scholarly journals Single-Cell Transcriptomics Analysis of Human Small Antral Follicles

2021 ◽  
Vol 22 (21) ◽  
pp. 11955
Author(s):  
Xueying Fan ◽  
Ioannis Moustakas ◽  
Monika Bialecka ◽  
Julieta S. del Valle ◽  
Arend W. Overeem ◽  
...  
Keyword(s):  
Single Cell ◽  
Tyrosine Kinases ◽  
Transforming Growth Factor ◽  
Integration Site ◽  
Transforming Growth Factor Β ◽  
Antral Follicle ◽  
Antral Follicles ◽  
Transcriptional Signature ◽  
Process Characterization ◽  
Ovulatory Follicles

Human ovarian folliculogenesis is a highly regulated and complex process. Characterization of follicular cell signatures during this dynamic process is important to understand follicle fate (to grow, become dominant, or undergo atresia). The transcriptional signature of human oocytes and granulosa cells (GCs) in early-growing and ovulatory follicles have been previously described; however, that of oocytes with surrounding GCs in small antral follicles have not been studied yet. Here, we have generated a unique dataset of single-cell transcriptomics (SmartSeq2) consisting of the oocyte with surrounding GCs from several individual (non-dominant) small antral follicles isolated from adult human ovaries. We have identified two main types of (healthy) follicles, with a distinct oocyte and GC signature. Using the CellphoneDB algorithm, we then investigated the bi-directional ligand–receptor interactions regarding the transforming growth factor-β (TGFβ)/bone morphogenetic protein (BMP), wingless-type (MMTV)-integration site (WNT), NOTCH, and receptor tyrosine kinases (RTK) signaling pathways between oocyte and GCs within each antral follicle type. Our work not only revealed the diversity of small antral follicles, but also contributes to fill the gap in mapping the molecular landscape of human folliculogenesis and oogenesis.

Fibroblasts positive for meflin have anti-fibrotic property in pulmonary fibrosis

2021 ◽  
pp. 2003397
Author(s):  
Yoshio Nakahara ◽  
Naozumi Hashimoto ◽  
Koji Sakamoto ◽  
Atsushi Enomoto ◽  
Taylor S. Adams ◽  
...  
Keyword(s):  
Pulmonary Fibrosis ◽  
Single Cell ◽  
Transforming Growth Factor ◽  
Transforming Growth Factor Β ◽  
Normal Lung ◽  
Potential Marker ◽  
Rna Hybridisation ◽  
Secretory Phenotype

The prognosis of elderly individuals with idiopathic pulmonary fibrosis (IPF) remains poor. Fibroblastic foci, in which aggregates of proliferating fibroblasts and myofibroblasts are involved, are the pathological hallmark lesions in IPF to represent focal areas of active fibrogenesis. Fibroblast heterogeneity in fibrotic lesions hampers the discovery of the pathogenesis of pulmonary fibrosis. Therefore, to determine of the pathogenesis of IPF, identification of functional fibroblasts is warranted. This study was aimed to determine the role of fibroblasts positive for meflin, identified as a potential marker for mesenchymal stromal cells, during the development of pulmonary fibrosis. We characterised meflin-positive cells in a single cell atlas established by single-cell RNA sequencing (scRNA-seq)-based profiling of 243 472 cells from 32 IPF lungs and 29 normal lung samples. scRNA-seq combined with in situ RNA hybridisation identified proliferating fibroblasts positive for meflin in fibroblastic foci, not dense fibrosis, of fibrotic lungs in IPF patients. We determined the role of fibroblasts positive for meflin using bleomycin (BLM)-induced pulmonary fibrosis. A BLM-induced lung fibrosis model for meflin-deficient mice showed that fibroblasts positive for meflin had anti-fibrotic property to prevent pulmonary fibrosis. Although transforming growth factor-β-induced fibrogenesis and cell senescence with senescence-associated secretory phenotype were exacerbated in fibroblasts via the repression or lack of meflin, these were inhibited in meflin-deficient fibroblasts with meflin reconstitution. These findings provide evidence to show the biological importance of meflin expression on fibroblasts and myofibroblasts in the active fibrotic region of pulmonary fibrosis.

scholarly journals Single cell lineage tracing reveals a role for TgfβR2 in intestinal stem cell dynamics and differentiation

2016 ◽  
Vol 113 (43) ◽  
pp. 12192-12197 ◽  
Author(s):  
Jared M. Fischer ◽  
Peter P. Calabrese ◽  
Ashleigh J. Miller ◽  
Nina M. Muñoz ◽  
William M. Grady ◽  
...  
Keyword(s):  
Single Cell ◽  
Transforming Growth Factor ◽  
Critical Role ◽  
Cell Lineage ◽  
Transforming Growth Factor Β ◽  
Lineage Tracing ◽  
Secretory Cells ◽  
Cancer Tissue ◽  
Tgfβ Signaling

Intestinal stem cells (ISCs) are maintained by a niche mechanism, in which multiple ISCs undergo differential fates where a single ISC clone ultimately occupies the niche. Importantly, mutations continually accumulate within ISCs creating a potential competitive niche environment. Here we use single cell lineage tracing following stochastic transforming growth factor β receptor 2 (TgfβR2) mutation to show cell autonomous effects of TgfβR2 loss on ISC clonal dynamics and differentiation. Specifically, TgfβR2 mutation in ISCs increased clone survival while lengthening times to monoclonality, suggesting that Tgfβ signaling controls both ISC clone extinction and expansion, independent of proliferation. In addition, TgfβR2 loss in vivo reduced crypt fission, irradiation-induced crypt regeneration, and differentiation toward Paneth cells. Finally, altered Tgfβ signaling in cultured mouse and human enteroids supports further the in vivo data and reveals a critical role for Tgfβ signaling in generating precursor secretory cells. Overall, our data reveal a key role for Tgfβ signaling in regulating ISCs clonal dynamics and differentiation, with implications for cancer, tissue regeneration, and inflammation.

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.

Transforming Growth Factor β Expression in the Porcine Ovary: Evidence that Theca Cells are the Major Secretory Source during Antral Follicle Development1

1996 ◽  
Vol 54 (2) ◽  
pp. 485-496 ◽  
Author(s):  
Jeffrey V. May ◽  
Lisa A. Stephenson ◽  
Craig J. Turzcynski ◽  
Hon W. Fong ◽  
Yun-Hwa Mau ◽  
...  
Keyword(s):  
Growth Factor ◽  
Transforming Growth Factor ◽  
Transforming Growth Factor Β ◽  
Antral Follicle ◽  
Theca Cells

182 IMMUNOHISTOCHEMISTRY LOCALIZATION OF GROWTH DIFFERENTIATION FACTOR 9 (GDF-9) AND BONE MORPHOGENETIC PROTEIN 15 (BMP-15) IN CANINE FOLLICLES THROUGHOUT ESTRUS CYCLE

2015 ◽  
Vol 27 (1) ◽  
pp. 182 ◽  
Author(s):  
D. Maupeu ◽  
J. Palomino ◽  
M. De los Reyes
Keyword(s):  
Transforming Growth Factor ◽  
Transforming Growth Factor Β ◽  
Negative Control ◽  
Oestrous Cycle ◽  
Follicle Cells ◽  
Rabbit Serum ◽  
Antral Follicles ◽  
Differentiation Factor ◽  
Morphogenetic Protein ◽  
Growth Differentiation Factor 9

Among different developmental proteins, growth differentiation factor-9 (GDF-9) and bone morphogenic protein 15 (BMP-15), members of the transforming growth factor β (TGFb) superfamily, have been described in many species as key mediators for folliculogenesis. The aim of this study was to analyse in canines the presence and localization of GDF-9 and BMP-15 proteins in follicle cells and oocytes during follicle development. To determine the immunolocalization of GDF-9 and BMP-15, sections of 4% paraformaldehyde in PBS ovaries obtained from 15 bitches at different stage of oestrous cycle (proestrus-oestrus n = 5; diestrus n = 5; anestrus n = 5) were dehydrated (xylene–ethanol solutions) and embedded into paraffin blocks. Individual paraffin sections (5 µm) were inactivated with methanol/H2O2. Sections were blocked at 37°C in the peroxidase reagent and incubated with rabbit anti-human GDF-9 (1 : 200) or goat anti-human BMP-15 (1 : 150) antibodies overnight at 4°C and with secondary antibodies goat anti-rabbit IgG and donkey anti-goat IgG, both HRP conjugated. Previously negative control sections received rabbit serum. Sections were stained with 1 mg mL–1 3,3-diaminobenzidine (DAB) and counterstained with haematoxylin. All sections were examined at magnification × 200 using an IX71 inverted microscope (Olympus, Center Valley, PA, USA), with an IX2-RFA lamp and a ProgRes CapturePro camera (Jenoptik AG, Jena, Germany). The same exposure time was applied to all samples and digital photos were taken and subsequently analysed. The intensity of immunostaining in both oocyte and follicular cells of each follicle was evaluated with Image J version 1.45s software (National Institutes of Health, Bethesda, MD, USA). The images were transformed to grey scale and the corrected total cell mark was calculated by subtracting the mean staining of background reading to the integrated density of each sample. The results showed that samples of all slices contained primordial, primary, secondary and pre-antral follicles as well as different sizes of antral follicles showed positive GDF-9 and BMP-15 immunoreactions in the oocyte cytoplasm and in the follicles cells. No such labelling was found in the negative controls. Follicle sizes differed in GDF-9 and BMP-15 immunoreactions according to oestrous cycle. The pattern of GDF-9 immunostaining in proestrus, oestrus, and diestrus was more intense within preantral rather than antral follicles/oocytes, whereas BMP-15 immunoreaction was prominent mainly in proestrus stage increasing to antral stage. These preliminary results indicated temporal pattern of GDF-9 and BMP-15 in canine ovarian follicles which might play important roles in follicular/oocyte development and ovulation during oestrous cycle. Research was funded by FONDECYT Grant 1140658.

scholarly journals Excessive exosome release is the pathogenic pathway linking a lysosomal deficiency to generalized fibrosis

2019 ◽  
Vol 5 (7) ◽  
pp. eaav3270 ◽  
Author(s):  
Diantha van de Vlekkert ◽  
Jeroen Demmers ◽  
Xinh-Xinh Nguyen ◽  
Yvan Campos ◽  
Eda Machado ◽  
...  
Keyword(s):  
Growth Factor ◽  
Connective Tissue ◽  
Signaling Pathway ◽  
Storage Disease ◽  
Transforming Growth Factor ◽  
Integration Site ◽  
Transforming Growth Factor Β ◽  
Fibrotic Disease ◽  
Disease Pathogenesis ◽  
Deregulated Pathway

Lysosomal exocytosis is a ubiquitous process negatively regulated by neuraminidase 1 (NEU1), a sialidase mutated in the glycoprotein storage disease sialidosis. In Neu1−/− mice, excessive lysosomal exocytosis is at the basis of disease pathogenesis. Yet, the tissue-specific molecular consequences of this deregulated pathway are still unfolding. We now report that in muscle connective tissue, Neu1−/− fibroblasts have features of myofibroblasts and are proliferative, migratory, and exocytose large amounts of exosomes. These nanocarriers loaded with activated transforming growth factor–β and wingless-related integration site (WNT)/β-catenin signaling molecules propagate fibrotic signals to other cells, maintaining the tissue in a prolonged transitional status. Myofibroblast-derived exosomes fed to normal fibroblasts convert them into myofibroblasts, changing the recipient cells’ proliferative and migratory properties. These findings reveal an unexpected exosome-mediated signaling pathway downstream of NEU1 deficiency that propagates a fibrotic disease and could be implicated in idiopathic forms of fibrosis in humans.

Perianale Fisteln bei CED: vom Mausmodell zur Klinik

2018 ◽  
Vol 75 (5) ◽  
pp. 287-294
Author(s):  
Michael Scharl
Keyword(s):  
Growth Factor ◽  
Morbus Crohn ◽  
Transforming Growth Factor ◽  
Transforming Growth Factor Β ◽  
Interleukin 13

Zusammenfassung. Fisteln stellen nach wie vor eine der wichtigsten Komplikationen bei Patienten mit Morbus Crohn dar. Bei mindestens einem Drittel aller Morbus Crohn Patienten treten im Laufe der Erkrankung Fisteln auf. Eine dauerhafte Heilung der Fistel wird jedoch, auch unter Ausschöpfung sämtlicher medikamentöser und chirurgischer Therapieoptionen, nur in rund einem Drittel dieser Patienten erreicht. Der genaue molekulare Mechanismus der Fistelentstehung ist bis heute nicht ganz klar. Aus histopathologischer Sichtweise stellen Fisteln eine röhrenartige Struktur dar, welche von flachen epithelartigen Zellen ausgekleidet ist. Als ursächlicher Entstehungsmechanismus wird dabei die sogenannte epitheliale-zu-mesenchymale Transition (EMT) angesehen und es kann eine starke Expression der Entzündungsmediatoren Tumor Nekrose Faktor, Interleukin-13 und Transforming Growth Factor β in den Fistelarealen nachgewiesen werden. Zusätzlich zu den bereits etablierten, medikamentösen Therapieoptionen, also Antibiotika, Immunmodulatoren und anti-TNF Antikörper, stellt insbesondere der Einsatz der mesenchymalen Stammzelltherapie einen erfolgversprechenden Therapieansatz für die Zukunft dar.

A Novel Missense Mutation in the Endoglin Gene in Hereditary Hemorrhagic Telangiectasia

1997 ◽  
Vol 77 (02) ◽  
pp. 243-247 ◽  
Author(s):  
Hiroshi Yamaguchi ◽  
Hiroyuki Azuma ◽  
Toshio Shigekiyo ◽  
Hideo Inoue ◽  
Shiro Saito
Keyword(s):  
Missense Mutation ◽  
Hereditary Hemorrhagic Telangiectasia ◽  
Transforming Growth Factor ◽  
Transforming Growth Factor Β ◽  
Autosomal Dominant Disorder ◽  
Her Family ◽  
Normal Individuals ◽  
Oligonucleotide Hybridization ◽  
Vascular Dysplasia ◽  
Exon 4

SummaryHereditary hemorrhagic telangiectasia (HHT) is an autosomal dominant disorder characterized by multisystem vascular dysplasia and recurrent hemorrhage. Recent investigation has mapped one of the responsible genes for HHT to chromosome 9q33-q34; subsequently, nine different mutations have been identified in the endoglin gene, which encodes a transforming growth factor β(TGF-β) binding protein, in nine unrelated families with HHT. We examined the endoglin gene in a Japanese patient with HHT and her family members. Using PCR-SSCP. analysis followed by sequencing, we identified a C to A missense mutation in exon 4 which changed an Ala160 codon(GCT) to an Asp160 codon (GAT). Since this mutation destroys one of three Fnu4H I sites in exon 4, the Fnu4H I digestion patterns of the PCR-amplified exon 4 fragments from each family member were analyzed. In affected members, the restriction patterns were all consistent with a phenotype of HHT. PCR-amplified exon 4 fragments from 150 normal individuals were also analyzed by allele-specific oligonucleotide hybridization analysis. As a result, the mutation was not found in any of them. We conclude that the C to A mutation in exon 4 of the endoglin gene in this proband is responsible for the occurrence of HHT in this family.

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