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Identification and functional analysis of CG3526 in spermatogenesis of Drosophila melanogaster

insect science 2024年第1期31卷 79-90页

作者： Qihao Hu Yanhong Xiao Runnan Wei Ting Tang Liang Wen Yuzhen Lu Xiao-Qiang Yu guangdong provincial key laboratory of insect developmental biology and applied technology Guangzhou Key Laboratory of Insect Development Regulation and Application ResearchInstitute of Insect Science and TechnologySchool of Life SciencesSouth China Normal UniversityGuangzhou510631 China

Spermatogenesis is a critical part of reproduction in insects;however,its molecular mechanism is still largely unknown.In this study,we identified a testis-specific gene CG3526 in Drosophila melanogaster.Bioinformatics analysis showed that CG3526 contains a zinc binding domain and 2 C2H2 type zinc fingers,and it is clustered to the vertebrate really interesting new gene(RING)family E3 ubiquitin-protein ligases.When CG3526 was knocked down by RNA interference(RNAi),the testis became much smaller in size,and the apical tip exhibited a sharp and thin end instead of the blunt and round shape in the control testis.More importantly,compared to the control flies,only a few mature sperm were present in the seminal vesicle of C587-Gal4>CG3526 RNAi flies.Immunofluorescence staining of the testis from CG3526 RNAi flies showed that the homeostasis of testis stem cell niche was disrupted,cell distribution in the apical tip was scattered,and the process of spermatogenesis was not completed.Furthermore,we found that the phenotype of CG3526 RNAi flies’testis was similar to that of testis of Stat92E RNAi flies,the expression level of CG3526 was significantly downregulated in the Stat92EF06346 mutant flies,and the promoter activity of CG3526 was upregulated by STAT92E.Taken together,our results indicated that CG3526 is a downstream effector gene in the JAK-STAT signaling pathway that plays a key role in the spermatogenesis of Drosophila.

关键词： CG3526 JAK-STAT spermatogenesis STAT92E testis

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Genome-wide identification and characterization of basic helix-loop-helix transcription factors in Spodoptera litura upon pathogen infection

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insect science 2022年第4期29卷 977-992页

作者： Jie Zhang Mengyao Zhu Qilin Li Ting Tang Liang Wen Jielai Zhong Ruonan Zhang Xiao-Qiang Yu Yuzhen Lu guangdong provincial key laboratory of insect developmental biology and applied technology Institute of Insect Science and TechnologySchool of Life SciencesSouth China Normal UniversityGuangzhouChina Guangzhou key laboratory of insect Development Regulation and Application Research Institute of Insect Science and TechnologySchool of Life SciencesSouth China Normal UniversityGuangzhouChina

Basic helix-loop-helix(bHLH)transcription factors play an important role in a wide range of metabolic and developmental processes in eukaryotes,and bHLH proteins also participate in immune responses,especially in plants.However,their roles in insects upon entomopathogen infection are unknown.In this study,54 bHLH genes in 41 families were identified in a polyphagous pest,Spodoptera litura,including a new bHLH gene in group B,which is specifically present in Lepidoptera and was thus named Lep.The conserved amino acids in the bHLH domain,structural architecture,and chromosomal distribution of bHLH genes in S.litura were analyzed.The bHLH genes in Plutella xylostella and Apis mellifera were also updated,and genome-wide comparison and phylogenetic analysis of bHLH members in 5 holometabolous insects were performed.The expression profiles of S.litura bHLH(SlbHLH)genes in 3 tissues at different developmental stages and their responses to S.litura nucleopolyhedrovirus(SpltNPV),Nomuraea rileyi(Nr),and Bacillus thuringiensis(Bt)infection were investigated.More SlbHLHs in group B were expressed and differentially expressed during pathogen infections,and SlbHLHs tended to be downregulated in the midgut of S.litura larvae after B.thuringiensis treatment.Our study provides an overview of bHLH family members in S.litura and their responses to different pathogens used for pest biocontrol.These findings on bHLH members may contribute to uncovering the mechanism of host-pathogen interaction.

关键词： basic helix-loop-helix entomopathogen infection pest biocontrol Spodoptera liura

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Melanin synthesis genes BgTH and BgDdc affect body color and cuticle permeability in Blattella germanica

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insect science 2022年第6期29卷 1552-1568页

作者： Tian-Tian Bai Xiao-Jin Pei Tong-Xian Liu Yong-Liang Fan Shi-Ze Zhang State key laboratory of Crop Stress biology for Arid Areas and key laboratory of lIntegrated Pest Management on Crops in Northwestern Loess Plateau Ministry of AgricultureNorthwest A&F UniversityYanglingShaanxiChina present address:guangdong provincial key laboratory of insect developmental biology and applied technology and institute of insect science and technology School of Life SciencesSouth China Normal UniversityGuangzhou510631China

Melanin is involved in cuticle pigmentation and sclerotization of insects,which is critical for maintaining structural integrity and functional completeness of insect cuti-cle.The 2 key enzymes of tyrosine hydroxylase(TH)and dopa decarboxylase(DDC)predicted in melanin biosynthesis are usually conserved in insects.However,it is unclear whether their function is related to epidermal permeability.In this study,we identified and cloned the gene sequences of BgTH and BgDdc from Blattella germanica,and revealed that they both showed a high expression at the molting,and BgTH was abundant in the head and integument while BgDdc was expressed highest in the fat body.Using RNA in-terference(RNAi),we found that knockdown of BgTH caused molting obstacles in some cockroaches,with the survivors showing pale color and softer integuments,while knock-down of BgDdc was viable and generated an abnormal light brown body color.Desiccation assay showed that the dsBgTH-injected adults died earlier than control groups under a dry atmosphere,but dsBgDdc-injected cockroaches did not.In contrast,when dsRNA-treated cockroaches were reared under a high humidity condition,almost no cockroaches died in all treatments.Furthermore,with eosin Y staining assay,we found that BgTH-RNAi resulted in a higher cuticular permeability,and BgDdc-RNAi also caused slight dye pen-etration.These results demonstrate that BgTH and BgDdc function in body pigmentation and affect the waterproofing ability of the cuticle,and the reduction of cuticular perme-ability may be achieved through cuticle melanization.

关键词： cuticular permeability desiccation tolerance dopa decarboxylase German cockroach pigmentation tyrosine hydroxylase

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Multiple microRNAs control ecdysone signaling in the midgut of Spodoptera litura

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insect science 2020年第6期27卷 1208-1223页

作者： Wei Luo Li-Xia Huang Shuang-Kang Qin Xian Zhang Qi-Li Feng Jun Gu Li-Hua Huang guangdong provincial key laboratory of insect developmental biology and applied technology Institute of Insect Science and TechnologySchool of Life SciencesSouth China Normal UniversityGuangzhouChina Guangzhou key laboratory of insect Development Regulation and Application Research Institute of Insect Science and TechnologySchool of Life SciencesSouth China Normal UniversityGuangzhouChina Hubei key laboratory of Application of Plant-Cancer Active Ingredients School of Chemistry and Life SciencesHubei University of EducationWuhanChina

Metamorphosis is one of the most important physiological processes in insects.It is regulated by a serial of ecdysone cascade genes.Recently,lots o f microRNAs(miRNAs)were investigated in insects;however,their function in metamorphosis is largely unknown.In the present study,the dynamics of a small RNA population was investigated by RNA sequencing from the midgut of a lepidopteran pest Spodoptera litura during larval-pupal metamorphosis.A total of 101 miRNAs were identified,and 75 miRNAs were differentially expressed during the metamorphic process.The relationship between these differentially expressed miRNAs and 12 ecdysone cascade genes was analyzed by four classical software programs,and a multiple-to-multiple regulatory network was found to exist between these miRNAs and their targets.Among them,miR-14-3p and its two targets(EcR and E75)were chosen for further validation.MiR-14-3p had higher expression level in the 6 th instar larvae as compared with either the prepupae or pupae,which was opposite to that of both EcR and E75,two ecdysone cascade genes.Luciferase reporter assay confirmed that both EcR and E75 were regulated by miR-14-3p.Interestingly,the 3'untranslated regions are nearly identical to each other among different transcript variants of the ecdysone cascade genes,including EcR,USP,E75,E74,E78,E93,Hr3,Hr4,Hr39,K rhl and F tzfl.Thus,different transcript variants of one ecdysone cascade gene could be regulated by the same mi RNA.The above data suggest that the ecdysone signaling pathway is under the tight control of miRNA.These findings expand our understanding of the mechanism of insect metamorphosis and may also provide a novel possibility for the control of pest insects in the future.

关键词： ecdysone EcR E75 metamorphosis microRNA midgut

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BmCH25H,a vertebrate interferon-stimulated gene(ISG)homolog,inhibits BmNPV infection dependent on its hydroxylase activity in Bombyx mori

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insect science 2023年第2期30卷 321-337页

作者： Hongyun Wu Junming Xia Shigang Fei Yeyuan Wang Mengmeng Zhang Yiyao Guo Xian Li Luc Swevers Jingchen Sun Min Feng guangdong provincial key laboratory of Agro-Animal Genomics and Molecular Breeding College of Animal ScienceSouth China Agricultural UniversityGuangzhouChina insect Molecular Genetics and Biotechnology National Center for Scientific Research DemokritosInstitute of Biosciences and ApplicationsAthensGreece

Cholesterol-25-hydroxylase(CH25H)has been identified as an interferon-stimulated gene(ISG)in mammals that exerts its antiviral effects by catalyzing the conversion of cholesterol to 25-hydroxycholesterol(25HC).However,invertebrates lack an antiviral system homologous to vertebrate interferons(IFNs)because the genomes of invertebrates do not encode IFN-like cytokines.Nevertheless,CH25H is present in insect genomes and it therefore deserves further study of whether and by which mechanism it could exert an antiviral effect in invertebrates.In this study,the Bombyx mori CH25H(BmCH25H)gene,of which the encoded protein has high homology with other lepidopteran species,was identified and located on chromosome 9.Interestingly,we found that the expression of BmCH25H was significantly upregulated in B.mori nucleopolyhedrovirus(BmNPV)-infected BmN cells and silkworm(B.mori)larvae at the early infection stage.The inhibitory effect of BmCH25H on BmNPV replication was further demonstrated to depend on its catalytic residues to convert cholesterol to 25HC.More importantly,we demonstrated that during BmNPV infection,BmCH25H expression was increased through the Janus kinase–signal transducer and activator of transcription(JAK–STAT)pathway,similar to the induction of ISGs following virus infection in vertebrates.This is the first report that CH25H has antiviral effects in insects;the study also elucidates the regulation of its expression and its mechanism of action.

关键词： Bombyx mori nucleopolyhedrovirus Bombyx mori Cholesterol 25-hydroxylase Interferon-stimulated genes Signal transducer and activator of transcription

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The kinase p38αfunctions in dendritic cells to regulate Th2-cell differentiation and allergic inflammation

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Cellular & Molecular Immunology 2022年第7期19卷 805-819页

作者： Miaomiao Han Jingyu Ma Suidong Ouyang Yanyan Wang Tingting Zheng Peishan Lu Zihan Zheng Weiheng Zhao Hongjin Li Yun Wu Baohua Zhang Ran Hu Kinya Otsu Xinguang Liu Ying Wan Huabin Li Gonghua Huang ENT institute and Department of Otorhinolaryngology Eye&ENT HospitalFudan University200031 ShanghaiChina Shanghai institute of Immunology Shanghai Jiao Tong University School of Medicine200025 ShanghaiChina guangdong provincial key laboratory of Medical Molecular Diagnostics Guangdong Medical University523808 DongguanChina Biomedical Analysis Center Army Medical University400038 ChongqingChina Department of Cardiovascular Medicine Graduate School of MedicineOsaka UniversityOsaka 565-0871Japan School of Cardiovascular Medicine and sciences King’s College LondonLondon SE59NUUK present address:Tongji Hospital Tongji Medical CollegeHuazhong University of Science and Technology430030 WuhanChina Department of Dermatology Yueyang Hospital of Integrated Traditional Chinese and Western MedicineShanghai University of Traditional Chinese Medicine200437 ShanghaiChina Department of Clinical laboratory Medicine Shanghai Tenth People’s Hospital of Tongji University200072 ShanghaiChina present address:Basic Department of Cancer Center Shanghai Tenth People’s Hospital of Tongji University200072 ShanghaiChina

Dendritic cells(DCs)play a critical role in controlling T helper 2(Th2)cell-dependent diseases,but the signaling mechanism that triggers this function is not fully understood.We showed that p38αactivity in DCs was decreased upon HDM stimulation and dynamically regulated by both extrinsic signals and Th2-instructive cytokines.p38α-specific deletion in cDC1s but not in cDC2s or macrophages promoted Th2 responses under HDM stimulation.Further study showed that p38αin cDC1s regulated Th2-cell differentiation by modulating the MK2−c-FOS−IL-12 axis.Importantly,crosstalk between p38α-dependent DCs and Th2 cells occurred during the sensitization phase,not the effector phase,and was conserved between mice and humans.Our results identify p38αsignaling as a central pathway in DCs that integrates allergic and parasitic instructive signals with Th2-instructive cytokines from the microenvironment to regulate Th2-cell differentiation and function,and this finding may offer a novel strategy for the treatment of allergic diseases and parasitic infection.

关键词： Allergy p38α dendritic cell Th2

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Regulation of hypoxic stress and oxidative stress in bone grafting: Current trends and future perspectives

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Journal of Materials science & technology 2023年第26期157卷 144-153页

作者： Hao Hu Xiao Liu Jun Chen Shangbin Cui Hualin Yi Gang Wang Renxian Wang Tiansheng Zheng Ben Wan Zhiyu Zhou Yong Wan Manman Gao Dafu Chen Xuenong Zou guangdong provincial key laboratory of Orthopaedics and Traumatology Department of Spinal SurgeryThe First Affiliated Hospital of Sun Yat-sen UniversityGuangzhou 510080China laboratory of Bone Tissue Engineering Beijing Laboratory of Biomedical MaterialsBeijing Research Institute of Traumatology and OrthopaedicsBeijing Jishuitan HospitalBeijing 100035China Department of Orthopedics The First Affiliated Hospital of Gannan Medical UniversityGanzhouChina Department of Oral and Maxillofacial Surgery/Pathology Amsterdam UMC and Academic Center for Dentistry Amsterdam(ACTA)Vrije Universiteit Amsterdam(VU)Amsterdam Movement Science(AMS)Amsterdamthe Netherlands

Tissue engineering aims to offer large-scale replacement of damaged organs using implants with the com-bination of cells,growth factors and scaffolds.However,the intra/peri-implant region is exposed to se-vere hypoxic stress and oxidative stress during the early stage of implantation with bone graft materials,which endangers the survival,proliferation and differentiation of seed cells within the implants as well as the host cells surrounding the implants.If the bone graft material could spontaneously and intelligently regulate the hypoxic stress and oxidative stress to a moderate level,it will facilitate the vascularization of the implants and the rapid regeneration of the bone tissue.In this review,we will first introduce the signaling pathways of cellular response under hypoxic stress and oxidative stress,then present the clas-sical material designs and examples in response to hypoxic stress and oxidative stress.And finally,we will address the important role of epigenetic mechanisms in the regulation of hypoxic stress and oxida-tive stress and describe the potential applications and prospective smart bone graft materials based on novel epigenetic factors against hypoxic stress and oxidative stress in bone repair.The main content of this review is summarized in the following graphical abstract.

关键词： Hypoxic stress Oxidative stress Bone repair Bone graft material Epigenetic regulation

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