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1.
Gene ; 913: 148371, 2024 Jun 30.
Article in English | MEDLINE | ID: mdl-38485034

ABSTRACT

The intestinal microbiota is increasingly recognized as playing an important role in aquatic animals. To investigate the functional roles and mechanisms of the intestinal microbial genes/enzymes responding to salinity stress or osmotic pressure in fish, metagenomic analysis was carried out to evaluate the response of intestinal microbiota and especially their functional genes/enzymes from freshwater (the control group) to acute high salinity stress (the treatment group) in Nile tilapia. Our results showed that at the microbial community level, the intestinal microbiota in Nile tilapia generally underwent significant changes in diversity after acute high salinity stress. Among them, the shift in the bacterial community (mainly from Actinobacteria to Proteobacteria) dominated and had a large impact, the fungal community showed a very limited response, and other microbiota, such as phages, likely had a negligible response. At the functional level, the intestinal bacteriadecreased the normal physiological demand and processes, such as those of the digestive system and nervous system, but enhanced energy metabolism. Furthermore, at the gene level, some gene biomarkers, such as glutathione S-transferase, myo-inositol-1(or 4)-monophosphatase, glycine betaine/proline transport system permease protein, and some families of carbohydrate-active enzymes (GT4, GT2), were significantly enriched. However, GH15, GH23 and so on were significantly reduced. Exploring the functional details of the intestinal microbial genes/enzymes that respond to salinity stress in Nile tilapia sheds light on the mechanism of action of the intestinal microbiota with respect to the salinity adaptation of fish.


Subject(s)
Cichlids , Animals , Cichlids/genetics , Salinity , Intestines , Osmotic Pressure , Salt Stress
3.
Amino Acids ; 55(4): 499-507, 2023 Apr.
Article in English | MEDLINE | ID: mdl-36715768

ABSTRACT

During trauma and surgery, bleeding is a major concern. One of the crucial strategies for hemostasis is the use of biological hemostatic material. Herein, we reported an amino acid-based hydrogel FmocF-ADP hydrogel, which consisted of N-[(9H-fluoren-9-ylmethoxy) carbonyl]-3-phenyl-L-alanine (FmocF) and adenosine diphosphate (ADP) sodium solution. The hydrogel was created by FmocF self-assembling to nanofiber in ADP sodium solution and then cross-linking to hydrogel. FmocF-ADP hydrogel showed good in vitro coagulation activity as measured by whole blood clotting assays, platelet clotting assays, platelet activation assays, and platelet adhesion assays. Further, it was noted to reveal an exceptional in vivo hemostatic effect in a mouse liver bleeding model. Together with the previous report of the good biocompatibility and antimicrobial activity of FmocF hydrogel, our study would extend the biomedical application of FmocF hydrogel. In conclusion, the present study would provide a constructive strategy for the development of new antimicrobial and hemostatic materials or develop a potential hemostatic material.


Subject(s)
Hemostatics , Animals , Mice , Hemostatics/pharmacology , Hydrogels/pharmacology , Hemostasis , Adenosine Diphosphate/pharmacology
4.
Front Genet ; 13: 998442, 2022.
Article in English | MEDLINE | ID: mdl-36437914

ABSTRACT

Background: There have been many studies on long non-coding RNAs (lncRNAs) as tumor markers. LINC00958 is a lncRNA that has been studied in a variety of tumor types. This meta-analysis aims to explore the relationship between LINC00958 and clinical prognosis and pathological characteristics in various cancers. Methods: We searched for related studies from PubMed, Web of Science, The Cochrane Library and Embase (up to October 2021). The association of LINC00958 expression with clinicopathological characteristics and prognosis was evaluated using the pooled odds ratios (ORs) or hazard ratios (HRs) with 95% confidence intervals (CIs). Results: 16 studies (1,121 patients) were included in this meta-analysis, we found that overexpression of LINC00958 was associated with poor overall survival (OS) (HR = 1.84; 95% CI: 1.36-2.49; p < 0.001). We also found that LINC00958 overexpression was correlated with positive lymph node metastasis (LNM) (OR = 1.91; 95% CI: 1.39-2.63; p < 0.001), advanced degree of infiltration (OR = 1.64; 95% CI: 1.11-2.41; p = 0.013), advanced tumor-node-metastasis (TNM) stage (OR = 2.80; 95% CI: 1.48-5.33; p = 0.002). Other clinicopathological characteristics have no obvious correlation, such as age, sex, tumor size, distant metastasis, and differentiation grade (p > 0.05). Conclusion: In summary, the overexpression of LINC00958 is significantly correlated with poor OS, positive LNM, advanced degree of infiltration, and advanced TNM stage. LINC00958 might serve as a potential prognostic biomarker and therapeutic target for a variety of cancers. However, rigorous studies with large sample sizes are still needed for further research and demonstration.

5.
Viruses ; 14(9)2022 08 25.
Article in English | MEDLINE | ID: mdl-36146680

ABSTRACT

Tomato spotted wilt orthotospovirus (TSWV) severely damaged agricultural production in many places around the world. It is generally believed that TSWV transmits among plants via their insect vector. In this study, we provide evidence on the seed-borne transmission of TSWV in pepper (Capsicum annuum L.) plants. RT-PCR, RT-qPCR, and transmission electron microscopy data demonstrate the seed transmission ability of TSWV in peppers. Endosperm, but not the embryo, is the abundant virus-containing seed organ. TSWV can also be detected in the second generation of newly germinated seedlings from virus-containing seed germination experiments. Our data are useful for researchers, certification agencies, the seed industry, and policy makers when considering the importance of TSWV in vegetable production all over the world.


Subject(s)
Capsicum , RNA Viruses , Solanum lycopersicum , Tospovirus , Plant Diseases , Plants , Seeds , Tospovirus/genetics
6.
Front Genet ; 13: 936292, 2022.
Article in English | MEDLINE | ID: mdl-35928447

ABSTRACT

Hereditary Spastic Paraplegia (HSP) is considered to be one of the common neurodegenerative diseases with marked genetic heterogeneity. Recently, the mutations in ubiquitin-associated protein 1 (UBAP1) have been described in patients with HSP, known as spastic paraplegias 80 (SPG80). Here, we reported a Chinese HSP family presenting a frameshift mutation in the UBAP1 gene leading to complex HSP. Their clinical features encompassed spastic paraparetic gait, exaggerated patellar tendon reflexes, bilateral Babinski signs, and hyperactive Achilles tendon reflex. The proband also had severe urinary incontinence and a dermoid cyst at the lumbar 4-5 spinal cord, which rarely occurs in HSP patients. Following whole-exome sequencing, a novel heterozygous mutation (c.437dupG, NM_016,525) was identified in the UBAP1 that segregated with the family's phenotype and resulted in truncating UBAP1 protein (p.Ser146ArgfsTer13). Moreover, we reviewed the genotypes of UBAP1 and the phenotypic variability in 90 HSP patients reported in the literature. We found that the age of onset in UBAP1-related patients was juvenile, and there were population differences in the age of onset. The main complications were lower extremity spasticity, hyperreflexia, and the Babinski sign. Exon 4 of UBAP1 was identified as a mutation hotspot region. Our study expands the knowledge of UBAP1 mutations, which will aid in HSP patient counseling. Further molecular biological research is needed to explore the genotype-phenotype correlations of UBAP1-related HSP.

7.
Sci Data ; 9(1): 310, 2022 06 16.
Article in English | MEDLINE | ID: mdl-35710651

ABSTRACT

As a unique geographical transition zone, the estuary is considered as a model environment to decipher the diversity, functions and ecological processes of microbial communities, which play important roles in the global biogeochemical cycle. Here we used surface water metagenomic sequencing datasets to construct metagenome-assembled genomes (MAGs) from 30 subtropical estuaries at a large scale along South China. In total, 500 dereplicated MAGs with completeness ≥ 50% and contamination ≤ 10% were obtained, among which more than one-thirds (n = 207 MAGs) have a completeness ≥ 70%. These MAGs are dominated by taxa assigned to the phylum Proteobacteria (n = 182 MAGs), Bacteroidota (n = 110) and Actinobacteriota (n = 104). These draft genomes can be used to study the diversity, phylogenetic history and metabolic potential of microbiota in the estuary, which should help improve our understanding of the structure and function of these microorganisms and how they evolved and adapted to extreme conditions in the estuarine ecosystem.


Subject(s)
Genome, Microbial , Metagenome , Microbiota , China , Estuaries , Metagenomics
8.
Reprod Toxicol ; 111: 49-58, 2022 08.
Article in English | MEDLINE | ID: mdl-35430282

ABSTRACT

Tributyltin (TBT) is extensively applied as an antifouling mediator, and it is well-known as an endocrine disrupting chemical. However, it remains elusive whether TBT can affect the development of pubertal Leydig cells (LCs), as a part of its endocrine mechanism of action. In this study, male Sprague Dawley rats (35-day-old) orally received TBT (0, 1, or 5 mg/kg) for 24 days. Immature LCs (ILCs) were separated from 35-day-old rats, and the cells were exposed to TBT at various concentrations (0, 0.05, 0.5, 1, or 5 µM) for 24 h. In vivo TBT treatment decreased the number of LCs and inhibited androgen production (2.92 ±â€¯0.44, 1.16 ±â€¯0.29, and 0.67 ±â€¯0.10 ng/ml after treatment with TBT at 0, 1, and 5 mg/kg, respectively). In vitro TBT treatment reduced androgen production, cell viability, and cell cycle progression, while increased the levels of reactive oxygen species (ROS) along with subsequent apoptosis, particularly at the concentration of 5 µM. According to in vitro and in vivo findings, TBT downregulated the expression levels of genes that could control steroidogenesis (Hsd17b3, Scarb1, Hsd3b1, and Star), and decreased protein levels due to potential reduction of NR5A1 (Nr5a1). In summary, TBT inhibited cholesterol transport and activities of certain steroidogenic enzymes, thereby leading to the reduction of androgen production.


Subject(s)
Leydig Cells , Trialkyltin Compounds , Androgens , Animals , Leydig Cells/metabolism , Male , Rats , Rats, Sprague-Dawley , Trialkyltin Compounds/toxicity
9.
Urol Int ; 106(3): 304-312, 2022.
Article in English | MEDLINE | ID: mdl-33789312

ABSTRACT

Bladder cancer (BC), as one of the most common cancers around the world, begins in the inner side of the bladder and is inclined to spread to the remaining parts of the body. Extensive documents have shown that long noncoding RNAs function as stimuli in various cancer types. With regard to LINC00649, there is limited investigation on its role previously. In our research, we discovered that LINC00649 was considerably highly expressed in BC cells and the lack of LINC00649 would cause inactivity in cellular proliferation, migration, and invasion. miR-16-5p turned out to be competitively incorporated by LINC00649 in the upstream or JARID2 downstream. In BC cells, LINC00649 was found to bind with miR-16-5p to increase the expression of JARID2. Overly expressed JARID2 was found to reverse the LINC00649 shortage-mediated suppressive impacts on the cellular process of BC cells. Concisely, it was the first study on the molecular mechanism of LINC00649 in BC. This work detected that LINC00649 enhanced cell proliferation, migration, and invasion of BC cells by acting as a sponge of miR-16-5p and upregulating JARID2, providing novel insight into understating BC.


Subject(s)
MicroRNAs , RNA, Long Noncoding , Urinary Bladder Neoplasms , Cell Line, Tumor , Cell Movement , Cell Proliferation , Gene Expression Regulation, Neoplastic , Humans , MicroRNAs/genetics , MicroRNAs/metabolism , Polycomb Repressive Complex 2/genetics , Polycomb Repressive Complex 2/metabolism , RNA, Long Noncoding/genetics , RNA, Long Noncoding/metabolism , Urinary Bladder/metabolism , Urinary Bladder Neoplasms/genetics , Urinary Bladder Neoplasms/metabolism
10.
Sci Total Environ ; 810: 152263, 2022 Mar 01.
Article in English | MEDLINE | ID: mdl-34896510

ABSTRACT

Estuaries are resistome hotspots owing to resistome accumulation and propagation at these locations from surrounding rivers, yet the large-scale biogeographic pattern of resistome, especially biocide and metal resistance genes (BMRGs) and its driving mechanisms in estuarine waters remains to be elucidated. Here, a metagenomics-based approach was firstly used to investigate resistome and mobilome profiles in waters from 30 subtropical estuaries, South China. The Pearl River estuaries had a higher diversity and abundance of antibiotic resistance genes (ARGs), BMRGs, and mobile genetic elements (MGEs) when compared with estuaries from east and west regions. Genes resistant to multiple antibiotics, metals, and biocides were the most abundant gene types in the resistome. The abundance of MGEs (e.g., intI1, IS91, and tnpA) was highly associated with the total abundance of resistance genes, suggesting their utility as potential indicators for quantitative estimations of the resistome contamination. Further, MGEs contributed more than bacterial communities in shaping the resistome in subtropical estuaries. Physicochemical factors (e.g., pH) regulated MGE composition and stochastic assembly, which mediated the co-selection of ARGs and BMRGs via horizontal gene transfer. Our findings have important implications and provide a reference on the management of ARGs and BMRGs in subtropical estuarine ecosystems.


Subject(s)
Estuaries , Metagenomics , Anti-Bacterial Agents/pharmacology , Drug Resistance, Microbial/genetics , Ecosystem , Genes, Bacterial
11.
Microorganisms ; 8(6)2020 May 29.
Article in English | MEDLINE | ID: mdl-32485891

ABSTRACT

River ecosystems are critical for human and environmental health, with bacterioplankton playing a vital role in biogeochemical cycles. Unveiling the spatial patterns of bacterioplankton communities in relation to environmental factors is important for understanding the processes of microbial variation and functional maintenance. However, our understanding of the correlations among bacterioplankton communities, physicochemical factors, and land use, especially in large rivers affected by intensive anthropogenic activities, remains relatively poor. Here, we investigated the bacterioplankton communities in July 2018 in three main tributaries of the Pearl River, i.e., Beijiang, Xijiang, and Pearl River Delta, based on 16S rRNA high-throughput sequencing. Results showed that the most dominant phyla, Proteobacteria, Actinobacteria, Cyanobacteria, and Planctomycetes accounted for 33.75%, 22.15%, 11.65%, and 10.48% of the total abundance, respectively. The bacterioplankton communities showed remarkable differences among the three tributaries in terms of composition, structure, diversity, and predictive functional profiles. Mantel and partial Mantel tests revealed that the bacterioplankton communities were affected by physicochemical variables (p < 0.01) and land use (p < 0.01). Redundancy analysis identified specific conductivity, dissolved oxygen, agricultural land, ammonium, urban land, and water transparency as the dominant environmental factors influencing the bacterioplankton communities in the Pearl River. Variation partitioning analysis indicated that both physicochemical factors and land use had direct effects on the bacterioplankton community, and that land use may also shape bacterioplankton communities through indirect effects of physicochemical factors on riverine ecosystems. This study provides fundamental information on the diversity, spatial patterns, and influencing factors of bacterioplankton communities in the Pearl River, which should enhance our understanding of how such communities change in response to environmental gradients and anthropogenic activities.

12.
Environ Pollut ; 264: 114683, 2020 Sep.
Article in English | MEDLINE | ID: mdl-32388300

ABSTRACT

Understanding the composition and assembly mechanism of waterborne pathogen is essential for preventing the pathogenic infection and protecting the human health. Here, based on 16S rRNA sequencing, we investigated the composition and spatial variation of potentially pathogenic bacteria from different sections of the Pearl River, the most important source of water for human in Southern China. The results showed that the potential pathogen communities consisted of 6 phyla and 64 genera, covering 11 categories of potential pathogens mainly involving animal parasites or symbionts (AniP), human pathogens all (HumPA), and intracellular parasites (IntCelP). Proteobacteria (75.87%) and Chlamydiae (20.56%) were dominant at the phylum level, and Acinetobacter (35.01%) and Roseomonas (8.24%) were dominant at the genus level. Multivariate analysis showed that the potential pathogenic bacterial community was significantly different among the four sections in the Pearl River. Both physicochemical factors (e.g., NO3-N, and suspended solids) and land use (e.g., urban land and forest) significantly shaped the pathogen community structure. However, spatial effects contributed more to the variation of pathogen community based on variation partitioning and path analysis. Null model based normalized stochasticity ratio analysis further indicated that the stochastic process rather than deterministic process dominated the assembly mechanisms by controlling the spatial patterns of potential pathogens. In conclusion, high-throughput sequencing shows great potential for monitoring the potential pathogens, and provided more comprehensive information on the potentially pathogenic community. Our study highlighted the importance of considering the influences of dispersal-related processes in future risk assessments for the prevention and control of pathogenic bacteria.


Subject(s)
Bacteria/genetics , Rivers , Animals , China , Humans , Proteobacteria , RNA, Ribosomal, 16S
13.
Microorganisms ; 8(3)2020 Mar 18.
Article in English | MEDLINE | ID: mdl-32197354

ABSTRACT

Investigations of gut microbial diversity among fish to provide baseline data for wild marine fish, especially the carnivorous coral reef fishes of the South China Sea, are lacking. The present study investigated the gut microbiota of four carnivorous coral reef fishes, including Oxycheilinus unifasciatus, Cephalopholis urodeta, Lutjanus kasmira, and Gnathodentex aurolineatus, from the South China Sea for the first time using high-throughput Illumina sequencing. Proteobacteria, Firmicutes, and Bacteroidetes constituted 98% of the gut microbiota of the four fishes, and 20 of the gut microbial genera recovered in this study represent new reports from marine fishes. Comparative analysis indicated that the four fishes shared a similar microbial community, suggesting that diet type (carnivorous) might play a more important role in shaping the gut microbiota of coral reef fishes than the species of fish. Furthermore, the genera Psychrobacter, Escherichia-Shigella, and Vibrio constituted the core microbial community of the four fishes, accounting for 61-91% of the total sequences in each fish. The lack of the genus Epulopiscium in the four fishes was in sharp contrast to what has been found in coral reef fishes from the Red Sea, in which Epulopiscium was shown to be the most dominant gut microbial genus in seven herbivorous coral reef fishes. In addition, while unique gut microbial genera accounted for a small proportion (8-13%) of the total sequences, many such genera were distributed in each coral reef fish species, including several genera (Endozoicomonas, Clostridium, and Staphylococcus) that are frequently found in marine fishes and 11 new reports of gut microbes in marine fishes. The present study expands our knowledge of the diversity and specificity of gut microbes associated with coral reef fishes.

14.
Food Chem ; 314: 126205, 2020 Jun 01.
Article in English | MEDLINE | ID: mdl-31968291

ABSTRACT

An 8-week feeding trial and transcriptome analysis were conducted to investigate the potential mechanism of muscle-hardening caused by faba bean in grass carp (Ctenopharyngodon idellus). Ordinary grass carp (fed with practical diet) and crisp grass carp (fed with faba bean meal) groups were designed. Lower water holding capacity and higher some texture parameters were observed in the muscle of crisp grass carp compared with another group. 19.62 GB clean reads were generated, and total 1354 genes exhibiting differentially expression were identified (FDR < 0.05). Genes function enrichment revealed up-regulated genes in crisp grass carp mainly in response to myofibroblast proliferation, while down-regulated genes in response to immune regulation. Consistent with this, the tight junction pathway and the NF-κB signaling pathway were likewise significantly enriched. In summary, this study identified several candidate genes and putative signaling pathways deserving further investigation to the mechanism of muscle-hardening in fish fed with faba bean.


Subject(s)
Animal Feed , Carps/genetics , Animals , Carps/metabolism , Fish Proteins/genetics , Gene Expression Profiling , Gene Expression Regulation , Muscles/metabolism , Sequence Analysis, RNA , Vicia faba
15.
Microorganisms ; 7(10)2019 Oct 17.
Article in English | MEDLINE | ID: mdl-31627338

ABSTRACT

Many reports of the intestinal microbiota of grass carp have addressed the microbial response to diet or starvation or the effect of microbes on metabolism; however, the intestinal microbiota of crisp grass carp has yet to be elucidated. Moreover, the specific bacteria that play a role in the crispiness of grass carp fed faba beans have not been elucidated. In the present study, 16S sequencing was carried out to compare the intestinal microbiota in the fore-, mid- and hind-intestine segments of grass carp following feeding with either faba beans or formula feed. Our results showed that (1) the hind-intestine presented significant differences in diversity relative to the fore- or midintestine and (2) faba beans significantly increased the diversity of intestinal microbiota, changed the intestinal microbiota structure (Fusobacteria was reduced from 64.26% to 18.24%, while Proteobacteria was significantly increased from 17.75% to 51.99%), and decreased the metabolism of energy, cofactors and vitamins in grass carp. Furthermore, at the genus and species levels, Acinetobacter accounted for 15.09% of the microbiota, and Acinetobacter johnsonii and Acinetobacter radioresistens constituted 3.41% and 2.99%, respectively, which indicated that Acinetobacter of the family Moraxellaceae contributed to changes in the intestinal microbiota structure and could be used as a potential biomarker. These results may provide clues at the intestinal microbiota level to understanding the mechanism underlying the crispiness of grass carp fed faba beans.

16.
Philos Trans R Soc Lond B Biol Sci ; 374(1767): 20180313, 2019 03 04.
Article in English | MEDLINE | ID: mdl-30967015

ABSTRACT

Whiteflies, Bemisia tabaci (Hemiptera), are pests causing economic damage to many crops, capable of transmitting hundreds of plant vector-borne viruses. They are believed to secrete salivary protein effectors that can improve vector colonization and reproductive fitness in host plants. However, little is known about effector biology and the precise mechanism of action of whitefly effectors. Here, we report a functional screening of B. tabaci salivary effector proteins (Bsp) capable of modulating plant innate immunity triggered by plant endogenous pattern peptide Pep1. Four immunity suppressors and two elicitors were identified. Bsp9, the most effective immunity suppressor, was further identified to directly interact with an immunity regulator WRKY33. We provide evidence that Bsp9 may suppress plant immune signalling by interfering with the interaction between WRKY33 and a central regulator in the MAPK cascade. The interference by Bsp9 therefore reduces plant resistance to whitefly by inhibiting activation of WRKY33-regulated immunity-related genes. Further detailed analysis based on transgenic plants found that whitefly effector Bsp9 could promote whitefly preference and performance, increasing virus transmission. This study enriches our knowledge on insect effector biology. This article is part of the theme issue 'Biotic signalling sheds light on smart pest management'.


Subject(s)
Hemiptera/physiology , Insect Proteins/genetics , Plant Immunity/immunology , Plant Proteins/genetics , Transcription Factors/genetics , Animals , Hemiptera/genetics , Herbivory , Insect Proteins/metabolism , Plant Immunity/genetics , Plant Proteins/metabolism , Transcription Factors/metabolism
17.
Virus Res ; 265: 166-171, 2019 05.
Article in English | MEDLINE | ID: mdl-30910699

ABSTRACT

Papaya ringspot virus watermelon strain (PRSV-W) causes huge economic losses to cucurbits production. Here, we constructed an infectious clone of PRSV-W, pCamPRSV-W, which can induce similar symptoms and accumulate to same levels as wild type virus in plants of Cucurbita pepo, Cucumis melo, Citrullus lanatus and Cucumis sativus. The green fluorescence protein gene gfp was cloned into pCamPRSV-W to produce pCamPRSV-W-GFP, which produced strong green fluorescence in systemic leaves of inoculated Cucurbita pepo, Cucumis melo, Citrullus lanatus and Cucumis sativus plants, indicating that pCamPRSV-W can be used to express foreign genes. Ten mutants of PRSV-W, obtained by site-directed mutagenesis in the RNA silencing suppressor helper-component proteinase encoding region, produced dramatically attenuated symptoms in plants of Cucumis melo. The Cucumis melo plants pre-infected with mutants K125D and G317 K showed effective protection against the challenge inoculation of wild type PRSV-W. The attenuated mutants generated in this study will be helpful for the eco-friendly control of PRSV-W.


Subject(s)
Cross Protection , Cucumis/virology , Plant Diseases/prevention & control , Potyvirus/genetics , RNA Interference , Citrullus/virology , Cucurbita/virology , Mutation , Phylogeny , Plant Diseases/virology , RNA, Viral/genetics , Sequence Analysis, DNA , Viral Proteins/genetics
18.
Front Pharmacol ; 9: 1320, 2018.
Article in English | MEDLINE | ID: mdl-30555322

ABSTRACT

Dibutyltin dichloride (DBTCl), widely used as plastic stabilizer, can cause comprehensive toxicity. The present study aims to investigate the effects of DBTCl on rat Leydig cell developmental regeneration and characterize the related mechanism. Adult male Sprague Dawley rats were randomly divided into four groups and gavaged with saline (control) or 5, 10, or 20 mg/kg/day of DBTCl consecutively for 10 days. At the end of the DBTCl treatment, all rats received a single intraperitoneal injection (i.p.,) of 75 mg/kg ethane dimethane sulfonate (EDS) to eliminate all the adult Leydig cells and to induce Leydig cell developmental regeneration. Leydig cell developmental regeneration was evaluated by measuring the levels of serum testosterone, luteinizing hormone, and follicle-stimulating hormone on days 7, 35, and 56 post-EDS. Leydig cell gene and protein expression levels, as well as cell morphology and cell counts were also carried out on day 56 post-EDS. The present study found that DBTCl significantly reduced serum testosterone levels on days 35 and 56 post-EDS, but increased serum luteinizing hormone (LH) and follicle-stimulating hormone (FSH) levels on day 56 at ≥ 5 mg/kg/day. The mRNA and protein levels of Leydig (Lhcgr, Scarb1, Star, Cyp11a1, Hsd17b3, and Hsd11b1) and Sertoli cells (Fshr, Amh, and Sox9) were significantly downregulated in the DBTCl-treated testes compared to the control. Immunohistochemical staining showed that DBTCl-treatment caused fewer regenerated Leydig cells and impaired Sertoli cell development and function in the testis on day 56 post-EDS. In conclusion, the present study demonstrates that DBTCl retards rat Leydig cell developmental regeneration by downregulating steroidogenesis-related enzymes at the gene and protein levels, inhibiting Leydig cell proliferation and impairing Sertoli cell function and development.

19.
Fish Shellfish Immunol ; 81: 108-112, 2018 Oct.
Article in English | MEDLINE | ID: mdl-30017925

ABSTRACT

Long noncoding RNAs (lncRNAs), once thought to be nonfunctional, have recently been shown to participate in the multilevel regulation of transcriptional, posttranscriptional and epigenetic modifications and to play important roles in various biological processes, including immune responses. However, the expression and roles of lncRNAs in invertebrates, especially nonmodel organisms, remain poorly understood. In this study, by comparing a transcriptome to the PfIRF-2 genomic structure, we identified lncIRF-2 in the PfIRF-2 genomic intron. The results of the RNA interference (RNAi) and the nucleus grafting experiments indicated that PfIRF-2 might have a negative regulatory effect on lncIRF-2, and PfIRF-2 and lncIRF-2 may have a positive regulatory effect on PfIL-17. Additionally, lncIRF-2, PfIRF-2 and PfIL-17 were involved in responses to the nucleus graft. These results will enhance the knowledge of lncIRF-2, IRF-2, and IL-17 functions in both pearl oysters and other invertebrates.


Subject(s)
Gene Expression Regulation , Interferon Regulatory Factor-2/genetics , Interleukin-17/genetics , Pinctada/genetics , RNA, Long Noncoding/genetics , Animals , Gene Expression Profiling , Introns , RNA Interference , RNA, Messenger/genetics , Sequence Analysis, DNA , Transcriptome
20.
Fish Shellfish Immunol ; 80: 63-70, 2018 Sep.
Article in English | MEDLINE | ID: mdl-29859309

ABSTRACT

Extracellular signal-regulated kinases (ERKs) are conserved and related with protein-serine/threonine kinases that participate in the regulation of multiple biological processes, such as cell survival, cell differentiation, proliferation, metabolism, and inflammation. However, little is known about the roles of this kinase in the pearl oyster. In this study, we cloned and identified an ERK homolog from Pinctada fucata (PfErk). Furthermore, we have unraveled its expressional kinetics after lipopolysaccharide (LPS) and polyinosinic-epolycytidylic acid (poly I:C) immune challenge. Pferk harbored a 5' untranslated region (UTR) of 12 bp, a coding sequence of 1074 bp, and a 3' UTR of 882 bp. The putative peptide comprised a predicted molecular mass of 41.19 kDa, with a theoretical pI of 6.15. Sequence analysis showed that it possesses one STK catalytic domain and a conserved His-Arg-Asp (HRD) domain. In addition, a canonical Thr-Glu-Tyr (TEY) dual phosphorylation motif and an ATRW substrate binding site were also identified in the coding protein. Homology assessment of PfErk showed high similarity to Homo sapiens ERK. Phylogenetic analysis supported a close evolutionary relationship with molluscan orthologs. The expression patterns of Pferk were observed in seven different tissues of pearl oyster, with highest expression in the mantle and lowest expression in the digestive gland. Pferk mRNA expression levels were detected at developmental stages, with the highest expression in D-shaped larvae, followed by the 32-cell stage. The mRNA expression of Pferk was upregulated significantly in P. fucata mantle primary cells and mantle tissue after LPS and poly (I:C) treatment, and PfErk phosphorylation levels were activated by LPS and poly (I:C) challenges. Overall, our results suggested that PfErk may play important roles in pearl oyster innate immunity, and provided a new understanding of mantle immunity in the pearl oyster.


Subject(s)
Mitogen-Activated Protein Kinases/genetics , Mitogen-Activated Protein Kinases/immunology , Pinctada/genetics , Pinctada/immunology , Amino Acid Sequence , Animals , Cells, Cultured , Cloning, Molecular , DNA, Complementary/genetics , Lipopolysaccharides/pharmacology , Open Reading Frames , Phylogeny , Poly I-C/pharmacology , RNA, Messenger/metabolism
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