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  1. Article ; Online: Sperm-specific proteins: new implications for diagnostic development and cancer immunotherapy.

    O'Donnell, Liza / Smith, Lee B / Rebourcet, Diane

    Current opinion in cell biology

    2022  Volume 77, Page(s) 102104

    Abstract: Spermatozoa are comprised of many unique proteins not expressed elsewhere. Sperm-specific proteins are first expressed at puberty, after the development of immune tolerance to self-antigens, and have been assumed to remain confined inside the ... ...

    Abstract Spermatozoa are comprised of many unique proteins not expressed elsewhere. Sperm-specific proteins are first expressed at puberty, after the development of immune tolerance to self-antigens, and have been assumed to remain confined inside the seminiferous tubules, protected from immune cell recognition by various mechanisms of testicular immune privilege. However, new data has shown that sperm-specific proteins are released by the tubules into the surrounding interstitial fluid; from here they can contact immune cells, potentially promote immune tolerance, and enter the circulation. These new findings have clinical implications for diagnostics and therapeutics targeted at a specific class of proteins known as cancer-testis antigens (CTA), the opportunity to identify new communication pathways in the testis, and to discover new ways to monitor testis function.
    MeSH term(s) Humans ; Immunotherapy ; Male ; Neoplasms/diagnosis ; Neoplasms/therapy ; Semen ; Seminiferous Tubules ; Spermatozoa
    Language English
    Publishing date 2022-06-04
    Publishing country England
    Document type Journal Article ; Review ; Research Support, Non-U.S. Gov't
    ZDB-ID 1026381-0
    ISSN 1879-0410 ; 0955-0674
    ISSN (online) 1879-0410
    ISSN 0955-0674
    DOI 10.1016/j.ceb.2022.102104
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  2. Article ; Online: New Insights into Testosterone Biosynthesis: Novel Observations from HSD17B3 Deficient Mice.

    Lawrence, Ben M / O'Donnell, Liza / Smith, Lee B / Rebourcet, Diane

    International journal of molecular sciences

    2022  Volume 23, Issue 24

    Abstract: Androgens such as testosterone and dihydrotestosterone (DHT) are essential for male sexual development, masculinisation, and fertility. Testosterone is produced via the canonical androgen production pathway and is essential for normal masculinisation and ...

    Abstract Androgens such as testosterone and dihydrotestosterone (DHT) are essential for male sexual development, masculinisation, and fertility. Testosterone is produced via the canonical androgen production pathway and is essential for normal masculinisation and testis function. Disruption to androgen production can result in disorders of sexual development (DSD). In the canonical pathway, 17β-hydroxysteroid dehydrogenase type 3 (HSD17B3) is viewed as a critical enzyme in the production of testosterone, performing the final conversion required. HSD17B3 deficiency in humans is associated with DSD due to low testosterone concentration during development. Individuals with
    Language English
    Publishing date 2022-12-08
    Publishing country Switzerland
    Document type Journal Article ; Review
    ZDB-ID 2019364-6
    ISSN 1422-0067 ; 1422-0067 ; 1661-6596
    ISSN (online) 1422-0067
    ISSN 1422-0067 ; 1661-6596
    DOI 10.3390/ijms232415555
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  3. Article ; Online: Development of peptides for targeting cell ablation agents concurrently to the Sertoli and Leydig cell populations of the testes: An approach to non-surgical sterilization.

    Fraser, Barbara / Wilkins, Alex / Whiting, Sara / Liang, Mingtao / Rebourcet, Diane / Nixon, Brett / Aitken, Robert John

    PloS one

    2024  Volume 19, Issue 4, Page(s) e0292198

    Abstract: The surgical sterilization of cats and dogs has been used to prevent their unwanted breeding for decades. However, this is an expensive and invasive procedure, and often impractical in wider contexts, for example the control of feral populations. A ... ...

    Abstract The surgical sterilization of cats and dogs has been used to prevent their unwanted breeding for decades. However, this is an expensive and invasive procedure, and often impractical in wider contexts, for example the control of feral populations. A sterilization agent that could be administered in a single injection, would not only eliminate the risks imposed by surgery but also be a much more cost-effective solution to this worldwide problem. In this study, we sought to develop a targeting peptide that would selectively bind to Leydig cells of the testes. Subsequently, after covalently attaching a cell ablation agent, Auristatin, to this peptide we aimed to apply this conjugated product (LH2Auristatin) to adult male mice in vivo, both alone and together with a previously developed Sertoli cell targeting peptide (FSH2Menadione). The application of LH2Auristatin alone resulted in an increase in sperm DNA damage, reduced mean testes weights and mean seminiferous tubule size, along with extensive germ cell apoptosis and a reduction in litter sizes. Together with FSH2Menadione there was also an increase in embryo resorptions. These promising results were observed in around a third of all treated animals. Given this variability, we discuss how these reagents might be modified in order to increase target cell ablation and improve their efficacy as sterilization agents.
    MeSH term(s) Male ; Mice ; Animals ; Cats ; Dogs ; Testis ; Leydig Cells ; Spermatogenesis ; Semen ; Sertoli Cells/metabolism ; Peptides/metabolism
    Chemical Substances Peptides
    Language English
    Publishing date 2024-04-04
    Publishing country United States
    Document type Journal Article
    ZDB-ID 2267670-3
    ISSN 1932-6203 ; 1932-6203
    ISSN (online) 1932-6203
    ISSN 1932-6203
    DOI 10.1371/journal.pone.0292198
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  4. Article ; Online: Sertoli cells as key drivers of testis function.

    O'Donnell, Liza / Smith, Lee B / Rebourcet, Diane

    Seminars in cell & developmental biology

    2021  Volume 121, Page(s) 2–9

    Abstract: Sertoli cells are the orchestrators of spermatogenesis; they support fetal germ cell commitment to the male pathway and are essential for germ cell development, from maintenance of the spermatogonial stem cell niche and spermatogonial populations, ... ...

    Abstract Sertoli cells are the orchestrators of spermatogenesis; they support fetal germ cell commitment to the male pathway and are essential for germ cell development, from maintenance of the spermatogonial stem cell niche and spermatogonial populations, through meiosis and spermiogeneis and to the final release of mature spermatids during spermiation. However, Sertoli cells are also emerging as key regulators of other testis somatic cells, including supporting peritubular myoid cell development in the pre-pubertal testis and supporting the function of the testicular vasculature and in contributing to testicular immune privilege. Sertoli cells also have a major role in regulating androgen production within the testis, by specifying interstitial cells to a steroidogenic fate, contributing to androgen production in the fetal testis, and supporting fetal and adult Leydig cell development and function. Here, we provide an overview of the specific roles for Sertoli cells in the testis and highlight how these cells are key drivers of testicular sperm output, and of adult testis size and optimal function of other testicular somatic cells, including the steroidogenic Leydig cells.
    MeSH term(s) Animals ; Humans ; Leydig Cells/metabolism ; Male ; Rats ; Sertoli Cells/metabolism ; Testis
    Language English
    Publishing date 2021-07-03
    Publishing country England
    Document type Journal Article ; Research Support, Non-U.S. Gov't ; Review
    ZDB-ID 1312473-0
    ISSN 1096-3634 ; 1084-9521
    ISSN (online) 1096-3634
    ISSN 1084-9521
    DOI 10.1016/j.semcdb.2021.06.016
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  5. Article ; Online: Development of a model for studying the developmental consequences of oxidative sperm DNA damage by targeting redox-cycling naphthoquinones to the Sertoli cell population.

    Fraser, Barbara Anne / Wilkins, Alexandra Louise / De Iuliis, Geoffry Nunzio / Rebourcet, Diane / Nixon, Brett / Aitken, Robert John

    Free radical biology & medicine

    2023  Volume 206, Page(s) 50–62

    Abstract: Oxidative stress can be induced in the testes by a wide range of factors, including scrotal hyperthermia, varicocele, environmental toxicants, obesity and infection. The clinical consequences of such stress include the induction of genetic damage in the ... ...

    Abstract Oxidative stress can be induced in the testes by a wide range of factors, including scrotal hyperthermia, varicocele, environmental toxicants, obesity and infection. The clinical consequences of such stress include the induction of genetic damage in the male germ line which may, in turn, have serious implications for the health and wellbeing of the progeny. In order to confirm the transgenerational impact of oxidative stress in the testes, we sought to develop an animal model in which this process could be analysed. Our primary approach to this problem was to induce Sertoli cells (robust, terminally differentiated, tissue-specific testicular cells whose radioresistance indicates significant resistance to oxidative stress) to generate high levels of reactive oxygen species (ROS) within the testes. To achieve this aim, six follicle-stimulating hormone (FSH) peptides were developed and compared for selective targeting to Sertoli cells both in vitro and in vivo. Menadione, a redox-cycling agent, was then conjugated to the most promising FSH candidate using a linker that had been optimised to enable maximum production of ROS in the targeted cells. A TM4 Sertoli cell line co-incubated with the FSH-menadione conjugate in vitro exhibited significantly higher levels of mitochondrial ROS generation (10-fold), lipid peroxidation (2-fold) and oxidative DNA damage (2-fold) than the vehicle control. Additionally, in a proof-of-concept study, ten weeks after a single injection of the FSH-menadione conjugate in vivo, injected male mice were found to exhibit a 1.6 fold increase in DNA double strand breaks and 13-fold increase in oxidative DNA damage to their spermatozoa while still retaining their ability to initiate a pregnancy. We suggest this model could now be used to study the influence of chronic oxidative stress on testicular function with emphasis on the impact of DNA damage in the male germ line on the mutational profile and health of future generations.
    MeSH term(s) Pregnancy ; Female ; Male ; Mice ; Animals ; Sertoli Cells/metabolism ; Reactive Oxygen Species/metabolism ; Naphthoquinones ; Vitamin K 3/metabolism ; Semen/metabolism ; Spermatozoa/metabolism ; Testis ; Oxidative Stress ; Follicle Stimulating Hormone/pharmacology ; Oxidation-Reduction ; DNA Damage
    Chemical Substances Reactive Oxygen Species ; Naphthoquinones ; Vitamin K 3 (723JX6CXY5) ; Follicle Stimulating Hormone (9002-68-0)
    Language English
    Publishing date 2023-06-23
    Publishing country United States
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 807032-5
    ISSN 1873-4596 ; 0891-5849
    ISSN (online) 1873-4596
    ISSN 0891-5849
    DOI 10.1016/j.freeradbiomed.2023.06.008
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  6. Article ; Online: Age-related DNA damage in stallions: an ongoing investigation

    Griffin, Roisin A / Harrison, Natasha / Swegen, Aleona / Miller, Kasey / DeIuliis, Geoffry / Rebourcet, Diane / Aitken, Robert J / Gibb, Zamira

    Journal of Equine Veterinary Science. 2023 June, v. 125 p.104590-

    2023  

    Abstract: Foals sired by younger stallions record significantly higher racing speeds (Sharman.et.al. Royal Society Open Science. 2022;9:220691), and more race wins than foals sired by older stallions (Brazil.et.al. Honours Dissertation. University of Limerick. ... ...

    Abstract Foals sired by younger stallions record significantly higher racing speeds (Sharman.et.al. Royal Society Open Science. 2022;9:220691), and more race wins than foals sired by older stallions (Brazil.et.al. Honours Dissertation. University of Limerick. 2016). We hypothesize that this may be due to an age-associated increase in sperm DNA damage, as has been demonstrated in other species. Therefore, this study aimed to investigate the relationship between stallion age and sperm DNA damage. Post-coital dismount semen samples (n=75) were collected weekly from 46 commercial Thoroughbred stallions in the Hunter Valley (Australia). Samples were immediately diluted (2:1, EquiPlus:semen), and the high-density spermatozoa were isolated using EquiPure centrifugation to reduce contaminants (somatic cells and other debris). Any samples that were contaminated with urine were excluded from the study. Sperm count and motilities were recorded onsite (iSperm™), and samples were snap frozen for DNA damage analyses. Management information, including pregnancy scan results, were also collected. For this study, stallions aged ≤9 years were categorized as “young” while those aged ≥12 were categorized as “old”. Stallions between 9 and 12 years were excluded to improve the likelihood of observing an age-related difference. Mare age and fertility status were similar across both groups. All data were checked for normality (Shapiro-Wilk) and analyzed using unpaired t-tests. Sperm count and motility did not differsignificantly between age cohorts. DNA strand breaks were significantly higher in spermatozoa from older stallions compared to younger stallions (alkaline comet assay: 19.7±0.55% vs 15.4±0.57% mean tail intensity; P≤0.05, and sperm chromatin dispersion or ‘Halo’ assay; 0.48±0.045in vs 0.70±0.074in ‘halo’ area; P≤0.05, respectively). These breaks may be attributed to poor chromatin packaging, as older stallions had a concomitant deficiency of sperm protamines compared to young stallions (11.6±1.53 vs 6.9±1.07 chromomycin A₃ positive cells; P≤0.05, respectively). Although younger stallions had lower levels of DNA strand breakage, they had more oxidized DNA adducts (8-hydroxy-2′-deoxyguanosine (8-OHdG) assay; P≤0.05). Interestingly, regardless of age, positive correlations between the proportion of oxidized DNA adducts and fertility were observed (R²=0.269 for younger, R²=0.483 for older stallions; P≤0.05). This is likely a reflection of the increased use of oxidative phosphorylation, and subsequent ROS production, in higher fertility ejaculates (Gibb.et.al. Biology of Reproduction. 2014;91(3):77). In conclusion, while stallion sperm DNA damage rises with age, it does not appear to influence fertility outcomes. However, further research is needed to determine if age-related sperm DNA damage compromises the next generation's genome. In addition, we must identify the most vulnerable genomic loci for sperm DNA damage and determine how these DNA lesions may contribute to reduced racing performance and health in offspring.
    Keywords DNA adducts ; DNA damage ; Thoroughbred ; centrifugation ; chromatin ; comet assay ; genome ; genomics ; oxidation ; oxidative phosphorylation ; pregnancy ; progeny ; protamines ; semen ; spermatozoa ; stallions ; t-test ; urine ; veterinary medicine ; Australia
    Language English
    Dates of publication 2023-06
    Publishing place Elsevier Inc.
    Document type Article ; Online
    ZDB-ID 2102631-2
    ISSN 1542-7412 ; 0737-0806
    ISSN (online) 1542-7412
    ISSN 0737-0806
    DOI 10.1016/j.jevs.2023.104590
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  7. Article: Leukemia inhibitory factor-receptor signalling negatively regulates gonadotrophin-stimulated testosterone production in mouse Leydig Cells

    Curley, Michael / Darbey, Annalucia / O'Donnell, Liza / Kilcoyne, Karen R. / Wilson, Kirsten / Mungall, Will / Rebourcet, Diane / Guo, Jingtao / Mitchell, Rod T. / Smith, Lee B.

    Molecular and cellular endocrinology. 2022 Mar. 15, v. 544

    2022  

    Abstract: Testicular Leydig cells (LCs) are the principal source of circulating testosterone in males. LC steroidogenesis maintains sexual function, fertility and general health, and is influenced by various paracrine factors. The leukemia inhibitory factor ... ...

    Abstract Testicular Leydig cells (LCs) are the principal source of circulating testosterone in males. LC steroidogenesis maintains sexual function, fertility and general health, and is influenced by various paracrine factors. The leukemia inhibitory factor receptor (LIFR) is expressed in the testis and activated by different ligands, including leukemia inhibitory factor (LIF), produced by peritubular myoid cells. LIF can modulate LC testosterone production in vitro under certain circumstances, but the role of consolidated signalling through LIFR in adult LC function in vivo has not been established. We used a conditional Lifr allele in combination with adenoviral vectors expressing Cre-recombinase to generate an acute model of LC Lifr-KO in the adult mouse testis, and showed that LC Lifr is not required for short term LC survival or basal steroidogenesis. However, LIFR-signalling negatively regulates steroidogenic enzyme expression and maximal gonadotrophin-stimulated testosterone biosynthesis, expanding our understanding of the intricate regulation of LC steroidogenic function.
    Keywords Adenoviridae ; adults ; alleles ; biosynthesis ; enzymes ; leukemia ; leukemia inhibitory factor ; ligands ; mice ; models ; steroidogenesis ; testosterone
    Language English
    Dates of publication 2022-0315
    Publishing place Elsevier B.V.
    Document type Article
    ZDB-ID 187438-x
    ISSN 1872-8057 ; 0303-7207
    ISSN (online) 1872-8057
    ISSN 0303-7207
    DOI 10.1016/j.mce.2022.111556
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  8. Article ; Online: Sertoli cell-enriched proteins in mouse and human testicular interstitial fluid.

    O'Donnell, Liza / Dagley, Laura F / Curley, Michael / Darbey, Annalucia / O'Shaughnessy, Peter J / Diemer, Thorsten / Pilatz, Adrian / Fietz, Daniela / Stanton, Peter G / Smith, Lee B / Rebourcet, Diane

    PloS one

    2023  Volume 18, Issue 9, Page(s) e0290846

    Abstract: Sertoli cells support the development of sperm and the function of various somatic cells in the interstitium between the tubules. Sertoli cells regulate the function of the testicular vasculature and the development and function of the Leydig cells that ... ...

    Abstract Sertoli cells support the development of sperm and the function of various somatic cells in the interstitium between the tubules. Sertoli cells regulate the function of the testicular vasculature and the development and function of the Leydig cells that produce testosterone for fertility and virility. However, the Sertoli cell-derived factors that regulate these cells are largely unknown. To define potential mechanisms by which Sertoli cells could support testicular somatic cell function, we aimed to identify Sertoli cell-enriched proteins in the testicular interstitial fluid (TIF) between the tubules. We previously resolved the proteome of TIF in mice and humans and have shown it to be a rich source of seminiferous tubule-derived proteins. In the current study, we designed bioinformatic strategies to interrogate relevant proteomic and genomic datasets to identify Sertoli cell-enriched proteins in mouse and human TIF. We analysed proteins in mouse TIF that were significantly reduced after one week of acute Sertoli cell ablation in vivo and validated which of these are likely to arise primarily from Sertoli cells based on relevant mouse testis RNASeq datasets. We used a different, but complementary, approach to identify Sertoli cell-enriched proteins in human TIF, taking advantage of high-quality human testis genomic, proteomic and immunohistochemical datasets. We identified a total of 47 and 40 Sertoli cell-enriched proteins in mouse and human TIF, respectively, including 15 proteins that are conserved in both species. Proteins with potential roles in angiogenesis, the regulation of Leydig cells or steroidogenesis, and immune cell regulation were identified. The data suggests that some of these proteins are secreted, but that Sertoli cells also deposit specific proteins into TIF via the release of extracellular vesicles. In conclusion, we have identified novel Sertoli cell-enriched proteins in TIF that are candidates for regulating somatic cell-cell communication and testis function.
    MeSH term(s) Humans ; Male ; Animals ; Mice ; Sertoli Cells ; Testis ; Extracellular Fluid ; Proteomics ; Semen
    Language English
    Publishing date 2023-09-01
    Publishing country United States
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 2267670-3
    ISSN 1932-6203 ; 1932-6203
    ISSN (online) 1932-6203
    ISSN 1932-6203
    DOI 10.1371/journal.pone.0290846
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  9. Article: Biocompatible Nanomaterials as an Emerging Technology in Reproductive Health; a Focus on the Male.

    Fraser, Barbara / Peters, Alexandra E / Sutherland, Jessie M / Liang, Mingtao / Rebourcet, Diane / Nixon, Brett / Aitken, Robert J

    Frontiers in physiology

    2021  Volume 12, Page(s) 753686

    Abstract: A growing body of research has confirmed that nanoparticle (NP) systems can enhance delivery of therapeutic and imaging agents as well as prevent potentially damaging systemic exposure to these agents by modifying the kinetics of their release. With a ... ...

    Abstract A growing body of research has confirmed that nanoparticle (NP) systems can enhance delivery of therapeutic and imaging agents as well as prevent potentially damaging systemic exposure to these agents by modifying the kinetics of their release. With a wide choice of NP materials possessing different properties and surface modification options with unique targeting agents, bespoke nanosystems have been developed for applications varying from cancer therapeutics and genetic modification to cell imaging. Although there remain many challenges for the clinical application of nanoparticles, including toxicity within the reproductive system, some of these may be overcome with the recent development of biodegradable nanoparticles that offer increased biocompatibility. In recognition of this potential, this review seeks to present recent NP research with a focus on the exciting possibilities posed by the application of biocompatible nanomaterials within the fields of male reproductive medicine, health, and research.
    Language English
    Publishing date 2021-11-11
    Publishing country Switzerland
    Document type Journal Article ; Review
    ZDB-ID 2564217-0
    ISSN 1664-042X
    ISSN 1664-042X
    DOI 10.3389/fphys.2021.753686
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  10. Article: A role for steroid 5 alpha-reductase 1 in vascular remodeling during endometrial decidualization.

    Shaw, Isaac W / Kirkwood, Phoebe M / Rebourcet, Diane / Cousins, Fiona L / Ainslie, Rebecca J / Livingstone, Dawn E W / Smith, Lee B / Saunders, Philippa T K / Gibson, Douglas A

    Frontiers in endocrinology

    2022  Volume 13, Page(s) 1027164

    Abstract: Decidualization is the hormone-dependent process of endometrial remodeling that is essential for fertility and reproductive health. It is characterized by dynamic changes in the endometrial stromal compartment including differentiation of fibroblasts, ... ...

    Abstract Decidualization is the hormone-dependent process of endometrial remodeling that is essential for fertility and reproductive health. It is characterized by dynamic changes in the endometrial stromal compartment including differentiation of fibroblasts, immune cell trafficking and vascular remodeling. Deficits in decidualization are implicated in disorders of pregnancy such as implantation failure, intra-uterine growth restriction, and pre-eclampsia. Androgens are key regulators of decidualization that promote optimal differentiation of stromal fibroblasts and activation of downstream signaling pathways required for endometrial remodeling. We have shown that androgen biosynthesis,
    MeSH term(s) Female ; Pregnancy ; Humans ; Mice ; Animals ; Vascular Remodeling ; Cholestenone 5 alpha-Reductase ; Vascular Endothelial Growth Factor A ; Dihydrotestosterone ; Androgens ; Endometrium
    Chemical Substances Cholestenone 5 alpha-Reductase (EC 1.3.1.22) ; Vascular Endothelial Growth Factor A ; Dihydrotestosterone (08J2K08A3Y) ; Androgens
    Language English
    Publishing date 2022-11-16
    Publishing country Switzerland
    Document type Journal Article
    ZDB-ID 2592084-4
    ISSN 1664-2392
    ISSN 1664-2392
    DOI 10.3389/fendo.2022.1027164
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