scholarly journals Proteomic analysis of follicular fluid in carriers and non-carriers of the Trio allele for high ovulation rate in cattle

2018 ◽  
Vol 30 (12) ◽  
pp. 1643 ◽  
Author(s):  
Mamat H. Kamalludin ◽  
Alvaro Garcia-Guerra ◽  
Milo C. Wiltbank ◽  
Brian W. Kirkpatrick
Keyword(s):  
Follicular Fluid ◽  
Transforming Growth Factor ◽  
Fluid Collection ◽  
Transforming Growth Factor Β ◽  
Initial Study ◽  
Ovulation Rate ◽  
Differentially Expressed ◽  
Masking Effect ◽  
Joint Analysis ◽  
Abundance Proteins

This study was conducted to characterise differences in follicular fluid proteins between carriers and non-carriers of a bovine allele for high ovulation rate. A total of four non-carrier and five carrier females were used in an initial study with four and six additional non-carriers and carriers respectively used in a validation study. Emergence of the follicular wave was synchronised and the ovaries containing the dominant follicle(s) were extracted by ovariectomy for follicular fluid collection. A hexapeptide ligand library was used to overcome the masking effect of high-abundance proteins and to increase detection of low-abundance proteins in tandem mass spectrometry. After correcting for multiple comparisons, only two proteins, glia-derived nexin precursor (SERPINE2) and inhibin β B chain precursor (INHBB), were significantly differentially expressed (false-discovery rate <0.05). In a replicate study of analogous design differential expression was confirmed (P < 0.05). Joint analysis of results from the two studies indicated that three additional proteins were consistently differentially expressed between genotypes. For three of these five, previous studies have indicated that expression is increased by transforming growth factor-β–bone morphogenetic protein signalling; their reduction in follicular fluid from carrier animals is consistent with the ~9-fold overexpression of SMAD family member 6 (SMAD6) in carriers that is inhibitory to this pathway.

scholarly journals Active immunization against the proregions of GDF9 or BMP15 alters ovulation rate and litter size in mice

Reproduction ◽  
2012 ◽  
Vol 143 (2) ◽  
pp. 195-201 ◽  
Author(s):  
C Joy McIntosh ◽  
Steve Lawrence ◽  
Peter Smith ◽  
Jennifer L Juengel ◽  
Kenneth P McNatty
Keyword(s):  
Litter Size ◽  
Transforming Growth Factor ◽  
Transforming Growth Factor Β ◽  
Cross Reactivity ◽  
Full Length ◽  
Ovulation Rate ◽  
Corpora Lutea ◽  
Immunization Group

The transforming growth factor β (TGFB) superfamily proteins bone morphogenetic protein 15 (BMP15) and growth differentiation factor 9 (GDF9), are essential for mammalian fertility. Recent in vitro evidence suggests that the proregions of mouse BMP15 and GDF9 interact with their mature proteins after secretion. In this study, we have actively immunized mice against these proregions to test the potential in vivo roles on fertility. Mice were immunized with either N- or C-terminus proregion peptides of BMP15 or GDF9, or a full-length GDF9 proregion protein, each conjugated to keyhole limpet hemocyanin (KLH). For each immunization group, ovaries were collected from ten mice for histology after immunization, while a further 20 mice were allowed to breed and litter sizes were counted. To link the ovulation and fertility data of these two experimental end points, mice were joined during the time period identified by histology as being the ovulatory period resulting in to the corpora lutea (CL) counted. Antibody titers in sera increased throughout the study period, with no cross-reactivity observed between BMP15 and GDF9 sera and antigens. Compared with KLH controls, mice immunized with the N-terminus BMP15 proregion peptide had ovaries with fewer CL (P<0.05) and produced smaller litters (P<0.05). In contrast, mice immunized with the full-length GDF9 proregion not only had more CL (P<0.01) but also had significantly smaller litter sizes (P<0.01). None of the treatments affected the number of antral follicles per ovary. These findings are consistent with the hypothesis that the proregions of BMP15 and GDF9, after secretion by the oocyte, have physiologically important roles in regulating ovulation rate and litter size in mice.

scholarly journals Differential expression of CART in ewes with differing ovulation rates

Reproduction ◽  
2017 ◽  
Vol 153 (4) ◽  
pp. 471-479 ◽  
Author(s):  
Jennifer L Juengel ◽  
Michelle C French ◽  
Laurel D Quirke ◽  
Alexia Kauff ◽  
George W Smith ◽  
...  
Keyword(s):  
Differential Expression ◽  
Expression Pattern ◽  
Follicular Fluid ◽  
Follicular Phase ◽  
Distinct Pattern ◽  
Ovulation Rate ◽  
Differentially Expressed ◽  
Antral Follicles ◽  
Cart Peptide

We hypothesised that cocaine- and amphetamine-regulated transcript (CARTPT) would be differentially expressed in ewes with differing ovulation rates. Expression of mRNA forCARTPT, as well asLHCGR,FSHR,CYP19A1andCYP17A1was determined in antral follicles ≥1 mm in diameter collected during the follicular phase in ewes heterozygous for the Booroola and Inverdale genes (I+B+; average ovulation rate 4) and ++ contemporaries (++; average ovulation rate 1.8). In ++ ewes (n = 6),CARTPTwas expressed in small follicles (1 to <3 mm diameter), where 18.8 ± 2.5% follicles expressedCARTPT. CART peptide was also detected in follicular fluid of some follicles of ++ ewes. In I+B+ ewes, 5/6 ewes did not have any follicles that expressedCARTPT, and no CART peptide was detected in any follicle examined. Expression pattern ofCYP19A1differed between I+B+ and ++ ewes with an increased percentage of small and medium follicles (3 to <4.5 mm diameter) but decreased percentage of large follicles (≥4.5 mm diameter) expressingCYP19A1in the I+B+ ewes. Many of the large follicles from the I+B+ ewes appeared non-functional and expression ofLHCGR,FSHR,CYP17A1andCYP19A1was less than that observed in ++ ewes. Expression ofFSHRandCYP17A1was not different between groups in small and medium follicles, butLHCGRexpression was approximately double in I+B+ ewes compared to that in ++ ewes. Thus, ewes with high ovulation rates had a distinct pattern of expression ofCARTPTmRNA and protein compared to ewes with normal ovulation rates, providing evidence for CART being important in the regulation of ovulation rate.

208 EFFECT OF GDF8 AND SB-431542 ON PORCINE OOCYTE DURING IN VITRO MATURATION AND SUBSEQUENT EMBRYONIC DEVELOPMENT

2016 ◽  
Vol 28 (2) ◽  
pp. 235
Author(s):  
J. D. Yoon ◽  
E. Lee ◽  
S.-H. Hyun
Keyword(s):  
Treatment Group ◽  
Embryonic Development ◽  
Follicular Fluid ◽  
In Vitro Maturation ◽  
Transforming Growth Factor ◽  
Formation Rate ◽  
Transforming Growth Factor Β ◽  
Nuclear Maturation ◽  
Porcine Oocyte

Growth differentiation factor-8 (GDF8) is a member of the transforming growth factor-β that has been identified as a strong physiological regulator. SB-431542 (SB) is a specific inhibitor of transforming growth factor-β superfamily type I activin receptor-like kinase (ALK) receptors such as ALK4, ALK5, and ALK7. The purpose of this study is investigation of the effects of GDF8 and SB on porcine oocytes during in vitro maturation and subsequent embryonic development. We first performed ELISA to detect GDF8 concentrations in follicular fluid for each size of follicle; sizes were as follows: small (<3 mm), medium (>3 mm and <6 mm), and large (>6 mm) follicle. After detection of the GDF8 concentration in follicular fluid, we investigated the effect of GDF8 and SB treatment during in vitro maturation (IVM) on nuclear maturation, intracellular glutathione (GSH), and reactive oxygen species (ROS) levels, and embryonic development after IVF and parthenogenetic activation (PA). Data were analysed by ANOVA followed by Duncan using SPSS (Statistical Package for Social Science, IBM, New York, NY, USA) mean ± SEM. The ELISA result showed different concentrations of GDF8 for each grade of follicular fluid: small, 0.479 ng mL–1; medium, 0.668 ng mL–1; and large, 1.318 ng mL–1. During the IVM process, 1.318 ng mL–1 of GDF8 and 5 ng mL–1 of SB were added to the maturation medium as control, SB, SB+GDF8, and GDF8 treatment groups. After 44 h of IVM, GDF8 group (90.4%) showed a significantly higher nuclear maturation rate than control and SB+GDF8 groups (85.4 and 81.7%). The SB group (78.9%) showed significantly reduced nuclear maturation rate compared with control (P < 0.05). The GDF8 treatment group showed a significant decreased intracellular ROS and increased GSH levels compared with other groups (P < 0.05). The SB+GBF8 treatment group showed a significantly better cytoplasmic maturation than the SB treatment group. In the PA embryonic development analysis, the GDF8 treatment group showed a significantly higher blastocyst formation rate compared with other groups (47.9, 37.2, 46.4, and 58.7% respectively; P < 0.05). In the IVF embryonic development analysis, the GDF8 treatment groups showed significantly higher blastocyst formation rate compared with the SB group (28.2 and 42.2%, respectively; P < 0.05). In conclusion, treatment with GDF8 during porcine oocyte IVM improved the embryonic developmental competence via increased cytoplasmic maturation and led to better oocyte maturation from the ALK receptor inhibition by SB.

scholarly journals Identification of novel genes and pathways in carotid atheroma using integrated bioinformatic methods

2016 ◽  
Vol 6 (1) ◽  
Author(s):  
Wenqing Nai ◽  
Diane Threapleton ◽  
Jingbo Lu ◽  
Kewei Zhang ◽  
Hongyuan Wu ◽  
...  
Keyword(s):  
Signaling Pathway ◽  
Transforming Growth Factor ◽  
Interaction Network ◽  
Enrichment Analysis ◽  
Functional Enrichment Analysis ◽  
Transforming Growth Factor Β ◽  
Functional Enrichment ◽  
Differentially Expressed ◽  
Pathway Enrichment Analysis ◽  
Atheromatous Plaques

Abstract Atherosclerosis is the primary cause of cardiovascular events and its molecular mechanism urgently needs to be clarified. In our study, atheromatous plaques (ATH) and macroscopically intact tissue (MIT) sampled from 32 patients were compared and an integrated series of bioinformatic microarray analyses were used to identify altered genes and pathways. Our work showed 816 genes were differentially expressed between ATH and MIT, including 443 that were up-regulated and 373 that were down-regulated in ATH tissues. GO functional-enrichment analysis for differentially expressed genes (DEGs) indicated that genes related to the “immune response” and “muscle contraction” were altered in ATHs. KEGG pathway-enrichment analysis showed that up-regulated DEGs were significantly enriched in the “FcεRI-mediated signaling pathway”, while down-regulated genes were significantly enriched in the “transforming growth factor-β signaling pathway”. Protein-protein interaction network and module analysis demonstrated that VAV1, SYK, LYN and PTPN6 may play critical roles in the network. Additionally, similar observations were seen in a validation study where SYK, LYN and PTPN6 were markedly elevated in ATH. All in all, identification of these genes and pathways not only provides new insights into the pathogenesis of atherosclerosis, but may also aid in the development of prognostic and therapeutic biomarkers for advanced atheroma.

Mechanisms of action of the principal prolific genes and their application to sheep production

2004 ◽  
Vol 16 (4) ◽  
pp. 395 ◽  
Author(s):  
C. J. H. Souza ◽  
A. González-Bulnes ◽  
B. K. Campbell ◽  
A. S. McNeilly ◽  
D. T. Baird
Keyword(s):  
Transforming Growth Factor ◽  
Ovarian Follicle ◽  
Transforming Growth Factor Β ◽  
Ovulation Rate ◽  
Follicle Growth ◽  
Sheep Industry ◽  
Morphogenetic Protein ◽  
Sheep Production ◽  
Ovulatory Follicles ◽  
On Farm

The prolificacy variation in sheep makes it an excellent animal model to understand the mechanisms regulating ovulation rate. Identification of mutations responsible for the increased prolificacy of the Inverdale, Booroola, Javanese, Cambridge and Belclare sheep open new avenues of investigation for the paracrine control of folliculogenesis. To date, all known mutations are in genes from ligands or receptors of the transforming growth factor β superfamily, and point to the bone morphogenetic protein family of peptides as local regulators of ovarian follicle growth. The mechanism of action of the mutated genes is not fully understood, but results in the ovulation of a higher number of follicles with smaller diameter and fewer granulosa cells than that of the wildtype, thus speeding the differentiation of ovulatory follicles. Comparisons of the performance of Booroola-crossed flocks in different countries showed that carriers of the prolificacy mutation have higher ewe productivity but also higher perinatal mortality and lighter weight lambs. Their economic impact on the sheep industry depends on farm environment and management. Nevertheless, the diagnostic tests now available to identify the genetic mutations resulting in increased ovulation rate, will simplify the introduction of these mutations and their monitoring in flocks for research and commercial purposes.

scholarly journals Transcriptomic analysis of vitamin D responses in uterine and peripheral NK cells

Reproduction ◽  
2019 ◽  
Vol 158 (2) ◽  
pp. 213-223 ◽  
Author(s):  
J A Tamblyn ◽  
L E Jeffery ◽  
R Susarla ◽  
D M Lissauer ◽  
S L Coort ◽  
...  
Keyword(s):  
Vitamin D ◽  
Natural Killer Cells ◽  
Natural Killer ◽  
Transforming Growth Factor ◽  
Active Form ◽  
Transforming Growth Factor Β ◽  
Differentially Expressed ◽  
Rna Seq ◽  
Killer Cells ◽  
Adverse Pregnancy

Vitamin D deficiency is prevalent in pregnant women and is associated with adverse pregnancy outcomes, in particular disorders of malplacentation. The active form of vitamin D, 1,25-dihydroxyvitamin D3 (1,25(OH)2D3), is a potent regulator of innate and adaptive immunity, but its immune effects during pregnancy remain poorly understood. During early gestation, the predominant immune cells in maternal decidua are uterine natural killer cells (uNK), but the responsivity of these cells to 1,25(OH)2D3 is unknown despite high levels of 1,25(OH)2D3 in decidua. Transcriptomic responses to 1,25(OH)2D3 were characterised in paired donor uNK and peripheral natural killer cells (pNK) following cytokine (CK) stimulation. RNA-seq analyses indicated 911 genes were differentially expressed in CK-stimulated uNK versus CK-stimulated pNK in the absence of 1,25(OH)2D3, with predominant differentially expressed pathways being associated with glycolysis and transforming growth factor β (TGFβ). RNA-seq also showed that the vitamin D receptor (VDR) and its heterodimer partner retinoid X receptor were differentially expressed in CK-stimulated uNK vs CK-stimulated pNK. Further analyses confirmed increased expression of VDR mRNA and protein, as well as VDR-RXR target in CK-stimulated uNK. RNA-seq analysis showed that in CK-stimulated pNK, 1,25(OH)2D3 induced 38 and suppressed 33 transcripts, whilst in CK-stimulated uNK 1,25(OH)2D3 induced 46 and suppressed 19 genes. However, multiple comparison analysis of transcriptomic data indicated that 1,25(OH)2D3 had no significant overall effect on gene expression in either CK-stimulated pNK or uNK. These data indicate that CK-stimulated uNK are transcriptionally distinct from pNK and, despite expressing abundant VDR, neither pNK nor uNK are sensitive targets for vitamin D.

scholarly journals The latent transforming growth factor-β binding protein 4, LTBP4, is differentially expressed in non-small cell lung cancer and associates with patient survival.

2020 ◽  
Author(s):  
Shahan Mamoor
Keyword(s):  
Lung Cancer ◽  
Small Cell Lung Cancer ◽  
Cell Lung Cancer ◽  
Transforming Growth Factor ◽  
Transforming Growth Factor Β ◽  
The United States ◽  
Small Cell ◽  
Differentially Expressed ◽  
Small Cell Lung ◽  
Lower Expression

Non-small cell lung cancer (NSCLC) is the leading cause of cancer death in the United States (1). We mined published microarray data (2, 3, 4) to identify differentially expressed genes in NSCLC. We found that LTBP4 was among the genes whose expression was most quantitatively different in tumors from patients with NSCLC as compared to the lung. LTBP4 expression was significantly decreased in NSCLC tumors as compared to the lung, and lower expression of LTBP4 in patient tumors was significantly associated with worse overall survival. LTBP4 may be important for initiation or progression of non-small cell lung cancer in humans.

scholarly journals Common fibroid-associated genes are differentially expressed in phenotypically dissimilar cell populations isolated from within human fibroids and myometrium

Reproduction ◽  
2014 ◽  
Vol 147 (5) ◽  
pp. 683-692 ◽  
Author(s):  
Sarah J Holdsworth-Carson ◽  
Marina Zaitseva ◽  
Jane E Girling ◽  
Beverley J Vollenhoven ◽  
Peter A W Rogers
Keyword(s):  
Hormone Receptors ◽  
Transforming Growth Factor ◽  
Uterine Fibroids ◽  
Transforming Growth Factor Β ◽  
Cell Types ◽  
Steroid Hormone Receptors ◽  
Differentially Expressed ◽  
Common Disease ◽  
New Paradigm ◽  
Progesterone Receptor B

Uterine fibroids are a prevalent gynaecological condition in reproductive-aged women and are the commonest reason for hysterectomy. The cellular composition of clonal fibroids are heterogeneous, with phenotypically dissimilar cells that include smooth muscle cells (SMC), vascular SMC (VSMC) and fibroblasts. The aim of our study was to investigate genes that are commonly differentially expressed between fibroid and myometrial whole tissues in phenotypically different sub-populations of cells isolated from fibroid and myometrium. Genes to be investigated by fluorescence-activated cell sorting, quantitative real-time PCR and immunocytochemistry include transforming growth factor β (TGFB) and retinoic acid (RA) signalling families and steroid hormone receptors. We hypothesised that each cell population isolated from fibroid and myometrium would differ in the expression of fibroid-associated genes. We demonstrated that phenotypically different cellular constituents of uterine fibroids differentially express cellular RA-binding protein 2 (CRABP2), progesterone receptor B (PRB) and TGFB receptor 2 mRNA in fibroid-derived cells of VSMC and SMC phenotype. CRABP2 mRNA was also differentially expressed in fibroblasts and VSMC sub-populations from within clonal fibroid tumours. We conclude that differential regulation of RA, TGFB and PR pathway transcription occurs in fibroid-associated SMC and -fibroblasts and that investigation of paracrine interactions between different cell types within the fibroid microenvironment provides an important new paradigm for understanding the pathophysiology of this common disease.

scholarly journals Proteomics Analysis of Colostrum Samples from Sows Housed under Different Conditions

Animals ◽  
2020 ◽  
Vol 10 (2) ◽  
pp. 355
Author(s):  
Guoan Yin ◽  
Lei Wang ◽  
Xiaoyu Zhao ◽  
Langchao Yu ◽  
Dapeng Huang
Keyword(s):  
Signaling Pathway ◽  
Transforming Growth Factor ◽  
Cholesterol Metabolism ◽  
Transforming Growth Factor Β ◽  
Differentially Expressed ◽  
Housing System ◽  
Glutathione Metabolism ◽  
Peroxisome Proliferator ◽  
Differentially Expressed Proteins ◽  
Peroxisome Proliferator Activated Receptor

This study investigated the proteomic characteristics of colostrum for sows housed under different conditions. Among 12 gilts, four were housed in a gestation-crate and farrowing-crate combined housing system (CC) as controls, four were housed in a gestation-pen and farrowing-pen combined housing system (PP), and four were housed in a gestation-pen and farrowing-crate combined housing system (PC). Differentially expressed proteins in the colostrum (PP versus CC, and PC versus CC) were screened by proteomics technology, and bioinformatics analysis was then performed. Results showed that 93 proteins were differentially expressed in PP versus CC, and that 126 proteins were differentially expressed in PC versus CC. The differentially expressed proteins in the PP versus CC comparison were mainly enriched in interleukin (IL)-17, transforming growth factor-β, and nuclear factor-κ B signaling pathways, and in metabolic pathways, including glutathione metabolism, peroxisome, and carbon metabolism. In contrast, differentially expressed proteins in the PC versus CC comparison were enriched in the IL-17 signaling pathway, cholesterol metabolism, and peroxisome proliferator-activated receptor signaling pathway. In conclusion, the housing environment appeared to affect the colostrum composition of sows by acting on their immune system and metabolic processes, particularly fat metabolism.

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