Aristolochic acid nephropathy (AAN) is a progressive form of kidney disease marked by acute tubular injury and interstitial fibrosis, ultimately leading to end-stage renal disease (ESRD). Despite regulatory restrictions, aristolochic acid (AA) remains a global health threat due to its presence in traditional herbal medicines. While mitochondria-mediated apoptosis is a hallmark of AA-induced tubular epithelial cell (TEC) injury, the upstream molecular mechanisms remain unclear. Here, we identify Z-DNA-binding protein 1 (ZBP1) as a key mediator of AA-induced kidney injury. Using Zbp1 knockout (Zbp1-/-) and Zα domain-mutant (ZαMut) mice, we show that loss of ZBP1 or its Z-form nucleic acid sensing capability protects against AA-induced renal dysfunction, apoptosis, and inflammation. Mechanistically, aristolochic acid I (AAI) induces mitochondrial oxidative stress and release of mitochondrial DNA (mtDNA), which adopts a Z-conformation and is recognized by ZBP1. The ZBP1 binding subsequently promotes RHIM-dependent interaction with RIPK1, culminating in caspase-8 activation and apoptotic cell death. Notably, ZBP1-mediated cell death was abolished by RIPK1 kinase inhibition or mutation, but unaffected by Ripk3 or Mlkl deletion, revealing a mechanism distinct from RIPK3/MLKL-dependent necroptosis. These findings uncover a previously unrecognized ZBP1-RIPK1-caspase-8 signaling axis driving non-canonical apoptosis in AAN and suggest that targeting this pathway may provide a novel therapeutic approach for nephrotoxin-induced kidney injury.

Tubulins undergo various posttranslational modifications (PTMs) that confer diverse functions to microtubules. Tubulin polyglutamylation is dynamically regulated by glutamylation and deglutamylation. AGBL5 is a deglutamylase that specifically removes glutamate at the branching point. Mutations in AGBL5 in humans are associated with retinitis pigmentosa, but the underlying mechanism remains undefined. Here, we generated an Agbl5 knockout mouse that showed tubulin hyperglutamylation in photoreceptors, resulting in progressive retinal degeneration. RNA-seq analysis revealed the altered ciliary function in the Agbl5 knockout. Transmission electron microscopy indicated an impaired inner scaffold in the connecting cilium (CC). Consequently, phototransduction proteins were mislocalized or downregulated in mutant rod and cone photoreceptors. Disk membranes in photoreceptor outer segments were disorganized. Immunofluorescence revealed that the recruitment of IFT88, kinesin-II, and dynein-2 to the CC was affected, implicating defective intraflagellar transport (IFT). Collectively, these findings demonstrate that tubulin glutamylation homeostasis regulated by AGBL5 is critical for photoreceptor survival by maintaining the structural integrity of the CC and normal protein trafficking through IFT.

Microbiota assembly during early ontogeny in teleost fish plays a central role in shaping immune maturation and establishing host-microbe homeostasis, yet the regulatory mechanisms driving microbial succession across key developmental windows remain poorly understood. In this study, Larimichthys crocea was used to delineate microbiota assembly dynamics and the impact of stochastic and deterministic processes. Results indicated that community assembly peaked at day 18 post-hatching (DPH18), coinciding with the highest neutral model fit (R 2=0.71) and migration rate (m=0.88). Alpha (α)-diversity exhibited a hump-shaped pattern, with Comamonas dominance inversely correlating with Vibrio at DPH18. Microbial source tracking indicated that host-associated taxa played a more prominent role than dietary or environmental sources. Transcriptomic profiling revealed pronounced immune modulation during early development. Pro-inflammatory signaling, including IL-17 pathway activation, was elevated prior to DPH18, while anti-inflammatory regulators, such as transforming growth factor beta 2 ( tgfb2), declined over time, consistent with a transient reduction in immune restraint. Immune constraints in dexamethasone-treated zebrafish produced intestinal barrier impairment and microbial dysbiosis, demonstrating functional consequences of compromised early immune regulation. Collectively, these patterns defined DPH3-DPH18 as a critical colonization window in L. crocea, during which reduced immune constraint facilitates niche establishment by early colonizers. This temporally restricted window optimizes microbial resilience and long-term resistance to dysbiosis, providing a mechanistic basis for early-life microbiota-directed strategies in teleost development.

Heterozygous variants in ARR3, encoding cone arrestin, have emerged as a frequent cause of early-onset high myopia with a unique X-linked female-limited inheritance pattern. However, the mechanistic basis for this unusual anti-X-linked pattern is still unclear. Developmental expression profiling in mice demonstrated robust Arr3 expression in the retina from postnatal day 14 onward, with localization confined predominantly to outer segments of cones marked by red/green opsins, including a subset co-labeled with both red/green and blue opsins. Retinal flatmounts from Arr3 mutation knock-in mice and Arr3 knockout rats revealed a mosaic pattern of Arr3 expression in heterozygous individuals. Retinal single-cell RNA sequencing revealed significant shifts in cone subtype proportions in Arr3 +/- rats, with a marked reduction in M/S cones and a corresponding increase in S cones. Among differentially expressed genes, Pde6h was the only transcript altered in M/S cones across both Arr3 +/+ vs. Arr3 +/- and Arr3 -/0 vs. Arr3 +/- comparisons but not in Arr3 +/+ vs. Arr3 -/0 . These findings suggest that heterozygous Arr3 deficiency induces cone mosaicism that may mimic retinal defocus-like signals during phototransduction, potentially driving the development of high myopia under this distinctive inheritance model.

Body shape evolution in vertebrates frequently involves modifications in vertebral number or patterns of vertebral fusion, with distinct lineages displaying divergent trajectories. This study investigated the morphological and genetic basis of body shape variation between Hebao red carp (HB, Cyprinus carpio wuyuanensis) and Yellow River carp (YR, Cyprinus carpio haematopterus). Although both subspecies share an identical vertebral count (35), the compressed morphology of HB was attributable to skeletal anomalies, including vertebral shortening and fusion. Genome-wide association and population genetic analyses were performed on F1 and F2 hybrid cohorts to identify loci associated with this phenotype. A total of 231 selective sweep regions were detected across chromosomes A06, A08, A16, B05, and B06, with a prominent locus on chromosome A08 (15.99-16.39 Mb) strongly correlated with body shape traits. Transcriptomic analysis revealed haplotype-dependent expression of rflna within this interval, implicating rflna in axial skeletal patterning. Functional validation using CRISPR/Cas9-mediated knockout of rfln in zebrafish ( Danio rerio) induced vertebral malformations, including axial shortening, kyphosis, fusion, and a rounded abdominal profile. These results delineate the morphological and molecular framework governing axial remodeling in HB and highlight a conserved regulatory role for rflna in teleost skeletal development.

Arrhythmogenic cardiomyopathy (ACM) confers elevated risk of ventricular arrhythmias and sudden cardiac death, yet limitations in early lesion sampling and model development continue to hinder mechanistic and translational research. Clinical, pathological, and mutational profiles were examined in 24 individuals with ACM harboring PKP2 variants. Among these, a patient carrying the c.1132C>T mutation exhibited the earliest onset and presented both structural cardiac abnormalities and major adverse cardiovascular events. To facilitate disease modeling, the c.1147C>T variant-a previously reported pathogenic substitution located proximal to position c.1132 in PKP2-was selected to enhance the feasibility of generating a porcine model. The BE3 gene editing system was used to induce C>T mutation. Two single guide RNAs targeting the PKP2 gene were designed (sgRNA1 for c.1132C>T and sgRNA2 for c.1147C>T), yielding editing efficiencies of 42.9% and 25.9%, respectively. SgRNA1 was used to generate PKP2 +/- porcine fetal fibroblasts. A total of 14 cloned piglets were produced, including 11 viable and three stillborn PKP2 +/- individuals. By 24 months of age, PKP2 +/- pigs developed premature ventricular contractions and right ventricular dilatation. Histological analysis revealed adipocyte infiltration within the right ventricular wall, and electron microscopy demonstrated reduced desmosomal length and electron density consistent with desmosomal dysfunction. Transcriptomic profiling showed high expression of genes associated with lipid catabolic processes. This study established the first PKP2 +/- porcine model of ACM using BE3-mediated base editing, providing a valuable platform for elucidating early pathogenic mechanisms and evaluating therapeutic interventions.

Trichoplax adhaerens, one of the simplest multicellular organisms belonging to the phylum Placozoa, occupies a basal position within Metazoa and serves as a critical model for investigating early multicellular evolution. Despite its significance, the absence of functional genetic tools has limited research to comparative genomics, ultrastructural characterization, and behavioral observation. Developing methodologies such as transgenesis and gene editing would substantially expand experimental capabilities. However, the absence of sexual reproduction precludes the use of conventional approaches like single-cell microinjection. To overcome these limitations, the present study established a transgenic method for expressing exogenous genes in T. adhaerens. Systematic evaluation of multiple transfection techniques identified electroporation as the most effective strategy for inducing ectopic gene expression in this species. Electroporation parameters were optimized to maximize efficiency while minimizing cytotoxicity. Using this approach, robust expression of green fluorescent protein (GFP) was achieved using various promoters. Ectopic expression of the puromycin acetyltransferase ( PAC) gene conferred increased resistance to puromycin exposure, suggesting a potential strategy for generating stable transgenic lines in the future. This work introduces a reliable framework for ectopic gene expression in T. adhaerens, enabling molecular and functional analyses in one of the earliest-diverging metazoan lineages.