scholarly journals Transcriptome analyses of ovarian stroma: tunica albuginea, interstitium and theca interna

Reproduction ◽  
2019 ◽  
Vol 157 (6) ◽  
pp. 545-565 ◽  
Author(s):  
Katja Hummitzsch ◽  
Nicholas Hatzirodos ◽  
Anne M Macpherson ◽  
Jeff Schwartz ◽  
Raymond J Rodgers ◽  
...  
Keyword(s):  
Tunica Albuginea ◽  
Rt Pcr ◽  
Western Immunoblotting ◽  
Pathway Network ◽  
Fibrous Matrix ◽  
Antral Follicles ◽  
Upstream Regulator ◽  
Ovarian Stroma ◽  
Molecular Determinants ◽  
Theca Interna

The ovary has specialised stromal compartments, including the tunica albuginea, interstitial stroma and theca interna, which develops concurrently with the follicular antrum. To characterise the molecular determinants of these compartments, stroma adjacent to preantral follicles (pre-theca), interstitium and tunica albuginea were laser microdissected (n = 4 per group) and theca interna was dissected from bovine antral follicles (n = 6). RNA microarray analysis showed minimal differences between interstitial stroma and pre-theca, and these were combined for some analyses and referred to as stroma. Genes significantly upregulated in theca interna compared to stroma includedINSL3,LHCGR,HSD3B1,CYP17A1,ALDH1A1,OGN,POSTNandASPN. Quantitative RT-PCR showed significantly greater expression ofOGNandLGALS1in interstitial stroma and theca interna versus tunica and greater expression ofACDin tunica compared to theca interna.PLNwas significantly higher in interstitial stroma compared to tunica and theca. Ingenuity pathway, network and upstream regulator analyses were undertaken. Cell survival was also upregulated in theca interna. The tunica albuginea was associated with GPCR and cAMP signalling, suggesting tunica contractility. It was also associated with TGF-β signalling and increased fibrous matrix. Western immunoblotting was positive for OGN, LGALS1, ALDH1A1, ACD and PLN with PLN and OGN highly expressed in tunica and interstitial stroma (eachn = 6), but not in theca interna from antral follicles (n = 24). Immunohistochemistry localised LGALS1 and POSTN to extracellular matrix and PLN to smooth muscle cells. These results have identified novel differences between the ovarian stromal compartments.

scholarly journals Automated Western immunoblotting detection of anti-SARS-CoV-2 serum antibodies

2021 ◽  
Author(s):  
Sophie Edouard ◽  
Rita Jaafar ◽  
Nicolas Orain ◽  
Philippe Parola ◽  
Philippe Colson ◽  
...  
Keyword(s):  
Negative Control ◽  
Cross Reactivity ◽  
Spike Protein ◽  
Rt Pcr ◽  
Substantial Agreement ◽  
Western Immunoblotting ◽  
Elisa Kit ◽  
Line Test ◽  
Serologic Assay ◽  
Control Samples

AbstractELISA and chemiluminescence serological assays for COVID-19 are currently incorporating only one or two SARS-CoV-2 antigens. We developed an automated Western immunoblotting as a complementary serologic assay for COVID-19. The JessTM Simple Western system, an automated capillary-based assay, was used, incorporating an inactivated SARS-CoV-2 lineage 20a strain as the source of antigen, and total immunoglobulins (IgG, IgM, IgA) detection. In total, 602 sera were tested including 223 from RT-PCR-confirmed COVID-19 patients, 76 from patients diagnosed with seasonal HCoVs and 303 from coronavirus-negative control sera. We also compared this assay with the EUROIMMUN® SARS-CoV-2 IgG ELISA kit. Among 223 sera obtained from RT-PCR-confirmed COVID-19 patients, 180/223 (81%) exhibited reactivity against the nucleocapsid and 70/223 (31%) against the spike protein. Nucleocapsid reactivity was further detected in 9/76 (14%) samples collected from patients diagnosed with seasonal HCoVs and in 15/303 (5%) coronavirus-negative control samples. In the subset of sera collected more than 2 weeks after the onset of symptoms, the sensitivity was 94% and the specificity 93%, the latter value probably reflecting cross-reactivity of SARS-CoV-2 with other coronaviruses. The automated Western immunoblotting presented a substantial agreement (90%) with the compared ELISA (Cohen’s Kappa=0.64). Automated Western immunoblotting may be used as a second line test to monitor exposure of people to HCoVs including SARS-CoV-2.

scholarly journals Hepatic Transcriptome Reveals That Fructose Downregulated Xenobiotics Metabolizing Enzymes Through Aryl Hydrocarbon Receptor Signaling Suppression in C57BL/6 N Mice (P15-011-19)

2019 ◽  
Vol 3 (Supplement_1) ◽  
Author(s):  
Jeong Hoon Pan ◽  
Jingsi Tang ◽  
Kaleigh Beane ◽  
Mersady Redding ◽  
Jiangchao Zhao ◽  
...  
Keyword(s):  
Aryl Hydrocarbon Receptor ◽  
Signaling Pathway ◽  
Bioinformatic Analysis ◽  
University Of Arkansas ◽  
Rt Pcr ◽  
Aryl Hydrocarbon ◽  
Fructose Intake ◽  
Upstream Regulator ◽  
Mice Liver ◽  
The University

Abstract Objectives For decades, fructose intake has been recognized as an environmental risk for metabolic syndromes and diseases. Thus, we comprehensively examined the effects of fructose intake on mice liver transcriptomes. Methods Fructose supplemented water (34%; wt/vol) was fed to both male and female C57BL/6 N mice at their free will for six weeks, followed by hepatic transcriptomics analysis. Based on our criteria, differentially expressed genes (DEGs) were selected and subjected to further computational analyses to predict key pathways and upstream regulator(s). Subsequently, predicted genes and pathways from the transcriptomics dataset were validated via quantitative RT-PCR analyses. Results As results, we identified 89 down-regulated and 88 up-regulated mRNAs in fructose-fed mice livers. These DEGs were subjected to bioinformatic analysis tools in which DEGs were mainly enriched in xenobiotic metabolic processes; further, in the Ingenuity Pathway Analysis software, it was suggested that the aryl hydrocarbon receptor (AhR) is an upstream regulator governing overall changes while fructose suppresses the AhR signaling pathway. In our quantitative RT-PCR validation, we confirmed that fructose suppressed AhR signaling through modulating expressions of transcription factor (arnt) and upstream regulators (ncor2, and rb1). Conclusions Altogether, we demonstrated that ad libitum fructose intake suppresses the canonical AhR signaling pathway in C57BL/6 N mice liver. Based on our current observations, further studies are warranted, especially with regard to the effects of co-exposure to fructose on 1) other types of carcinogens and 2) inflammation inducing agents (or even diets such as a high-fat diet), to find implications of fructose induced-AhR suppression. Funding Sources This work was supported by the University of Arkansas, VPRED Start-up fund and Dale Bumpers College of Agricultural, Food and Life Sciences. Support has been also provided in part by the Arkansas Biosciences Institute, a partnership of scientists from Arkansas Children's Hospital, Arkansas State University, the University of Arkansas-Division of Agriculture, the University of Arkansas, Fayetteville, and the University of Arkansas for Medical Sciences. The Arkansas Biosciences Institute is the major research component of the Arkansas Tobacco Settlement Proceeds Act of 2000. Supporting Tables, Images and/or Graphs

scholarly journals Human Chorionic Gonadotropin-Dependent Up-Regulation of Genes Responsible for Estrogen Sulfoconjugation and Export in Granulosa Cells of Luteinizing Preovulatory Follicles

Endocrinology ◽  
2006 ◽  
Vol 147 (9) ◽  
pp. 4222-4233 ◽  
Author(s):  
Kristy A. Brown ◽  
Monique Doré ◽  
Jacques G. Lussier ◽  
Jean Sirois
Keyword(s):  
Chorionic Gonadotropin ◽  
Granulosa Cells ◽  
Human Chorionic Gonadotropin ◽  
Cell Layer ◽  
Rt Pcr ◽  
Expression Of Genes ◽  
Preovulatory Follicles ◽  
Steady State Levels ◽  
17Β Estradiol ◽  
Theca Interna

Estrogen sulfotransferase (EST) is responsible for the sulfoconjugation of estrogens, thereby changing their physical properties and preventing their action via the estrogen receptors. These sulfoconjugated steroids no longer diffuse freely across the lipid bilayer; instead, they are exported by members of the ATP-binding cassette family, such as ABCC1. The objective of this study was to investigate the regulation of EST and ABCC1 during human chorionic gonadotropin (hCG)-induced ovulation/luteinization. The transcripts for EST and ABCC1 were cloned by RT-PCR, and the regulation of their mRNAs was studied in preovulatory follicles obtained during estrus at 0, 12, 24, 30, 33, 36, and 39 h after hCG. Results obtained from RT-PCR/Southern blot analyses showed significant changes in steady-state levels of both EST and ABCC1 mRNA after hCG treatment (P < 0.05). In granulosa cells, a significant increase in EST transcript was observed 30–39 h after hCG. Similarly, ABCC1 transcript levels were induced in granulosa cells 12–39 h after hCG. In contrast, no significant changes in either EST or ABCC1 were detected in theca interna samples after hCG. The increase in EST and ABCC1 transcripts observed in granulosa cells was reflected in preparations of intact follicle walls, suggesting that the granulosa cell layer contributes the majority of EST and ABCC1 expression in preovulatory follicles. The present study demonstrates that follicular luteinization is accompanied not only by a decrease in 17β-estradiol biosynthesis but also by an increase in expression of genes responsible for estrogen inactivation and elimination from granulosa cells, such as EST and ABCC1, respectively.

Functional ion channels in mouse bone marrow mesenchymal stem cells

2007 ◽  
Vol 293 (5) ◽  
pp. C1561-C1567 ◽  
Author(s):  
Rong Tao ◽  
Chu-Pak Lau ◽  
Hung-Fat Tse ◽  
Gui-Rong Li
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Mesenchymal Stem Cells ◽  
Ion Channel ◽  
Mouse Bone Marrow ◽  
Bone Marrow Mscs ◽  
Rt Pcr ◽  
Western Immunoblotting ◽  
Immunoblotting Analysis ◽  
Marrow Mesenchymal Stem Cells

Bone marrow mesenchymal stem cells (MSCs) are used as a cell source for cardiomyoplasty; however, the cellular electrophysiological properties are not fully understood. The present study was to investigate the functional ionic channels in undifferentiated mouse bone marrow MSCs using whole cell patch-voltage clamp technique, RT-PCR, and Western immunoblotting analysis. We found that three types of ionic currents were present in mouse MSCs, including a Ca2+-activated K+ current ( IKCa), an inwardly rectifying K+ current ( IKir), and a chloride current ( ICl). IKir was inhibited by Ba2+, and IKCa was activated by the Ca2+ ionophore A-23187 and inhibited by the intermediate-conductance IKCa channel blocker clotrimazole. ICl was activated by hyposmotic (0.8 T) conditions and inhibited by the chloride channel blockers DIDS and NPPB. The corresponding ion channel genes and proteins, KCa3.1 for IKCa, Kir2.1 for IKir, and Clcn3 for ICl, were confirmed by RT-PCR and Western immunoblotting analysis in mouse MSCs. These results demonstrate that three types of functional ion channel currents (i.e., IKir, IKCa, and ICl) are present in mouse bone marrow MSCs.

The effect of growth hormone on rat pre-antral follicles in vitro

Zygote ◽  
2000 ◽  
Vol 8 (3) ◽  
pp. 275-283 ◽  
Author(s):  
J. Zhao ◽  
H.T.A. van Tol ◽  
M.A.M. Taverne ◽  
G.C. van der Weijden ◽  
M.M. Bevers ◽  
...  
Keyword(s):  
Growth Hormone ◽  
Growth Hormone Receptor ◽  
Antral Follicle ◽  
High Dose ◽  
Cortical Granules ◽  
Rt Pcr ◽  
Lower Survival Rate ◽  
Antral Follicles ◽  
Before And After

The aim of the present study was to investigate whether growth hormone (GH) has any effect on the development of cultured rat pre-antral follicles. Pre-antral follicles with a diameter between 120 μm and 160 μm were mechanically isolated from 10-day-old rat ovaries and cultured in groups for 6 days in serum-free medium without GH or with GH supplemented at concentrations of 1, 10 and 100 ng/ml, respectively. DNA content of the follicles before and after culture was measured to determine whether possible growth is due to proliferation of follicular cells. To investigate the quality of follicles cultured under different conditions, the ultrastructure of the cultured follicles was studied with transmission electron microscopy. Furthermore, reverse transcriptase polymerase chain reaction (RT-PCR) was used to assess the expression of growth hormone receptor (GHR) in pre-antral follicles. GH, regardless of the concentration, stimulated the growth of pre-antral follicles. However, follicles cultured in medium supplemented with high-dose GH (100 ng/ml) showed a significantly lower survival rate compared with the other groups. Follicles cultured in GH-containing medium showed a better ultrastructure in comparison with those cultured in medium without GH. Remarkably, scattered cortical granules were observed in oocytes of follicles cultured in the presence of GH. With RT-PCR, the presence of the mRNA of GHR was demonstrated in pre-antral follicles. It can be concluded that GH promotes rat pre-antral follicle development in vitro and better supports the morphology of cultured pre-antral follicles. The gene expression of GHR suggests that the action of GH could be mediated by its receptors present in pre-antral follicles.

scholarly journals 206.Cell death of the theca interna during bovine ovarian follicular atresia

2004 ◽  
Vol 16 (9) ◽  
pp. 206
Author(s):  
L. J. Clark ◽  
H. F. Irving-Rodgers ◽  
A. M. Dharmarajan ◽  
R. J. Rodgers
Keyword(s):  
Endothelial Cells ◽  
Cell Death ◽  
Side Chain ◽  
Numerical Density ◽  
Follicular Atresia ◽  
Antral Follicles ◽  
Chain Cleavage ◽  
Significant Difference ◽  
Steroidogenic Cells ◽  
Theca Interna

It is generally accepted that death of cells within the theca interna occurs late during ovarian follicular atresia. Histological classifications of atresia are usually based solely upon the morphology of the membrana granulosa. Atresia of bovine antral follicles less than 5 mm in diameter has been redefined as either antral or basal atresia depending on where in the membrana granulosa cell death is initiated. The aim of present study was to investigate changes within the theca interna during both antral and basal atresia. Bovine ovaries were collected and processed for light microscopy and immunohistochemistry. Each follicle less then 5 mm was classified as either healthy, antral atretic or basal atretic, with antral atresia being further classified either early-mid or late stage. Sections were labelled by TUNEL to identify dead cells combined with lectin from Bandeiraea simplificifolia to identify endothelial cells or with an antibody to cytochrome P450 cholesterol side-chain cleavage to identify steroidogenic cells. The numerical density of steroidogenic cells within the theca interna was significantly reduced (P < 0.001) in basal atretic follicles compared to healthy and antral atretic follicles. In both antral and basal atresia there was death of endothelial cells and steroidogenic cells. However cell death was greater in endothelial cells (P < 0.05) and steroidogenic cells (P < 0.001) of the theca interna of basal atretic follicles. There was no significant difference in the amount of cell death in the membrana granulosa between early-mid antral atresia and basal atresia while death of the membrana granulosa was significantly increased in late antral atresia compared to basal atresia (P < 0.01). Therefore we conclude that basal atresia is not a progression of antral atresia and that the theca interna can be susceptible to cell death early in atresia in basal atretic follicles.

scholarly journals Stromal ovarian tecomatosis

1996 ◽  
Vol 42 (3) ◽  
pp. 40-45
Author(s):  
M. F. Maltseva ◽  
A. A. Pishchulin ◽  
M. E. Bronstein
Keyword(s):  
Polycystic Ovary ◽  
Ovarian Tissue ◽  
Treatment Options ◽  
Interstitial Tissue ◽  
Morphological Form ◽  
Ovarian Stroma ◽  
Morphological Picture ◽  
The Subject ◽  
Theca Interna ◽  
First Time

In 1942, S. Geist and J. Geines were the first to publish the unusual results of a histological examination of ovarian tissue in patients with Stein-Leventhal syndrome. They found foci of "luteinized" tecal cells, similar to theca interna folliculi cells, scattered along the ovarian "stroma" outside the follicles. The following year, L. Fraenkel in his work called these histopathological changes in ovarian hypertosis. In 1949, A. Culiner and S. Shipel used the term "hypertension syndrome" to refer to conditions in which foci of "tekomatosis", consisting of 10-30-60 individual hypertrophic and luteinized tecal cells in the ovarian "stroma", combined with the clinical picture of defemini ation and virilization. In subsequent years, due to the lack of a unified terminology in the literature, synonyms such as hypertosis, cortical stromal hyperplasia, stromal proliferation, stromal tecosis, and tecomatosis were used. R. Feinberg proposed dividing hypertosis into 3 groups, based on the predominance of cystic follicles and a thickened white membrane (mixed tecosis) with increased proliferation of the actual cells of the interstitial tissue or without it (internal tecosis). The presence of "stromal" proliferation in the absence of phenomena of cystic atresia of the follicles, he defined the term "stromal tecosis" (tekomatosis). I.V. Golubeva for the first time in our country drew attention to the clinical and hormonal features of stromal tecomatosis (ST), which was previously considered in the framework of polycystic ovary syndrome, and after 3 years M.E. Bronstein proved that this is an independent morphological form and gave a definition concepts of ST. Ovarian CT is understood as hyperplasia of the interstitial tissue of the ovaries and the appearance in it of groups of hypertrophied epithelioid cells that form foci of various sizes without any connection with follicles. The subject of this review is the consideration of the clinical and morphological picture of this disease, the features of hormonal homeostasis, modern hypotheses of the pathogenetic mechanisms of its development, as well as possible treatment options.

Smooth Muscular Cells in the Mammalian Ovary

1971 ◽  
Vol 29 ◽  
pp. 508-509
Author(s):  
Z. Fumagalli ◽  
P. Motta ◽  
S. Calvieri
Keyword(s):  
Endoplasmic Reticulum ◽  
Plasma Membrane ◽  
Electron Microscope ◽  
Small Groups ◽  
Cortical Area ◽  
Lipid Droplets ◽  
Corpora Lutea ◽  
Ovarian Stroma ◽  
Theca Interna ◽  
Glycogen Particles

The presence of smooth muscular cells was demonstrated with the electron microscope in different areas of the ovary of cats, mice and rabbits. The myocytes were arranged in fascicles, small groups, or most frequently appeared isolated. They were scattered in the ovarian stroma, related to the interstitial cells, in the periphery of the corpora lutea (rarely between luteal cells) in the middle of the gland. Smooth muscular cells were seldom observed between cells of the theca interna and externa of developing follicles and in the middle of atresic follicles. Some smooth muscular cells were found in the cortical area of the ovaries.Each smooth muscular cell showed typical filaments, free ribosomes, lipid droplets and at times glycogen particles. Mitochondria were vesicular; the (Golgi) vesicular complex was often related to two centrioles (frequently in a process of ciliogenesis). The granular endoplasmic reticulum was moderately developed. The plasma membrane presented invaginations and micropinocytotic vesicles as well as tight junctions between adjacent cells. The nucleus was elongated and its envelope formed wide perinuclear cisternae.

scholarly journals The ovarian stroma as a new frontier

Reproduction ◽  
2020 ◽  
Vol 160 (3) ◽  
pp. R25-R39 ◽  
Author(s):  
Hadrian M Kinnear ◽  
Claire E Tomaszewski ◽  
Faith L Chang ◽  
Molly B Moravek ◽  
Min Xu ◽  
...  
Keyword(s):  
Stromal Cell ◽  
Polycystic Ovary ◽  
Tunica Albuginea ◽  
Lymphatic Vessels ◽  
Ovarian Follicles ◽  
Surface Epithelium ◽  
Hormone Production ◽  
Ovarian Stroma ◽  
Extracellular Matrix Components ◽  
New Frontier

Historically, research in ovarian biology has focused on folliculogenesis, but recently the ovarian stroma has become an exciting new frontier for research, holding critical keys to understanding complex ovarian dynamics. Ovarian follicles, which are the functional units of the ovary, comprise the ovarian parenchyma, while the ovarian stroma thus refers to the inverse or the components of the ovary that are not ovarian follicles. The ovarian stroma includes more general components such as immune cells, blood vessels, nerves, and lymphatic vessels, as well as ovary-specific components including ovarian surface epithelium, tunica albuginea, intraovarian rete ovarii, hilar cells, stem cells, and a majority of incompletely characterized stromal cells including the fibroblast-like, spindle-shaped, and interstitial cells. The stroma also includes ovarian extracellular matrix components. This review combines foundational and emerging scholarship regarding the structures and roles of the different components of the ovarian stroma in normal physiology. This is followed by a discussion of key areas for further research regarding the ovarian stroma, including elucidating theca cell origins, understanding stromal cell hormone production and responsiveness, investigating pathological conditions such as polycystic ovary syndrome (PCOS), developing artificial ovary technology, and using technological advances to further delineate the multiple stromal cell types.

scholarly journals Capillary angiogenesis and degeneration in bovine ovarian antral follicles

Reproduction ◽  
2003 ◽  
pp. 211-223 ◽  
Author(s):  
JY Jiang ◽  
G Macchiarelli ◽  
BK Tsang ◽  
E Sato
Keyword(s):  
Ovarian Follicle ◽  
Apical Part ◽  
Follicular Growth ◽  
Functional Changes ◽  
Antral Follicles ◽  
Theca Interna ◽  
Vascular Plexuses ◽  
Capillary Degeneration ◽  
Capillary Angiogenesis

Angiogenesis and capillary degeneration are both evident during ovarian follicle growth. However, the characteristics and distribution of thecal capillary proliferative and degenerative structures have not been fully defined. Indeed, the role of thecal microvasculature changes in follicular atresia is still a matter of debate. The present study examined the distribution of thecal capillary changes occurring during follicular growth and related the changes to capillary morphology (by scanning electron microscopy, SEM, on bovine ovarian corrosion casts) with the incidence of capillary apoptosis (TdT-mediated dUTP nick end-labelling, TUNEL) and follicular status (as confirmed by follicular fluid steroid concentrations). SEM demonstrated well-perfused vascular plexuses of small to large antral follicles with structural and functional changes to capillaries. Angiogenesis was evident mainly in the apical part of the inner capillary layer of medium follicles and the middle or basal part of the inner capillary layer of dominant follicles that exhibited high oestradiol:progesterone ratios. Degenerative capillaries were observed mainly in the outer vascular layers of small follicles, and in the inner and outer vascular layers of medium antral follicles. Although apoptotic structures were present only in the outer capillaries of the theca interna of morphologically healthy antral follicles, atretic follicles showed apoptotic structures in both the outer and inner thecal capillary layers. These results show that angiogenesis increases during bovine follicular growth and occurs unevenly in different inner theca regions of the follicles. The differential angiogenic and degenerative response of theca interna capillaries may reflect differences in the microenvironment of the follicles, which in turn determine the fate of the follicles (continued growth versus atresia).

Sign in / Sign up

Export Citation Format

Share Document