. Here, we show that adenovirus-driven cyclin D3 replacement in Hoxa-10(-/-) mice improves stromal cell decidualization. To address our question of whether cyclin D3 replacement in Hoxa-10(-/-) mice can improve decidualization, both in vitro and in vivo studies were completed after the addition of cyclin D3 or empty (control) viral vectors. Immunostaining demonstrated increased proliferation and decidualization in both in vitro and in vivo studies, and in situ hybridization confirmed increased expression of decidualization markers in vivo. Placentation was demonstrated as well in vivo in the cyclin D3-replaced animals. However, fertility was not restored in Hoxa-10(-/-) mice after d 10 of pregnancy. Finally, we identified several downstream targets of cyclin D3 during decidualization in vitro via proteomics experiments, and these were confirmed using in situ hybridization in vivo. Collectively, these results demonstrate that cyclin D3 expression influences a host of genes involved in decidualization and can improve decidualization in Hoxa-10(-/-) mice.

Hmgn1 acts downstream of C/EBPβ to regulate the decidualization of uterine stromal cells in mice

“…2,23 The present study showed that Hmgn1 could stimulate the expression of Prl8a2 and Prl3c1 which were 2 well-known markers for decidual cell differentiation. Taken together, these results revealed an important role of Hmgn1 in maintaining the proliferation and differentiation of uterine stromal cells during decidualization.…”

Egr2 mediates the differentiation of mouse uterine stromal cells responsiveness to HB‐EGF during decidualization

“…Further analysis demonstrated that Egr2 was required for the HB‐EGF function in stromal cell differentiation by this evidence that siRNA‐mediated knockdown of Egr2 impeded the stimulation of rHB‐EGF on the expression of Prl8a2 , Prl3c1 , and Pgr . Previous studies have found that HB‐EGF could facilitate uterine decidualization by upregulating Ccnd3 , which was a key cell cycle regulator in stromal cell proliferation and differentiation (Sroga, Gao, Ma, & Das, ; Tan et al., ). Attenuation of Egr2 expression in stromal cells abrogated the induction of Ccnd3 elicited by rHB‐EGF, suggesting that Egr2 may serve as an intermediate to mediate the regulation of HB‐EGF on Ccnd3 .…”

Rbbp7 Is Required for Uterine Stromal Decidualization in Mice1

“…Among various cell cycle related molecules including various cyclins, CDKs and cell cycle inhibitors, cyclin D3, a well-known regulator of mammalian cell proliferation [46], has been shown to be highly expressed in the decidualizing stromal cells within implantation chambers and essential for normal postimplantation uterine decidualization [47,48]. Previous studies demonstrated that many key signaling and regulatory molecules such as heparin-binding EGF-like growth factor (HB-EGF) [49], interleukin 11-interleukin 11 receptor alpha (IL11-IL11Rα) [50,51], Hox family transcription factor HOXA10 [52], Kruppel-like factor 9 (KLF9, previously known as BTEB1) [53], death effector domain-containing protein (DEDD) [54] were considered to regulate decidualization at least partially through cyclin D3. In this respect, we added a new line of evidence showing that Rbbp7 is critical for normal cyclin D3 expression in mESCs during decidualization.…”