The Min pig, a representative northern Chinese indigenous breed, carries a unique ancestral background shaped by the historical phylogeography of Northeast Asia. This study aimed to dissect the population structure, temporal genetic divergence, and ancestral composition of Min pigs, trace their evolutionary origin, and identify trait-linked functional genes, providing information regarding their evolutionary history and conservation. We analyzed 61 Min pigs sampled across nearly 20 years and 701 reference pigs comprising other Chinese indigenous breeds, Western commercial lines, and Chinese wild boars, using PCA, NJ phylogenetic analysis, Admixture, TreeMix, D-statistic, f4-ratio, and combined selection signature scans (sliding-window FST, XP-EHH, and π-ratio). Clear genetic stratification was observed among Min pig subpopulations, reflecting long-term divergence under natural and artificial selection. PCA and Admixture (K = 2-4) separated East Asian indigenous and Western ancestral components, verifying an admixed Northeast Asian origin with a dominant ancient East Asian component and a Western component. Compared with early-2000s Min pigs, contemporary individuals are genetically closer to Western breeds and exhibit a more scattered structure due to shifted ancestral component proportions, further confirmed by D-statistic and f4-ratio values. We identified 321 differentiated SNP loci based on the Animal QTL database, corresponding to core candidate genes (AKT3, ACACA, MAP3K5, FGFR4, C3, and SERPINC1) enriched for meat quality, growth, reproduction, immunity, energy metabolism, and MAPK/PI3K-Akt/AMPK pathways. This study reveals Min pigs' admixed origin and temporal divergence, clarifying their Northeast Asian evolution and providing molecular markers for genetic monitoring and conservation.

As an important reproductive trait, teat number plays a vital role in the pork industry, which is related to the ability of sows to host more piglets. However, the genetic mechanism of the teat number in pigs is still not very clear. We collected phenotype data, including the total teat number (TTN), left teat number (LTN), and right teat number (RTN), from 300 Jishen Black (JSB) sows and performed genotyping using a porcine SNP 50K panel. Most JSB sows exhibited 14, 15, or 16 TTN (259, 86%). Only 41 pigs (14%) had TTN values outside this range. Moreover, the LTN and TTN exhibited approximate frequency distribution in the studied population. Seven was the most common LTN and RTN in JSB sows, accounting for 181 (60%) and 173 (57%), respectively. Additionally, these traits exhibited low-to-medium heritability (0.14-0.22). Using the BLINK and FarmCPU models, three, six, and three key loci were identified in the GWAS of TTN, LTN, and RTN, respectively. Additionally, five key genes were detected to play vital roles in teat number traits, including ME1, SCN8A, EVC, UBC, and PDE4D. Our research provided efficient molecular markers and new insights into further improvement and understanding for teat number traits in Jishen Black pigs.

BACKGROUND:Heat stress can induce skeletal muscle injury. Typical characteristics of heat-exposed muscle tissues include apoptosis, oxidative stress and autophagy. The understanding of molecular mechanisms underlying heat stress-induced muscle injury is limited, especially at the single-cell transcription level. METHODS:We collected skeletal muscles from 12-week-old female C57BL/6J mice in control (NC) and four heat stress groups. The experimental scheme comprised five groups: NC (25.5°C ± 0.5°C), HS0 (after ~3-h heat exposure, 41.5°C ± 0.5°C), HS8, HS16 and HS24 group (recovery at 25.5°C ± 0.5°C for 8, 16 and 24 h, respectively). Skeletal muscles were subjected to HE staining (n = 6), TUNEL staining (n = 3) and transmission electron microscopy (n = 3). Transcripts were measured at the tissue (n = 5 or 6) and single-nucleus levels (n = 2). RESULTS:Histologically, myofibrillar structure deformation, mitochondrial swelling and fusion, intramuscular triglyceride accumulation and autophagy occurred in the muscles subjected to heat stress. At the tissue level, a gene cluster associated with the response to heat exhibited an increasing trend of transcription in the HS0 versus NC groups, and the levels decreased to those in the NC group after 16 h. At the single-cell level, 134 320 high-quality myonuclei were collected from the muscles and annotated as seven cell types, including myonuclei, muscle stem cells (MuSCs) and immune cells. We identified the larger number of differentially expressed genes in the myonuclei. After heat stress, new cell clusters appeared in type IIa/IIx (HS8 group) and IIb (HS0 group) myonuclei but not in type I myonuclei. Generally, immediate early genes, highly expressed genes and transcription factor regulons identified in new cell clusters induced by heat were related to responses to heat, heat shock, oxidative stress and antioxidative stress. To repair the injured muscles, MuSCs highly expressed the development-related genes, such as Atp2a1, Ckm, Myh1, Aldoa, Pde4d and Pdlim5. Analysis of cell-cell communication showed that Dag1 and Egf signals associated with myonuclei were the underlying pathways that participated in repair tissues. CONCLUSIONS:We constructed the largest transcriptomic dataset, to date, for heat-exposed skeletal muscles. At tissue resolution, the response of muscles to heat stress was eliminated after a recovery period of 16 h. At single-nucleus resolution, the myofibre was the most heat-sensitive cell type, and type IIb myonuclei were the most heat-sensitive subtype of myonuclei. To our knowledge, this is the first study to reveal the molecular mechanisms underlying heat-induced muscle injury and repair at the single-cell transcriptional level.

A primary anti-predator adaptation is crypsis,in which animals reduce their visibility to predators,often through camouflage,to lower the risk of predation.Alternatively,prey species may employ strategies such as aposematism,which advertises unpalatability,or mimicry,which involves imitating other organisms.Because domestic animals typically experience reduced predation pressure compared with their wild counterparts,they are expected to exhibit less developed cryptic coloration.To test this prediction,we compared dorsal and ventral crypsis between domestic and wild black-spotted frogs(Pelophylax nigromaculatus)using reflectance-measurement methods.Our results showed that domestic males displayed significantly weaker dorsal crypsis than wild males.In contrast,no statistically significant difference in dorsal crypsis was observed between domestic and wild females.These findings suggest that the reduction in dorsal crypsis among domestic males may be attributable to relaxed predation pressure and shifts in dietary resources within human-modified environments.

Testis morphology may act as a determinant of sperm size, and the evolutionary pressure to produce longer sperm may impose functional constraints on testicular architecture. Postcopulatory sexual selection, particularly sperm competition, is widely recognised as a primary driver of the remarkable interspecific variation in sperm size and number across animals. Theoretical hypotheses predict that longer sperm require wider seminiferous tubules and that intense sperm competition favours an increased proportion of sperm-producing (seminiferous) tissue relative to non-spermatogenic components, independent of overall testis size. Using a phylogenetic comparative approach across 78 anuran species, we tested these two predictions. Our analyses revealed that the cross-sectional area of the seminiferous tubules was significantly and positively correlated with sperm length, thereby supporting the proposed morphogenetic constraint. Furthermore, relative testis size, which was used as a validated proxy for sperm competition intensity, was positively associated with the proportion of seminiferous tissue. In contrast, absolute testis size did not predict this tissue proportion. Our findings suggest that sperm competition shapes anuran testicular architecture through two distinct evolutionary pathways: indirect selection for wider tubules mediated by longer sperm, and direct selection for increased investment in seminiferous tissue. This tissue optimisation extends beyond simple testicular enlargement and reveals sophisticated evolutionary adaptations to postcopulatory sexual selection.

Against the backdrop of the global trend toward delayed childbearing, elucidating the mechanisms underlying uterine aging has emerged as a critical biomedical priority for addressing age-related implantation failure. Through unbiased global metabolomic profiling of peri-implantation uteri across different ages in mice, we identified nicotinamide adenine dinucleotide (NAD+) depletion as a hallmark metabolic feature of endometrial aging. Single-cell RNA sequencing further revealed an expansion of senescent stromal cell populations, which was accompanied by a decline in NAD+ levels. Supplementation with NAD+ precursors alleviated age-related stromal senescence and endometrial dysfunction, thereby restoring the uterus' implantation competence. Mechanically, we demonstrate that CD38 derived from myeloid serves as a principal driver of uterine NAD+ depletion; this process accelerates stromal senescence and impairs uterine receptivity. These findings establish CD38 as a central physiological integrator that links NAD+ metabolism to uterine function and highlight it as a promising target for rejuvenation strategies aimed at improving reproductive outcomes in women of advanced maternal age.

As delayed childbearing becomes increasingly common, elucidating the mechanisms of age-related placental senescence is critical for the development of effective therapeutic strategies. Here, we integrate metabolomics, single-cell RNA sequencing, and spatial transcriptomics to uncover conserved features of aged placentas in humans, mice, and pigs, namely, increased macrophage CD38 expression and NAD⁺ deficiency. Through pharmacological and genetic approaches, we demonstrate that macrophage CD38 depletes NAD⁺ in decidual stromal cells, thereby promoting placental senescence. Mechanistically, early-onset inflammation and senescence-associated secretory phenotype activity drive CD38 expression in macrophages via the IRF5 pathway. Importantly, treatment with NAD⁺ precursors or CD38 inhibitors attenuates age-related placental senescence, rescues intrauterine growth restriction, and improves long-term metabolic outcomes in offspring. These findings reveal a critical role for macrophage-driven metabolic dysregulation in reproductive aging and establish CD38 as a potential therapeutic target for age-associated pregnancy complications. The authors show that CD38-activated macrophages deplete NAD+ in decidual stromal cells, driving placental senescence and fetal growth restriction during aging. NAD+ restoration in animal models improves pregnancy outcomes and offspring health.

The Body Mass Index (BMI), integrating body weight and length, is a widely used metric for obesity assessment in humans. As pigs serve as crucial biomedical models, the application of BMI in swine and its genetic basis remain poorly explored. This study aimed to investigate the genetic architecture of pig BMI and compare two carcass-based BMI metrics (BMI-S and BMI-O) for breeding applicability. A total of 439 Landrace × Yorkshire crossbred pigs were genotyped with a 50 K SNP chip; heritability was estimated via a mixed linear model, and genome-wide association study (GWAS) was performed using the BLINK model. BMI-S and BMI-O exhibited moderate-to-high heritability of 0.55 and 0.47, respectively, with 17 genome-wide significant SNPs detected—including the top associated SNP rs81382440 on chromosome 4 and rs80898583 on chromosome 7. Key candidate genes (GPHN, ADAM33, KCNH8, PDCD4) and 5 SNP-trait associations validated in PigQTLdb were linked to lipid/energy metabolism and muscle development. Carcass-based BMI improved phenotypic accuracy, and our findings provide core genetic markers and a theoretical basis for molecular breeding of pig body conformation and lipid deposition traits.

Porcine gastric epithelial cells (PGECs) serve as a valuable model for studying the molecular and pathogenic mechanisms of the stomach. However, PGECs face limitations such as isolation challenges, short lifespan, and restricted proliferation. To address this, we established an immortalized PGECs (i-PGECs) to enable in vitro investigation of pathogen infection mechanisms. Primary PGECs were isolated from the acid-secreting glands using stepwise digestion with multiple enzymes (dispase II/collagenase I/hyaluronidase). Immortalization was achieved via lentiviral vectors expressing simian virus 40 large T antigen (SV40T) and human telomerase reverse transcriptase (hTERT), with successful expression confirmed by qRT-PCR (P < 0.05). Epithelial identity of i-PGECs was confirmed by stable expression of CK18, EpCAM, and E-cadherin, as shown by qRT-PCR and immunofluorescence. i-PGECs retained the morphological and ultrastructural features of PGECs and exhibited enhanced proliferation, as demonstrated by WST-8 assays, apoptosis and cell cycle analysis, karyotyping, and transmission electron microscopy (TEM). Telomere length analysis and scratch wound assays demonstrated stable telomere maintenance and consistent migration capacity unaffected by passaging. RNA-sequencing and differential expressed genes (DEGs) analysis revealed significantly upregulating of genes involved in cell proliferation pathways (P < 0.01). Following aflatoxin B1 (AFB1) exposure, i-PGECs significantly upregulated immune-related factors, such as NPC1 and PLAUR (P < 0.01). CRISPR/Cas9-mediated knockout of NPC1 in i-PGECs conferred increased resistance to AFB1-induced cytotoxicity, as shown by WST-8 assay. The i-PGECs remained stable after more than 50 passages, supporting their use as a reliable for in vitro model investigating the mechanisms of toxicity infection in the porcine gastric epithelium.

Oxybenzone (OBZ), a widely used ultraviolet filter detected in human bodily fluids, has been associated with reproductive toxicity, yet its effects on preimplantation development remain unclear. Using mouse embryos, human tripronuclear embryos, and blastoids, we investigated whether melatonin (MT) could mitigate OBZ-induced developmental defects. OBZ exposure impaired blastocyst formation, disrupted redox homeostasis, increased oxidative stress, and reduced developmental potential. MT supplementation restored glucose metabolism, elevated NAD+ levels, re-established redox balance, and enhanced histone acetylation, thereby preserving pluripotency and normal gene expression. This integrated metabolic-redox-epigenetic axis underlies the protective effects of MT and also improved embryo quality and reproductive outcomes in vivo. Our findings reveal a previously unrecognized mechanism by which MT safeguards early embryonic development through coordinated regulation of energy metabolism, oxidative status, and chromatin modification, providing new mechanistic insight for improving assisted reproduction.

Mountain ecosystems offer valuable opportunities to study species distribution and diversity along altitudinal gradients, particularly for amphibians. This research examined amphibian species distribution, diversity, and conservation across an elevational gradient in the Tangjiahe National Nature Reserve, part of the Hengduan Mountains in southwestern China. A total of 25 amphibian species, encompassing 8 families and 2 orders, were documented, including three newly recorded species: Fejervarya kawamurai, Polypedates braueri, and Boulenophrys minor. Among these, eight species were designated as "threatened" under IUCN criteria and are listed on China's Biodiversity Red List. Caijiaba exhibited the highest species diversity, whereas Shuichiping had the lowest. Fourteen species were found in terrestrial-farmland and aquatic-lotic habitats, which supported the greatest species richness. The distribution pattern along the altitudinal gradient showed peaks in species richness at 900-1100 m and 1900-2100 m elevation bands, with higher elevations displaying reduced richness. These findings highlighted the spatial characteristics of amphibian distribution and diversity across altitudinal ranges in the Tangjiahe National Nature Reserve and provide insights for formulating conservation policies and adaptive habitat management strategies.

Genetic mutation and drift, coupled with natural and human-mediated selection and migration, have produced a wide variety of genotypes and phenotypes in farmed animals. We here introduce the Farm Animal Genotype-Tissue Expression (FarmGTEx) Project, which aims to elucidate the genetic determinants of gene expression across 16 terrestrial and aquatic domestic species under diverse biological and environmental contexts. For each species, we aim to collect multiomics data, particularly genomics and transcriptomics, from 50 tissues of 1,000 healthy adults and 200 additional animals representing a specific context. This Perspective provides an overview of the priorities of FarmGTEx and advocates for coordinated strategies of data analysis and resource-sharing initiatives. FarmGTEx aims to serve as a platform for investigating context-specific regulatory effects, which will deepen our understanding of molecular mechanisms underlying complex phenotypes. The knowledge and insights provided by FarmGTEx will contribute to improving sustainable agriculture-based food systems, comparative biology and eventual human biomedicine.

Sexual dimorphism, a widespread phenomenon across the animal kingdom, encompasses differences between sexes in size, morphology, and physiological traits. In this study, we investigated sexual dimorphism in the flexor carpi radialis (FCR) muscle, which is critical for amplexus in Asiatic toads (Bufo gargarizans), using integrated transcriptomic and metabolomic approaches. Male toads exhibited significantly larger FCR muscles, reflecting enhanced muscle function required for sustained amplexus. Transcriptomic analysis identified 818 differentially expressed genes (DEGs) between sexes, with 389 upregulated and 429 downregulated in males, predominantly associated with muscle contraction, sarcomere organization, and energy metabolism. Metabolomic profiling revealed 69 differentially expressed metabolites (DEMs), with male-biased enrichment in pathways involved in protein synthesis and degradation, energy metabolism, and material transport. Integrated analysis pinpointed key metabolic pathways—such as glycine, serine, and threonine metabolism; alanine, aspartate, and glutamate metabolism; fatty acid degradation; and the tricarboxylic acid (TCA) cycle—as central to the observed sexual dimorphism. Among these, the genes AGXT, ACADL, ACAT1, MDH2, and SUCLG2 emerged as pivotal regulators. Collectively, these findings provide novel insights into the genetic and metabolic basis of sexual dimorphism in B. gargarizans, offering a deeper understanding of the evolutionary mechanisms driving sex-specific traits in vertebrates.

IntroductionTibetan pigs, native to the Qinghai-Tibet Plateau, have adapted over millennia to extreme conditions such as low oxygen, harsh cold, and high UV radiation, impacting their muscle characteristics and digestive tract microbiota. The quality of pork from Tibetan pigs (TP) and black pigs (BP) is influenced by various factors, including genetics, diet, and environmental adaptation. However, the specific influence of digestive tract microbiota metabolites on muscle traits remains poorly understood. Our goal was to correlate omic variations with meat quality traits and identify potential biomarkers predictive of superior meat quality, elucidate the regulatory effects of digestive tract microbial metabolites on Tibetan pig muscle characteristics, and reveal the genetic and nutritional mechanisms that promote adaptation to extreme environmental conditions.MethodsThis analysis encompassed metabolomic profiling of the entire digestive tract-including the stomach, jejunum, cecum, colon, and rectum-as well as histological, amino acid, fatty acid composition, and transcriptomic assessments of the longissimus dorsi muscle tissues to investigate how digestive tract microbial metabolites influence muscle adaptation to high altitudes.ResultsAnalyses revealed that Tibetan pig muscles contain smaller, more oxidative fibers enriched with flavor-enhancing amino acids. This was accompanied by a more favorable n-6/n-3 fatty acid ratio. Distinct patterns of microbial metabolites were observed in the digestive tract, influencing protein digestion and purine metabolism, and correlating with muscle glycine levels. Transcriptomic data showed varied gene expression in metabolic pathways related to salivary and pancreatic secretion, as well as carbohydrate and fatty acid metabolism. Integrated multi-omics approaches linked stomach metabolism, particularly through bile secretion pathways influenced by acetylcholine, to muscle functionality, highlighting the important role played by the ATP1B4 gene in enabling muscle physiology in Tibetan pigs.DiscussionThis study highlights the importance of targeted dietary interventions in improving meat quality for specific pig breeds. It also provides a theoretical foundation for precision agriculture strategies aimed at enhancing the meat quality of both TP and BP pigs.

Oxybenzone (2-Hydroxy-4-methoxybenzophenone, OBZ) is an organic ultraviolet filter extensively used in sunscreens and cosmetics, and has become a pervasive contaminant in aquatic environments. While its endocrine-disrupting properties and adverse reproductive effects are recognized, potential impacts on oocyte mitochondrial physiology remain incompletely understood. In this study, we investigated whether melatonin (MT), a pineal-derived antioxidant that regulates mitochondrial dynamics, can counteract OBZ-induced oocyte damage. Key indicators were evaluated in both in vivo and in vitro models, including mitochondrial dynamics, Ca²⁺ homeostasis, distribution and membrane potential (ΔΨm), electron transport chain (ETC) gene expression, reactive oxygen species (ROS) generation, antioxidant enzyme activity, and spindle organization. MT supplementation restored mitochondrial homeostasis by promoting fusion gene expression (Mfn1/2, Opa1) and suppressing fission genes (Drp1, Fis1, Mff), while reducing OBZ-induced cytosolic and mitochondrial Ca²⁺ overload. These protective effects reversed OBZ-related abnormalities such as disrupted mitochondrial distribution, spindle disorganization, decreased ΔΨm, downregulated ETC genes, and pronounced oxidative stress characterized by elevated ROS and diminished GSH. Microtranscriptomic analysis further suggested involvement of enhanced Ca²⁺ signaling and ATP-dependent chromatin remodeling in MT-mediated oocyte protection. Overall, our findings indicate that mitochondrial dysfunction is a critical component of OBZ-induced oocyte impairment, and support MT as a potential intervention to preserve oocyte quality under environmental toxicant exposure.

To explore the relationship between intestinal mucin 2 (MUC2) and weaning-induced diarrhoea in piglets, we analysed Min and Landrace piglets. The piglets were divided into a healthy weaned group, a weaned diarrhoea group, and a healthy unweaned control group. Intestinal tissues were collected, and goblet cell numbers, sizes, and degrees of intestinal injury were observed and recorded. Intestinal tissue MUC2 mRNA and protein expression were analysed via quantitative real-time PCR (qRT–PCR) and Western blotting. Min pigs presented significantly lower diarrhoea rates and intestinal injury scores than Landrace pigs (p < 0.01). The intestinal injury scores in the weaned diarrhoea group were significantly greater than those in the unweaned groups (p < 0.05), with Min pigs consistently exhibiting lower injury scores than Landrace pigs. Specifically, unweaned Min pigs presented significantly greater duodenal MUC2 mRNA (p < 0.05), and weaned healthy Min pigs presented notably greater expression in both the duodenum and jejunum (p < 0.01). These findings reveal enhanced intestinal protection against weaning stress and diarrhoea in Min pigs, with elevated MUC2 levels likely contributing to lower injury scores and milder symptoms, thus highlighting the influence of genetic differences.

Domestication is considered one of the most powerful evolutionary forces driving brain size reduction in animals, as observed when domestic species are compared with their wild conspecifics. This pattern is often attributed to the relaxed selective pressures associated with foraging and predator avoidance in domestic environments. In this study, we compared relative brain size between domestic and wild conspecific frogs, including the American bullfrog (Lithobates catesbeianus) and the black spotted frog (Pelophylax nigromaculatus), while controlling for body size. We found that domestic individuals exhibited significantly reduced brain sizes compared with their wild counterparts in both species. These findings align with domestication theory, which posits that reduced predation risk and human-provided resources in captive environments contribute to brain size reduction. We also found that domestic bullfrogs tended to show greater brain size reduction than domestic black spotted frogs. To our knowledge, this is the first empirical evidence demonstrating that domestication is a major factor driving brain reduction in frogs.

The Tibetan pig, as a typical highland breed, has a higher newborn-to-sow weight ratio than low-altitude breeds, which may be associated with better offspring survival in high-altitude environments. This suggests Tibetan pigs need a healthier physiological state during gestation. Understanding physiological changes during pregnancy is thus crucial for improving their reproductive performance. The ovaries secrete pregnancy-related hormones, while the spleen regulates immunity as a key immune organ. Both are vital for maintaining maternal health, yet their physiological changes during pregnancy in Tibetan pigs remain unclear. This study compared the transcriptome changes of the ovaries and spleens in non-pregnant and pregnant Tibetan pigs. The ovaries showed more pronounced transcriptional changes, with 757 differentially expressed PCGs (466 upregulated, e.g., ISM1, PRKAA2, STAR, CEBPB; 291 downregulated, e.g., BMP15, MOS) and 53 differentially expressed lncRNAs (34 upregulated, 19 downregulated). In the spleen, 229 PCGs were differentially expressed (76 upregulated, e.g., ISM1, PRKAA2, ESR1, CXCL8; 153 downregulated, e.g., NFATC4), along with 14 differentially expressed lncRNAs (5 upregulated, 9 downregulated). Functional analysis revealed enhanced ovarian hormone synthesis and suppressed oocyte activity, alongside reduced splenic inflammation during pregnancy. Notably, both organs showed increased lipid metabolism, and inflammation-related pathways were activated in the ovaries. Key candidate genes and regulatory factors potentially involved in pregnancy-related changes were also predicted. Overall, this study highlights transcriptional shifts in Tibetan pig ovaries and spleens during pregnancy, offering insights into plateau animal reproduction and strategies to improve Tibetan pig fertility.

Pigs on high-fat diets maintaining metabolic homeostasis and are resistant to hepatic steatosis, differing from humans and mice. Obesity-induced metabolic dysregulation and inflammation in skeletal muscle are well-studied in humans and mice, but less is known about pig skeletal muscle responses. This study constructs the skeletal muscle transcriptome of obese pigs and integrates it with publicly available transcriptional profiles from obese humans and mice, and ATAC-seq data from lean individuals across species. We systematically characterized transcriptional changes in skeletal muscle under stress of obesity, focusing on the evolution of gene families, orthologous genes, and epigenetic regulation. Our results show that obesity activates lipid catabolism genes and inhibits immune response genes in pig skeletal muscle, contrasting with humans and mice. We identify expanding gene families in pigs, such as olfactory receptors, α-amylase, and ABC transporters, which are upregulated in obesity. While oxidative metabolism-related gene families are contracted in the human and mouse genomes and are downregulated with obesity. By comparing orthologous genes, we identify a set of divergently changing genes induced by obesity across species, which primarily participate in lipid metabolism, inflammation, and immune cell activation. High-divergence genes show conserved coding and promoter sequences, and exhibit greater chromatin accessibility in promoter regions, compare with low-divergence genes. These findings suggest that gene dosage and transcriptional plasticity contribute to species-specific expression divergent responses to obesity. Identifying rapidly evolving gene families, divergently expressed genes, and potential transcription factor binding sites may reveal new insights into obesity-related metabolic disorders and therapeutic targets.

High-protein (HP) diets are widely adopted in Western societies for body-weight management; yet, they exacerbate senescence-associated metabolic deterioration, posing an unresolved pathophysiological conundrum. Here, we demonstrate that long-term HP intake mediates adipocyte-specific NAD+ depletion and mitochondrial dysfunction in white adipose tissue (WAT). Single-nucleus transcriptomic analyses revealed adipocyte-restricted senescence signatures in HP-fed mice. Mechanistically, HP intake triggers macrophage-specific upregulation of CD38 (a key NAD+ hydrolase), which depletes adipocyte NAD+ pools and thereby accelerates cellular senescence. Restoration of NAD+ levels, either via supplementation with NAD+ precursor or pharmacological inhibition of CD38 activity, alleviated the senescence-associated metabolic sequelae induced by HP diets. Our findings establish macrophage-adipocyte NAD+ crosstalk as a central axis linking dietary protein excess to WAT aging, providing actionable targets for the prevention and treatment of age-related metabolic disorders.