332. GYCOMIC ANALYSES OF THE GRANULOSA AND THECA OF BOVINE OVARIAN FOLLICLES

2010 ◽  
Vol 22 (9) ◽  
pp. 132
Author(s):  
N. Hatzirodos ◽  
J. Nigro ◽  
A. V. Vashi ◽  
H. F. Irving-Rodgers ◽  
B. Caterson ◽  
...  
Keyword(s):  
Granulosa Cells ◽  
Cell Function ◽  
Chondroitin Sulphate ◽  
Heparan Sulphate ◽  
Cell Types ◽  
Ovarian Follicles ◽  
Cell Type ◽  
Thecal Cells ◽  
Theca Externa

Development of ovarian follicles involves changes in cell function and remodelling of the follicular wall. Remodelling necessitates changes in the extracellular matrix, including proteoglycans (PGs). PGs contain glycosaminoglycans (GAGs) covalently bound to a protein core. The length of GAG chains in PGs and the degree and pattern of sulphation differs between cell types and change as cells alter their phenotype. PGs that have been identified in follicles include the chondroitin sulphate (CS) PG, versican and inter-a trypsin inhibitor, and the heparan sulphate (HS) PGs, perlecan and type XVIII collagen. The latter two are found in focimatrix, the follicular and the thecal subendothelial basal laminas. To examine GAGs composition in follicles, bovine antral follicles of various sizes were collected. Follicles were dissected and a biopsy taken for histological classification of health. Theca layers and granulosa cells were collected separately and analysed by fluorophore-assisted carbohydrate (FACE) analysis of GAGs following digestion to disaccharides with chondroitinase ABC, hyaluronidase, heparinase, and heparitinases I and II. Four non GAG sugars and 12 different GAG derived disaccharides were identified and quantitated on a per DNA basis. Healthy versus atretic follicles for each cell type were compared and correlation analyses were also undertaken. Immunohistochemistry using CS specific antibodies was also conducted. There was no effect of size on the GAG content for either granulosa or theca cells. The 4- and 6- sulphated CS sugars were the most abundant following digestion in all tissues. Theca had higher levels than granulosa cells of HS derived disaccharides and also of un- or 4- or 6- N-sulphated CS derived disaccharides. Some sulphated CS moieties localised uniquely to the stroma surrounding blood vessels in the theca externa. Atretic follicles had lower amounts of disaccharides derived from HS in both granulosa and thecal cells, suggesting that heparanase may be activated upon atresia.

scholarly journals Localization of binding sites for IGF-I, insulin and GH in the sow ovary

1999 ◽  
Vol 163 (2) ◽  
pp. 363-372 ◽  
Author(s):  
H Quesnel
Keyword(s):  
Granulosa Cells ◽  
Stromal Cells ◽  
Binding Sites ◽  
Binding Proteins ◽  
Cell Function ◽  
Type I ◽  
Thecal Cells ◽  
Igf I ◽  
Theca Externa ◽  
Theca Interna

Binding sites for IGF-I, insulin, and GH were localized by in situ binding of (125)I-labelled hormones to the different compartments of the sow ovary. Binding sites for IGF-I were detected in oocytes, granulosa and thecal cells of healthy and atretic follicles as well as in the antrum and the stroma. Competition of (125)I-labelled IGF-I with IGF-I, insulin and an analogue of IGF-I (Long R(3)IGF-I), which allowed discrimination between binding to binding proteins from binding to type-I receptors, suggested that type-I receptors were present in granulosa cells of healthy follicles, whilst binding in other compartments was mainly due to binding proteins. Binding of insulin was revealed in oocytes, granulosa and theca interna cells of healthy preantral and antral follicles, and, to a lesser extent, in theca externa and stromal cells, and was still observed in granulosa cells of atretic follicles. Labelling with (125)I-labelled bovine GH was demonstrated in oocytes, granulosa cells, theca interna cells, and, although less intense, in theca externa and stromal cells. It disappeared in granulosa cells during atresia. Binding sites for GH were detected at all follicular stages, from preantral to preovulatory stages, but the intensity of labelling in granulosa cells was more intense in preantral than in large follicles. These data support the participation of insulin, GH and IGF-I in oocyte maturation, follicular growth and stromal cell function in swine.

scholarly journals Northstar enables automatic classification of known and novel cell types from tumor samples

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Fabio Zanini ◽  
Bojk A. Berghuis ◽  
Robert C. Jones ◽  
Benedetta Nicolis di Robilant ◽  
Rachel Yuan Nong ◽  
...  
Keyword(s):  
Cell Types ◽  
Neoplastic Cell ◽  
Computational Approach ◽  
Pancreatic Tumors ◽  
Published Data ◽  
Cell Type ◽  
Disease Heterogeneity ◽  
Manual Task ◽  
Insight Into

Abstract Single cell transcriptomics is revolutionising our understanding of tissue and disease heterogeneity, yet cell type identification remains a partially manual task. Published algorithms for automatic cell annotation are limited to known cell types and fail to capture novel populations, especially cancer cells. We developed northstar, a computational approach to classify thousands of cells based on published data within seconds while simultaneously identifying and highlighting new cell states such as malignancies. We tested northstar on data from glioblastoma, melanoma, and seven different healthy tissues and obtained high accuracy and robustness. We collected eleven pancreatic tumors and identified three shared and five private neoplastic cell populations, offering insight into the origins of neuroendocrine and exocrine tumors. Northstar is a useful tool to assign known and novel cell type and states in the age of cell atlases.

scholarly journals Bone morphogenetic protein-6 enhances gonadotrophin-dependent progesterone and inhibin secretion and expression of mRNA transcripts encoding gonadotrophin receptors and inhibin/activin subunits in chicken granulosa cells

Reproduction ◽  
2007 ◽  
Vol 134 (2) ◽  
pp. 293-306 ◽  
Author(s):  
Sara L Al-Musawi ◽  
Richard T Gladwell ◽  
Philip G Knight
Keyword(s):  
Bone Morphogenetic Protein ◽  
Granulosa Cell ◽  
Granulosa Cells ◽  
Cell Function ◽  
Dependent Manner ◽  
Cell Type ◽  
Inhibin A ◽  
Theca Cells ◽  
Subunit Mrna ◽  
Morphogenetic Protein

The aims were to examine ovarian expression of bone morphogenetic protein (BMP) ligands/receptor mRNAs in the chicken and to test the hypothesis that theca-derived BMP(s) modulates granulosa cell function in a paracrine manner. RT-PCR revealed expression of multiple BMPs in granulosa and theca cells from prehierarchical and preovulatory follicles with greater expression in theca cells; both cell types expressed BMP receptors-IA, -IB and -II consistent with tissue responsiveness. Preovulatory granulosa cells (F1, F2 and F3/4) were cultured with BMP-6 (expressed by theca but not granulosa) in the presence/absence of LH, FSH or 8-Br-cAMP. BMP-6 increased ‘basal’ and gonadotrophin-induced inhibin-A and progesterone secretion by each cell type but did not enhance the effect of 8-Br-cAMP. This indicates that the observed synergism between BMP-6 and gonadotrophin might involve BMP-induced up-regulation of gonadotrophin receptors. In support of this, BMP-6 alone increased LH-receptor (LHR) mRNA in F1 cells and FSH-receptor (FSHR) mRNA in F1, F2 and F3/4 cells. BMP-6 also enhanced LH/FSH-induced LHR transcript amount in each cell type but did not raise FSHR transcript amounts above those induced by BMP-6 alone. To further explore BMP-6 action on inhibin-A secretion, we quantified inhibin/activin subunits (α, βA, βB) mRNAs. Consistent with its effect on inhibin-A secretion, BMP-6 enhanced ‘basal’ expression of α- and βA-subunit mRNA in F1, F2 and F3/4 cells, and βB-subunit mRNA in F3/4 cells. BMP-6 markedly enhanced FSH/LH-induced expression of α-subunit in all follicles and FSH-induced βA-subunit in F2 and F3/4 follicles but not in F1 follicles. Neither BMP-6 alone, nor FSH/LH alone, affected ‘basal’ βB mRNA abundance. However, co-treatment with gonadotrophin and BMP-6 greatly increased βB-subunit expression, the response being lowest in F1 follicles and greatest in F3/4 follicles. Collectively, these results support the hypothesis that intraovarian BMPs of thecal origin have a paracrine role in modulating granulosa cell function in the chicken in a preovulatory stage-dependent manner.

scholarly journals Cell-based therapy technology classifications and translational challenges

2015 ◽  
Vol 370 (1680) ◽  
pp. 20150017 ◽  
Author(s):  
Natalie M. Mount ◽  
Stephen J. Ward ◽  
Panos Kefalas ◽  
Johan Hyllner
Keyword(s):  
Cell Types ◽  
Clinical Translation ◽  
Diverse Group ◽  
Patient Access ◽  
Cell Type ◽  
Commercial Development ◽  
Cell Therapies ◽  
Cell Based Therapy ◽  
Technology Area

Cell therapies offer the promise of treating and altering the course of diseases which cannot be addressed adequately by existing pharmaceuticals. Cell therapies are a diverse group across cell types and therapeutic indications and have been an active area of research for many years but are now strongly emerging through translation and towards successful commercial development and patient access. In this article, we present a description of a classification of cell therapies on the basis of their underlying technologies rather than the more commonly used classification by cell type because the regulatory path and manufacturing solutions are often similar within a technology area due to the nature of the methods used. We analyse the progress of new cell therapies towards clinical translation, examine how they are addressing the clinical, regulatory, manufacturing and reimbursement requirements, describe some of the remaining challenges and provide perspectives on how the field may progress for the future.

scholarly journals EnClaSC: A novel ensemble approach for accurate and robust cell-type classification of single-cell transcriptomes

2019 ◽  
Author(s):  
Xiaoyang Chen ◽  
Shengquan Chen ◽  
Rui Jiang
Keyword(s):  
Single Cell ◽  
Rapid Development ◽  
Scale Up ◽  
Cell Types ◽  
Cell Type ◽  
Species Classification ◽  
Transcriptomic Data ◽  
General Data ◽  
Type Classification

AbstractBackgroundIn recent years, the rapid development of single-cell RNA-sequencing (scRNA-seq) techniques enables the quantitative characterization of cell types at a single-cell resolution. With the explosive growth of the number of cells profiled in individual scRNA-seq experiments, there is a demand for novel computational methods for classifying newly-generated scRNA-seq data onto annotated labels. Although several methods have recently been proposed for the cell-type classification of single-cell transcriptomic data, such limitations as inadequate accuracy, inferior robustness, and low stability greatly limit their wide applications.ResultsWe propose a novel ensemble approach, named EnClaSC, for accurate and robust cell-type classification of single-cell transcriptomic data. Through comprehensive validation experiments, we demonstrate that EnClaSC can not only be applied to the self-projection within a specific dataset and the cell-type classification across different datasets, but also scale up well to various data dimensionality and different data sparsity. We further illustrate the ability of EnClaSC to effectively make cross-species classification, which may shed light on the studies in correlation of different species. EnClaSC is freely available at https://github.com/xy-chen16/EnClaSC.ConclusionsEnClaSC enables highly accurate and robust cell-type classification of single-cell transcriptomic data via an ensemble learning method. We expect to see wide applications of our method to not only transcriptome studies, but also the classification of more general data.

scholarly journals Fibroblasts – the neglected cell type in peripheral sensitization and chronic pain? - A systematic view on the current state of the literature

2021 ◽  
Author(s):  
Naomi Shinotsuka ◽  
Franziska Denk
Keyword(s):  
Chronic Pain ◽  
Cell Function ◽  
Immune Cell ◽  
Data Extraction ◽  
Cell Types ◽  
Extraction Methods ◽  
Peripheral Sensitization ◽  
Cell Type ◽  
Systematic Screening ◽  
Pain Research

AbstractChronic pain and its underlying biological mechanisms have been studied for many decades, with a myriad of molecules, receptors and cell types known to contribute to abnormal pain sensations. We now know that besides an obvious role for neuronal populations in the peripheral and central nervous system, immune cells like microglia, macrophages and T cells are also important drivers of persistent pain. While neuroinflammation has therefore been widely studied in pain research, there is one cell-type that appears to be rather neglected in this context: the humble fibroblast.Fibroblasts may seem unassuming, but actually play a major part in regulating immune cell function and driving chronic inflammation. What is known about them in the context chronic pain?Here we set out to analyze the literature on this topic – using systematic screening and data extraction methods to obtain a balanced view on what has been published. We found that there has been surprisingly little research in this area: 134 articles met our inclusion criteria, only a tiny minority of which directly investigated interactions between fibroblasts and peripheral neurons. We categorized the articles we included – stratifying them according to what was investigated, the estimated quality of results, and any common conclusions.Fibroblasts are a ubiquitous cell type and a prominent source of many pro-algesic mediators in a wide variety of tissues. We think that they deserve a more central role in pain research and propose a new, testable model of how fibroblasts might drive peripheral neuron sensitization.

scholarly journals Quantitative imaging of intracellular density with ratiometric stimulated Raman scattering microscopy

2021 ◽  
Author(s):  
Benjamin Figueroa ◽  
Fiona Xi Xu ◽  
Ruoqian Hu ◽  
Shuaiqian Men ◽  
Dan Fu
Keyword(s):  
Light Scattering ◽  
Raman Scattering ◽  
Cell Size ◽  
Cell Density ◽  
Cell Function ◽  
Stimulated Raman Scattering ◽  
Cell Types ◽  
Tissue Homeostasis ◽  
Cell Type ◽  
Intact Tissue

AbstractCell size and density impact a wide range of physiological functions, including tissue homeostasis, growth regulation, and osmoregulation. Both are tightly regulated in mammalian cells. In comparison, density variation of a given cell type is much smaller than cell size, indicating that maintenance of cell type-specific density is important for cell function. Despite this importance, little is known about how cell density affects cell function and how it is controlled. Current tools for intracellular cell density measurements are limited either to suspended cells or cells growing on 2D substrates, neither of which recapitulate the physiology of single cells in intact tissue. While optical measurements have the potential to measure cell density in situ and noninvasively, light scattering in multicellular systems prevents direct quantification. Here, we introduce an intracellular density imaging technique based on ratiometric stimulated Raman scattering microscopy (rSRS). It quantifies intracellular drymass density through vibrational imaging of macromolecules. Moreover, water is used as an internal standard to correct for aberration and light scattering. We demonstrate real-time measurement of intracellular density quantification and show that density is tightly regulated across different cell types and can be used to differentiate cell types as well as cell states. We further demonstrate dynamic imaging of density change in response to osmotic challenge as well as intracellular density imaging of a 3D tumor spheroid. Our technique has the potential for imaging intracellular density in intact tissue and understanding density regulation and its role in tissue homeostasis.

scholarly journals Characteristic of factors influencing the proper course of folliculogenesis in mammals

2018 ◽  
Vol 6 (1) ◽  
pp. 33-38 ◽  
Author(s):  
Marta Rybska ◽  
Sandra Knap ◽  
Maurycy Jankowski ◽  
Michal Jeseta ◽  
Dorota Bukowska ◽  
...  
Keyword(s):  
Granulosa Cells ◽  
Ovarian Follicle ◽  
Follicular Development ◽  
Ovarian Follicles ◽  
Postnatal Life ◽  
Metabolic Hormones ◽  
Corona Radiata ◽  
Mural Cells ◽  
Thecal Cells ◽  
Reproductive Processes

AbstractFolliculogenesis is the process of ovarian follicle formation,, taking presence during foetal period. During the follicular development, oogoniums undergo meiosis and oocytes are formed. In the ovaries of new born sows, primary and secondary follicles are present and, 90 days after birth, tertiary follicles appear. During development in the ovarian follicles growth of granulosa cells and differentiation of the thecal cells can be observed. A cavity filled with follicular fluid appears. Granulosa cells are divided into: mural cells and corona radiata, which together with the oocyte form the cumulus oophorus. Corona radiata cells, mural layers and oolemma contact each other by a network of gap junctions. Secreted from the pituitary gland, FSH and LH gonadotropin hormones act on receptors located in granular and follicular cells. In the postnatal life tertiary follicles and Graafian follicles are formed. When the follicle reaches a diameter of 1 mm, further growth depends on the secretion of gonadotropins. Mature ovarian follicles produce: progestins, androgens and oestrogens. The growth, differentiation and steroidogenic activity of ovarian follicles, in addition to FSH and LH, is also affected by prolactin, oxytocin, steroid and protein hormones, numerous proteins from the cytokine and interleukin family, metabolic hormones like insulin, glucocorticoids, leptin, thyroid hormones and growth hormones. Despite numerous studies, many processes related to folliculogenesis have not been discovered Learning the mechanisms regulating reproductive processes would allow to easily distinguish pathological processes and discover more and more genes and mechanisms of their expression in cells that build ovarian follicles.

scholarly journals Oestrogen receptor α and β, androgen receptor and progesterone receptor mRNA and protein localisation within the developing ovary and in small growing follicles of sheep

Reproduction ◽  
2006 ◽  
Vol 131 (1) ◽  
pp. 81-92 ◽  
Author(s):  
Jennifer L Juengel ◽  
Derek A Heath ◽  
Laurel D Quirke ◽  
Kenneth P McNatty
Keyword(s):  
Granulosa Cells ◽  
Ovarian Development ◽  
Cell Types ◽  
Surface Epithelium ◽  
Fetal Life ◽  
Follicular Growth ◽  
Early Stages ◽  
Thecal Cells ◽  
Protein Localisation ◽  
Type 3

A first step to elucidating the roles that steroids may play in the processes of ovarian development and early follicular growth is to identify the cell types that are likely to be receptive to steroids. Thus, cell types expressing receptors for oestrogen (α and β form; ERα and ERβ respectively), androgen (AR) and progesterone (PR) were determined by in situ hybridisation and immunohistochemistry in ovine ovarian tissues collected during ovarian development and follicular formation (days 26–75 of fetal life) as well as during the early stages of follicular growth. Expression of ERβ was observed early during ovarian development and continued to be expressed throughout follicular formation and also during the early stages of follicular growth. ERβ was identified in germ cells as well as in the granulosa cells. At the large preantral stage of follicular growth, expression of ERα was also consistently observed in granulosa cells. AR was first consistently observed at day 55 of fetal life in stroma cells throughout the ovary. Within the follicle, expression was observed in granulosa and thecal cells from the type-2 to -3 stage of follicular growth. PR mRNA did not appear to be expressed during ovarian development (days 26–75 of gestation). However, PR (mRNA and protein) was observed in the theca of type-3 (small preantral) and larger follicles, with mRNA – but not protein – observed in granulosa cells of some type-4 and 5 follicles. Expression of ERβ, ERα and AR, as well as PR, was also observed in the surface epithelium and ovarian stroma of the fetal, neonatal and adult ovary. Thus, in sheep, steroid hormones have the potential to regulate the function of a number of different ovarian cell types during development, follicular formation and early follicular growth.

scholarly journals Chronic stress induces co-ordinated cortical microcircuit cell type transcriptomic changes consistent with altered information processing

2020 ◽  
Author(s):  
Dwight F. Newton ◽  
Hyunjung Oh ◽  
Rammohan Shukla ◽  
Keith Misquitta ◽  
Corey Fee ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Information Processing ◽  
Stress Response ◽  
Cell Function ◽  
Cell Types ◽  
Brain Regions ◽  
Oxidative Stress Response ◽  
Dendritic Morphology ◽  
Mild Stress ◽  
Cell Type

ABSTRACTMajor depressive disorder (MDD) is associated with altered GABAergic and glutamatergic signalling, suggesting altered excitation-inhibition balance (EIB) in cortical mood- and cognition-regulating brain regions. Information processing in cortical microcircuits involves regulation of pyramidal (PYR) cells by Somatostatin-(SST), Parvalbumin-(PV), and Vasoactive intestinal peptide-(VIP) expressing interneurons. Human and rodent studies suggest that impaired PYR-cell dendritic morphology and decreased SST-cell function may mediate altered EIB in MDD. However, knowledge of co-ordinated changes across microcircuit cell types is virtually absent. We thus investigated the co-ordinated transcriptomic effects of UCMS on microcircuit cell types in the medial prefrontal cortex. C57Bl/6 mice, exposed to unpredictable chronic mild stress (UCMS) or control housing for five weeks were assessed for anxiety- and depressive-like behaviours. Microcircuit cell types were laser-microdissected and processed for RNA-sequencing. UCMS-exposed mice displayed predicted elevated behavioural emotionality. Each microcircuit cell type showed a unique transcriptional signature after UCMS. Pre-synaptic functions, oxidative stress response, metabolism, and translational regulation were differentially dysregulated across cell types, whereas nearly all cell types showed down-regulated post-synaptic gene signatures. At the microcircuit level, we observed a shift from distributed transcriptomic co-ordination across cell types in controls towards UCMS-induced increased co-ordination between PYR-, SST- and PV-cells, and a hub-like role for PYR-cells. Lastly, we identified a microcircuit-wide co-expression network enriched in synaptic, bioenergetic, and oxidative stress response genes that correlated with UCMS-induced behaviours. Together, these findings suggest cell-specific deficits, microcircuit-wide synaptic reorganization, and a shift in cortical EIB mediated by increased co-ordinated regulation of PYR-cells by SST- and PV-cells.

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