Inhibition of Small Ubiquitin-Related Modifier-1 Expression by Luteinizing Hormone Receptor Stimulation is Linked to Induction of Progesterone Receptor during Ovulation in Mouse Granulosa Cells

Shao, Ruijin; Zhang, Fuping; Rung, Emilia; Palvimo, Jorma J.; Huhtaniemi, Ilpo; Billig, Håkan

doi:10.1210/en.2003-0527

Cited by 31 publications

(38 citation statements)

References 39 publications

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“…Whole cell protein extract and Western blot were done essentially as described previously (64). Tissue extracts were homogenized at 4°C and incubated on ice for 30 min for a high-salt lysis buffer that effectively extracts all DNA-binding proteins.…”

Section: Treatment With P4 and Pr Antagonists And Pharmacological Promentioning

confidence: 99%

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Nuclear progesterone receptor A and B isoforms in mouse fallopian tube and uterus: implications for expression, regulation, and cellular function

Shao

Weijdegård²,

Ljungström³

et al. 2006

American Journal of Physiology-Endocrinology and Metabolism

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Progesterone and its interaction with nuclear progesterone receptors (PR) PR-A and PR-B play a critical role in the regulation of female reproductive function in all mammals. However, our knowledge of the regulation and possible cellular function of PR protein isoforms in the fallopian tube and uterus in vivo is still very limited. In the present study, we revealed that equine chorionic gonadotropin (eCG) treatment resulted in a time-dependent increase in expression of both isoforms, reaching a maximal level at 48 h in the fallopian tube. Regulation of PR-A protein expression paralleled that of PR-B protein expression. However, in the uterus PR-B protein levels increased and peaked earlier than PR-A protein levels after eCG treatment. With prolonged exposure to eCG, PR-B protein levels decreased, whereas PR-A protein levels continued to increase. Furthermore, subsequent treatment with human (h)CG decreased the levels of PR protein isoforms in both tissues in parallel with increased endogenous serum progesterone levels. To further elucidate whether progesterone regulates PR protein isoforms, we demonstrated that a time-dependent treatment with progesterone (P4) decreased the expression of PR protein isoforms in both tissues, whereas decreases in p27, cyclin D2, and proliferating cell nuclear antigen protein levels were observed only in the uterus. To define the potential PR-mediated effects on apoptosis, we demonstrated that the PR antagonist treatment increased the levels of PR protein isoforms, induced mitochondrial-associated apoptosis, and decreased in epidermal growth factor (EGF) and EGF receptor protein expression in both tissues. Interestingly, immunohistochemistry indicated that the induction of apoptosis by PR antagonists was predominant in the epithelium, whereas increase in PR protein expression was observed in stromal cells of both tissues. Taken together, these observations suggest that 1) the tissue-specific and hormonal regulation of PR isoform expression in mouse fallopian tube and uterus, where they are potentially involved in regulation of mitochondrial-mediated apoptosis depending on the cellular compartment; and 2) a possible interaction between functional PR protein and growth factor signaling may have a coordinated role for regulating apoptotic process in both tissues in vivo.

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Section: Treatment With P4 and Pr Antagonists And Pharmacological Promentioning

confidence: 99%

“…They were dewaxed by two 5-min washes in xylene and rehydrated through graded alcohol by 5-min washes with a final wash in water. A standard citrate boiling antigen retrieval procedure was used (62,64). Sections were blocked in 0.3% H2O2 in TBS and 10% normal goat serum.…”

Section: Treatment With P4 and Pr Antagonists And Pharmacological Promentioning

confidence: 99%

Nuclear progesterone receptor A and B isoforms in mouse fallopian tube and uterus: implications for expression, regulation, and cellular function

Shao

Weijdegård²,

Ljungström³

et al. 2006

American Journal of Physiology-Endocrinology and Metabolism

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“…SUMO modifications have been associated with many disease conditions, ranging from neurodegeneration to diabetes and inflammation (9, 16), as well as linked to the pathogenesis of several disorders, including Alzheimer's disease, Huntington's disease, and cancer (1, 9, 13, 21). Ubiquitin and ubiquitin-related proteins, such as SUMO paralogues, are important in diverse reproductive functions, including gametogenesis, ovulation, and steroid receptor activity (7,8,11,36,40). In C. elegans, the reproductive system is a major SUMO target during postembryonic development, and SUMO is required for gonadal and uterine-vulval morphogenesis (8).…”

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confidence: 99%

Sumo-1 Function Is Dispensable in Normal Mouse Development

Zhang

Mikkonen

Toppari

et al. 2008

Molecular and Cellular Biology

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158

121

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To elucidate SUMO-1 functions in vivo, we targeted by homologous recombination the last three exons of the murine Sumo-1 gene. Sumo-1 mRNA abundance was reduced to one-half in heterozygotes and was undetectable in Sumo-1 ؊/؊ mice, and SUMO-1-conjugated RanGAP1 was detectable in wild-type mouse embryo fibroblasts (MEFs) but not in Sumo-1 ؊/؊ MEFs, indicating that gene targeting yielded Sumo-1-null mice. Sumo-1 mRNA is expressed in all tissues of wild-type mice, and its abundance is highest in the testis, brain, lungs, and spleen. Sumo-2 and Sumo-3 mRNAs are also expressed in all tissues, but their abundance was not upregulated in Sumo-1-null mice. The development and function of testis are normal in the absence of Sumo-1, and Sumo-1 Small ubiquitin-related modifier 1 (SUMO-1) is a member of the ubiquitin-related protein family (15,16,19,23,24). SUMO proteins are ubiquitously expressed throughout the eukaryotic kingdom. The yeast Caenorhabditis elegans and Drosophila melanogaster have a single SUMO gene, but other organisms, such as vertebrates and plants, have several SUMO genes. Four SUMO family members, SUMO-1 to SUMO-4, have been found in mammals. SUMO-4 precursor cannot be processed for substrate conjugation in vivo, and it may function exclusively through noncovalent interactions (27). SUMO-2 to SUMO-4 share greater amino acid sequence identity with each other than with SUMO-1. SUMO-1 to SUMO-3 have a wide tissue distribution, whereas SUMO-4 expression is limited to the kidneys, lymph nodes, and spleen (10, 21). SUMO-1, a 97-amino-acid-residue mature polypeptide, shares 18% sequence identity with ubiquitin, and the two proteins have similar three-dimensional structures. Like ubiquitin, SUMO is covalently conjugated to substrates by an isopeptide bond through the carboxyl terminus.A consensus SUMO acceptor site, comprising the sequence ⌿KXE (where "⌿" is a large hydrophobic amino acid and K is the SUMO attachment residue) has been identified, although nonconsensus acceptor sites are also used for SUMO conjugation (15, 16). In addition, biological functions have been assigned to SUMO paralogues that are not covalently attached to substrate proteins (13). Many nuclear receptors, including androgen, progesterone, and glucocorticoid receptors, contain SUMO acceptor sites, suggesting a role for sumoylation in their signaling (18). Sumoylation is an evolutionarily conserved pathway from yeast to humans. SUMO is first activated by an E1 activating enzyme, the Aos1/Uba2 heterodimer, subsequently transferred to the unique E2 conjugating enzyme Ubc9, and conjugated to substrates in a reaction facilitated by different E3 ligases, including members of the PIAS family, RanBP2 nucleoporin, and polycomb protein Pc2. SUMO modification is a dynamic, reversible process, and distinct enzymes are responsible for SUMO conjugation and deconjugation. Several SUMO-specific proteases, including SENP1 (SuPr-2), SENP2 (SuPr-1), SENP3 (SMT3IP1), SENP5, and SENP6 (SUSP1), have been identified and localized (16,17).Sumoylation is an impor...

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“…Treatment of immature female mice with eCG results in follicular steroid secretion (17␤-estradiol in granulosa cells and androgen in theca cells), whereas subsequent treatment with hCG mimics the stimulatory effect of luteinizing hormone to induce P 4 production during luteinization of follicular cells (52,55). To examine the effect of gonadotropins on AR protein expression, 26-day-old mice were randomly divided into two groups.…”

Section: Hormone Treatment and Tissue Preparationmentioning

confidence: 99%

“…Immunohistochemical analysis was performed as described previously (54,55), with minor modification. Paraffin sections were deparaffinized and rehydrated through a graded alcohol series followed by antigen retrieval in 10 mM sodium citrate buffer (pH 6.0, microwave 10 min at full power).…”

Section: Immunohistochemistrymentioning

confidence: 99%

Estrogen-induced upregulation of AR expression and enhancement of AR nuclear translocation in mouse fallopian tubes in vivo

Shao

Ljungström²,

Weijdegård³

et al. 2007

American Journal of Physiology-Endocrinology and Metabolism

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To gain an insight into potential roles of AR in this tissue, we demonstrated that eCG treatment increased AR expression in a timedependent manner and subsequent treatment with hCG decreased AR expression in mouse fallopian tubes. This expression pattern was positively associated with 17␤-estradiol and testosterone levels in vivo. Immunohistochemical analysis of fallopian tube epithelial cells revealed that nuclear localization of AR increased in parallel with decreased AR in the cytoplasm following eCG treatment. Moreover, we found that treatment with flutamide upregulated AR expression in immature mice in association with a decrease in serum testosterone levels, whereas the same treatment resulted in downregulation of AR expression in gonadotropin-stimulated mice with concomitant decreases in serum 17␤-estradiol concentrations, suggesting that androgen differs from estrogen in the regulation of AR expression. Furthermore, we demonstrated that DES increased both AR protein expression and nuclear location over a 48-h time course. DHT had rapid effects, with induction of AR expression and translocation at 6 h after injection, but unlike DES it had prolonged efficacy. In addition, we provided direct in vivo evidence that nuclear protein interaction between AR and p21 Cip1 , a previously reported AR-regulated gene, was enhanced by gonadotropin stimulation. To our knowledge, this study provides the first demonstration to illustrate that estrogen as a principal regulator may contribute to regulate and activate AR in the fallopian tubes in vivo. androgen receptor; gonadotropins THE PHYSIOLOGICAL FUNCTIONS of androgen receptor (AR) in the male, in particular, regulating the development and function of reproduction, have been well characterized (5, 14, 25), but its regulation and biological roles in the female reproductive tissues still remains to be fully understood (15,20). The expression of AR protein has been investigated in human (2, 7, 17, 35, 60, 71) and rat (23, 44, 61, 65) ovaries and uteri, and a role for AR in the ovulatory process has been demonstrated. For example, hydroxyflutamide, a specific inhibitor of AR, suppresses ovulation rate in rats (8). Furthermore, female AR knockout mice (72) have longer estrous cycles and display reduced fertility with increased age due to lost follicle numbers (24, 58). These observations are consistent with studies in the testicular feminized mice, which have dysfunctional ARs in vivo (33). In addition, activated AR has been shown to modulate uterine growth in rats (48), and AR knockout mice lack exogenous gonadotropin-induced endometrial growth in uteri in vivo (24,58). These studies indicate a potential relationship between AR expression and normal female reproductive function.

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Inhibition of Small Ubiquitin-Related Modifier-1 Expression by Luteinizing Hormone Receptor Stimulation is Linked to Induction of Progesterone Receptor during Ovulation in Mouse Granulosa Cells

Cited by 31 publications

References 39 publications

Nuclear progesterone receptor A and B isoforms in mouse fallopian tube and uterus: implications for expression, regulation, and cellular function

Nuclear progesterone receptor A and B isoforms in mouse fallopian tube and uterus: implications for expression, regulation, and cellular function

Sumo-1 Function Is Dispensable in Normal Mouse Development

Estrogen-induced upregulation of AR expression and enhancement of AR nuclear translocation in mouse fallopian tubes in vivo

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