Zoological Research最新文献

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, HNRNPA2B1 functioned as a critical mediator of METTL3-dependent m6A modification, modulating NORHA expression and activity in sGCs. This study highlights an important m6A-dependent regulatory mechanism governing NORHA expression in sGCs.

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 cetacean-specific 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.

Photodynamic therapy (PDT) is an emerging minimally invasive therapeutic modality that relies on the activation of a photosensitizing agent by light of a specific wavelength in the presence of molecular oxygen, leading to the generation of reactive oxygen species (ROS). This mechanism facilitates selective cytotoxic effects within pathological tissues and has demonstrated therapeutic potential across diverse disease contexts. However, the broader clinical applications remain limited by photosensitizer selectivity, shallow light penetration, and the risk of off-target cytotoxicity. Recent advancements in PDT have focused on the development of next-generation photosensitizers, the integration of nanotechnology for enhanced delivery and targeting, and the strategic combination of PDT with complementary therapeutic approaches. Experimental animal models play a crucial role in validating the efficacy and safety of PDT, optimizing its therapeutic parameters, and determining its mechanisms of action. This review provides a comprehensive overview of PDT applications in various disease models, including oncological, infectious, and nonconventional indications. Special emphasis is placed on the importance of large animal models in PDT research, such as rabbits, pigs, dogs, and non-human primates, which provide experimental platforms that more closely resemble human physiological and pathological states. The use of these models for understanding the mechanisms of PDT, optimizing therapeutic regimens, and evaluating clinical outcomes is also discussed. This review aims to inform future directions in PDT research and emphasizes the importance of selecting appropriate preclinical animal models to facilitate successful clinical translation.

Ectothermic organisms may expand their thermal tolerance by producing multiple protein isoforms with differing thermal sensitivities. While such isoforms commonly originate from allelic variation at a single locus (allozymes) or from gene duplication that gives rise to paralogs with distinct thermal responses, this study investigated mRNA editing as an alternative, post-transcriptional mechanism for generating mRNA variants. Cytosolic malate dehydrogenase (cMDH) was examined in foot tissue of two congeners of the marine mussel genus Mytilus, which occupy different thermal environments. Multiple editing events were detected within the mRNA coding region in both species. Editing sites were species-specific, with no shared positions identified. In M. coruscus, editing occurred at 117, 123, 135, 190, 195, 204, 279, and 444, while in M. galloprovincialis, editing was detected at 216 and 597. Each species exhibited multiple edited mRNA variants, and these isoforms were associated with differential protein expression. These findings suggest that mRNA editing may contribute an additional layer of molecular variation. The generation of diverse mRNA isoforms from a single DNA coding sequence may enhance enzymatic flexibility across temperature ranges, supporting eurythermal physiological performance and mitigating thermal stress. Moreover, the presence of multiple edited transcripts within individual organisms raises important caveats about the limitations of approaches that deduce amino acid sequences or estimate adaptive variation solely from genomic data.

Ischemic heart disease (IHD) remains a leading contributor to cardiovascular disease (CVD) worldwide. Despite advances in diagnostic and therapeutic approaches, translational research demands robust large animal models to bridge the gap between experimental interventions and clinical application. Among these, porcine models have gained prominence due to their anatomical, physiological, immunological, and genomic similarities to humans. This review provides a comprehensive overview of current methodologies for establishing porcine IHD models, critically assesses emerging rehabilitative strategies, and outlines innovative therapeutic technologies, with the goal of guiding model selection and fostering the development of novel treatment strategies.

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.

Chronic pancreatitis (CP) is a progressive and irreversible fibroinflammatory disease that markedly increases susceptibility to pancreatic cancer and remains without effective targeted therapies. Among the genetic contributors to CP, the carboxypeptidase A1 p.Ser282Pro ( CPA1 ^S282P ) variant has been proposed to promote disease through misfolding-induced endoplasmic reticulum stress (ERS), although the broader pathogenic landscape remains incompletely defined. This study generated a rabbit model mimicking the human CPA1 ^S282P mutation using the SpRY-ABE-8.17 system. Homozygous CPA1 ^S282P rabbits exhibited characteristic human CP phenotypes following alcohol induction, including visceral pain, elevated serum lipase and amylase, inflammatory cell infiltration, and extensive pancreatic fibrosis. Biochemical analyses confirmed that the p.S282P mutation induced CPA1 misfolding and elevated the expression of ERS markers GRP78 and CHOP in both transfected HEK293T cells and homozygous mutant rabbits. Notably, the CPA1 ^S282P mutation markedly disrupted intra-pancreatic lipid homeostasis, contributing to the development of CP in mutant rabbits. This study successfully established the first rabbit model of CP that accurately recapitulates CP caused by a defined human point mutation. Additionally, this study provides insights into a previously unrecognized link between CPA1 and intra-pancreatic lipid metabolism, offering a foundation for identifying novel therapeutic targets for human CP.

Flight feathers represent a hallmark innovation of avian evolution. Recent comparative genomic analyses identified a 284 bp avian-specific highly conserved element (ASHCE) located within the eighth intron of the SIM bHLH transcription factor 1 ( Sim1) gene, postulated to act as a cis-regulatory element governing flight feather morphogenesis. To investigate its functional significance, genome-edited (GE) primordial germ cell (PGC) lines carrying targeted ASHCE deletions were generated using CRISPR/Cas9-mediated editing, with germline chimeric males subsequently mated with wild-type (WT) hens to obtain GE progeny. The resulting GE chickens harbored 257-260 bp deletions, excising approximately half of the Sim1-ASHCE sequence. Reverse transcription-quantitative real-time polymerase chain reaction (RT-qPCR) analysis showed an average 0.32-fold reduction in Sim1 expression in the forelimbs of GE embryos at day 8 (E8) compared to WT counterparts. Despite this, GE chickens developed structurally normal flight and tail feathers. In situ hybridization localized Sim1 expression to the posterior mesenchyme surrounding flight feather buds in E8 WT embryos, but not within the buds themselves. These results suggest that partial deletion of Sim1-ASHCE, despite diminishing Sim1 expression, does not disrupt flight feather formation. The excised region appears to possess enhancer activity toward Sim1 but is dispensable for flight feather development. Complete ablation of the ASHCE will be necessary to fully resolve the regulatory role of Sim1 in avian feather morphogenesis.

The immunomodulatory function of estrogen within the ovary remains a subject of ongoing debate, and the neonatal ovarian immune microenvironment, particularly its modulation by estrogen, has not been comprehensively characterized. In this study, the effects of 17β-estradiol (E ₂), a key regulator of immune function, were investigated using single-cell transcriptomic profiling of C57BL/6J neonatal mouse ovaries after E ₂ treatment. Results revealed dynamic alterations in the proportion of immune cell types after E ₂ treatment, accompanied by changes in cytokine and chemokine expression. Detailed analyses of gene expression, cell states, and developmental trajectories across distinct cell types indicated that E ₂ treatment influenced cell differentiation and development. Notably, E ₂ treatment reduced the abundance of macrophages and promoted a phenotypic transition from M1 to M2 macrophages. These findings demonstrate that the neonatal mouse ovarian immune microenvironment is sensitive to estrogenic modulation, which governs both the distribution and functional specialization of resident immune cells, offering novel mechanistic insights into the immunomodulatory roles of estrogen across various immune cell types.