scholarly journals Peroxiredoxin 1 Controls Ovulation and Ovulated Cumulus–Oocyte Complex Activity through TLR4-Derived ERK1/2 Signaling in Mice

2021 ◽  
Vol 22 (17) ◽  
pp. 9437
Author(s):  
Hyo-Jin Park ◽  
Bokyung Kim ◽  
Deog-Bon Koo ◽  
Dong-Seok Lee
Keyword(s):  
Protein Expression ◽  
Gene Deletion ◽  
Cumulus Cell ◽  
Mrna Levels ◽  
Wild Type ◽  
Complex Activity ◽  
Cumulus Expansion ◽  
Peroxiredoxin 1 ◽  
Cumulus Oocyte Complex ◽  
Serum Gonadotropin

Peroxiredoxins (PRDXs) are expressed in the ovary and during ovulation. PRDX1 activity related to the immuno-like response during ovulation is unknown. We investigated the roles of Prdx1 on TLR4 and ERK1/2 signaling from the ovulated cumulus–oocyte complex (COC) using Prdx1-knockout (K/O) and wild-type (WT) mice. Ovulated COCs were collected 12 and 16 h after pregnant mare serum gonadotropin/hCG injection. PRDX1 protein expression and COC secretion factors (Il-6, Tnfaip6, and Ptgs2) increased 16 h after ovulated COCs of the WT mice were obtained. We treated the ovulated COCs in mice with LPS (0.5 μg/mL) or hyaluronidase (Hya) (10 units/mL) to induce TLR4 activity. Intracellular reactive oxygen species (ROS), cumulus cell apoptosis, PRDX1, TLR4/P38/ERK1/2 protein expression, and COC secretion factors’ mRNA levels increased in LPS- and Hya-treated COCs. The ERK inhibitor (U0126) and Prdx1 siRNA affected TLR4/ERK1/2 expression. The number and cumulus expansion of ovulated COCs by ROS were impaired in Prdx1 K/O mice but not in WT ones. Prdx1 gene deletion induced TLR4/P38/ERK1/2 expression and cumulus expansion genes. These results show the controlling roles of PRDX1 for TLR4/P38/ERK1/2 signaling activity in ovulated mice and the interlink of COCs with ovulation.

scholarly journals The effect of calnexin deletion on the expression level of PDI in Saccharomyces cerevisiae under heat stress conditions

2008 ◽  
Vol 13 (1) ◽  
Author(s):  
Huili Zhang ◽  
Jianwei He ◽  
Yanyan Ji ◽  
Akio Kato ◽  
Youtao Song
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Growth Rate ◽  
Heat Stress ◽  
Protein Expression ◽  
Molecular Chaperone ◽  
Mrna Level ◽  
Stress Conditions ◽  
Mrna Levels ◽  
Wild Type ◽  
Wild Type Yeast

AbstractWe cultured calnexin-disrupted and wild-type Saccharomyces cerevisiae strains under conditions of heat stress. The growth rate of the calnexin-disrupted yeast was almost the same as that of the wild-type yeast under those conditions. However, the induced mRNA level of the molecular chaperone PDI in the ER was clearly higher in calnexin-disrupted S. cerevisiae relative to the wild type at 37°C, despite being almost the same in the two strains under normal conditions. The western blotting analysis for PDI protein expression in the ER yielded results that show a parallel in their mRNA levels in the two strains. We suggest that PDI may interact with calnexin under heat stress conditions, and that the induction of PDI in the ER can recover part of the function of calnexin in calnexin-disrupted yeast, and result in the same growth rate as in wild-type yeast.

scholarly journals ChREBP-Knockout Mice Show Sucrose Intolerance and Fructose Malabsorption

2018 ◽  
Author(s):  
Takehiro Kato ◽  
Katsumi Iizuka ◽  
Ken Takao ◽  
Yukio Horikawa ◽  
Tadahiro Kitamura ◽  
...  
Keyword(s):  
Plasma Glucose ◽  
Knockout Mice ◽  
Gene Deletion ◽  
Body Weight Loss ◽  
Mrna Levels ◽  
Wild Type ◽  
Glucose Levels ◽  
Fructose Malabsorption ◽  
Sucrose Diet ◽  
Alpha Glucosidase

We have previously reported that 60% sucrose diet-fed ChREBP knockout mice (KO) showed body weight loss resulting in lethality. We aimed to elucidate whether sucrose and fructose metabolism are impaired in KO. Wild type mice (WT) and KO were fed a diet containing 30% sucrose with/without 0.08% miglitol, an α-glucosidase inhibitor, and these effects on phenotypes were tested. Furthermore, we compared metabolic changes of oral and peritoneal fructose injection. Thirty percent sucrose diet feeding did not affect phenotypes in KO. However, miglitol induced lethality in 30% sucrose-fed KO. Thirty percent sucrose plus miglitol diet-fed KO showed increased cecal contents, increased fecal lactate contents, increased growth of lactobacillales and Bifidobacterium and decreased growth of clostridium cluster XIVa. ChREBP gene deletion suppressed the mRNA levels of sucrose and fructose related genes. Next, oral fructose injection did not affect plasma glucose levels and liver fructose contents; however, intestinal sucrose and fructose related mRNA levels were increased only in WT. In contrast, peritoneal fructose injection increased plasma glucose levels in both mice; however, the hepatic fructose content in KO was much higher owing to decreased hepatic Khk mRNA expression. Taken together, KO showed sucrose intolerance and fructose malabsorption owing to decreased gene expression.

Distribution patterns of leucocyte subpopulations expressing different cell markers in the cumulus - oocyte complexes of pregnant and pseudopregnant mice

2003 ◽  
Vol 15 (7) ◽  
pp. 389 ◽  
Author(s):  
Gökhan Akkoyunlu ◽  
Emin Türkay Korgun ◽  
Çiler Çelik-Özenci ◽  
Yasemin Seval ◽  
Ramazan Demir ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
Cumulus Cell ◽  
Cumulus Cells ◽  
Distribution Patterns ◽  
Mature Female ◽  
Cell Markers ◽  
Cumulus Oocyte Complex ◽  
Transmission Electron ◽  
Serum Gonadotropin

The nature of leucocyte subpopulations expressing different cell markers around the cumulus–oocyte complex (COC) of pregnant and pseudopregnant mice was investigated in the present study. Immunolabelling for CD4, CD8, CD14, CD45 and CD163 and transmission electron microscopy were used to determine whether leucocytes differ between pregnant and pseudopregnant mice. Sexually mature female BALB/c mice (n = 36; 18 pregnant, 18 pseudopregnant) were stimulated to superovulate with pregnant mare’s serum gonadotropin and human chorionic gonadotrophin, then mated with either fertile or vasectomised males. Postovulatory oocytes were collected after mating. The cumulus cell masses of the pregnant group contained spermatozoa between cells and were more variable than COCs of the pseudopregnant group. Streptavidin–biotin–peroxidase immunohistochemical labelling of the cell markers CD4, CD8, CD14, CD45 and CD163 showed that there were fewer leucocytes in the COCs of the pseudopregnant group compared with the pregnant group. Transmission electron microscopy revealed that often there were macrophage-like cells containing spermiophagic bodies between the cumulus cells. These observations suggest that, together with other cumulus cells and oviducal cells, these macrophage-like cells may be involved in removing unsuitable or excess spermatozoa and, therefore, in maintaining a suitable microenvironment for normal fertilisation.

scholarly journals Oocyte-specific ablation of N- and O-glycans alters cumulus cell signalling and extracellular matrix composition

2019 ◽  
Vol 31 (3) ◽  
pp. 529 ◽  
Author(s):  
Belinda K. M. Lo ◽  
Agnes Archibong-Omon ◽  
Panayiota Ploutarchou ◽  
Anthony J. Day ◽  
Caroline M. Milner ◽  
...  
Keyword(s):  
Cell Signalling ◽  
Cumulus Cell ◽  
Cumulus Cells ◽  
Oocyte Quality ◽  
Matrix Composition ◽  
Cumulus Expansion ◽  
Heavy Chains ◽  
Cumulus Oocyte Complex ◽  
Morphogenetic Protein ◽  
Necrosis Factor

Cumulus–oocyte complex (COC) expansion is essential for ovulation and fertilisation and is linked to oocyte quality. Hyaluronan (HA), the major matrix constituent, is cross-linked via inter-α-inhibitor heavy chains (HCs), pentraxin 3 (PTX3) and tumour necrosis factor-stimulated gene 6 (TSG-6). All except HCs are secreted by cumulus cells in response to oocyte-secreted factors, which signal via SMAD pathways. The double mutant (DM) mouse generates oocytes lacking complex N- and O-glycans due to oocyte-specific deletion of core 1 β1,3-galactosyltransferase (C1galt1) and N-acetylglucosaminyltransferase I (Mgat1) and has modified cumulus expansion. We compared COCs before expansion (48 h-post-pregnant mare serum gonadotrophin (PMSG)) and at late-stage expansion (9 h-post-human chorionic gonadotrophin (hCG); control n=3 mice, DM n=3 per group). Using histochemistry the levels of HA, HCs, PTX3, TSG-6 and phosphorylated-SMAD1/5/8 and -SMAD2 (12–25 COCs per group) were assessed. DM COCs did not differ from Controls in cumulus size or cell density at 9 h-post-hCG; however, HA and HC levels and phosphorylated-SMAD1/5/8 were reduced. Furthermore, no correlations were found between the levels of matrix molecules and cumulus area in DM or Control samples. These data suggest that HA and HCs can support cumulus expansion provided that they are present above minimum threshold levels. We propose that oocyte-specific ablation of C1galt1 and Mgat1 may affect bone morphogenetic protein 15 synthesis or bioactivity, thereby reducing SMAD1/5/8 phosphorylation and HA production.

249. Regulated expression of KIT protein in juvenile and adult germ cells of the rodent testis

2008 ◽  
Vol 20 (9) ◽  
pp. 49
Author(s):  
S. Mithraprabhu ◽  
C. W. Brown ◽  
M. M. Matzuk ◽  
K. L. Loveland
Keyword(s):  
Protein Expression ◽  
Germ Cells ◽  
Wild Type Mouse ◽  
Erythroid Differentiation ◽  
Activin A ◽  
Mrna Levels ◽  
Wild Type ◽  
Kit Receptor ◽  
Murine Erythroleukemia

KIT receptor is an established marker of differentiating spermatogonia and its activation is required to trigger spermatogonial maturation. KIT mRNA, however, can be detected in undifferentiated spermatogonia in the absence of protein expression, as previously established by us in the irradiated adult rat testes [1]. This differential regulation of mRNA and protein is presumably modulated by either local hormone action or by cues from the adult testicular microenvironment. Endogenous regulatory factors known to stimulate KIT synthesis in juvenile male germ cells in vitro are bone morphogenetic protein 4 (BMP4) and retinoic acid (RA), while factors known to suppress KIT at the onset of spermatogenesis have not yet been identified. Activin A, implicated in KIT downregulation in a murine erythroleukemia cell line [2], is produced within the juvenile mammalian testis and influences activities of spermatogonia and Sertoli cells. We hypothesised that activin acts to repress KIT expression in spermatogonia and therefore modulate spermatogonial behaviour. Evidence for this was first derived from Sertoli and germ cell co-cultures of day 8 wild type mouse testes in which exogenous activin addition caused a dose-dependent reduction of KIT mRNA. Whole testes mRNA analyses of two activin transgenic mouse models, the newborn Inhba−/− (lacking activin A) and postnatal InhbaBK/BK (decreased bioactive activin), revealed a significant elevation in KIT expression relative to wild type littermates. In the postnatal day 7 InhbaBK/BK testes, an elevated proportion of differentiated spermatogonia, increased cell surface KIT protein levels, enhanced mRNA levels of a known downstream target of KIT signalling pathway, cyclind3 and a meiotic marker, Sycp3, were observed. These data provide the first comprehensive evidence for activin modulation of KIT expression at spermatogenesis onset, in germ cells of the juvenile testis. This finding is of fundamental importance to other KIT-dependent processes. (1) Prabhu, S.M., et al. Expression of c-Kit receptor mRNA and protein in the developing, adult and irradiated rodent testis. Reproduction, 2006. 131(3): p. 489–99. (2) Hino, M., et al. Down-modulation of c-kit mRNA and protein expression by erythroid differentiation factor/activin A. FEBS Lett, 1995. 374(1): p. 69–71.

scholarly journals Impact of Ovarian Sex Steroids on Ovulation and Ovulatory Gene Induction in Aromatase-Null Mice

Endocrinology ◽  
2012 ◽  
Vol 153 (1) ◽  
pp. 386-394 ◽  
Author(s):  
Katsumi Toda ◽  
Yoshihiro Hayashi ◽  
Masafumi Ono ◽  
Toshiji Saibara
Keyword(s):  
Chorionic Gonadotropin ◽  
Human Chorionic Gonadotropin ◽  
Sex Steroids ◽  
Mrna Levels ◽  
Wild Type ◽  
Sequential Administration ◽  
Serum Gonadotropin ◽  
17Β Estradiol ◽  
Related Proteins

Female mice deficient in the aromatase gene [aromatase knockout (ArKO)] fail to ovulate owing to an inability to produce estrogens. Here, we demonstrated that sequential administration of adequate amounts of 17β-estradiol (E2), pregnant mare serum gonadotropin, and human chorionic gonadotropin could induce ovulation in immature ArKO mice; nevertheless, significantly fewer oocytes were released into the oviducts in ArKO mice than in wild-type mice. Analysis of ovarian steroids by liquid chromatography coupled with electrospray ionization-tandem mass spectrometry identified a trace amount of E2 in the untreated immature ArKO ovary. The analysis further detected significant increases and decreases in progesterone and testosterone contents, respectively, in addition to an increase of E2 in the ovulation-induced ArKO ovaries compared with the levels in untreated ArKO ovaries. Gene expression analysis demonstrated marked elevation in the mRNA levels of members of the epidermal growth factor family and extracellular matrix-related proteins at 4 h after human chorionic gonadotropin injection in the ovaries of ArKO mice treated for ovulation, as observed in the ovulation-induced wild-type ovaries. Collectively, these findings suggest the vital contribution of the intraovarian milieu of sex steroids to ovulatory regulation in vivo.

Intestinal Na-Pi cotransporter adaptation to dietary Pi content in vitamin D receptor null mice

2004 ◽  
Vol 287 (1) ◽  
pp. F39-F47 ◽  
Author(s):  
Hiroko Segawa ◽  
Ichiro Kaneko ◽  
Setsuko Yamanaka ◽  
Mikiko Ito ◽  
Masahi Kuwahata ◽  
...  
Keyword(s):  
Vitamin D ◽  
Protein Expression ◽  
Vitamin D Receptor ◽  
Brush Border Membrane ◽  
Phosphate Transport ◽  
Mrna Levels ◽  
Wild Type ◽  
Intestinal Brush Border Membrane ◽  
Intestinal Brush Border ◽  
Diet Type

Recent studies suggest that vitamin D may play a role in intestinal Na+-dependent phosphate transport adaptation to variable levels of dietary Pi. Therefore, the goal of the current study was to assess Na+-dependent Pi cotransport activity in transgenic mice to determine whether vitamin D is an essential mediator of this process. Intestinal brush-border membrane (BBM), Na+-dependent Pi cotransport activity was significantly decreased in vitamin D receptor (VDR) null [VDR (−/−)] mice compared with wild-type (VDR+/+) mice. While intestinal Na-Pi cotransporter (type IIb) mRNA levels were similar in VDR (−/−) and VDR (+/+) mice, type IIb Na-Pi cotransporter protein expression was markedly suppressed in VDR (−/−) mice compared with VDR (+/+) mice. Furthermore, Na-Pi cotransport activity in renal BBM was similar in VDR (−/−) and VDR (+/+) mice, but type IIa Na-Pi cotransporter protein expression was decreased in VDR (−/−) mice. After administration of a low-Pi diet, type IIb protein expression was significantly increased in VDR (+/+) and VDR (−/−) mice, and type IIb protein expression was present in the intestinal BBM of VDR (−/−) mice. These data demonstrate that intestinal Na-Pi cotransport adaptation to a low-Pi diet occurs independently of vitamin D.

scholarly journals Multiple Signaling Defects in the Absence of RIP140 Impair Both Cumulus Expansion and Follicle Rupture

Endocrinology ◽  
2005 ◽  
Vol 146 (9) ◽  
pp. 4127-4137 ◽  
Author(s):  
Jennifer M. A. Tullet ◽  
Victoria Pocock ◽  
Jennifer H. Steel ◽  
Roger White ◽  
Stuart Milligan ◽  
...  
Keyword(s):  
Extracellular Matrix ◽  
Cell Attachment ◽  
Expression Profiles ◽  
Ovarian Tissue ◽  
Cumulus Cell ◽  
Gene Expression Profiles ◽  
Mural Cell ◽  
Cumulus Expansion ◽  
Serum Gonadotropin ◽  
Cell Cell

Abstract The nuclear receptor corepressor RIP140 is essential in the ovary for ovulation, but is not required for follicle growth and luteinization. To identify genes that may be subject to regulation by RIP140 or play a role in ovulation, we compared ovarian gene expression profiles in untreated immature wild-type and RIP140 null mice and after treatment with pregnant mare serum gonadotropin and human chorionic gonadotropin. Many genes involved in signaling, extracellular matrix formation, cell-cell attachment, and adhesion were aberrantly regulated in the absence of RIP140, varying according to the hormone status of the mice. Notable among these was the reduced expression of a number of genes that encode components of signaling pathways and matrix proteins required for cumulus expansion, a key remodeling process necessary for ovulation. Histological analysis confirmed that cumulus expansion in RIP140 null mice is reduced, oocyte detachment from the mural cell wall is impaired, and follicles fail to rupture in response to LH. Although the expression of many genes involved in cumulus cell expansion was reduced, there was a subset of genes involved in extracellular matrix formation and cell-cell interactions that was up-regulated and may interfere with ovarian tissue remodeling. We propose that widespread gene dysregulation in ovarian tissues in the absence of RIP140 leads to the anovulatory phenotype. This helps to define an important role for RIP140 in the regulation of multiple processes leading to ovulation.

scholarly journals Silent Point Mutation in an Avian Retrovirus RNA Processing Element Promotes c-myb-Associated Short-Latency Lymphomas

2003 ◽  
Vol 77 (17) ◽  
pp. 9378-9387 ◽  
Author(s):  
Tatjana S. Polony ◽  
Sandra J. Bowers ◽  
Paul E. Neiman ◽  
Karen L. Beemon
Keyword(s):  
Rna Processing ◽  
High Frequency ◽  
Gene Deletion ◽  
Point Mutations ◽  
Short Latency ◽  
Negative Regulator ◽  
Mrna Levels ◽  
Wild Type ◽  
The Matrix ◽  
Myb Protein

ABSTRACT The avian leukosis virus ΔLR-9 causes a high frequency of B-cell lymphomas within weeks after injection into 10-day-old chicken embryos. These lymphomas result from proviral integrations into the oncogene c-myb. In contrast, LR-9, which lacks the 42-nucleotide gag gene deletion of ΔLR-9, does not cause a high frequency of c-myb-associated short-latency lymphomas. Although viral replication rates and spliced env mRNA levels were found to be similar for both viruses, ΔLR-9 exhibited an increase in readthrough transcription compared to LR-9. The ΔLR-9 deletion is located in the region of the gag gene corresponding to the matrix (MA) protein as well as in the negative regulator of splicing (NRS) element. To test whether disruption of the NRS or of the MA protein was responsible for inducing short-latency lymphomas, we generated viruses with NRS point mutations that maintained the wild-type Gag amino acid sequence. One of the mutant viruses induced an even higher incidence than ΔLR-9 of short-latency lymphomas with viral integrations into c-myb. Thus, we propose that disruption of the NRS sequence promotes readthrough transcription and splicing to the downstream myb gene, causing overexpression of a slightly truncated Myb protein, which induces short-latency tumors.

MiR-493 Induces Cytotoxic Autophagy in Prostate Cancer Cells through Regulation on PHLPP2

2020 ◽  
Vol 21 (14) ◽  
pp. 1451-1456 ◽  
Author(s):  
Jun Deng ◽  
Ming Ma ◽  
Wei Jiang ◽  
Liangliang Zheng ◽  
Suping Cui
Keyword(s):  
Prostate Cancer ◽  
Protein Expression ◽  
Cell Function ◽  
Mrna Levels ◽  
Prostate Cancer Cells ◽  
Potent Inducer ◽  
Qrt Pcr ◽  
Autophagy Gene ◽  
The Relationship ◽  
Cancer Inhibition

Background: MiR-493 promotes the proliferation of prostate cancer (PC) cells by targeting PHLPP2. We aimed to explore the relationship between miR-493 and autophagy in PC. Methods: qRT-PCR and western blotting were used to determine the mRNA levels and protein expression of miR-493, PHLPP2, autophagy gene BECN1 and ATG7 in PC cells. The autophagy gene expression was determined after PC cells transfected with miR-493 precursor or PHLPP2 precursor. Corresponding changes of autophagy phenotype and PC cell function were also studied. Results: The mRNA levels and protein expression of miR-493, PHLPP2, BECN1 and ATG7 in PC cells were significantly decreased in PC cells. Overexpression of miR-493 or PHLPP2 markedly upregulated the expression levels of BECN1 and ATG7 in PC cells. Overexpression of miR-493 and PHLPP2 markedly promoted autophagy, and inhibited the invasion and cloning formation of PC cells. Conclusion: MiR-493 is a potent inducer of cytotoxic autophagy that leads to prostate cancer inhibition by regulating on PHLPP2.

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