Acute toxicity effects of perfluorooctane sulfonate on sperm vitality,kinematics and fertilization success in zebrafish

Perfluorooctane sulfonate(PFOS) has emerged as one of the most concerning contaminants in recent years.This study aimed to investigate the acute toxicity effect of PFOS on sperm viability,kinematics and fertilization success in zebrafish(Danio rerio).Sperm were activated in aqueous media containing a range of PFOS concentrations(0,0.09,0.9 and 9 mg/L).Viabilities and kinematics of the sperm exposed to different PFOS treatments were assessed via computer-assisted sperm analysis(CASA) at 20,40,60,and 80 s after activation.PFOS exposure decreased the percentage of motile sperm,the curvilinear velocity(VCL),and the mean angular displacement(MAD) of spermatozoa,but showed no influence on the straightline velocity(VSL) or the angular path velocity(VAP).Furthermore,a significant decrease in fertiUzation success was observed in spermatozoa that were exposed to 0.9 mg/L PFOS or more.These findings indicate that PFOS pollution in natural aquatic environment may be a potential threaten to successful reproduction offish.

Discovery of males of Simocephalus himalayensis (Cladocera, Simocephalus), with the re-description of females

The lack of data concerning the morphology of male cladocerans greatly impedes our understanding of the systematics of cladocerans. This is because the differentiation of males is of significant importance for distinguishing closely related species. Simocephalus himalayensis himalayensis Chiang & Chen, 1974 (Cladocera, Simocephalus) is an endemic cladoceran species in the Tibetan plateau, China. To date, no information has been available regarding the morphology and biology of its males. In this study, we provide the morphological and biological data of the males that were produced in laboratory populations. The male individuals are about 1.01–1.20 mm long; the margin of antennulae possess two sensory papilla; dorsal valve margin smooth. Two vas deferens are located between the supra-anal angle and the proximal anal teeth. Sperm is round with a diameter of two micrometers. An updated morphological description and reproductive data of the females of this species are also provided.

Maternal-effect Floped gene is essential for the derivation of embryonic stem cells in mice

Floped(official name Ooep)is specifically and abundantly expressed in mouse oocytes and embryonic stem cells(ESCs).Depletion of Floped from oocytes leads to early embryonic arrest at the 2-cell stage.Although crucial in cleavage stage development,its roles in early embryos as well as in ESCs remain completely unknown.Here,we compared the efficiency of mouse ESC derivation from inner cell mass(ICM)with and without Floped to study its possible roles in mESCs.Derivation rates of mESC from wild-type,heterozygous,and homozygous blastocysts were 33.3%,21.43%,and 3.85%,respectively,indicating that Floped-/-blastocysts had significantly decreased derivation rates.Respective outgrowth appearing rate five days after blastocyst attachment were 83.3%,85.7%,and 15.4%.Morphologically,the outgrowth of ICM from Floped-/-blastocysts appeared severely death three to five days after blastocyst attachment,and the respective derived stem cells showed long-term instability with long-standing epithelial-like colonies.This result suggests a possible role of Floped in the course of ICM-ESCs transition.

Cloning,Expression and Bioinformatic Analysis of Dorsal Gene in Delia antique

[ Objective ] The aim was to explore the relationship between the Dorsal and diapause development of Delia araiqua. [ Method ] The full-length cDNA of Dorsal in D. antiqua was cloned through RACE. The similarity among deduced amino acid sequence of Dorsal cDNA and the Dorsals of other 14 insect species were compared, and the phylogenetic analysis of these Dorsals was conducted. The expression level of Dorsal gene in winter-, summer- and non-diapausing pupae was analyzed. [ Result] The full-length of Dorsal cDNA sequence, 2 412 bp, was obtained with ORF 1974 bp, which coded 657 amino acids with predicted Mw 72.9 kDa and PI 8.5. The result of similarity comparison indicated that the DaDorsal was most related to those of Drosophlia melanogaster and Drosophlia pseudoobscura. The result of semi-quantitative RT-PCR analysis showed the expression level of Dorsal gene increased in characteristic duration of winter-, summer- and non- diapansing pupae, especially at the late diapanse, which might imply its relationship to D. antiqua diapause and development. [ Conclusion] The study lays the foundation for further study on gene function of Dorsal in insect diapause and development.

Cloning of Immune System-related Gene ApTOLL1 and Its Expression in Nosema-infected Antheraea pernyi

[ Objective ] This study aimed to identify the immune system-related TOLL-like receptor family gene of Aherea pernyi, to lay the foundation for further investigating the immune mechanism of Antherea perny/. [ Method] Immune system-related TOLL-like receptor family gene of Antherea perny/was cloned for se- quencing and phylogenetic analysis ; in addition, expression variations of TOLL-like receptor family gene in Antherea pernyi infected with Nosema pernyi and Nosema bombycis were detected to analyze the differences in immunological reactivity of Antherea pernyi to Nosema infection. [ Result] Based on cDNA cloning and sequen- cing, an immune system-related gene fragment was isolated from Antherea pernyi, which is the most homologous to the ToUl gene in Toll signaling pathway of Bom- byx mot/according to the sequencing result and phylogenetic analysis, which is named ApTolll gene. Subsequently, Antherea pernyi pupae were injected respective- ly with Nosema bombycis and Nosema pernyi, fluorescent real-time quantitative PCR analysis showed that ApTolll was abundantly expressed in Antherea pernyi pupa 2 h after Nosema bombycis injection, which began to express in Antherea pernyi pupa 11 h after Nosema pernyi injection, indicating that the immune response time induced by various Nosema strains varies in Toll signaling pathway of Antherea perny/. [ Conclusion ] ApTolll gene of Antherea perny/was first cloned in this study, which provides reference for further investigating the immune mechanism of Antherea pernyi.

Maternal-effect Floped gene is essential for the derivation of embryonic stem cells in mice

Floped （official name Ooep） is specifically and abundantly expressed in mouse oocytes and embryonic stem cells （ESCs）. Depletion of Floped from oocytes leads to early embryonic arrest at the 2-cell stage. Although crucial in cleavage stage development, its roles in early embryos as well as in ESCs remain completely unknown. Here, we compared the efficiency of mouse ESC derivation from inner cell mass （ICM） with and without Floped to study its possible roles in mESCs. Derivation rates of mESC from wild-type, heterozygous, and homozygous blastocysts were 33.3%, 21.43%, and 3.85%, respectively, indicating that Floped-/- blastocysts had significantly decreased derivation rates. Respective outgrowth appearing rate five days after blastocyst attachment were 83.3%, 85.7%, and 15.4%. Morphologically, the outgrowth of ICM from Floped-/- blastocysts appeared severely death three to five days after blastocyst attachment, and the respective derived stem cells showed long-term instability with long-standing epithelial-like colonies. This result suggests a possible role of Floped in the course of ICM-ESCs transition.

Hypoxia Inhibits Proliferation of Human Dermal Lymphatic Endothelial Cells via Downregulation of Carcinoembryonic Antigen-related Cell Adhesion Molecule 1 Expression

Objective:Lymphatic endothelial cell(LEC)proliferation is essential for lymphangiogenesis.Hypoxia induces lymphangiogenesis,but it directly inhibits LEC proliferation and the underlying mechanisms have not been fully understood.The aim of this study was to investigate the role of carcinoembryonic antigen-related cell adhesion molecule 1(CEACAM1)in hypoxia-repressed LEC proliferation.Methods:Human dermal lymphatic endothelial cells(HDLECs)were cultured under normoxic or hypoxic conditions,and cell proliferation was determined using MTT or CCK-8 assays.CEACAM1 expression was silenced by siRNA transfection.Activation of mitogen-activated protein kinases(MAPKs)was examined by Western blotting and blocked by specific inhibitors.Results:Under hypoxia,HDLECs proliferation was suppressed and CEACAM1 expression was downregulated.Silence of CEACAM1 in normoxia inhibited HDLECs proliferation and did not further decrease proliferation in HDLECs in response to hypoxia,suggesting that CEACAM1 may mediate hypoxia-induced inhibition of HDLECs proliferation.In addition,silence of CEACAM1 increased phosphorylation of MAPK molecules:extracellular signal-regulated kinase(ERK),p38 MAPK and Jun N-terminal kinase(JNK)in HDLECs.However,only inhibition of the JNK pathway rescued the reduction of HDLEC proliferation induced by CEACAM1 silence.Conclusion:Our results suggested that hypoxia downregulates CEACAM1 expression by activation of the JNK pathway,leading to inhibition of HDLEC proliferation.These findings may help to understand the mechanisms of LEC-specific response to hypoxia and develop novel therapies for pathological lymphangiogenesis.

From beasts to bytes:Revolutionizing zoological research with artificial intelligence

Since the late 2010s,Artificial Intelligence(AI)including machine learning,boosted through deep learning,has boomed as a vital tool to leverage computer vision,natural language processing and speech recognition in revolutionizing zoological research.This review provides an overview of the primary tasks,core models,datasets,and applications of AI in zoological research,including animal classification,resource conservation,behavior,development,genetics and evolution,breeding and health,disease models,and paleontology.Additionally,we explore the challenges and future directions of integrating AI into this field.Based on numerous case studies,this review outlines various avenues for incorporating AI into zoological research and underscores its potential to enhance our understanding of the intricate relationships that exist within the animal kingdom.As we build a bridge between beast and byte realms,this review serves as a resource for envisioning novel AI applications in zoological research that have not yet been explored.

Primary cilia in corneal development and disease

As a transparent avascular tissue located at the front of the eyeball, the cornea is an important barrier to external damage. Both epithelial and endothelial cells of the cornea harbor primary cilia, which sense changes in the external environment and regulate intracellular signaling pathways. Accumulating evidence suggests that the primary cilium regulates corneal development in several ways, including participation in corneal epithelial stratification and maintenance of corneal endothelial cell morphology.In addition, the primary cilium has been implicated in the pathogenesis of several corneal diseases. In this review, we discuss recent findings that demonstrate the critical role of the primary cilium in corneal development. We also discuss the link between ciliary dysfunction and corneal diseases, which suggests that the primary cilium could be targeted to treat these diseases.

Peripheral and central control of obesity by primary cilia

Primary cilia are hair-like structures that protrude from the cell surface.They are capable of sensing external cues and conveying a vast array of signals into cells to regulate a variety of physiological activities.Mutations in cilium-associated genes are linked to a group of diseases with overlapping clinical manifestations,collectively known as ciliopathies.A significant proportion of human ciliopathy cases are accompanied by metabolic disorders such as obesity and type 2 diabetes.Nevertheless,the mechanisms through which dysfunction of primary cilia contributes to obesity are complex.In this review,we present an overview of primary cilia and highlight obesity-related ciliopathies.We also discuss the potential role of primary cilia in peripheral organs,with a focus on adipose tissues.In addition,we emphasize the significance of primary cilia in the central regulation of obesity,especially the involvement of ciliary signaling in the hypothalamic control of feeding behavior.This review therefore proposes a framework of both peripheral and central regulation of obesity by primary cilia,which may benefit further exploration of the ciliary role in metabolic regulation.

Decreased expression of m6A demethylase ALKBH5 in decidua contributes to preeclampsia via m6A-CORIN-HuR pathway

Preeclampsia(PE)is a pregnancy-specific hypertensive disorder which poses a severe threat to maternal and fetal health.1 Defective decidualization may contribute to PE.^(2) N6-methyladenosine(m6A)is associated with various diseases.The regulatory mechanism of m6A in PE is not well established to date.We aimed to identify differentially expressed m6A and explore its regulatory role in the pathogenesis of PE.

Altered Neuronal Activity in the Central Nucleus of the Amygdala Induced by Restraint Water-Immersion Stress in Rats

Restraint water-immersion stress(RWIS), a compound stress model, has been widely used to induce acute gastric ulceration in rats. A wealth of evidence suggests that the central nucleus of the amygdala(CEA) is a focal region for mediating the biological response to stress. Different stressors induce distinct alterations of neuronal activity in the CEA; however, few studies have reported the characteristics of CEA neuronal activity induced by RWIS. Therefore, we explored this issue using immunohistochemistry and in vivo extracellular single-unit recording. Our results showed that RWIS and restraint stress(RS) differentially changed the c-Fos expression and firing properties of neurons in the medial CEA. In addition,RWIS, but not RS, induced the activation of corticotropinreleasing hormone neurons in the CEA. These findings suggested that specific neuronal activation in the CEA is involved in the formation of RWIS-induced gastric ulcers.This study also provides a possible theoretical explanation for the different gastric dysfunctions induced by different stressors.

Comparative analyses of simple sequence repeats(SSRs)in 23 mosquito species genomes:Identification,characterization and distribution(Diptera:Culicidae)

Simple sequence repeats (SSRs) exist in both eukaryotic and prokaryotic genomes and are the most popular genetic markers, but the SSRs of mosquito genomes are still not well understood. In this study, we identified and analyzed the SSRs in 23 mosquito species using Drosophila melanogaster as reference at the whole-genome level. The results show that SSR numbers (33 076-560 175/genome) and genome sizes (574.57-1342.21 Mb) are significantly positively correlated (R~= 0.8992, P < 0.01), but the correlation in individual species varies in these mosquito species. In six types of SSR, mono- to trinucleotide SSRs are dominant with cumulative percentages of 95.14%-99.00% and densities of 195.65/Mb-787.51/Mb, whereas tetra- to hexanucleotide SSRs are rare with 1.12%-4.22% and 3.76/Mb-40.23/Mb. The (A/T)n,(AC/GT)n and (AGC/GCT)n are the most frequent motifs in mononucleotide, dinucleotide and trinucleotide SSRs, respectively, and the motif frequencies of tetra- to hexanucleotide SSRs appear to be species-specific. The 10-20 bp length of SSRs are dominant with the number of 11() 561 ± 93 482 and the frequency of 87.25%± 5.73% on average, and the number and frequency decline with the increase oflength. Most SSRs(83.34%± 7.72%) are located in intergenic regions, followed by intron regions (11.59%± 5.59%), exon regions (3.74%± 1.95%), and untranslated regions (1.32%± 1.39%). The mono-, di- and trinucleotide SSRs are the main SSRs in both gene regions (98.55%± 0.85%) and exon regions (99.27%± 0.52%). An average of 42.52% of total genes contains SSRs, and the preference for SSR occurrenee in different gene subcategories are species-specific. The study provides useful insights into the SSR diversity, characteristics and distribution in 23 mosquito species of genomes.

The organization,regulation,and biological functions of the synaptonemal complex

The synaptonemal complex(SC)is a meiosis-specific proteinaceous macromolecular structure that assembles between paired homologous chromosomes during meiosis in various eukaryotes.The SC has a highly conserved ultrastructure and plays critical roles in controlling multiple steps in meiotic recombination and crossover formation,ensuring accurate meiotic chromosome segregation.Recent studies in different organisms,facilitated by advances in super-resolution microscopy,have provided insights into the macromolecular structure of the SC,including the internal organization of the meiotic chromosome axis and SC central region,the regulatory pathways that control SC assembly and dynamics,and the biological functions exerted by the SC and its substructures.This review summarizes recent discoveries about how the SC is organized and regulated that help to explain the biological functions associated with this meiosis-specific structure.

Identification and characterization of a gene encoding a UBX domain-containing protein in the migratory locust, Locusta migratoria manilensis

Ubiquitin regulatory X （UBX） domain-containing proteins are believed to func- tion as cofactors for p97/CDC48, an adenosine triphosphatase shown to be involved in mul- tiple cellular processes. In the present study, a full-length complementary DNA （cDNA） of UBX domain-containing gene, termed LmUBX1, was cloned from Locusta migratoria manilensis and characterized, using random amplification of cDNA ends polymerase chain reaction （RACE PCR）, sequence analysis and quantitative real-time PCR. LmUBX1, 1 600 bp in length, is predicted to encode a 446-amino acid protein with a predicted molec- ular weight of 51.18 kDa that contains a central PUB domain and a carboxy-terminal UBX domain. Homology analysis revealed that LmUBX1 has higher similarity to the known UBX domain-containing proteins from insects than from other species. Moreover, based on sequence characteristics and phylogenetic relationships, it is suggested that LmUBX1 can be classified into the UBXD1 subfamily. Expression analysis founded that LmUBX1 exhibited significant expression variations at different developmental stages and in differ- ent tissues, suggesting that the expression of LmUBX1 was highly regulated. Interestingly, its messenger RNA transcript was more abundant in ovary and testis than in other tissues examined, suggesting that it may have more important roles in the reproductive system. In addition, LmUBX1 was differentially expressed in gregarious and solitary locusts and was significantly up-regulated in third and fifth instars of gregarious locusts, implying that LmUBX1 was also likely involved in the phase polyphenisms in L. migratoria manilen- sis. To our knowledge, this is the first report of cloning of a full-length cDNA of UBX domain-containing gene from L. migratoria manilensis.

Ecotoxicological effects of waterborne PFOS exposure on swimming performance and energy expenditure in juvenile goldfish (Carassius auratus)

The potential risks of perfluorooctane sulfonate （PFOS） are of increasing ecological concern. Swimming performance is linked to the fitness and health of fish. However, the impacts of PFOS on swimming performance remain largely unknown. We investigated the ecotoxicological effects of acute exposure to PFOS on the swimming performance and energy expenditure of juvenile goldfish （Carassius auratus）. The fish were exposed to a range of PFOS concentrations （0, 0.5, 2, 8 and 32 mg/L） for 48 hr. The spontaneous swimming activity, fast-start swimming performance, critical swimming speed （Ucrit） and active metabolic rate （AMR） of the goldfish were examined after exposure to PFOS. PFOS exposure resulted in remarkable effects on spontaneous activity. Motion distance was reduced, and the proportion of motionless time increased with increasing concentrations of PFOS. However, no significant alterations in the fast-start performance-related kinematic parameters, such as latency time, maximum linear velocity, maximum linear acceleration or escape distance during the first 120 msec after stimulus, were observed after PFOS exposure. Unexpectedly, although PFOS exposure had marked influences on the swimming oxygen consumption rates and AMR of goldfish, the U crit of the goldfish was not significantly affected by PFOS. This may result in a noteworthy increase in the energetic cost of transport. The overall results indicate that, in contrast to spontaneous activity, underlying swimming capabilities are maintained in goldfish after short-term exposure to PFOS, but energy expenditure during the process of swimming is dramatically aggravated.

Physicochemical properties, molecular structure, antioxidant activity, and biological function of extracellular melanin from Ascosphaera apis

Ascosphaera apis spores containing a dark-colored pigment infect honeybee larvae,resulting in a large-scale collapse of the bee colony due to chalkbrood disease.However,little is known about the pigment or whether it plays a role in bee infection caused by A.apis.In this study,the pigment was isolated by alkali extraction,acid hydrolysis,and repeated precipitation.Ultraviolet(UV)analysis revealed that the pigment had a color value of 273,a maximum absorption peak at 195 nm,and a high alkaline solubility(7.67%)and acid precipitability.Further chemical structure analysis of the pigment,including elemental composition,Fourier transform infrared(FTIR)spectroscopy,Raman spectroscopy,mass spectrometry,and nuclear magnetic resonance(NMR),proved that it was a eumelanin with a typical indole structure.The molecular formula of melanin is C10H6O4N2,and its molecular weight is 409 Da.Melanin has hydroxyl,carboxyl,amino,and phenolic groups that can potentially chelate to metal ions.Antioxidant function analyses showed that A.apis melanin had a high scavenging activity against superoxide,hydroxyl,and 2,2-diphenyl-1-picrylhydrazyl(DPPH)radicals,and a high reducing ability to Fe3+.Indirect immunofluorescence assay(IFA),scanning electron microscopy(SEM),and transmission electron microscopy(TEM)analyses showed that A.apis melanin was located on the spore wall.The spore wall localization,antioxidant activity,and metal ion chelating properties of fungal melanin have been suggested to contribute to spore pathogenicity.However,further infection experiments showed that melanin-deficient spores did not reduce the mortality of bee larvae,indicating that melanin does not increase the virulence of A.apis spores.This study is the first report on melanin produced by A.apis,providing an important background reference for further study on its role in A.apis.