103 MASS SPECTROMETRY ANALYSIS OF UTERUS ENDOMETRIUM DURING PREGNANCY IN THE PIG

2009 ◽  
Vol 21 (1) ◽  
pp. 152
Author(s):  
J.-I. Chae ◽  
Y. K. Cho ◽  
S.-K. Cho ◽  
K.-K. Lee ◽  
D.-B. Koo
Keyword(s):  
Growth Factor ◽  
Western Blotting ◽  
Reproductive Tract ◽  
Female Reproductive Tract ◽  
Late Gestation ◽  
Mass Spectrometry Analysis ◽  
Spectrometry Analysis ◽  
Molecular Events ◽  
Associated Proteins ◽  
Endothelial Growth Factor

In general, many important molecular events occur within the female reproductive tract, especially within the uterine endometrium, during periods of pregnancy. The endometrium includes a mucosal lining of the uterus, which functions to provide a suitable site for implantation and development of a fertilized egg and fetus. To date, developmental integrity involves molecular cascades whose interrelationships are not fully understood within the endometrium during pregnancy in pigs. In this study, we explored the functional regulated proteins in endometrium during periods of pregnancy periods (Day 40, n = 6; Day 70, n = 7; Day 90, n = 6 of pregnancy; and nonpregnancy, n = 1) using 2-dimensional gel electrophoresis (2-DE) and Western blotting. The functional regulated proteins were identified and discovered from differentially expressed proteins in the endometrium during pregnancy. In the proteomic analysis of the endometrium, 820 protein spots were matched on 2-DE gels. With 98 proteins regulated differentially among nonpregnant and pregnant tissues (matched and unmatched spots), 63 up- or down-regulated proteins have been identified. Interestingly, 6 of these 63 proteins were endothelial growth factor-associated proteins such as transgelin, transferrin, galectin-1, tropomyosin alpha, protein DJ-1, and beta-defensin. We also confirmed the expression levels of these proteins in the endometrium during pregnancy by using Western blotting. Our results suggest that the expressions of these genes involved in endometrial function and development from early to late gestation are associated with the regulation of endothelial growth factor.

scholarly journals Relaxin (RLX) and estrogen affect estrogen receptor α, vascular endothelial growth factor, and RLX receptor expression in the neonatal porcine uterus and cervix

Reproduction ◽  
2008 ◽  
Vol 135 (5) ◽  
pp. 705-712 ◽  
Author(s):  
Wenbo Yan ◽  
Joseph Chen ◽  
Anne A Wiley ◽  
Bethany D Crean-Harris ◽  
Frank F Bartol ◽  
...  
Keyword(s):  
Vascular Endothelial Growth Factor ◽  
Estrogen Receptor ◽  
Growth Factor ◽  
Protein Content ◽  
Reproductive Tract ◽  
Female Reproductive Tract ◽  
Receptor Expression ◽  
Vascular Endothelial ◽  
Wet Weight ◽  
Endothelial Growth Factor

The porcine female reproductive tract undergoes estrogen receptor (ER) α-dependent development after birth (postnatal day=PND 0), the course of which can determine adult uterine function. Uterotrophic effects of relaxin (RLX) in the porcine neonate are age specific and may involve ER activation. Here, objectives were to determine effects of RLX and estrogen administered from birth on uterine and cervical growth and expression of ERα, vascular endothelial growth factor (VEGF), and the RLX receptor (RXFP1). On PND 0, gilts were treated with the antiestrogen ICI 182 780 (ICI) or vehicle alone and, 2 h later, were given estradiol-17β (E) or porcine RLX for 2 days. Neither RLX nor E affected uterine wet weight or protein content on PND 2. However, RLX, but not E, increased cervical wet weight and protein content when compared with controls. Pretreatment with ICI did not inhibit RLX-stimulated cervical growth. Uterine and cervical ERα increased in response to RLX, but not E. Both RLX and E increased VEGF in the uterus and cervix on PND 2. Pretreatment with ICI increased VEGF in both tissues and increased RLX-induced cervical VEGF. In the uterus E, but not RLX, increased RXFP1 mRNA. In the cervix, E increased RXFP1 gene expression whereas RLX decreased it. Results indicate that the neonatal uterus and cervix are sensitive to E and RLX and that growth responses to RLX in these tissues differ by PND 2. Effects of RLX on uterine and cervical ERα and VEGF expression may be important for neonatal reproductive tract development.

178. THE CHAPERONIN CONTAINING TCP-1 (CCT/TRiC) MULTISUBUNIT COMPLEX IS INVOLVED IN MEDIATING SPERM - OOCYTE INTERACTIONS

2010 ◽  
Vol 22 (9) ◽  
pp. 96
Author(s):  
M. D. Dun ◽  
R. Aitken ◽  
B. Nixon
Keyword(s):  
Zona Pellucida ◽  
Reproductive Tract ◽  
Protein Complexes ◽  
Female Reproductive Tract ◽  
Mass Spectrometry Analysis ◽  
Spectrometry Analysis ◽  
Sperm Surface ◽  
Collective Data ◽  
Concomitant Reduction ◽  
Future Work

Mammalian spermatozoa only express their capacity for fertilization following capacitation, a process characterized by a suite of biophysical and biochemical changes that occurs as the cells ascend the female reproductive tract. A key event associated with the attainment of a capacitated state is a dramatic reorganization of the sperm surface architecture to render these cells competent to bind to the protective matrix of the oocyte, the zona pellucida. Our previous analysis of these remodeling events has provided compelling evidence that they include the assembly and/or presentation of multimeric protein complexes on the sperm surface. In addition, we have demonstrated that at least two of these complexes possess strong affinity for solubilized zona pellucida. In our current study we have utilised mass spectrometry analysis to reveal that one of these complexes comprises the eight subunits that form a composite, multimeric structure known as the chaperonin containing TCP-1 (CCT/TRiC) complex. Our collective data suggest that this complex participates indirectly in zona pellucida interaction, possibly through the conveyance of key zona adhesion molecules to the sperm surface during capacitation. Consistent with this notion, we were able to demonstrate that the sperm CCT/TRiC complex releases its bound substrates upon exposure to ATP, and this treatment induced a significant, concomitant reduction in the ability of capacitated sperm to bind to the zona pellucida. Furthermore, the use of immunoprecipitation assays confirmed the interaction of the CCT/TRiC complex with at least one putative zona pellucida receptor candidate, namely zona pellucida binding protein 2 (ZPBP2). Future work is now aimed at identifying additional zona receptors that may reside within this complex and the pathways that regulate its functional assembly.

scholarly journals LYPD4, mouse homolog of a human acrosome protein, is essential for sperm fertilizing ability and male fertility†

2020 ◽  
Vol 102 (5) ◽  
pp. 1033-1044
Author(s):  
Dan Wang ◽  
Liping Cheng ◽  
Wenjuan Xia ◽  
Xiaofei Liu ◽  
Yueshuai Guo ◽  
...  
Keyword(s):  
Male Fertility ◽  
Reproductive Tract ◽  
Female Reproductive Tract ◽  
Mass Spectrometry Analysis ◽  
Small Subset ◽  
Vitro Fertilization ◽  
Spectrometry Analysis ◽  
Human Fertilization ◽  
Sperm Migration

Abstract Fertilization is one of the fundamental biological processes, but so far, we still do not have a full understanding of the underlying molecular mechanism. We have identified a human acrosome protein, LY6/PLAUR domain containing 4 (LYPD4), expressed specifically in human testes and sperm, and conserved within mammals. Mouse Lypd4, also specific to the testis and sperm, is essential for male fertility. LYPD4 protein first appeared in round spermatids during acrosome biogenesis and became part of acrosomes during spermatogenesis and in mature sperm. Lypd4 knockout mice are infertile with normal sperm number and motility. Mutant sperm, however, failed to reach oviduct during sperm migration inside the female reproductive tract, leading to fertilization failure and infertility. In addition, Lypd4 mutant sperms were unable to fertilize denuded egg via IVF (in vitro fertilization) but could fertilize eggs within intact Cumulus-Oocyte Complex, supporting an additional role in sperm-zona interaction. Out of more than five thousand spermatozoa proteins identified by mass spectrometry analysis, only a small subset of proteins (26 proteins) was changed in the absence of LYPD4, revealing a whole proteome picture of mutant sperm defective in sperm migration and sperm-zona binding. ADAM3, a key component of fertilization complex, as well as other sperm ADAM proteins are significantly reduced. We hence propose that LYPD4 plays an essential role in mammalian fertilization, and further investigation of its function and its interaction with other sperm membrane complexes may yield insights into human fertilization and novel strategy to improve IVF success.

scholarly journals Estradiol modulation of hepatocyte growth factor by stromal fibroblasts in the female reproductive tract

2009 ◽  
Vol 92 (3) ◽  
pp. 1107-1109 ◽  
Author(s):  
Kimberly D. Coleman ◽  
Jacqueline A. Wright ◽  
Mimi Ghosh ◽  
Charles R. Wira ◽  
John V. Fahey
Keyword(s):  
Growth Factor ◽  
Hepatocyte Growth Factor ◽  
Reproductive Tract ◽  
Female Reproductive Tract ◽  
Stromal Fibroblasts ◽  
Hepatocyte Growth

scholarly journals Proteomic Identification of an Endogenous Synaptic SUMOylome in the Developing Rat Brain

2021 ◽  
Vol 14 ◽  
Author(s):  
Marie Pronot ◽  
Félicie Kieffer ◽  
Anne-Sophie Gay ◽  
Delphine Debayle ◽  
Raphaël Forquet ◽  
...  
Keyword(s):  
Subcellular Fractionation ◽  
Mass Spectrometry Analysis ◽  
Mammalian Brain ◽  
Synaptic Organization ◽  
Post Translational Modifications ◽  
Spectrometry Analysis ◽  
Functional Changes ◽  
Neuronal Communication ◽  
Associated Proteins ◽  
And Function

Synapses are highly specialized structures that interconnect neurons to form functional networks dedicated to neuronal communication. During brain development, synapses undergo activity-dependent rearrangements leading to both structural and functional changes. Many molecular processes are involved in this regulation, including post-translational modifications by the Small Ubiquitin-like MOdifier SUMO. To get a wider view of the panel of endogenous synaptic SUMO-modified proteins in the mammalian brain, we combined subcellular fractionation of rat brains at the post-natal day 14 with denaturing immunoprecipitation using SUMO2/3 antibodies and tandem mass spectrometry analysis. Our screening identified 803 candidate SUMO2/3 targets, which represents about 18% of the synaptic proteome. Our dataset includes neurotransmitter receptors, transporters, adhesion molecules, scaffolding proteins as well as vesicular trafficking and cytoskeleton-associated proteins, defining SUMO2/3 as a central regulator of the synaptic organization and function.

scholarly journals Involvement and Targeted Intervention of Mortalin-Regulated Proteome Phosphorylated-Modification in Hepatocellular Carcinoma

2021 ◽  
Vol 11 ◽  
Author(s):  
Ye Yang ◽  
Ming Jin ◽  
Yi Dai ◽  
Wenqi Shan ◽  
Shuai Chen ◽  
...  
Keyword(s):  
Hepatocellular Carcinoma ◽  
Vascular Endothelial Growth Factor ◽  
Risk Factor ◽  
Growth Factor ◽  
Caffeic Acid ◽  
Molecular Mechanisms ◽  
Vascular Endothelial ◽  
Huh7 Cells ◽  
Associated Proteins ◽  
Endothelial Growth Factor

ObjectivesTo reveal the mechanisms of the effects of mortalin in hepatocellular carcinoma (HCC) and to identify potential novel chemical inhibitors of mortalin.Materials and MethodsFor the experiments, three HCC cell lines (HepG2 cells, Hep3B cells, and sorafenib-resistant HuH7 cells) and xenografted nude mice were used. For the clinical analysis, cohorts of 126 patients with HCC and 34 patients with advanced recurrent HCC receiving sorafenib therapy were examined.ResultsMortalin regulated the phosphorylation-modification of cancer-associated proteins and also regulated angiogenesis-related secretome to cause angiogenesis and sorafenib resistance in HCC cells. Two molecular mechanisms were identified. In one, via phosphatidylinositol 3-kinase (PI3K)/Akt signaling, mortalin regulated nuclear factor (NF)-κB and then activated vascular endothelial growth factor (VEGF)/vascular endothelial growth factor receptor (VEGFR)2 and granulocyte-macrophage colony-stimulating factor (GM-CSF), leading to neovascularization. In the other, mortalin regulated PI3K/Akt/β-catenin and then regulated Bcl-XL and Bcl-2, leading to the antiapoptosis effect of HCC. Treatment of the sorafenib-resistant xenografts with sorafenib in combination with mortalin knockdown facilitated the sorafenib-mediated inhibition of tumor growth and angiogenesis and increased apoptosis. Mortalin was a potential risk factor for HCC, predicting poor prognosis and sorafenib resistance. Finally, we showed that caffeic acid (C9H8O4) could bind to and induce the ubiquitination-mediated degradation of mortalin, which in turn blocked the abovementioned signaling pathways, leading to the inhibition of angiogenesis and the reversal of sorafenib resistance.ConclusionsMortalin, which regulates the phosphorylation of cancer-associated proteins, caused angiogenesis and sorafenib resistance, and was a competitive risk factor for HCC. Caffeic acid can therefore be considered a novel chemical inhibitor that targets the action of mortalin and a potential treatment for HCC.

scholarly journals Synergic Neuroprotection Between Ligusticum Chuanxiong Hort and Borneol Against Ischemic Stroke by Neurogenesis via Modulating Reactive Astrogliosis and Maintaining the Blood–Brain Barrier

2021 ◽  
Vol 12 ◽  
Author(s):  
Bin Yu ◽  
Yao Yao ◽  
Xiaofeng Zhang ◽  
Ming Ruan ◽  
Zhennian Zhang ◽  
...  
Keyword(s):  
Vascular Endothelial Growth Factor ◽  
Growth Factor ◽  
Neurotrophic Factor ◽  
Vascular Endothelial ◽  
Neurological Severity Score ◽  
Associated Proteins ◽  
Von Willebrand ◽  
Endothelial Growth Factor ◽  
Willebrand Factor ◽  
Complement Component 3

Background:Ligusticum chuanxiong Hort (LCH) is a famous ethnomedicine in Asia known for its excellent output on stroke treatment, and borneol usually acts as an assistant for its reducing permeability of the blood–brain barrier (BBB) after stroke. Although their synergy against brain ischemia was verified in previous studies, the potential mechanism is still unknown.Methods: The research aimed to explore the exact synergic mechanisms between LCH and borneol on neurogenesis within the areas of the dentate gyrus and subventricular zone. After treating middle cerebral artery occlusion rats with LCH (0.1 g/kg) and/or borneol (0.08 g/kg), the neurological severity score, brain infarct ratio, Nissl staining, Evans blue permeability, BBB ultrastructure, and expressions of von Willebrand factor and tight junction–associated proteins were measured. Co-localizations of Nestin+/BrdU+ and doublecortin+/BrdU+, and expressions of neuronal nuclei (NeuN) and glial fibrillary acidic protein (GFAP) were observed under a fluorescence microscope. Moreover, astrocyte polarization markers of complement component 3 and pentraxin 3, and relevant neurotrophins were also detected by immunoblotting.Results: Basically, LCH and borneol had different focuses, although both of them decreased infarct areas, and increased quantity of Nissl bodies and expression of brain-derived neurotrophic factor. LCH increased the neurological severity score, NeuN+ cells, and the ratios of Nestin+/BrdU+ and doublecortin+/BrdU+, and decreased GFAP+ cells and ciliary neurotrophic factor expression. Additionally, it regulated the expressions of complement component 3 and pentraxin 3 to transform astrocyte phenotypes. Borneol improved BBB ultrastructure and increased the expressions of von Willebrand factor, tight junction–associated proteins, vascular endothelial growth factor, and vascular endothelial growth factor receptor 2. Unexpectedly, their combined therapy showed more obvious regulations on the Nissl score, Evans blue permeability, doublecortin+/BrdU+, NeuN+ cells, brain-derived neurotrophic factor, and vascular endothelial growth factor than both of their monotherapies.Conclusions: The results indicated that LCH and borneol were complementary to each other in attenuating brain ischemia by and large. LCH mainly promoted neural stem cell proliferation, neurogenesis, and mature neuron preservation, which was probably related to the transformation of reactive astrocytes from A1 subtype to A2, while borneol preferred to maintain the integrity of the BBB, which provided neurogenesis with a homeostatic environment.

scholarly journals Human Podocytes Express Angiopoietin 1, a Potential Regulator of Glomerular Vascular Endothelial Growth Factor

2002 ◽  
Vol 13 (2) ◽  
pp. 544-550 ◽  
Author(s):  
Simon C. Satchell ◽  
Steve J. Harper ◽  
John E. Tooke ◽  
Dontscho Kerjaschki ◽  
Moin A. Saleem ◽  
...  
Keyword(s):  
Vascular Endothelial Growth Factor ◽  
Endothelial Cells ◽  
Growth Factor ◽  
Western Blotting ◽  
Reverse Transcription ◽  
Vascular Endothelial ◽  
Glomerular Capillary ◽  
Reverse Transcription Pcr ◽  
Endothelial Growth Factor ◽  
Angiopoietin 1

ABSTRACT. Vascular endothelial growth factor (VEGF) is abundantly expressed by podocytes, but its role in glomeruli is unknown. Angiopoietins are endothelial cell growth factors that function in concert with VEGF but have not previously been observed in human glomeruli. Angiopoietin 1 (Ang1) acts via the endothelial receptor Tie2 to promote maturation and stabilization of blood vessels, resisting angiogenesis and opposing some actions of VEGF. Ang1, Ang2, Tie2, and VEGF expression in normal human renal cortex was examined with immunofluorescence and immunohistochemical analyses. High-power, multiple-color, immunofluorescence analyses and additional antibodies (specific for particular components of the glomerular filtration barrier) were used to determine the exact locations of Ang1 and Tie2 in the glomerular capillary wall. Immuno-electron-microscopic analysis of rat glomeruli was used to further localize endothelial Tie2 expression. RNA and protein extracted from human glomeruli, cultured human podocytes, and cultured human endothelial cells were analyzed for Ang1, Ang2, and Tie2 by using reverse transcription-PCR and Western blotting. Ang1 was detected in podocytes in normal glomeruli and, with VEGF, was concentrated in podocyte foot processes. Tie2 was demonstrated on glomerular capillary endothelial cells, particularly on the abluminal surface. Reverse transcription-PCR and Western blotting analyses confirmed the expression of Ang1 and Tie2 in glomeruli and of Ang1 in cultured podocytes. These findings suggest a role for Ang1 in the maintenance of the glomerular endothelium and make it a good candidate to be a regulator of the actions of VEGF on glomerular permeability, resisting angiogenesis while allowing the induction of high permeability to water and small solutes.

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