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Sperm function, mitochondrial activity and in vivo fertility are associated to their mitochondrial DNA content in pigs

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Journal of Animal Science and Biotechnology 2024年第4期15卷 1550-1558页

作者： Marc Llavanera Yentel Mateo‑Otero Estel Vinolas‑Vergés Sergi Bonet Marc Yeste Unit of Cell biology Department of BiologyFaculty of SciencesUniversity of GironaGirona ES‑17003Spain Biotechnology of Animal and Human reproduction(TechnoSperm) Institute of Food and Agricultural TechnologyUniversity of GironaGirona ES‑17003Spain Catalan Institution for Research and Advanced Studies(ICREA) Barcelona ES‑08010Spain

Background Despite their low abundance in sperm, mitochondria have diverse functions in this cell type, includ-ing energy production, signalling and calcium regulation. In humans, sperm mitochondrial DNA content(mtDNAc) has been reported to be negatively linked to sperm function and fertility. Yet, the association between mtDNAc and sperm function in livestock remains unexplored. For this reason, this study aimed to shed some light on the link between mtDNAc and sperm function and fertilising potential in pigs. A qPCR method for mtDNAc quantification was optimised for pig sperm, and the association of this parameter with sperm motility, kinematics, mitochondrial activity, and fertility was subsequently *** First, the q PCR method was found to be sensitive and efficient for mtDNAc quantification in pig sperm. By using this technique, mtDNAc was observed to be associated to sperm motility, mitochondrial activity and in vivo, but not in vitro, fertility outcomes. Specifically, sperm with low mtDNAc were seen to exhibit greater motility but decreased mitochondrial activity and intracellular reactive oxygen species. Interestingly, samples with lower mtD-NAc showed higher conception and farrowing rates, but similar in vitro fertilisation rates and embryo development, when compared to those with greater *** These findings enrich our comprehension of the association of mtDNAc with sperm biology, and lay the foundation for future research into employing this parameter as a molecular predictor for sperm function and fer-tility in livestock.

关键词： Fertility Mitochondria Motility mtDNA copy number qPCR Sperm

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Chromatin condensation but not DNA integrity of pig sperm is greater in the sperm-rich fraction

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Journal of Animal Science and Biotechnology 2024年第1期15卷 171-181页

作者： Estel Viñolas-Vergés Jordi Ribas-Maynou Isabel Barranco Camila Peres Rubio Sergi Bonet Jordi Roca Marc Yeste Unit of Cell biology Department of BiologyFaculty of SciencesUniversity of GironaGironaSpain Biotechnology of Animal and Human reproduction(TechnoSperm) Institute of Food and Agricultural TechnologyUniversity of GironaGironaSpain department of Medicine and Animal Surgery Faculty of Veterinary ScienceUniversity of MurciaMurciaSpain Catalan Institution for Research and Advanced Studies(ICREA) S08010 BarcelonaSpain

Background Protamination and condensation of sperm chromatin as well as DNA integrity play an essential role during fertilization and embryo *** some mammals,like pigs,ejaculates are emitted in three separate fractions:pre-sperm,sperm-rich(SRF)and post sperm-rich(PSRF).These fractions are known to vary in volume,sperm concentration and quality,as well as in the origin and composition of seminal plasma(SP),with differences being also observed within the SRF ***,whether disparities in the DNA integrity and chromatin condensation and pro-tamination of their sperm exist has not been *** This study determined chromatin protamination(Chromomycin A3 test,CMA_(3)),condensation(Dibromobi-mane test,DBB),and DNA integrity(Comet assay)in the pig sperm contained in the first 10 m L of the SRF(SRF-P1),the remaining portion of the sperm-rich fraction(SRF-P2),and the post sperm-rich fraction(PSRF).While chromatin protamination was found to be similar between the different ejaculate fractions(P>0.05),chromatin condensation was seen to be greater in SRF-P1 and SRF-P2 than in the PSRF(P=0.018 and P=0.004,respectively).Regarding DNA integrity,no differences between fractions were observed(P>0.05).As the SRF-P1 has the highest sperm concentra-tion and ejaculate fractions are known to differ in antioxidant composition,the oxidative stress index(OSi)in SP,calcu-lated as total oxidant activity divided by total antioxidant capacity,was tested and confirmed to be higher in the SRF-P1 than in SRF-P2 and PSRF(0.42±0.06 vs.0.23±0.09 and 0.08±0.00,respectively;P<0.01);this index,in addition,was observed to be correlated to the sperm concentration of each fraction(Rs=0.973;P<0.001).Conclusion While sperm DNA integrity was not found to differ between ejaculate fractions,SRF-P1 and SRF-P2 were observed to exhibit greater chromatin condensation than the *** could be related to the OSi of each fraction.

关键词： Chromatin Condensation DNA integrity Ejaculate fractions Pig Protamination Sperm

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A mutation in TBXT causes congenital vertebral malformations in humans and mice

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Journal of Genetics and Genomics 2024年第4期51卷 433-442页

作者： Shuxia Chen Yunping Lei Yajun Yang Chennan Liu Lele Kuang Li Jin Richard HFinnell Xueyan Yang Hongyan Wang Obstetrics and Gynecology Hospital State Key Laboratory of Genetic Engineering at School of Life SciencesKey Laboratory of Reproduction Regulation of NPFPCInstitute of Reproduction and DevelopmentFudan UniversityShanghai 200438China Shanghai Key Laboratory of Metabolic Remodeling and Health Institute of Metabolism and Integrative BiologyFudan UniversityShanghai 200438China Center for Precision Environmental Health Department of Molecular and Cellular BiologyBaylor College of MedicineHoustonTX 77030USA MOE Key Laboratory of Contemporary Anthropology School of Life SciencesFudan UniversityShanghai 200438China department of Assisted reproduction Xinhua HospitalSchool of MedicineShanghai Jiao Tong UniversityShanghai 200092China Children’s Hospital Fudan University399 Wanyuan RoadShanghai 201102China

T-box transcription factor T(TBXT;T)is required for mesodermal formation and axial skeletal *** it has been extensively studied in various model organisms,human congenital vertebral malformations(CVMs)involving T are not well ***,we report a family with 15 CVM patients distributed across 4 *** affected individuals carry a heterozygous mutation,T c.596A>G(p.Q199R),which is not found in unaffected family members,indicating co-segregation of the genotype and *** vitro assays show that T p.Q199R increases the nucleocytoplasmic ratio and enhances its DNA-binding affinity,but reduces its transcriptional activity compared to the *** determine the pathogenicity of this mutation in vivo,we generated a Q199R knock-in mouse model that recapitulates the human CVM *** heterozygous Q199R mice show subtle kinked or shortened tails,while homozygous mice exhibit tail filaments and severe vertebral ***,we show that the Q199R mutation in T causes CVM in humans and mice,providing previously unreported evidence supporting the function of T in the genetic etiology of human CVM.

关键词： Congenital vertebral malformation TBXT T gene Loss-of-function mutation

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Design of an automated robotic microinjection system for batch injection of zebrafish embryos and larvae

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Microsystems & Nanoengineering 2024年第1期10卷 89-103页

作者： Zhongyi Guo Nana Ai Wei Ge Qingsong Xu department of Electromechanical Engineering Faculty of Science and TechnologyUniversity of MacaoMacaoChina department of Biomedical Sciences and Centre of reproduction Development and Aging(CRDA)Faculty of Health SciencesUniversity of MacaoMacaoChina

This paper presents the design of a vision-based automated robotic microinjection system for batch injection of both zebrafish embryos and larvae.A novel visual recognition algorithm based on an automatic threshold and excessive dilatation is introduced to accurately identify the center of zebrafish embryos and larval yolks.A corresponding software system is developed using the producer-consumer model as the framework structure,and a friendly user interface is designed to allow operators to choose from a range of desired functions according to their different *** addition,a novel microstructural agarose device is designed and fabricated to simultaneously immobilize mixed batches of embryos and ***,a prototype microinjection system is fabricated by integrating hardware devices with visual *** experimental study is conducted to verify the performance of the robotic microinjection *** results show that the reported system can accurately identify zebrafish embryos and larvae and efficiently complete batch microinjection tasks of the mixtures with an injection success rate of 92.05%in 13.88 s per *** with manual and existing microinjection systems,the proposed system demonstrates the merits of versatility,excellent efficiency,high success rate,high survival rate,and sufficient stability.

关键词： zebra automated hardware

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Extracellular vesicles-coupled miRNAs from oviduct and uterus modulate signaling pathways related to lipid metabolism and bovine early embryo development

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Journal of Animal Science and Biotechnology 2024年第4期15卷 1528-1549页

作者： Rosane Mazzarella Karina Canon‑Beltran Yulia N.Cajas Meriem Hamdi Encina M.Gonzalez Juliano C.da Silveira Claudia L.V.Leal D.Rizos department of Animal reproduction INIA-CSICMadridSpain department of Biochemistry and Molecular biology Veterinary FacultyComplutense University of Madrid(UCM)MadridSpain department Agrarian Production Technical University of MadridUPMMadridSpain Departamento de Ciencias Biologicas Universidad Tecnica Particular de LojaUTPLLojaEcuador department of Anatomy and Embryology FV-UCMMadridSpain department of Veterinary Medicine FZEA-USPPirassunungaBrazil

Background Extracellular vesicles(EVs) present in oviductal(OF) and uterine fluid(UF) have been shown to enhance bovine embryo quality during in vitro culture by reducing lipid contents and modulating lipid metabolism-related genes(LMGs), while also influencing cell proliferation, suggesting their involvement on the regulation of different biological pathways. The regulation of signaling pathways related to cell differentiation, proliferation, and metabo-lism is crucial for early embryo development and can determine the success or failure of the pregnancy. Bioactive molecules within EVs in maternal reproductive fluids, such as micro RNAs(miRNAs), may contribute to this regulatory process as they modulate gene expression through post-transcriptional *** This study evaluated miRNA cargo in OF-EVs from the early luteal phase and UF-EVs from the mid-luteal phase, coinciding with embryo transit within oviduct and uterus in vivo, and its possible influence on LMGs and sign-aling pathways crucial for early embryo development. A total of 333 miRNAs were detected, with 11 exclusive to OF, 59 to UF, and 263 were common between both groups. From the 20 differentially expressed miRNAs, 19 up-regulated in UF-EVs(bta-miR-134, bta-miR-151-3p, bta-miR-155, bta-miR-188, bta-miR-181b, bta-miR-181d, bta-miR-224, bta-miR-23b-3p, bta-miR-24-3p, bta-miR-27a-3p, bta-miR-29a, bta-miR-324, bta-miR-326, bta-miR-345-3p, bta-miR-410, bta-miR-652, bta-miR-677, bta-miR-873 and bta-miR-708) and one(bta-miR-148b) in OF-EVs. These miRNAs were predicted to modulate several pathways such as Wnt, Hippo, MAPK, and lipid metabolism and degradation. Differ-ences in miRNAs found in OF-EVs from the early luteal phase and UF-EVs from mid-luteal phase may reflect differ-ent environments to meet the changing needs of the embryo. Additionally, miRNAs may be involved, particularly in the uterus, in the regulation of embryo lipid metabolism, immune system, and *** Our study su

关键词： Embryo-maternal interaction Epigenetic Exosomes Mammalian Preimplantation Reproductive fluids

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Short neuropeptide F in integrated insect physiology

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Journal of Zhejiang University-Science B(Biomedicine & Biotechnology) 2024年第5期25卷 389-409页

作者： Marcin CHOLEWINSKI Szymon CHOWANSKI Jan LUBAWY Arkadiusz URBANSKI Karolina WALKOWIAK-NOWICKA Paweł MARCINIAK department of Animal Physiology and Developmental biology Adam Mickiewicz UniversityPoznan 61-614Poland

The short neuropeptide F(sNPF)family of peptides is a multifunctional group of neurohormones involved in the regulation of various physiological processes in *** have been found in a broad spectrum of species,but the number of isoforms in the precursor molecule varies from one to *** receptor for sNPF(sNPFR),which belongs to the G protein-coupled receptor family,has been characterized in various insect orders and was shown to be an ortholog of the mammalian prolactin-releasing peptide receptor(PrPR).The sNPF signaling pathway interacts with other neurohormones such as insulin-like peptides,SIFamide,and pigment-dispersing factors(PDFs)to regulate various *** main physiological function of sNPF seems to be involved in the regulation of feeding,but the observed effects are *** is also connected with the regulation of foraging behavior and the olfactory *** influence of sNPF on feeding and thus energy metabolism may also indirectly affect other vital processes,such as reproduction and *** addition,these neurohormones are involved in the regulation of locomotor activity and circadian rhythm in *** review summarizes the current state of knowledge about the sNPF system in insects.

关键词： Insect neuropeptide Invertebrate neurobiology Short neuropeptide F(sNPF) Feeding Neurohormone

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RNA m6A dynamic modification mediated by nucleuslocalized FTO is involved in follicular reserve

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Zoological Research 2024年第2期45卷 415-428页

作者： Jiao-Na Zhang Rui-Ting Wang Francesca-Gioia Klinger Shun-Feng Cheng Wei Shen Xiao-Feng Sun College of Life Sciences Key Laboratory of Animal Reproduction and Germplasm Enhancement in Universities of ShandongQingdao Agricultural UniversityQingdaoShandong 266109China department of Biomedicine and Prevention University of Rome Tor VergataRome 00133Italy

In eukaryotic organisms,the most common internal modification of messenger RNA(m RNA)is N6-methyladenosine(m6A).This modification can be dynamically and reversibly controlled by specific enzymes known as m6A writers and *** fat-mass and obesity-associated protein(FTO)catalyzes RNA demethylation and plays a critical role in various physiological and pathological *** research identified dynamic alterations in both m6A and FTO during the assembly of primordial follicles,with an inverse relationship observed for m6A levels and nuclear-localized FTO *** of Fto small interfering RNA(si RNA)altered the expression of genes related to cell proliferation,hormone regulation,and cell chemotaxis,and affected RNA alternative *** of the full-length Fto gene led to changes in m6A levels,alternative splicing of Cdk5,cell proliferation,cell cycle progression,and proportion of primordial ***,overexpression of Fto lacking a nuclear localization signal(NLS)did not significantly alter m6A levels or primordial follicle *** findings suggest that FTO,localized in the nucleus but not in the cytoplasm,regulates RNA m6A demethylation and plays a role in cell proliferation,cell cycle progression,and primordial follicle *** results highlight the potential of m6A and its eraser FTO as possible biomarkers and therapeutic targets.

关键词： m6A FTO Ovary Primordial follicle assembly Alternative splicing

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Human CD56+CD39+ dNK cells support fetal survival through controlling trophoblastic cell fate:immune mechanisms of recurrent early pregnancy loss

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National Science Review 2024年第6期11卷 129-145页

作者： Wentong Jia Liyang Ma Xin Yu Feiyang Wang Qian Yang Xiaoye Wang Mengjie Fan Yan Gu Ran Meng Jian Wang Yuxia Li Rong Li Xuan Shao Yan-Ling Wang State Key Laboratory of Stem cell and Reproductive biology Key Laboratory of Organ Regeneration and ReconstructionInstitute of ZoologyChinese Academy of Sciences Beijing Institute for Stem Cell and Regenerative Medicine University of the Chinese Academy of Sciences NHC Key Lab of reproduction Regulation Shanghai Engineering Research Center of Reproductive Health Drug and Devices Shanghai Institute for Biomedical and Pharmaceutical Technologies National Clinical Center for Obstetrics and Gynecology Peking University Third Hospital department of Family Planning The Second Hospital of Tianjin Medical University department of Prenatal Screening Haidian Maternal and Child Health Hospital

Decidual natural killer（dNK） cells are the most abundant immune cells at the maternal-fetal interface during early pregnancy in both mice and humans, and emerging single-cell transcriptomic studies have uncovered various human dNK subsets that are disrupted in patients experiencing recurrent early pregnancy loss（RPL） at early gestational stage, suggesting a connection between abnormal proportions or characteristics of dNK subsets and RPL pathogenesis. However, the functional mechanisms underlying this association remain unclear. Here, we established a mouse model by adoptively transferring human dNK cells into pregnant NOG（NOD/Shi-scid/IL-2Rγnull） mice, where human dNK cells predominantly homed into the uteri of recipients. Using this model, we observed a strong correlation between the properties of human dNK cells and pregnancy outcome. The transfer of dNK cells from RPL patients（dNK-RPL）remarkably worsened early pregnancy loss and impaired placental trophoblast cell differentiation in the recipients. These adverse effects were effectively reversed by transferring CD56+CD39+dNK *** studies revealed that CD56+CD39+dNK subset facilitates early differentiation of mouse trophoblast stem cells（m TSCs） towards both invasive and syncytial pathways through secreting macrophage colony-stimulating factor（M-CSF）. Administration of recombinant M-CSF to NOG mice transferred with dNK-RPL efficiently rescued the exacerbated pregnancy outcomes and fetal/placental development. Collectively, this study established a novel humanized mouse model featuring functional human dNK cells homing into the uteri of recipients and uncovered the pivotal role of M-CSF in fetal-supporting function of CD56+CD39+dNK cells during early pregnancy, highlighting that M-CSF may be a previously unappreciated therapeutic target for intervening RPL.

关键词： recurrent pregnancy loss CD56+CD39+ dNK subset NOG mice adoptive transfer placental trophoblast cell fate M-CSF

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Is the speed of adjusting to environmental change condition dependent?An experiment with house mice(Mus musculus)

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Current Zoology 2024年第3期70卷 350-360页

作者： Karem Lopez-Hervas Neelam Porwal Mathilde Delacoux Alexandros Vezyrakis Anja Guenther RG Behavioural Ecology of Individual Differences Max Planck Institute for Evolutionary Biology24306 PlönGermany department of Evolutionary biology Faculty of BiologyAdam Mickiewicz UniversityWieniawskiego 161-712 PoznańPoland department for Collective Behaviour Max Planck Institute of Animal Behaviour78464 ConstanceGermany Centre for the Advanced Study of Collective Behaviour University of Konstanz78464 ConstanceGermany Animal Ecology Institute for Biochemistry and BiologyUniversity of Potsdam14469 PotsdamGermany

Environmental conditions change constantly either by anthropogenic perturbation or naturally across space and ***,a change in behavior is the frst response to changing *** fexibility can potentially improve an organism’s chances to survive and ***,we lack an understanding on the time-scale such behavioral adjustments need,how they actually affect reproduction and survival and whether behavioral adjustments are suffcient in keeping up with changing *** used house mice(Mus musculus)to test whether personality and life-history traits can adjust to an experimentally induced food-switch fexibly in adulthood or by intergenerational plasticity,that is,adjustments only becoming visible in the offspring *** lived in 6 experimental populations of semi-natural environments either on high or standard quality food for 4 *** showed previously that high-quality food induced better conditions and a less risk-prone ***,we tested whether the speed and/or magnitude of adjustment shows condition-dependency and whether adjustments incur ftness ***-history but not personality traits reacted fexibly to a food-switch,primarily by a direct reduction of reproduction and sloweddown *** whose parents received a food-switch developed a more active stress-coping personality and gained weight at a slower rate compared with their respective ***,the modulation of most traits was condition-dependent,with animals previously fed with high-quality food showing stronger *** study highlights that life-history and personality traits adjust at different speed toward environmental change,thus,highlighting the importance of the environment and the mode of response for evolutionary models.

关键词： animal personality context dependence environmental change Match-mismatch non-genetic inheritance phenotypic flexibility phenotypic plasticity

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Physiological phenotypes differ among color morphs in introduced common wall lizards(Podarcis muralis)

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Integrative Zoology 2024年第3期19卷 505-523页

作者： Ali AMER Sierra SPEARS Princeton L.VAUGHN Cece COLWELL Ethan H.LIVINGSTON Wyatt MCQUEEN Anna SCHILL Dustin G.REICHARD Eric J.GANGLOFF Kinsey M.BROCK department of Biological Sciences Ohio Wesleyan UniversityDelawareOhioUSA department of Ecology and Evolutionary biology Princeton UniversityPrincetonNew JerseyUSA department of biology Idaho State UniversityPocatelloIdahoUSA department of Environmental Science Policyand ManagementCollege of Natural ResourcesUniversity of CaliforniaBerkeleyUSA Museum of Vertebrate Zoology University of CaliforniaBerkeleyUSA

Many species exhibit color polymorphisms which have distinct physiological and behavioral ***,the consistency of morph trait covariation patterns across species,time,and ecological contexts remains *** trait covariation is especially relevant in the context of invasion biology and urban ***,physiological traits pertaining to energy maintenance are crucial to fitness,given their immediate ties to individual reproduction,growth,and population *** investigated the physiological traits of Podarcis muralis,a versatile color polymorphic species that thrives in urban environments(including invasive populations in Ohio,USA).We measured five physiological traits(plasma corticosterone and triglycerides,hematocrit,body condition,and field body temperature),which compose an integrated multivariate *** then tested variation among co-occurring color morphs in the context of establishment in an urban *** found that the traits describing physiological status and strategy shifted across the active season in a morph-dependent manner—the white and yellow morphs exhibited clearly different multivariate physiological phenotypes,characterized primarily by differences in plasma *** suggests that morphs have different strategies in physiological regulation,the flexibility of which is crucial to urban *** white-yellow morph exhibited an intermediate phenotype,suggesting an intermediary energy maintenance *** morphs also exhibited distinct phenotypes,but the low prevalence of this morph in our study populations precludes clear *** work provides insight into how differences among stable polymorphisms exist across axes of the phenotype and how this variation may aid in establishment within novel environments.

关键词： color polymorphism physiological status seasonal variation thermoregulation urban habitat

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