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PloS One 2013Babies with intra-uterine growth restriction (IUGR) are at increased risk for experiencing negative neonatal outcomes due to their general developmental delay. The...
Babies with intra-uterine growth restriction (IUGR) are at increased risk for experiencing negative neonatal outcomes due to their general developmental delay. The present study aimed to investigate the effects of a short postnatal leptin supply on the growth, structure, and functionality of several organs at weaning. IUGR piglets were injected from day 0 to day 5 with either 0.5 mg/kg/d leptin (IUGRLep) or saline (IUGRSal) and euthanized at day 21. Their organs were collected, weighed, and sampled for histological, biochemical, and immunohistochemical analyses. Leptin induced an increase in body weight and the relative weights of the liver, spleen, pancreas, kidneys, and small intestine without any changes in triglycerides, glucose and cholesterol levels. Notable structural and functional changes occurred in the ovaries, pancreas, and secondary lymphoid organs. The ovaries of IUGRLep piglets contained less oogonia but more oocytes enclosed in primordial and growing follicles than the ovaries of IUGRSal piglets, and FOXO3A staining grade was higher in the germ cells of IUGRLep piglets. Within the exocrine parenchyma of the pancreas, IUGRLep piglets presented a high rate of apoptotic cells associated with a higher trypsin activity. In the spleen and the Peyer's patches, B lymphocyte follicles were much larger in IUGRLep piglets than in IUGRSal piglets. Moreover, IUGRLep piglets showed numerous CD79(+) cells in well-differentiated follicle structures, suggesting a more mature immune system. This study highlights a new role for leptin in general developmental processes and may provide new insight into IUGR pathology.
Topics: Animals; Animals, Newborn; CD79 Antigens; Female; Fetal Growth Retardation; Fetus; Forkhead Transcription Factors; Gene Expression Regulation, Developmental; Gonads; Humans; Immune System; Infant, Newborn; Injections, Intramuscular; Leptin; Liver; Male; Organ Size; Organogenesis; Pancreas; Swine
PubMed: 23741353
DOI: 10.1371/journal.pone.0064616 -
Frontiers in Endocrinology 2021Vitellogenesis-inhibiting hormone (VIH) negatively regulates reproduction in shrimp and other decapod crustaceans. In order to assess the effects of transcriptional...
Vitellogenesis-inhibiting hormone (VIH) negatively regulates reproduction in shrimp and other decapod crustaceans. In order to assess the effects of transcriptional silencing by multiple VIH subtype I sinus gland peptides (SGPs) on ovarian maturation in female whiteleg shrimp, , we synthesized five dsRNAs targeting , -, -, -, and - and injected them into subadults. The following treatments were employed: sgpG-dsRNA (targeting ), sgpC-dsRNA (targeting ), and mixed-dsRNA (targeting , , and ). The expression of in eyestalks was significantly decreased at 10, 20, and 30 days after the injection of sgpG-dsRNA In addition, it was significantly decreased at 10 and 30 days after the injection of mixed-dsRNA. The expression of vitellogenin (Vg) gene expression in the ovaries, and concentrations of Vg protein in the hemolymph, were not changed by the administration of any dsRNA treatment (the ovaries remained immature in all treated individuals and contained mostly oogonia and previtellogenic oocytes). Although the administration of dsRNAs corresponding to multiple VIHs did not promote ovarian maturation, this is the first report of the co-transcriptional repression of by the injection of dsRNA for homologous genes (, -, and ). These results indicate that subadults can respond to the techniques of transcriptional silencing.
Topics: Animals; Female; Hemolymph; Invertebrate Hormones; Penaeidae; Peptides; RNA, Double-Stranded; Vitellogenesis; Vitellogenins
PubMed: 33737908
DOI: 10.3389/fendo.2021.594001 -
Plant Disease Nov 2003Phytophthora ramorum causes shoot and foliar blight on Rhododendron spp., Viburnum spp. (4), Pieris spp. (2), Kalmia latifolia, and Camellia spp. in several European...
Phytophthora ramorum causes shoot and foliar blight on Rhododendron spp., Viburnum spp. (4), Pieris spp. (2), Kalmia latifolia, and Camellia spp. in several European countries (1-4). In December 2002, we received diseased C. japonica growing in containers from several nurseries in Galicia (northwestern Spain). These young camellia plants had leaves with brown-to-black, water-soaked lesions with diffuse borders that expanded into larger blotches resulting in dead leaves and necrotic lesions on the petioles. Eventually the entire plant wilted and died. Tissue from the leading edge of the lesions was transferred to a selective medium (V8 agar supplemented with pimaricin (10 μg/ml), rifampicin (25 μg/ml), hymexazol (5 μg/ml), and benomyl (5 μg/ml)) and incubated for 3 to 4 days at 20°C in the dark. A Phytophthora sp. was isolated, transferred to carrot piece agar (CPA) (4), and incubated in alternating light. Isolates exhibited coralloid mycelium with concentric rings and a radial growth of 2.5 to 3 mm per day at 20°C. The hyaline-to-yellowish chlamydospores were terminal and intercalary, occasionally lateral, and 24 to 74 μm in diameter. The caducous, sympodial, semipapillate sporangia had a length/breadth ratio of 1.8 to 2.1 and a short pedicel (<5 μm) or no pedicel. Oogonia, antheridia, and oospores were produced by pairing the isolates with P. cryptogea A2 tester BBA 63651 (3,4) provided by S. Werres. Oogonia were subspherical and smooth-walled, antheridia were amphyginous, and oospores were plerotic. The internal transcribed spacer-rDNA polymerase chain reaction (PCR) product obtained by using DNA extracted from mycelium and nested PCR with P. ramorum-specific primers was the size reported for P. ramorum (1). Pathogenicity of the isolates was confirmed by inoculating detached leaves of C. japonica. Five isolates were tested on leaves from 15 young plants growing in containers. Three leaves of each plant were detached and inoculated with each isolate. Leaves were dipped for 5 min into a suspension of sporangia and mycelial fragments and maintained at 20°C and 95% humidity. After 15 days, lesions that developed from the petiole base exceeded 25 mm, and the pathogen was consistently reisolated from the lesions. Leaves inoculated with water from sterile CPA plates did not develop symptoms. A C. japonica isolate has been deposited in the Spanish Type Culture Collection (CECT 20519). To our knowledge, this is the first report of P. ramorum on C. japonica in Spain, though the pathogen has been isolated from Rhododendron spp. and Viburnum tinus growing in several nurseries in Galicia. References: (1) J. M. Davidson et al. On-line publication doi:10.1094. Plant Health Progress, PHP, 2003-0707-01-DG, Plant Management Network. (2) A. J. Inman et al. First Report of Ramorum Dieback (Phytophthora ramorum) on Pieris in England. On-line publication. New Dis. Rep. Vol. 7, British Society for Plant Pathology, 2003. (3) E. Moralejo et al. Plant Dis. 86, 9:1052, 2002. (4) S. Werres et al. Mycol.Res.105:1155, 2001.
PubMed: 30812563
DOI: 10.1094/PDIS.2003.87.11.1396A -
Cell Death Discovery Jul 2023A faithful reconstitution of the complete process of oogenesis in vitro is helpful for understanding the molecular mechanisms, genetics, and epigenetic changes related...
A faithful reconstitution of the complete process of oogenesis in vitro is helpful for understanding the molecular mechanisms, genetics, and epigenetic changes related to gametogenesis; it can also be useful for clinical drug screening, disease research, and regenerative medicine. To this end, given the consensus that murine female germ cells initiate meiosis at E13.5, substantial works have reported the successful generation of fertile oocytes using E12.5 female gonads as starting materials. Nevertheless, our data demonstrated that murine germ cells at E12.5 have heterogeneously initiated a meiotic transcriptional program based on a measurement of pre-mRNAs (unspliced) and mature mRNAs (spliced) at a single-cell level. Therefore, to establish a platform that faithfully recapitulates the entire process in vitro (from premeiotic murine germ cells to fully developed oocytes), we here report a novel three-dimensional organoid culture (3-DOC) system, which successfully induced fully developed oocytes from E11.5 premeiotic female germ cells (oogonia). Compared with 2D culture and other 3D culture methods, this new culture system is more cost-effective and can create high-quality oocytes similar to in vivo oocytes. In summary, our new culture platform provides an experimental model for future research in regenerative medicine and reproductive biology.
PubMed: 37518361
DOI: 10.1038/s41420-023-01577-w -
Plant Disease Jan 2021Camelina sativa, an herbaceous annual plant in the family Brassicaceae, is especially well known for its oilseed crop that produce camelina oil (Hovsepyan et al. 2008)....
Camelina sativa, an herbaceous annual plant in the family Brassicaceae, is especially well known for its oilseed crop that produce camelina oil (Hovsepyan et al. 2008). In April 2016, white blister rust disease on C. sativa were observed in a cultivated farmland with an incidence of about 60% in Xinyuan County (43°33'39.17"N, 83°14'54.04"E), Xinjiang, China. Symptoms appeared as light-yellow chlorotic spots on the upper surface of the leaves and white blister on the corresponding lower surface. Blister sori were white, oval to ellipsoidal, scattered or coalesce, and 1.8 to 4 mm in diameter. Two representative voucher specimens were deposited in the Mycological Herbarium of Tarim University (HMUT 2527 and HMUT 2528), Aral, China. Sporangiophores hyaline, clavate or cylindrical, straight to slightly curved, (23.7 to) 27.9 to 37.9 (to 42.1) (av. 31) × (7.9 to) 9.6 to 13.7 (to 15.1) (av. 11.4) μm (n = 30), thick-walled on their lower parts, bearing sporangia in chains. Primary sporangia were globose to subglobose, wall equal thickness, and (9.5 to) 10.6 to 13.2 (to 14.3) (av. 11.9) μm in diameter (n = 50). Secondary sporangia were mostly subglobose to ovoid, with a subtruncated base, and (12.1 to) 13.2 to 16.9 (to 18) (av. 15.1) μm × (11 to) 12.1 to 15 (to 16.1) (av. 13.4) μm in size (n = 50). Oogonia were globose to subglobose, (39.7 to) 42.7 to 51.7 (to 54.1) (av. 48.3) μm in diameter (n = 30), irregular. Oospores were globose to subglobose, brown, (34.5 to) 37 to 42.7 (to 45.2) (av. 41.1) μm in diameter (n = 30), 3 to 5 μm wall in thickness, with single warts, 1.5 to 4 × 2 to 3.5 μm (n = 30). The morphological characteristics of specimens were consistent with those of Albugo koreana (Choi et al. 2007). To confirm the identification, genomic DNA were extracted directly from sori on diseased leaves from isolates HMUT 2527 and HMUT 2528, respectively. The internal transcribed spacer (ITS) rDNA and cytochrome oxidase II (cox2) mtDNA were amplified with primers DC6/LR-0 described by Choi et al. (2006) and cox2-F/cox2-R described by Hudspeth et al. (2000), respectively. A BLASTn search revealed that the ITS rDNA sequences (GenBank accession Nos. MW135444 and MW135445) were 99% (838/844 nucleotides)identical to that of A. koreana from Capsella bursa-pastoris (AY929829), and the cox2 sequences (GenBank accession Nos. MW147150 and MW147151) were 100% (567/567 nucleotides) identical to that of A. koreana from C. bursa-pastoris (AY927048). Based on the concatenated ITS and cox2 sequences, Maximum Likelihood and Bayesian analysis showed that pathogen from C. sativa with the reference isolate of A. koreana (ex C. bursa-pastoris) with high bootstrap support values and maximum posterior probability (100 ML BS and 1.00 BPP, respectively). For pathogenicity, sporangia collected from the infected leaves were suspended in sterile water at 4°C for 2 hours to improve zoospore release, and the zoospore suspension obtained from sporangial suspension (1×105 sporangia/ml) was inoculated to the lower surface of six healthy potted plants. Three non-inoculated plants were served as controls. Each plant was kept in a separate plastic humid chamber in a greenhouse with 25°C and 80% humidity for 15 days. Typical symptoms of white rust pustules developed on the inoculated plants were identical to that observed on the originally infected leaves. Control plants remained symptomless.. Based on morphological characteristics, molecular data, as well as pathogenicity tests, the pathogen on C. sativa was identified as Albugo koreana. A. koreana aslo is reported only on C. bursa-pastoris in Korea (Choi et al. 2007; Farr and Rossman 2020). To our knowledge, this is the first record of white rust disease caused by A. koreana on C. sativa, and the species is new to China. This report represents a new host plant association and a new geographical expansion for this species, presenting a potential threat to camelina production in northwest China.
PubMed: 33496607
DOI: 10.1094/PDIS-11-20-2332-PDN -
Scientific Reports Jul 2014The existence of mammalian female germline stem cells (FGSCs) indicates that mammalian ovaries possess germline stem cells analogous to testis, and continue to produce...
The existence of mammalian female germline stem cells (FGSCs) indicates that mammalian ovaries possess germline stem cells analogous to testis, and continue to produce gametes postnatally, which provides new insights into female fertility. In this study, we compared the morphological and molecular characteristics between FGSCs and spermatogonial stem cells (SSCs) by analysis of morphology, immunofluorescence, alkaline phosphatase activity assay, reverse transcription polymerase chain reaction (RT-PCR) and microarray hybridization. The results demonstrated that the morphology and growth patterns of FGSCs are similar to those of SSCs. Microarray analysis of global gene expression profiles of FGSCs and SSCs showed similar signatures in the transcriptome level. A list of 853 co-highly expressed genes (CEG) in female and male germline stem cells may be responsible for the morphological and molecular similarity. We constructed a continuous network of the CEG based on I2D protein-protein interaction database by breadth first search. From the network, we could observe the interactions of the CEG may be responsible for maintaining the properties of germline stem cells. This study was the first attempt to compare morphological and molecular characteristics between FGSCs and SSCs. These findings would provide some clues for further research on mammalian FGSCs.
Topics: Adult Stem Cells; Animals; Cell Proliferation; Cell Shape; Cells, Cultured; Female; Gene Ontology; Gene Regulatory Networks; Male; Mice; Oligonucleotide Array Sequence Analysis; Oogonia; Transcription Factors; Transcriptome
PubMed: 24993338
DOI: 10.1038/srep05580 -
Frontiers in Endocrinology 2023SOX17 has been identified as a critical factor in specification of human primordial germ cells, but whether SOX17 regulates development of germ cells after sex...
BACKGROUND
SOX17 has been identified as a critical factor in specification of human primordial germ cells, but whether SOX17 regulates development of germ cells after sex differentiation is poorly understood.
METHODS
We collected specimens of gonadal ridge from an embryo (n=1), and ovaries of foetuses (n=23) and adults (n=3). Germ cells were labelled with SOX17, VASA (classic germ cells marker), phosphohistone H3 (PHH3, mitosis marker) and synaptonemal complex protein 3 (SCP3, meiosis marker).
RESULTS
SOX17 was detected in both cytoplasm and nucleus of oogonia and oocytes of primordial and primary follicles from 15 to 28 gestational weeks (GW). However, it was exclusively expressed in cytoplasm of oogonia at 7 GW, and in nucleus of oocytes in secondary follicles. Co-expression rates of SOX17 in VASA germ cells ranged from 81.29% to 97.81% in foetuses. Co-staining rates of SOX17 and PHH3 or SCP3 were 0%-34% and 0%-57%, respectively. Interestingly, we distinguished a subpopulation of SOX17VASA germ cells in fetal ovaries. These cells clustered in the cortex and could be co-stained with the mitosis marker PHH3 but not the meiosis marker SCP3.
CONCLUSIONS
The dynamic expression of SOX17 was detected in human female germ cells. We discovered a population of SOX17 VASA germ cells clustering at the cortex of ovaries. We could not find a relationship between mitosis or meiosis and SOX17 or VASA staining in germ cells. Our findings provide insight into the potential role of SOX17 involving germ cells maturation after specification, although the mechanism is unclear and needs further investigation.
Topics: Humans; Female; Adult; Ovary; Germ Cells; Oocytes; Oogonia; Fetus; SOXF Transcription Factors
PubMed: 37576970
DOI: 10.3389/fendo.2023.1124143 -
Aging Jul 2018In the present paper, we found that human fetal ovaries (at ~16 weeks) express the transcripts for several subunits of the nicotinic acetylcholine receptor (nAChR)....
In the present paper, we found that human fetal ovaries (at ~16 weeks) express the transcripts for several subunits of the nicotinic acetylcholine receptor (nAChR). Exposure to the drug resulted in the marked increase of apoptosis in the ovaries in a time and dose-dependent manner. Evidence that adverse nicotine effects are potentially due to an increased level of reactive oxygen species (ROS) and consequent DNA damage, both in the ovarian somatic cells and germ cells, are reported. After 4 days of culture, exposure to 1 mM and 10 mM nicotine caused a 50% and 75% decrease, respectively, in the number of oogonia/oocytes present in the fetal ovaries. These results represent the first indication that nicotine may directly cause apoptosis in cells of the fetal human ovary and may lead to a reduction of the ovarian reserve oocytes and consequent precocious menopause in mothers smoking during pregnancy.
Topics: Apoptosis; Dose-Response Relationship, Drug; Female; Fetus; Germ Cells; Humans; Nicotine; Nicotinic Agonists; Ovary; Tissue Culture Techniques
PubMed: 30001218
DOI: 10.18632/aging.101492 -
Biology of Reproduction Apr 2018Following proliferation of oogonia in mammals, great numbers of germ cells are discarded, primarily by apoptosis, while the remainder form primordial follicles (the...
Following proliferation of oogonia in mammals, great numbers of germ cells are discarded, primarily by apoptosis, while the remainder form primordial follicles (the ovarian reserve) that determine fertility and reproductive lifespan. More massive, rapid, and essentially total loss of oocytes, however, occurs when the transcription factor Lhx8 is ablated-though the cause and mechanism of germ cell loss from the Lhx8-/- ovaries has been unknown. We found that Lhx8-/- ovaries maintain the same number of germ cells throughout embryonic development; rapid decrease in the pool of oocytes starts shortly before birth. The loss results from activation of autophagy, which becomes overwhelming within the first postnatal week, with extracellular matrix proteins filling the space previously occupied by follicles to produce a fibrotic ovary. Associated with this process, as early as a few days before birth, Lhx8-/- oocytes failed to repair DNA damage-which normally occurs when meiosis is initiated during embryonic development; and DNA damage repair genes were downregulated throughout the oocyte short lifespan. Based on gene expression analyses and morphological changes, we propose a model in which lineage-restricted failure of DNA repair triggers germ cell autophagy, causing premature depletion of the ovarian reserve in Lhx8-/- mice.
Topics: Animals; Apoptosis; Autophagy; DNA Damage; Female; LIM-Homeodomain Proteins; Meiosis; Mice; Mice, Knockout; Oocytes; Oogenesis; Oogonia; Ovarian Reserve; Transcription Factors
PubMed: 29329412
DOI: 10.1093/biolre/iox184 -
The EMBO Journal Nov 2020Human germ cells perpetuate human genetic and epigenetic information. However, the underlying mechanism remains elusive, due to a lack of appropriate experimental...
Human germ cells perpetuate human genetic and epigenetic information. However, the underlying mechanism remains elusive, due to a lack of appropriate experimental systems. Here, we show that human primordial germ cell-like cells (hPGCLCs) derived from human-induced pluripotent stem cells (hiPSCs) can be propagated to at least ~10 -fold over a period of 4 months under a defined condition in vitro. During expansion, hPGCLCs maintain an early hPGC-like transcriptome and preserve their genome-wide DNA methylation profiles, most likely due to retention of maintenance DNA methyltransferase activity. These characteristics contrast starkly with those of mouse PGCLCs, which, under an analogous condition, show a limited propagation (up to ~50-fold) and persist only around 1 week, yet undergo cell-autonomous genome-wide DNA demethylation. Importantly, upon aggregation culture with mouse embryonic ovarian somatic cells in xenogeneic-reconstituted ovaries, expanded hPGCLCs initiate genome-wide DNA demethylation and differentiate into oogonia/gonocyte-like cells, demonstrating their germline potential. By creating a paradigm for hPGCLC expansion, our study uncovers critical divergences in expansion potential and the mechanism for epigenetic reprogramming between the human and mouse germ cell lineage.
Topics: Animals; Cell Line; DNA Demethylation; DNA Methylation; Embryonic Stem Cells; Epigenesis, Genetic; Epigenomics; Female; Genome; Germ Cells; Humans; Induced Pluripotent Stem Cells; Mice; Ovary; Pluripotent Stem Cells
PubMed: 32954504
DOI: 10.15252/embj.2020104929