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Journal of the Chinese Medical... Apr 2021Kisspeptin (KISS1) and kisspeptin receptor (KISS1R) are essential gatekeepers of the reproductive system. The functions of KISS1 and KISS1R in corpus luteal cells remain...
BACKGROUND
Kisspeptin (KISS1) and kisspeptin receptor (KISS1R) are essential gatekeepers of the reproductive system. The functions of KISS1 and KISS1R in corpus luteal cells remain ambiguous. The objective was to observe normal physiologic functions of corpus luteal cells in vivo and clarify the functions of KISS1 in vitro.
METHODS
We conducted an in vivo observation of cellular patterns as well as the levels of steroidogenic enzymes and KISS1/KISS1R in corpus luteal cells obtained from female crossbred Taiwan native goats in the estrous cycle; the observation was performed using hematoxylin and eosin and immunohistochemistry staining. Subsequently, we used kisspeptin-10 (Kp-10) to stimulate temperature sensitive-caprine luteal cell line (ts-CLC-D) cells to investigate the progesterone (P4) levels, steroidogenic messenger RNA (mRNA)/protein levels, cell survival rate, intracellular Ca2+ concentration, and cell proliferation-related mRNA/protein levels in the mitogen-activated protein kinase pathway in vitro by applying immunofluorescence staining, Western blotting, 2,3-bis (2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide assay, and real-time polymerase chain reaction.
RESULTS
We observed the presence of proteins and mRNAs for STAR, CYP11A1, HSD3B, KISS1, and KISS1R in the corpus luteal cells from goats in vivo. In vitro, the addition of Kp-10 reduced the P4 levels (p < 0.01) and increased cell proliferation (p < 0.05) of the ts-CLC-D cells. Furthermore, we found that the levels of proteins and mRNA for STAR, CYP11A1, and HSD3B decreased significantly when Kp-10 was added (p < 0.05). However, adding Kp-10 did not affect the mRNA levels for PLCG2, DAG1, PRKCA, KRAS, RAF1, MAP2K1, MAP2K2, MAPK3, MAPK1, and MAPK14.
CONCLUSION
We determined that KISS1 could affect the P4 levels, steroidogenesis, and cell proliferation in luteal cells. However, further research is required to clarify how KISS1 regulates proliferation and steroid production in luteal cells.
Topics: Animals; Cell Proliferation; Cell Survival; Female; Gene Expression; Goats; Kisspeptins; Luteal Cells; Polymerase Chain Reaction; RNA, Messenger; Taiwan
PubMed: 33784266
DOI: 10.1097/JCMA.0000000000000508 -
Reproductive Biomedicine Online May 2022Increased granulosa cell division is associated with abnormal folliculogenesis in polycystic ovary syndrome (PCOS). Lethal-7i microRNA (let-7i) may play an important...
RESEARCH QUESTION
Increased granulosa cell division is associated with abnormal folliculogenesis in polycystic ovary syndrome (PCOS). Lethal-7i microRNA (let-7i) may play an important role in the follicular development and granulosa cell growth; therefore is let-7i involved in PCOS pathogenesis?
DESIGN
The expression of let-7i was measured in granulosa-luteal cells (GLC) from women with or without PCOS. A human granulosa cell line, KGN, was used for the functional study. Mimics and inhibitors of let-7i, lentiviruses expressing insulin-like growth factor 2 mRNA binding protein (IMP2), and small-interfering RNAs were transfected into KGN cells. KGN cell proliferation was determined by 5-ethynyl-2'-deoxyuridine (EdU) and Cell Counting Kit-8 (CCK-8) assays. The cell cycle and apoptosis were assessed by propidium iodide-annexin V (PI-A) staining and fluorescence-activated cell sorting. Oestradiol concentration was determined by enzyme-linked immunoassay. Bioinformatics analysis and luciferase reporter assay were applied to confirm the let-7i target genes.
RESULTS
The study showed that let-7i was down-regulated in PCOS GLC (P = 0.001). Mimics of let-7i inhibited KGN proliferation (P = 0.001), and decreased aromatase expression (P = 0.030) and oestradiol production (P = 0.029), whereas let-7i inhibitors had the opposite effect. Bioinformatics analysis and quantitative real-time (qRT) PCR identified IMP2 as a target of let-7i (P = 0.021). qRT-PCR and western blot analysis indicated that IMP2 was up-regulated in GLC in women with PCOS (P = 0.001 and P = 0.044), and IMP2 expression was suppressed by let-7i in KGN cells (P < 0.001). Luciferase reporter assay results (P = 0.002), combined with the rescue assay, confirmed that let-7i inhibited KGN cell proliferation and reduced oestradiol concentration by directly targeting IMP2.
CONCLUSIONS
let-7i was down-regulated in PCOS GLC. Overexpression of let-7i inhibited KGN cell proliferation and decreased oestradiol production in an IMP2-dependent manner, providing a new molecular mechanism for PCOS.
Topics: Female; Humans; Apoptosis; Cell Proliferation; Estradiol; Granulosa Cells; Luteal Cells; MicroRNAs; Polycystic Ovary Syndrome
PubMed: 35339367
DOI: 10.1016/j.rbmo.2022.01.016 -
Seminars in Reproductive Endocrinology 1997The corpus luteum is an extremely important reproductive endocrine gland that influences numerous physiological processes and is therefore perhaps the most widely... (Review)
Review
The corpus luteum is an extremely important reproductive endocrine gland that influences numerous physiological processes and is therefore perhaps the most widely studied reproductive endocrine structure to date. The corpus luteum and its major product, progesterone, are central to any discussion of impaired reproductive performance. It is generally agreed that our best chances for reducing reproductive losses lie in a thorough understanding of the mechanisms controlling luteal function during reproductive cycles and pregnancy. Normal corpus luteum function is dependent on numerous regulatory factors, such as prostaglandins, oxytocin, steroids, growth factors, cytokines, etc. Recently, increased interest in studying luteal cell interactions (steroidogenic large and small luteal cells as well as nonsteroidogenic "accessory" cells of the corpus luteum) has produced new evidence of communication within the corpus luteum that influences its function. The exact means of communication among the luteal cells are not known; however, this interaction seems to involve both luteotropic and luteolytic actions and can occur through either contact-dependent and contact-independent mechanisms.
Topics: Animals; Cell Communication; Corpus Luteum; Female; Humans; Luteal Cells; Pregnancy; Progesterone; Steroids
PubMed: 9580949
DOI: 10.1055/s-2008-1068378 -
Biology of Reproduction Oct 2019Transient receptor potential cation channel, mucolipin subfamily, member 1 (TRPML1) (MCOLN1/Mcoln1) is a lysosomal counter ion channel. Mutations in MCOLN1 cause...
Transient receptor potential cation channel, mucolipin subfamily, member 1 (TRPML1) (MCOLN1/Mcoln1) is a lysosomal counter ion channel. Mutations in MCOLN1 cause mucolipidosis type IV (MLIV), a progressive and severe lysosomal storage disorder with a slow onset. Mcoln1-/- mice recapitulate typical MLIV phenotypes but roles of TRPML1 in female reproduction are unknown. Despite normal mating activities, Mcoln1-/- female mice had reduced fertility at 2 months old and quickly became infertile at 5 months old. Progesterone deficiency was detected on 4.5 days post coitum/gestation day 4.5 (D4.5). Immunohistochemistry revealed TRPML1 expression in luteal cells of wild type corpus luteum (CL). Corpus luteum formation was not impaired in 5-6 months old Mcoln1-/- females indicated by comparable CL numbers in control and Mcoln1-/- ovaries on both D1.5 and D4.5. In the 5-6 months old Mcoln1-/- ovaries, histology revealed less defined corpus luteal cord formation, extensive luteal cell vacuolization and degeneration; immunofluorescence revealed disorganized staining of collagen IV, a basal lamina marker for endothelial cells; Nile Red staining detected lipid droplet accumulation, a typical phenotype of MLIV; immunofluorescence of heat shock protein 60 (HSP60, a mitochondrial marker) and in situ hybridization of steroidogenic acute regulatory protein (StAR, for the rate-limiting step of steroidogenesis) showed reduced expression of HSP60 and StAR, indicating impaired mitochondrial functions. Luteal cell degeneration and impaired mitochondrial functions can both contribute to progesterone deficiency in the Mcoln1-/- mice. This study demonstrates a novel function of TRPML1 in maintaining CL luteal cell integrity and function.
Topics: Animals; Corpus Luteum; Disease Models, Animal; Female; Infertility; Luteal Cells; Lysosomal Storage Diseases; Male; Mice; Mice, Inbred C57BL; Mice, Knockout; Mucolipidoses; Progesterone; Transient Receptor Potential Channels
PubMed: 31317194
DOI: 10.1093/biolre/ioz126 -
The Journal of Clinical Endocrinology... Jul 2017Inadequate progesterone production from the corpus luteum is associated with pregnancy loss. Data available in model species suggest important roles of microRNAs...
CONTEXT
Inadequate progesterone production from the corpus luteum is associated with pregnancy loss. Data available in model species suggest important roles of microRNAs (miRNAs) in luteal development and maintenance.
OBJECTIVE
To comprehensively investigate the involvement of miRNAs during the ovarian follicle-luteal transition.
DESIGN
The effects of specific miRNAs on survival and steroid production by human luteinized granulosa cells (hLGCs) were tested using specific miRNA inhibitors. Candidate miRNAs were identified through microarray analyses of follicular and luteal tissues in a bovine model.
SETTING
An academic institution in the United Kingdom associated with a teaching hospital. hLGCs were obtained by standard transvaginal follicular-fluid aspiration from 35 women undergoing assisted conception.
INTERVENTION(S)
Inhibition of candidate miRNAs in vitro.
MAIN OUTCOME MEASURE(S)
Levels of miRNAs, mRNAs, FOXO1 protein, apoptosis, and steroids were measured in tissues and/or cultured cells.
RESULTS
Two specific miRNA clusters, miR-183-96-182 and miR-212-132, were dramatically increased in luteal relative to follicular tissues. miR-96 and miR-132 were the most upregulated miRNAs within each cluster. Database analyses identified FOXO1 as a putative target of both these miRNAs. In cultured hLGCs, inhibition of miR-96 increased apoptosis and FOXO1 protein levels, and decreased progesterone production. These effects were prevented by small interfering RNA-mediated downregulation of FOXO1. In bovine luteal cells, miR-96 inhibition also led to increases in apoptosis and FOXO1 protein levels.
CONCLUSIONS
miR-96 targets FOXO1 to regulate luteal development through effects on cell survival and steroid production. The miR-183-96-182 cluster could provide a novel target for the manipulation of luteal function.
Topics: Animals; Apoptosis; Cattle; Cell Survival; Cells, Cultured; Female; Forkhead Box Protein O1; Gene Expression Profiling; Gene Expression Regulation; Granulosa Cells; Humans; Luteal Cells; MicroRNAs; Ovarian Follicle; Progesterone; Species Specificity; Up-Regulation
PubMed: 28368475
DOI: 10.1210/jc.2017-00259 -
Domestic Animal Endocrinology Jan 2022The corpus luteum (CL) plays a vital role in regulating the reproductive cycle, fertility, and in maintaining pregnancy. Interferon-tau (IFNT) is the maternal... (Review)
Review
The corpus luteum (CL) plays a vital role in regulating the reproductive cycle, fertility, and in maintaining pregnancy. Interferon-tau (IFNT) is the maternal recognition of a pregnancy signal in domestic ruminants; its uterine, paracrine actions, which extend the CL lifespan, are widely established. However, considerable evidence also suggests a direct, endocrine role for IFNT. The purpose of this review is to highlight the importance of IFNT in CL maintenance, acting directly and in a cell-specific manner. A transcriptomic study revealed a distinct molecular profile of IFNT-exposed day 18, pregnant bovine CL, compared to the non-pregnant gland. A substantial fraction of the differentially expressed genes was downregulated, many of which are known to be elevated by prostaglandin F2A (PGF2A). In vitro, IFNT was found to mimic changes observed in the luteal transcriptome of early pregnancy. Key luteolytic genes such as endothelin-1 (EDN1), transforming growth factor-B1 (TGFB1), thrombospondins (THBSs) 1&2 and serpine-1 (SERPINE1) were downregulated in luteal endothelial cells. Luteal steroidogenic large cells (LGCs) were also found to be a target for the antilutelotytic actions of IFNT. IFNT-treated LGCs showed a significant reduction in the expression of the proapoptotic, antiangiogenic THBS1&2, as well as TGFBR1 and 2. Furthermore, IFNT was shown to be a potent survival factor for luteal cells in vivo and in vitro, activating diverse pathways to promote cell survival while suppressing cell death signals. Pentraxin 3 (PTX3), robustly upregulated by IFNT in various luteal cell types, mediated many of the prosurvival effects of IFNT in LGCs. A novel reciprocal inhibitory crosstalk between PTX3 and THBS1 lends further support to their respective survival and apoptotic actions in the CL. Even though IFNT did not directly regulate progesterone synthesis, it could maintain its concentrations, by increasing luteal cell survival and by supporting vascular stabilization. The direct effects of IFNT in the CL, enhancing cell survival and vasculature stabilization while curbing luteolytic activities, may constitute an important complementary branch leading to the extension of the luteal lifespan during early pregnancy.
Topics: Animals; Cattle; Corpus Luteum; Dinoprost; Endothelial Cells; Female; Luteal Cells; Luteolysis; Pregnancy
PubMed: 34509740
DOI: 10.1016/j.domaniend.2021.106671 -
Scientific Reports Jul 2021In mammalian ovaries, mitochondria are integral sites of energy production and steroidogenesis. While shifts in cellular activities and steroidogenesis are well...
In mammalian ovaries, mitochondria are integral sites of energy production and steroidogenesis. While shifts in cellular activities and steroidogenesis are well characterized during the differentiation of large luteal cells in folliculogenesis and luteal formation, mitochondrial dynamics during this process have not been previously evaluated. In this study, we collected ovaries containing primordial follicles, mature follicles, corpus hemorrhagicum, or corpus luteum from goats at specific times in the estrous cycle. Enzyme histochemistry, ultrastructural observations, and 3D structural analysis of serial sections of mitochondria revealed that branched mitochondrial networks were predominant in follicles, while spherical and tubular mitochondria were typical in large luteal cells. Furthermore, the average mitochondrial diameter and volume increased from folliculogenesis to luteal formation. In primordial follicles, the signals of cytochrome c oxidase and ATP synthase were undetectable in most cells, and the large luteal cells from the corpus hemorrhagicum also showed low enzyme signals and content when compared with granulosa cells in mature follicles or large luteal cells from the corpus luteum. Our findings suggest that the mitochondrial enlargement could be an event during folliculogenesis and luteal formation, while the modulation of mitochondrial morphology and respiratory enzyme expressions may be related to tissue remodeling during luteal formation.
Topics: Animals; Female; Goats; Lipogenesis; Luteal Cells; Mitochondrial Dynamics
PubMed: 34330986
DOI: 10.1038/s41598-021-95161-w -
Androgen production by monkey luteal cell subpopulations at different stages of the menstrual cycle.The Journal of Clinical Endocrinology... Feb 1996Androgens produced by the primate corpus luteum (CL) serve as precursors for estrogen synthesis; moreover, detection of androgen receptors in luteal tissue suggests a...
Androgens produced by the primate corpus luteum (CL) serve as precursors for estrogen synthesis; moreover, detection of androgen receptors in luteal tissue suggests a regulatory role within the CL. To determine the cellular source(s) and agonist regulation of androgen production during the lifespan of the primate CL, luteal tissues were collected from rhesus monkeys in the early (days 3-5 post-LH surge), mid (days 7-8), mid-late (days 11-12), and late (days 14-15) luteal phase of the menstrual cycle. Collagenase-dispersed cells (i.e., mixed cells) were analyzed by flow cytometry based on light scatter properties and sorted into populations of small (< or = 15 microns) and large (> 20 microns) luteal cells. Cells (n = 4 animals/stage) were incubated in Ham's F-10 and 0.1% BSA for 3 h at 37 C with or without hCG (100 ng/mL), PGE2 (14 mumol/L), or dibutyryl cAMP (dbcAMP; 5 mmol/L), and androstenedione (A4) and testosterone were measured. Basal A4 production by large cells was markedly higher (P < 0.05) than that by small cells (e.g. mid-late luteal phase, 821 +/- 188 vs. 69 +/- 25 pg/mL.5 x 10(4) cells/3 h; mean +/- SEM), whereas that by mixed cells was intermediate (317 +/- 205 pg/mL). In the early luteal phase, hCG stimulated A4 synthesis by mixed (1.6-fold; P < 0.05) and large (3.1-fold; P < 0.05) luteal cells, but not by small cells (1.3-fold). By the mid-late luteal phase, hCG did not increase A4 production by any cell type, although hCG responsiveness returned to large cells (2.0-fold increase; P < 0.05) by the late luteal phase. PGE2 responsiveness by cell types was similar to that of hCG, except large cell responsiveness did not return in the late luteal phase. In all cell types, dbcAMP stimulated the largest increase in A4 levels; in the mid-late luteal phase, small and large cells responded to dbcAMP with 8.2- and 3.0-fold increases (P < 0.05) in A4 production, respectively. When luteal cells were incubated with the steroidogenic substrates, 17 alpha-hydroxyprogesterone or 17 alpha-hydroxypregnenolone (1 mumol/L), large cells produced much more (P < 0.05) A4, testosterone, estrone, and estradiol than small cells. Both substrates elicited similar patterns of androgen production, with A4 synthesis predominant in all luteal cell types. Thus, cell subpopulations in the primate CL can be distinguished by their ability to produce androgen and estrogen. Changes in agonist-responsive androgen production may influence the local steroid milieu and function of the CL during the menstrual cycle.
Topics: Androgens; Androstenedione; Animals; Bucladesine; Chorionic Gonadotropin; Dinoprostone; Estradiol; Estrone; Female; Flow Cytometry; Luteal Cells; Luteal Phase; Macaca mulatta; Menstrual Cycle; Serum Albumin, Bovine; Testosterone
PubMed: 8636273
DOI: 10.1210/jcem.81.2.8636273 -
Journal of Animal Science Jul 1994This article reviews literature on the highly differentiated nature of the luteal cell types and their regulation in the mid-cycle corpus luteum of ruminants. The blood... (Review)
Review
This article reviews literature on the highly differentiated nature of the luteal cell types and their regulation in the mid-cycle corpus luteum of ruminants. The blood vessels of the corpus luteum are composed primarily of endothelial cells with few intraluteal arterioles or arteries. Blood flow to the corpus luteum does not seem to be regulated by tropic hormones, metabolic activity, or autonomic nerves, but it is dependent on maintenance of high blood pressure. The density of luteal capillaries and lack of luteal arterioles allows an elevated, relatively unregulated luteal blood flow. Steroidogenesis in the ruminant corpus luteum manifests both dependence on and independence from the actions of LH. Initial luteinization, growth, and development of the corpus luteum is dependent on LH action, but progesterone production by the mid-cycle corpus luteum of ruminants is relatively independent of acute LH stimulation. This apparent enigma may be due to distinct functional properties of the steroidogenic cell types in the corpus luteum. Progesterone production by the large luteal cells is relatively independent of LH action, but the small luteal cells respond to LH stimulation. Much of the progesterone produced by the mid-cycle corpus luteum is produced by the large luteal cells and may not be acutely dependent on LH stimulation. Progesterone production by both cell types is dependent on lipoproteins delivered by the abundant luteal blood supply. Thus, distinct cellular features of the mid-cycle corpus luteum produce considerable steroidogenic capacity.
Topics: Animals; Animals, Domestic; Corpus Luteum; Estrus; Female; Hormones; Lipoproteins; Luteal Cells; Luteinizing Hormone; Progesterone; Regional Blood Flow
PubMed: 7928767
DOI: 10.2527/1994.7271873x -
Molecular and Cellular Endocrinology Jan 2022Glucose uptake increases in canine luteal cells under insulin treatment. We hypothesize that insulin also increases luteal cell steroidogenesis. Dogs underwent elective...
Glucose uptake increases in canine luteal cells under insulin treatment. We hypothesize that insulin also increases luteal cell steroidogenesis. Dogs underwent elective ovariohysterectomy from days 10-60 post ovulation and their corpora lutea (CL) and blood samples were collected. Deep RNA sequencing determined differentially expressed genes in CL; those related to insulin signaling and steroidogenesis were validated in vivo by qPCR and their respective proteins by Western blotting and immunofluorescence. Next, luteal cell cultures were stimulated with insulin with or without inhibition of MAPK14, MAP2K1 and PI3K. Studied proteins except P450 aromatase showed the same expression pattern of coding genes in vivo. The expression of HSD3B and CYP19A1 was higher in insulin-treated cells (P < 0.005). Following respective pathway blockades, the culture medium had decreased concentrations of progesterone (P4) and 17b-estradiol (E2) (P < 0.01). Our results indicate that insulin increases HSD3B and CYP19A1 expression via MAPK and PI3K, and contributes to the regulation of P4 and E2 production in canine luteal cells.
Topics: Animals; Cells, Cultured; Corpus Luteum; Dogs; Estradiol; Female; Glucose; Insulin; Luteal Cells; MAP Kinase Signaling System; Mitogen-Activated Protein Kinase Kinases; Phosphatidylinositol 3-Kinases; Progesterone; Signal Transduction; Steroids
PubMed: 34808277
DOI: 10.1016/j.mce.2021.111518