Search Results (436)

Intramuscular fat (IMF) is a crucial determinant of pork quality, influencing tenderness, flavor, and consumer preferences, yet selective breeding has reduced its levels in modern pigs. This review explores the molecular and cellular mechanisms of IMF deposition, including progenitor cell differentiation via pathways like Wnt/β-catenin and PPARγ, and advances in non-invasive detection methods such as hyperspectral imaging and Raman spectroscopy. It highlights correlations and causal links between the gut microbiota composition and IMF, established through omics analyses, fecal microbiota transplantation, and germ-free models. Key microbial metabolites, including short-chain fatty acids (SCFAs) and bile acids, modulate lipid metabolism bidirectionally via signaling receptors like GPR43, FXR, and TGR5. Future research should integrate multi-omics and develop probiotics to enhance IMF efficiency for sustainable pork production. Full article

Spermatogenesis is a tightly coordinated differentiation program that sustains male fertility while transmitting genetic and epigenetic information to the next generation. This review consolidates mechanistic evidence showing how RNA-centered regulation integrates with the epitranscriptome and three-dimensional (3D) genome architecture to orchestrate germ-cell fate transitions from spermatogonial stem cells through meiosis and spermiogenesis. Recent literature is critically surveyed and synthesized, with particular emphasis on human and primate data and on stage-resolved maps generated by single-cell and multi-omics technologies. Collectively, available studies support a layered regulatory model in which RNA-binding proteins and RNA modifications coordinate transcript processing, storage, translation, and decay; small and long noncoding RNAs shape post-transcriptional programs and transposon defense; and dynamic chromatin remodeling and 3D reconfiguration align transcriptional competence with recombination, sex-chromosome silencing, and genome packaging. Convergent nodes implicated in spermatogenic failure are highlighted, including defects in RNA metabolism, piRNA pathway integrity, epigenetic reprogramming, and nuclear architecture, and the potential of these frameworks to refine molecular phenotyping in male infertility is discussed. Finally, key gaps and priorities for causal testing in spatially informed, stage-specific experimental systems are outlined. Full article

The Transforming Growth Factor-β (TGF-β) superfamily comprises highly conserved cytokines that orchestrate key cellular functions, including proliferation, differentiation, and apoptosis. Within the ovary, TGF-β family members serve as pivotal regulators of folliculogenesis, exerting stage-specific actions from embryonic germ cell development to advanced follicular maturation. During fetal development, activins and SMAD-dependent signaling pathways are essential for primordial germ cell proliferation, survival, and the breakdown of germ cell cysts, enabling the establishment of the primordial follicle pool. Throughout folliculogenesis, TGF-β supports follicle activation, promotes the transition from dormant to growing follicles, stimulates granulosa cell proliferation, sustains follicular viability, and modulates steroidogenesis through theca cell regulation. Notably, anti-müllerian hormone, a TGF-β family member, plays a central role in inhibiting premature follicle recruitment and serves as a key biomarker of ovarian reserve. Dysregulation of TGF-β signaling contributes to various ovarian disorders, including polycystic ovary syndrome and premature ovarian insufficiency. A deeper understanding of these complex signaling networks is critical for identifying novel therapeutic targets and advancing clinical interventions in female reproductive pathologies. This review provides an integrated overview of the roles of the TGF-β superfamily in ovarian physiology and its contributions to disease development. Full article

Background and Objectives: Radiomics and artificial intelligence (AI) offer emerging quantitative tools for enhancing the diagnostic evaluation of testicular cancer. Conventional imaging—ultrasound (US), magnetic resonance imaging (MRI), and computed tomography (CT)—remains central to management but has limited ability to characterize tumor subtypes, detect occult nodal disease, or differentiate necrosis, teratoma, and viable tumor in post-chemotherapy residual masses. This systematic review summarizes current advances in radiomics and AI for both primary tumors and retroperitoneal disease. Materials and Methods: A systematic search of PubMed, Scopus, and Web of Science identified studies applying radiomics or AI to testicular tumors, retroperitoneal lymph nodes and post-chemotherapy residual masses. Eligible studies included quantitative imaging analyses performed on ultrasound, MRI, and CT, with optional integration of clinical or molecular biomarkers. Eighteen studies met inclusion criteria and were evaluated with respect to methodological design, diagnostic performance, and translational readiness. Results: Across modalities, radiomics demonstrated encouraging discriminatory capacity, with accuracies of 74–82% for ultrasound, 80.7–97.9% for MRI, and 71.7–85.3% for CT. CT-based radiomics for post-chemotherapy residual masses showed moderate ability to distinguish necrosis/fibrosis, teratoma, and viable germ-cell tumor, though heterogeneous methodologies and limited external validation constrained generalizability. The strongest performance was observed in multimodal approaches: integrating radiomics with clinical variables or circulating microRNAs improved accuracy by up to 12% and 15%, respectively, mirroring gains reported in other oncologic radiomics applications. Persisting variability in segmentation practices, acquisition protocols, feature extraction, and machine-learning methods highlights ongoing barriers to reproducibility. Conclusions: Radiomics and AI-enhanced frameworks represent promising adjuncts for improving the noninvasive evaluation of testicular cancer, particularly when combined with clinical or molecular biomarkers. Future progress will depend on standardized imaging protocols, harmonized radiomics pipelines, and multicenter prospective validation. With continued methodological refinement and clinical integration, radiomics may support more precise risk stratification and reduce unnecessary interventions in testicular cancer. Full article

Tooth germ development is a precisely orchestrated process dependent on integrated cellular interactions and molecular signals, yet its regulatory mechanisms remain incompletely defined. Here, we constructed a high-resolution cellular atlas of miniature pig tooth germs using 10× single-cell RNA sequencing to investigate the molecular mechanisms underlying tooth mineralization. By leveraging cellular heterogeneity and dynamic gene expression trajectories in epithelial and mesenchymal populations, we identified microfibril-associated protein 5 (MFAP5) as a previously unrecognized regulator of the odontogenic program. Functional assays demonstrate that MFAP5, an extracellular matrix component, is indispensable for mesenchymal differentiation and matrix mineralization in vitro. Mechanistically, MFAP5 engages Integrin alpha-5 (ITGA5) to activate Extracellular Signal-Regulated Kinase/Mitogen-Activated Protein Kinase (ERK/MAPK) signaling in odontoblast-lineage cells, thereby promoting odontoblast differentiation and dentin deposition. Collectively, our single-cell–resolved analyses uncovered a MFAP5–ITGA5–ERK/MAPK signaling axis that operates in a cell-state–specific manner during tooth germ mineralization, providing new mechanistic insights into odontogenic differentiation and a potential molecular basis for dental tissue regeneration strategies. Full article

Arginine vasotocin (AVT) has recently emerged as a local regulator of testicular function in fish. Using ex vivo culture system, it was demonstrated that AVT directly stimulates androgen-dependent basal spermatogenesis in zebrafish. In the presence of gonadotropins, AVT enhanced FSH-induced development of early phases of spermatogonial proliferation while blocking FSH-mediated spermiogenesis. However, AVT promoted proliferation of LH-induced pre-meiotic and meiotic germ cell populations without affecting the final stages of spermiogenesis. These findings led to the hypothesis that AVT plays a role by promoting early germ cell proliferation and differentiation while simultaneously inhibiting premature progression through spermiogenesis. To test this hypothesis, we investigated the chronic effects of AVT on adult zebrafish testes, in vivo. Prolonged AVT treatment for 21 days led to dose-dependent accumulation of undifferentiated type A spermatogonia and reduced post-meiotic germ cells and spermatozoa. We also observed decreased plasma 11-ketotestosterone (11-KT) levels and downregulation of fshr. This was accompanied by a basal suppression of avt and its receptors, avpr1aa, avpr1ab, avpr2aa, avpr2ab, avpr2l, in both brain and testis during the pre-spawning phase. The present findings, along with those of previously published studies, collectively demonstrate that AVT presence during the early stages of testicular development promotes spermatogonia proliferation while diminishing FSH-induced premature progress toward spermatogenesis. This occurs until later stages, when AVT expression is diminished, allowing for optimal LH-induced spermiogenesis in zebrafish. Full article

The localization and remodeling of mRNPs is inextricably linked to translational control. In recent years there has been great progress in the field of mRNA translational control due to the characterization of the proteins and small RNAs that compose mRNPs. But our initial assumptions about the physical nature and participation of germ cell granules/condensates in mRNA regulation may have been misguided. These “granules” were found to be non-membrane-bound liquid–liquid phase-separated (LLPS) condensates that form around proteins with intrinsically disordered regions (IDRs) and RNA. Their macrostructures are dynamic as germ cells differentiate into gametes and subsequently join to form embryos. In addition, they segregate translation-repressing RNA-binding proteins (RBPs), selected eIF4 initiation factors, Vasa/GLH-1 and other helicases, several Argonautes and their associated small RNAs, and frequently components of P bodies and stress granules (SGs). Condensate movement, separation, fusion, and dissolution were long conjectured to mediate the translational control of mRNAs residing in contained mRNPs. New high-resolution microscopy and tagging techniques identified order in their organization, showing the segregation of similar mRNAs and the stratification of proteins into distinct mRNPs. Functional transitions from repression to activation seem to corelate with the overt granule dynamics. Yet increasing evidence suggests that the resident mRNPs, and not the macroscopic condensates, exert the bulk of the regulation. Full article

Background/Objectives: Germ Cell Neoplasia In Situ (GCNIS) is considered the precursor lesion for the majority of testicular germ cell tumors (TGCTs). The aim of this study was to evaluate whether first-order radiomics features derived from volumetric diffusion tensor imaging (DTI) metrics—specifically apparent diffusion coefficient (ADC) and fractional anisotropy (FA) histogram parameters—can detect GCNIS. Methods: This study included 15 men with TGCTs and 10 controls. All participants underwent scrotal MRI, including DTI. Volumetric ADC and FA histogram metrics were calculated for the following tissues: group 1, TGCT; group 2: testicular parenchyma adjacent to tumor, histologically positive for GCNIS; and group 3, normal testis. Non-parametric statistics were used to assess differences in ADC and FA histogram parameters among the three groups. Pearson’s correlation analysis was followed by ordinal regression analysis to identify key predictive histogram parameters. Results: Widespread distributional differences (p < 0.05) were observed for many ADC and FA variables, with both TGCTs and GCNIS showing significant divergence from normal testes. Among the ADC statistics, the 10th percentile and skewness (p = 0.042), range (p = 0.023), interquartile range (p = 0.021), total energy (p = 0.033), entropy and kurtosis (p = 0.027) proved the most significant predictors for tissue classification. FA_energy (p = 0.039) was the most significant fingerprint of the carcinogenesis among the FA metrics. These parameters correctly characterized 88.8% of TGCTs, 87.5% of GCNIS tissues and 100% of normal testes. Conclusion: Radiomics features derived from volumetric ADC and FA histograms have promising potential to differentiate TGCTs, GCNIS, and normal testicular tissue, aiding early detection and characterization of pre-cancerous lesions. Full article

The Nile tilapia (Oreochromis niloticus), a key aquaculture species, displays marked sexual growth dimorphism, with males growing faster than females. This process is governed by intricate interactions between antagonistic regulators, including transcription factors, growth factors, and steroid hormones, operating through sex-specific developmental pathways. While circular RNAs (circRNAs) are known to modulate gene expression by sponging microRNAs (miRNAs), their role in teleost sex differentiation remains poorly understood. To address this gap, we profiled circRNA expression in tilapia gonads by constructing six circRNA libraries from testes and ovaries of 180 days after hatching (dah) fish, followed by high-throughput sequencing. We identified 6564 gonadal circRNAs distributed across all 22 linkage groups, including 226 differentially expressed circRNAs (DECs; 108 testis-biased, 118 ovary-biased). Functional enrichment analysis linked their host genes to critical pathways such as cAMP signaling, cell adhesion molecules, and—notably—sexual differentiation processes (e.g., estrogen signaling, oocyte meiosis, and steroid hormone biosynthesis). Furthermore, we deciphered competing endogenous RNA (ceRNA) networks, uncovering circRNA–miRNA–mRNA interactions targeting germ cell determinants, sex-specific transcription factors, and steroidogenic enzymes. This study provides the first systematic exploration of circRNA involvement in tilapia sex differentiation and gonadal differentiation, offering novel insights into the post-transcriptional regulation of sexual dimorphism. Our findings advance the understanding of circRNA biology in fish and establish a framework for future studies on aquaculture species with similar reproductive strategies. Full article

Low energy N⁺ ion beam implantation has been used to create the novel rice mutant “shuangli”, which produces partially fertile spikelets containing double grains. Abnormal ovule development is a major cause of partial fertility and grain diversity in rice mutants. To elucidate the developmental mechanism of ovule diversity in shuangli, ovules undergoing development were stained using eosin Y and H33342 and observed using confocal laser scanning microscopy. Different developmental abnormalities were observed in the ovary, embryo sac, and ovule. Abnormal development was observed in 35.18% of the ovary structures, primarily manifesting as “tumor” like cell clusters, “false ovaries”, stamen degeneration, and double ovaries. In the embryo sac, abnormal development occurred in about 17.35% of the megaspore cells, including the formation of three nuclei, two daughter cells of asynchronously divided dyads, multiple megaspore tetrads, and “narrow and elongated” cavities. At the female gametogenesis stage, the abnormal development rate was 27.53%, mainly involving the degeneration of the central polar nucleus, egg apparatus, antipodal cell mass, or female germ unit. In shuangli, abnormal development occurred in 28.06% of the ovule structures, including lateral tissue, nucellar tissue, double ovules and double embryo sacs. Of the observed lateral tissues, 8.27% did not differentiate into sexual reproductive tissue, which affected the fertilization of the embryo sac, leading to atrophy and degeneration. A new abnormal tissue similar to the inner integument was found on both sides of the nucellar tissue, and the two specialized nucellar tissues appeared to have “staggered” growth within a single ovary. Of the examined ovules, 10.79% exhibited different types of double ovules, including heart-shaped, “anatropous”, “conjoined” structures. However, the double ovules typically developed synchronously, explaining the production of different sizes of the two grains in shuangli. In addition, “double” embryo sacs from two “twinborn” nucelli were found in one ovule, and the frequency of “double” embryo sacs was 3.60%. Therefore, ovule development diversity may result in fertilization or gradual degeneration after fertilization, explaining the lower fertility of shuangli at the embryological level. Full article

X chromosome inactivation (XCI) is a fundamental epigenetic process that balances X-linked gene expression between females and males by silencing one X chromosome in female cells. Variability or skewing of XCI can influence the clinical presentation of X-linked disorders. Bain type X-linked intellectual disability syndrome (MRXSB), caused by mutations in the X-linked HNRNPH2 gene, is characterized by intellectual disability, developmental delay, and neurological abnormalities. In female patients, XCI heterogeneity complicates disease modeling and therapeutic development. Induced pluripotent stem cells (iPSCs) offer a unique platform to study patient-specific disease mechanisms, but the dynamics of XCI during iPSC reprogramming, maintenance, and differentiation are not fully understood. In this study, we generated 12 iPSC clones from fibroblasts of a female MRXSB patient heterozygous for the HNRNPH2 c.340C > T mutation. Four clones expressed the mutant HNRNPH2 allele and eight expressed the wild-type allele, indicating X chromosome reactivation (XCR) followed by random XCI during reprogramming. Importantly, these XCI patterns remained stable during long-term iPSC propagation and subsequent differentiation into the three germ layers and neural stem cells. Our findings provide new insights into XCI and XCR dynamics in the context of X-linked neurodevelopmental disorders and emphasize the importance of careful clone selection for accurate disease modeling using iPSC-based approaches. Full article

Male infertility is a global health concern, and many cases are idiopathic in nature. The development and differentiation of germ cells (Gcs) are supported by Sertoli cells (Scs). Differentiated Scs support the development of Gcs into sperm, and hence, male fertility. We previously reported on a developmental switch in Scs around 12 days of age onwards in rats. During the process of the differentiation of Scs, the differential expression of mitophagy-related genes and its role in male fertility are poorly understood. To address this gap, we evaluated the microarray dataset GSE48795 to identify 12 mitophagy-related hub genes, including B-Cell Leukemia/Lymphoma 2 (Bcl2) and FBJ murine osteosarcoma viral oncogene homolog (Fos). We identify Neuron-derived orphan receptor 1 (Nor1) as a potential mitophagy-related gene of interest due to its strong regulatory association with two hub genes, Bcl2 and Fos, which were differentially expressed during Sc maturation. To validate this finding, we generated a transgenic rat model with the Sc-specific knockdown of Nor1 during puberty. A functional analysis showed impaired spermatogenesis with reduced fertility in these transgenic rats. Our findings suggest that Nor1 may be an important mitophagy-related gene regulating the function of Scs and thereby regulating male fertility. Full article

Background/Objectives: Inactivation of the dnd gene involved in the development of primordial germ cells (PGCs) leads to the loss of gametes and halts reproductive development. Studies on sterile fish allow for the identification of genes and processes associated with GC differentiation. Methods: Atlantic salmon with GC-ablated testes were produced by temporal silencing of dnd. Gene expression was analyzed in sterile and fertile testes using 44k microarray and qPCR. Results: In sterile testes, transcripts of several GC markers were detected at low levels, suggesting the presence of cells with a GC-related expression profile that failed to initiate spermatogenesis. Expression of 260 genes was undetectable in the gonads of sterile males and females, and 61.5% of these were also inactivated during first maturation of fertile testes. This group was enriched with genes highly expressed in the brain, including those involved in endocrine and paracrine regulation, synaptic transmission, and numerous genes critical for brain development; among them, 45 genes encoding homeobox proteins. Another group of 229 genes showed increased expression in developing testes and included genes involved in neurosecretion and brain development regulation. GC-ablated testes showed increased expression of reproductive regulators such as amh and sdy and numerous immune genes, suggesting a reprogramming of GC-depleted testes. Temporal silencing of dnd indicated common developmental processes in the brains and gonads of Atlantic salmon testis that become inactive in testes at first maturation. These processes may play roles in PGC homing, the creation of a specific environment required for spermatogenesis, or facilitating communication between the gonads. Full article

iPSCs and their derivatives are used to investigate the molecular genetic mechanisms of human diseases, to identify therapeutic targets, and to screen for small molecules. Combining technologies for generating patient-specific iPSC lines and genome editing allows us to create cell models with unique characteristics. We obtained and characterized three iPSC lines by reprogramming peripheral blood mononuclear cells of a patient with Huntington’s disease (HD) using episomal vectors encoding Yamanaka factors. iPSC lines expressed pluripotency marker genes, had normal karyotypes and were capable of differentiating into all three germ layers. The obtained iPSC lines are useful for modeling disease progression in vitro and studying pathological mechanisms of HD, such as ER stress. A transgene of genetically encoded biosensor XBP1-TagRFP was introduced into the iPSCs to visualize ER stress state of cells. The study demonstrated that iPSC-derived medium spiny neurons develop ER stress, though the IRE1-mediated pathway does not seem to be involved in the process. Full article

Type 1 interferons (IFN-Is) exhibit significant antiviral, antitumor, and immunoregulatory properties, demonstrating substantial therapeutic potential. However, IFN-Is are pleiotropic cytokines, and the available data on their effect under specific pathological conditions are inconclusive. Furthermore, the systemic administration of IFN-Is can result in side effects. Generating cells that can migrate to the pathological focus and provide regulated local production of IFN-Is could overcome this limitation and provide a model for an in-depth analysis of the biological and therapeutic effects of IFN-Is. Induced pluripotent stem cells (iPSCs) are a valuable source of various differentiated cell types, including human immune cells. In this study, we describe the generation of genetically modified human iPSCs with doxycycline-controlled overexpression of interferon β (IFNB1). Three IFNB1-overexpressing iPSC lines (IFNB-iPSCs) and one control line expressing the transactivator M2rtTA (TA-iPSCs) were generated using the CRISPR/Cas9 technology. The pluripotency of the generated cell lines has been confirmed by the following: (i) cell morphology; (ii) the expression of the pluripotency markers OCT4, SOX2, TRA 1-60, and NANOG; and (iii) the ability to spontaneously differentiate into the derivatives of the three germ layers. Upon the addition of doxycycline, all IFNB-iPSCs upregulated IFNB1 expression at RNA (depending on the iPSC line, 126-816-fold) and protein levels. The IFNB-iPSCs and TA-iPSCs generated here represent a valuable cellular model for studying the effects of IFN-β on the activity and differentiation trajectories of different cell types, as well as for generating different types of cells with controllable IFN-β expression. Full article