摘要: The yellow boxfish(Ostracion cubicus) exhibits a combination of derived morphological traits specialized for coral reef environments and ancestral characteristics,including a fused dermal plate. Contradictory evolutionary evidence hinders true classification of O. cubicus. To clarify its evolutionary position within Tetraodontiformes, a chromosome-level genome assembly was generated,representing the most contiguous and complete genome to date for this lineage. Notably, O. cubicus possessed the largest genome within the order Tetraodontiformes,primarily due to extensive transposable element expansion. Phylogenetic analysis based on 19 whole genomes and 131 mitochondrial genomes resolved Tetraodontiformes into three major sister groups(Ostraciidae-Molidae, Tetraodontidae, and BalistidaeMonacanthidae). Comparative genomic evidence indicated that O. cubicus diverged early from the common ancestor of modern Tetraodontiformes and retained the highest number of HOX genes among surveyed taxa. Although overall genomic architecture was largely conserved,certain genetic and environmental changes may have contributed to its phenotypic adaptations, including climate cooling during the Miocene-Pliocene Transition, recent DNA and long interspersed nuclear element(LINE)transposon bursts, lineage-specific chromosomal rearrangements, and gene family expansion. Many positively selected genes and rapidly evolving genes were associated with skeletal development, including bmp7,egf7, and bmpr2. Transcriptomic comparisons between carapace and tail skin revealed various candidate genes and pathways related to carapace formation, such as postn, scpp1, and components of the TGF-β signaling pathway. A derived amino acid substitution in eda, coupled with protein structural modeling, suggested potential molecular convergence in dermal plate formation among teleosts. These findings provide novel insights into the genomic and developmental basis of carapace evolution and coral reef-adaptation in O. cubicus, offering a strong case for evolutionary balance between genomic conservation with regulatory innovation to achieve coral reef specialization.

摘要: Vertebrate limbs have undergone profound morphological diversification, enabling adaptations to a broad spectrum of ecological niches. In marine mammals, the evolution of highly specialized flipper-like forelimbs represents a profound structural transformation associated with aquatic habitats. This adaptation has been hypothesized to result,in part, from the inhibition of interphalangeal cell apoptosis during limb development, although the underlying genetic mechanism remains poorly understood. This study investigated the evolutionary dynamics and functional consequences of three key bone morphogenetic protein genes, BMP2, BMP4, and BMP7, which regulate apoptosis in interphalangeal mesenchymal stromal cells during embryonic limb development to ensure proper differentiation of interphalangeal tissues. Comparative genomic analysis revealed significantly accelerated evolution for BMP4 and BMP7 in the cetacean ancestral lineage, with two positively selected sites(V79I and H247R) involved in cetacean-specific amino acid substitutions located in the TGF-β propeptide functional domain in BMP4. In vitro assays confirmed that cetaceanspecific BMP4 mutations significantly disrupted normal cell apoptosis and proliferation and altered the transcription and protein expression of downstream apoptosis-related factors, including cytochrome c(Cyt c), BCL2 associated X, and B-cell lymphoma 2, within the BMP signaling pathway. The significant influence of BMP4 mutations on apoptotic inhibition highlights a potential role in the development of limb bud mesenchymal tissue and the emergence of the flipper forelimb phenotype in cetaceans.

摘要: Pentadactyl limbs represent a conserved morphological feature among tetrapods, with anterior digits considered more important than posterior digits for refined movement.While posterior digit formation is governed by graded expression of the Shh and 5'Hox genes, the regulatory mechanisms underlying anterior digit development,especially digit I(DI), remain poorly defined. This study identified an anterior expression pattern of Zic3 in the limb buds of representative tetrapods, including humans, which exerted an inhibitory effect on skeletal development. Zic3 was highly expressed in the anterior region of limb buds at early developmental stages, with species-specific divergence emerging during later development.Overexpression of Zic3 significantly delayed chondrogenesis and ossification, leading to bone shortening but not loss. Furthermore, RNA sequencing demonstrated that Zic3 down-regulated key genes associated with skeletal development, including Cytl1,Sox9, Ihh, Ptch1, Runx2, and Wnt16. These findings demonstrate that Zic3 acts as a conserved inhibitor of anterior skeletal maturation and contributes to the molecular asymmetry of tetrapod limb development.

摘要: NORHA, a long non-coding RNA(lncRNA), serves as a key inducer of follicular atresia in sows by triggering granulosa cells(GCs) apoptosis. However, its regulation by N6-methyladenosine(m6A)—the most abundant RNA modification—remains unresolved. This study identified NORHA as a functional target of the m6A reader HNRNPA2B1 in sow GCs(sGCs). Transcriptome-wide mapping of RNA modification sites revealed extensive m6A enrichment on NORHA, with HNRNPA2B1 binding directly to the transcript and enhancing its stability via modification of multiple m6A sites, including A261, A441,and A919. HNRNPA2B1 suppressed 17β-estradiol(E2)biosynthesis and promoted sGC apoptosis by activating the NORHA-FoxO1 axis. FoxO1 subsequently repressed expression of cytochrome P450 family 19 subfamily A member 1(CYP19A1), which encodes the enzyme essential for E2 biosynthesis. Additionally, HNRNPA2B1functioned as a critical mediator of METTL3-dependent m6A modification, modulating NORHA expression and activity in sGCs. This study highlights an important m6Adependent regulatory mechanism governing NORHA expression in sGCs.

摘要: Automated behavior monitoring of macaques offers transformative potential for advancing biomedical research and animal welfare. However, reliably identifying individual macaques in group environments remains a significant challenge. This study introduces ACE-YOLOX, a lightweight facial recognition model tailored for captive macaques. ACE-YOLOX incorporates Efficient Channel Attention(ECA), Complete Intersection over Union loss(CIoU), and Adaptive Spatial Feature Fusion(ASFF) into the YOLOX framework, enhancing prediction accuracy while reducing computational complexity. These integrated approaches enable effective multiscale feature extraction.Using a dataset comprising 179 400 labeled facial images from 1 196 macaques, ACE-YOLOX surpassed the performance of classical object detection models,demonstrating superior accuracy and real-time processing capabilities. An Android application was also developed to deploy ACE-YOLOX on smartphones, enabling on-device,real-time macaque recognition. Our experimental results highlight the potential of ACE-YOLOX as a non-invasive identification tool, offering an important foundation for future studies in macaque facial expression recognition,cognitive psychology, and social behavior.

摘要: N6-methyladenosine（m⁶A） modification of mRNA is a critical post-transcriptional regulatory mechanism that modulates mRNA metabolism and neuronal function. The m⁶A reader YTHDF1 has been shown to enhance the translational efficiency of m⁶A-modified mRNAs in the brain and is essential for learning and memory. However, its role in the mature retina remains unclear. Herein, we report a novel role of Ythdf1 in the maintenance of retinal function using a genetic knockout model. Loss of Ythdf1 resulted in impaired scotopic electroretinogram（ERG） responses and progressive retinal degeneration. Detailed analyses of rod photoreceptors confirmed substantial degenerative changes in the absence of ciliary defects. Single-cell RNA sequencing revealed comprehensive molecular alterations across all retinal cell types in Ythdf1-deficient retinas.Integrative analysis of methylated RNA immunoprecipitation（MeRIP） sequencing and RIP sequencing identified Tulp1 and Dhx38, two inheritable retinal degeneration disease-associated gene homologs,as direct targets of YTHDF1 in the retina. Specifically,YTHDF1 recognized and bound m⁶A-modified Tulp1 and Dhx38 mRNA at the coding sequence（CDS）, enhancing their translational efficiency without altering mRNA levels.Collectively, these findings highlight the essential role of YTHDF1 in preserving visual function and reveal a novel regulatory mechanism of m⁶A reader proteins in retinal degeneration, identifying potential therapeutic targets for severe retinopathies.

摘要: Cytoplasmic accumulation of TDP-43 is a pathological hallmark of amyotrophic lateral sclerosis(ALS) and other neurodegenerative diseases. While current studies have primarily focused on gene regulation mediated by full-length nuclear TDP-43, the potential effects of cytoplasmic TDP-43 fragments remain less explored. Our previous findings demonstrated that primate-specific cleavage of TDP-43 contributes to its cytoplasmic localization,prompting further investigation into its pathological effects.In the cynomolgus monkey brain, we observed that mutant or truncated TDP-43 was transported onto the ribosome organelle. Ribosome-associated transcriptomic analysis revealed dysregulation of apoptosis-and lysosome-related genes, indicating that cytoplasmic TDP-43 induces neurotoxicity by binding to ribosomes and disrupting mRNA expression. These findings provide mechanistic insights into the gain-of-function effects of pathological TDP-43.

摘要: Congenital disorders of glycosylation（CDG） are a cluster of monogenic disorders resulting from defects in glycosylation. FCSK encodes fucokinase, an enzyme that catalyzes the phosphorylation of L-fucose to generate fucose-1-phosphate, an important step in fucosylation.Mutations in FCSK lead to CDG with an autosomal recessive inheritance pattern, primarily manifesting as developmental delay, hypotonia, and brain abnormalities.However, no fcsk mutant animal models have yet been established. This study constructed the first fcsk knockout(fcsk^-/-) zebrafish model using CRISPR/Cas9 technology.Notably, fcsk^-/- zebrafish exhibited impaired growth,characterized by delayed epiboly and DNA accumulation during early embryonic development, as well as brain atrophy in adulthood. Larval-stage fcsk^-/- zebrafish displayed locomotor deficits and increased susceptibility to pentylenetetrazole-induced seizures. In adulthood, fcsk^-/- zebrafish showed neurodevelopmental abnormalities,including increased anxiety, decreased aggression,reduced social preference, and impaired memory.Additionally, total protein fucosylation was markedly reduced in fcsk^-/- zebrafish, accompanied by decreased expression of pofut2, which encodes protein Ofucosyltransferase 2, an enzyme involved in the fucosylation salvage pathway. Apoptotic activity was elevated in the midbrain-hindbrain boundary（MHB） of fcsk^-/- zebrafish. Supplementation with GDP-L-fucose or the human FCSK gene restored developmental defects and total protein fucosylation in fcsk^-/- zebrafish. RNA sequencing revealed dysregulated gene expression associated with glycosylation, apoptosis, and neurodegenerative diseases. These findings suggest that fcsk^-/- zebrafish exhibit neurodevelopmental disorders,providing the first fcsk gene knockout animal model and offering a platform for investigating the molecular underpinnings of the disease and facilitating drug screening efforts.

摘要: The kidney is essential for maintaining fluid, electrolyte, and metabolite homeostasis, and for regulating blood pressure. The pig serves as a valuable biomedical model for human renal physiology, offering insights across different physiological states. In this study, single-cell RNA sequencing was used to profile 138 469 cells from 12 pig kidney samples collected during the embryonic(E), fattening(F), and pregnancy(P) periods, identifying 29 cell types. Proximal tubule(PT) cells exhibited elevated expression of metabolism-related transcription factors(TFs), including GPD1, ACAA1, and AGMAT, with validation across multiple individuals, periods, and species.Fluorescence homologous double-labeling of paraffin sections further confirmed the expression of ACAA1 and AGMAT in PT cells. Comparative analysis of pig and human kidneys revealed a high degree of similarity among corresponding cell types. Analysis of cell-type heterogeneity highlighted the diversity of thick ascending limb(TAL) cells, identifying a TAL subpopulation related to immune function. Additionally, the functional heterogeneity of kidney-resident macrophages(KRM) was explored across different anatomical sites. In the renal medulla,KRM were implicated in phagocytosis and leukocyte activation, whereas in the renal pelvis, they functioned as ligands, recruiting neutrophils with bactericidal activity to the renal pelvis to combat urinary tract infections.

摘要: Haemaphysalis longicornis serves as the primary tick vector for severe fever with thrombocytopenia syndrome virus(SFTSV), the etiological agent responsible for severe fever with thrombocytopenia syndrome(SFTS).Understanding alterations in tick salivary gland microbiota during SFTSV transmission to vertebrate hosts is essential for developing novel control strategies. However, microbial shifts in tick salivary glands during pathogen transmission to hosts have not been reported for any tick-borne pathogens. In this study, SFTSV transmission from H. longicornis to vertebrate hosts was confirmed using a tickrabbit transmission model. Salivary gland microbiota profiling via 16S rRNA gene sequencing identified significant changes in bacterial composition associated with viral transmission. The relative abundance of three genera(Serratia, Bifidobacterium, and Akkermansia)increased, whereas five genera(Flavobacterium,Staphylococcus, Enhydrobacter, Massilia, and Stenotrophomonas) decreased. Correlation network analysis revealed a negative association between Akkermansia and Flavobacterium. These findings demonstrated that SFTSV transmission alters the salivary gland microbiota of H. longicornis, providing insights for future functional studies and the development of targeted strategies for SFTS control.