Biological Research最新文献

{"title":"A successful stacking of multiple biotic stress resistance in rice facilitated by genes and precision breeding tools.","authors":"Muthukumarasamy Sriram, Kailappan Amudha, Swaminathan Manonmani, Chellapan Gopalakrishnan, Venugopal Sheela, John Jesu Bonipas Antony, Selvaraj Vignesh, Ramalingam Suresh","doi":"10.1186/s40659-026-00710-0","DOIUrl":"https://doi.org/10.1186/s40659-026-00710-0","url":null,"abstract":"<p><strong>Background: </strong>Genetic improvement of rice for biotic stress is a major and continuous breeding objective owing to changing pest and disease scenarios. In response to climate change, an outbreak of new pathotypes and biotypes results in huge yield loss which directly impacts the economic stability and food security. The development of varieties with multiple resistance genes for a particular disease is the most effective approach to the combating mechanism against evolving pathogens and insects. Multiple resistance genes against blast (Pi54), bacterial leaf blight (xa5, xa13, Xa21), and brown plant hopper (Bph17, Bph3, bph2) were pyramided through marker-assisted forward breeding by attempting multiple crosses involving six parental lines. The F₄ lines were screened and the resistant lines were further reconfirmed at the F₅ stage through precise phenotyping and also by genotyping with trait-specific markers. Finally, the selected lines were evaluated for the agronomic performance.</p><p><strong>Results: </strong>Four lines were selected from the population which carrying seven resistance genes against BL, BB, BPH with superior agronomic performance. Another line, x21302-239, harboring all the resistant genes and showing a resistant response in screening experiments for all three stresses, with marginal agronomic performance (single-plant yield: 33 g), can be utilized as a desirable donor to develop elite rice cultivars with multiple biotic stress resistance.</p><p><strong>Conclusions: </strong>Homozygous genetic background is more favorable for epidemic outbreak in a short period of time in comparison with the population of multiple genetic background. Since this complex genetic background disturbs the infectious cycle of the pathogen. So that a variety or a hybrid which developed from multiple parental lines are notable for their durable resistance than the monogenic resistance variety.</p>","PeriodicalId":9084,"journal":{"name":"Biological Research","volume":"59 1","pages":""},"PeriodicalIF":5.4,"publicationDate":"2026-08-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13491621/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148788002","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Targeting oxidative stress-NLRP3-apoptotic-steroidogenic axis in doxorubicin-induced testicular toxicity: protective efficacy of ferulic acid niosomes.","authors":"Mahran Mohamed Abd El-Emam, Aya Elhady Atia, Mona A Bakry, Rabab A Husseini, Eman Elrefaei, Noha Osama El-Shaer, Safaa I Khater, Ola Ali, Ahmed El-Sayed Yousef, Hussein I Elbelbesy","doi":"10.1186/s40659-026-00707-9","DOIUrl":"10.1186/s40659-026-00707-9","url":null,"abstract":"<p><p>Doxorubicin (DOX) is a commonly prescribed chemotherapeutic agent whose clinical application is limited by its toxicity to rapidly dividing organs, particularly the testes.This study explored the protective potential of ferulic acid-loaded niosomes (FA-NIO) against DOX-induced testicular dysfunction in rats and elucidated the underlying molecular mechanisms. Forty male rats were allocated into four groups: Group 1 was given saline, Group 2 was given DOX, Group 3 was given DOX + free FA, and Group 4 was given DOX + FA-NIO. Serum reproductive hormone levels, testicular oxidative stress indices, and the expression of genes and proteins relevant to NLRP3 inflammasome signaling, apoptosis (Bax and Bcl-2), and steroidogenesis (StAR, CYP11A1, and 3β-HSD) were assessed, along with histopathological changes in the testicular tissue. DOX administration significantly increased MDA (malondialdehyde) while suppressing antioxidant defences, accompanied by hormonal imbalance, severe histopathological damage, upregulation of Bax, NLRP3, and IL-1β, downregulation of Bcl-2, and inhibition of steroidogenic gene expression. On the other hand, these changes were partially mitigated in the DOX + free FA group. Notably, FA-NIO treatment reduced histopathological damage, normalised reproductive hormone levels, markedly increased steroidogenesis-related gene expression, suppressed apoptotic and inflammasome-related gene expression, and greatly restored antioxidant capacity. Compared with DOX + free FA, FA-NIO consistently demonstrated greater protective efficacy. In summary, FA-NIO targets oxidative stress-mediated apoptosis, attenuates NLRP3 inflammasome-related signaling, and restores steroidogenic function, thereby successfully attenuating DOX-induced testicular toxicity. These results suggest that FA-NIO may represent a promising nanotherapeutic strategy for attenuating chemotherapy-induced reproductive toxicity in a rat model.</p>","PeriodicalId":9084,"journal":{"name":"Biological Research","volume":"59 1","pages":""},"PeriodicalIF":5.4,"publicationDate":"2026-07-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13422117/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148629651","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Mycophenolate mofetil induces angiogenesis disorders in zebrafish embryos by inhibiting the mitogen-activated protein kinase signaling pathway.","authors":"Mengqi Wan, Zhonghao Xiao, Xiaowen Shi, Zhijun Ye, Jiejun Liu, Xuyang Liu, Huimin Li, Xinjun Liao, Shouhua Zhang, Juhua Xiao, Zigang Cao","doi":"10.1186/s40659-026-00717-7","DOIUrl":"10.1186/s40659-026-00717-7","url":null,"abstract":"<p><p>As a commonly used immunosuppressant, mycophenolate mofetil (MMF) is widely prescribed after organ transplantation because it is not toxic to the liver and kidney. However, its use is associated with certain gastrointestinal toxicity and increased risk of opportunistic infections. Meanwhile, whether it affects vascular development is unclear. Herein, a zebrafish model was utilized to evaluate whether MMF affects vascular development, and the results showed that MMF exhibited teratogenicity, shortened the body length, caused pericardial edema, and delayed yolk sac absorption in zebrafish. In addition, although Intersegmental vessels showed hyperbranching, opposite changes were observed in cerebrovascular, with cerebrovascular inhibition. Moreover, MMF exposure upregulated oxidative stress levels, inhibited notch signaling and mitogen-activated protein kinase (MAPK) signaling, and affected the expression of vascular endothelial growth factor and receptor tyrosine kinase. Treatment with the antioxidant astaxanthin and the notch signaling pathway activator sodium valproate did not rescue the vascular development defects, indicating that MMF affects multiple signal pathways. However, the treatment of MMF-exposed zebrafish with guanosine and the MAPK signaling activators diprovocim significantly rescued the angiogenesis-related defects, suggesting that MMF induces ISV hyperbranching in zebrafish by inhibiting MAPK signaling and inosine 5'-monophosphate dehydrogenase (IMPDH).</p>","PeriodicalId":9084,"journal":{"name":"Biological Research","volume":" ","pages":""},"PeriodicalIF":5.4,"publicationDate":"2026-07-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148468960","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Reshaping the immune landscape: next-generation microglia-targeted therapies for Alzheimer's disease.","authors":"Jun Wu, Jueying Zhao, Shen Chen, Fangyi Xu","doi":"10.1186/s40659-026-00716-8","DOIUrl":"https://doi.org/10.1186/s40659-026-00716-8","url":null,"abstract":"<p><strong>Background: </strong>Alzheimer's disease (AD) is a significant global health challenge characterized as a multifactorial neurodegenerative disorder, involving amyloid-β (Aβ) and Tau aggregation, neuroinflammation and progressive neuronal injury. While Amyloid-targeted therapies have achieved a breakthrough in prevention of Aβ aggregation, the strategies face notable limitations in achieving curative outcomes and management of amyloid-independent central nervous system (CNS) dysfunction. Consequently, targeting microglia, the central immune cells of the brain, has emerged as a promising strategy to enhance the specificity and efficacy of AD interventions.</p><p><strong>Main body: </strong>Accumulating evidence indicates microglial dysfunction is not a passive immune bystander of AD, but serves as a critical mechanistic nexus linking Aβ accumulation and AD symptomatic phenotype. This review critically examines the \"next generation\" of microglial therapeutics, moving beyond broad immunosuppression to precision phenotype modulation. We highlight breakthrough strategies in recent years including immune reconstitution, metabolic reprogramming, nanomaterial-mediated drug delivery, and the revolutionary potential of iPSC-derived microglia replacement. By elucidating the rationale underlying the specific strategies based on microglial biofunction and potential molecular mechanism in AD pathology, we provide an overview of current development of clinical trials and cutting-edge modalities aimed at restoring microglial homeostasis, affording an opportunity to alter the AD trajectory.</p><p><strong>Conclusion: </strong>This review aims to delineate the path from bench to bedside and propose promising pathways to overcome current bottlenecks in AD drug development.</p>","PeriodicalId":9084,"journal":{"name":"Biological Research","volume":" ","pages":""},"PeriodicalIF":5.4,"publicationDate":"2026-07-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148418661","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"FOXQ1 promotes endometriosis development by enhancing mitochondrial function.","authors":"Yu Liu, Mei Chi, Yan Lei, Jing Jin, Xiaojing Lei, Shu Zhang, Qingmei Guan, Ling Zhang, Xin Du","doi":"10.1186/s40659-026-00698-7","DOIUrl":"https://doi.org/10.1186/s40659-026-00698-7","url":null,"abstract":"<p><strong>Background: </strong>Endometriosis (EMs) is a pathological condition characterized by the ectopic growth of endometrial tissue outside the uterine cavity. This study aimed to investigate the role of FOXQ1 in the pathogenesis of EMs, particularly through its regulation of mitochondrial function.</p><p><strong>Methods: </strong>Eutopic endometrial, ovarian EMs, pelvic sidewall EMs, and bowel EMs samples were obtained from 76 patients undergoing laparoscopic or hysteroscopic surgery. The expression levels of FOXQ1 in EMs tissues were determined using reverse transcription-quantitative PCR (RT-qPCR) and western blot analysis. Furthermore, the mRNA expression levels of pro-inflammatory cytokines, including interleukin (IL)-6, interleukin (IL)-1β, tumor necrosis factor (TNF)-α, and interleukin (IL)-8, were quantified in FOXQ1 overexpression (OE) and FOXQ1 knockdown (siFOXQ1) groups using RT-qPCR. Mitochondrial function in these groups, using primary human endometrial epithelial cells (hEECs), was evaluated by assessing mitochondrial DNA (mtDNA) content, ATP production, reactive oxygen species (ROS) levels, oxygen consumption rate (OCR), and MitoTracker staining. The in vitro effects of FOXQ1 on EMs were further examined in hEECs by analyzing cellular apoptosis, proliferation, invasion, and migration.</p><p><strong>Results: </strong>The expression of FOXQ1 was markedly elevated in endometriotic lesions, such as those on the pelvic sidewall and bowel, compared to control samples, with localization specifically observed in endometrial epithelial cells. Functional assays conducted on hEECs demonstrated that overexpression of FOXQ1 inhibited apoptosis, upregulated critical inflammatory cytokines (IL-6, IL-1β, TNF-α, IL-8), and promoted cellular proliferation, invasion, and migration. These effects appear to be mediated by enhanced mitochondrial function, as indicated by increased mtDNA content, ATP production, ROS levels, OCR, and MitoTracker signal. Conversely, the group treated with siFOXQ1 exhibited the opposite effects. The receiver operating characteristic curve analysis revealed that the area under the curve for FOXQ1 was 0.8784.</p><p><strong>Conclusions: </strong>FOXQ1 facilitates the progression of endometriosis by enhancing mitochondrial function, which subsequently stimulates inflammatory responses and promotes critical cellular processes such as proliferation, invasion, and migration.</p>","PeriodicalId":9084,"journal":{"name":"Biological Research","volume":" ","pages":""},"PeriodicalIF":5.4,"publicationDate":"2026-07-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148403566","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Expression and functional characterization of the TRPV3 channel in domestic cat (Felis catus) mature oocytes.","authors":"Sebastián Vergara, José Duarte-Zúñiga, Claudio Hidalgo, Constanza Suárez-Suárez, Karen Castillo, Matthias Piesche, Felipe Aguilera, Rafael A Fissore, Fernando Hinostroza, Daniel Veraguas-Dávila, Ingrid Carvacho","doi":"10.1186/s40659-026-00709-7","DOIUrl":"https://doi.org/10.1186/s40659-026-00709-7","url":null,"abstract":"<p><strong>Background: </strong>Intracellular calcium (Ca²⁺) signaling is essential for oocyte maturation, activation, and fertilization, with repetitive Ca²⁺ transients elicited at fertilization being critical for egg activation and early embryonic development. Mouse oocytes express several non-selective cation channels to support these oscillations, including TRPV3, a member of the transient receptor potential (TRP) channel family. In addition to Ca²⁺, TRPV3 mediates zinc (Zn²⁺) influx, which is a modulator of cortical granules (CGs) distribution and actin organization. In mammals, CG exocytosis mediates the fertilization-induced block to polyspermy, and pharmacological activation of TRPV3 in mouse oocytes elicits Ca²⁺ influx sufficient to trigger activation and parthenogenesis. Despite these critical roles in mice, the expression and function of TRPV3 in other mammals remain unexplored. Here, we evaluate the functional expression of TRPV3 channels in cat oocytes.</p><p><strong>Methods: </strong>Ovaries from domestic cats were obtained during ovariohysterectomies, and oocytes were isolated and matured in vitro. Trpv3 expression was assessed by RT-PCR from ovaries and germinal vesicle (GV) and metaphase II (MII) oocytes, while TRPV3 localization was evaluated by immunocytochemistry. Mouse WT and TRPV3-knockout oocytes were used as controls for antibody specificity. Functional channel activity was examined using Ca²⁺ imaging following addition of the TRPV3 agonist 2-APB. Three-dimensional modelling, molecular docking, and comparative sequence analysis of feline, mouse, and human TRPV3 proteins were aligned with MAFFT, focusing on identical and biochemically similar residues within the 2-APB-binding sites as well as pore-forming and temperature-sensing domains to assess potential species-specific functional differences.</p><p><strong>Results: </strong>We detected Trpv3 transcripts in cat ovaries and in GV and MII oocytes. Immunocytochemistry confirmed TRPV3 protein localization at the oocyte membrane in cats, consistent with reports in mouse oocytes. Additionally, addition of 2-APB elicited robust intracellular increase in Ca²⁺ in MII cat eggs, demonstrating functional TRPV3 channel activity. Comparative analyses revealed non-conservative substitutions in feline TRPV3 compared to mouse and human TRPV3, particularly within the pore-forming, channel gating, and temperature sensing regions, offering a molecular explanation for species-specific differences in TRPV3 function.</p><p><strong>Conclusions: </strong>Our results demonstrate functional TRPV3 expression in domestic cat oocytes. We find distinctive features in feline TRPV3 compared to rodent and human orthologs. These insights support the development of tailored artificial oocyte activation protocols in cats, with potential applications to Assisted Reproductive Technologies (ART) for endangered felids.</p>","PeriodicalId":9084,"journal":{"name":"Biological Research","volume":" ","pages":""},"PeriodicalIF":5.4,"publicationDate":"2026-07-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148395854","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Molecular insights into the protective role of nano-naringin in mancozeb-induced testicular damage and infertility.","authors":"Eman I Hassanen, Sara S Elbagwry, Zienab E Eldin, Rehab A Azouz, Marwa A Ibrahim, Rawhia Doghaim","doi":"10.1186/s40659-026-00712-y","DOIUrl":"10.1186/s40659-026-00712-y","url":null,"abstract":"<p><p>Mancozeb (MZ) is an extensively used fungicide with well-documented reproductive toxicity induced by oxidative stress and hormonal disruption. Antioxidants such as naringin (NAR) have been studied as protective agents, but their therapeutic application is limited by poor bioavailability. Nano-formulated approaches, including chitosan-coated nanoparticles (NARNPs), offer a promising strategy to enhance delivery and efficacy. The present study explores this concept by evaluating the protective effects of NAR and NARNPs against MZ-induced testicular damage and infertility in rats. 42 adult male albino Wistar rats were divided into six groups (n = 7) as follows: (1) control, (2) NAR (20 mg/kg bwt), (3) NARNPs (20 mg/kg bwt), (4) MZ (250 mg/kg bwt), (5) NAR + MZ, and (6) NARNPs + MZ. Data revealed that the exposure of rats to MZ caused a significant decrease in the body weight gain, sperm viability and motility, serum testosterone level, and testicular antioxidant enzyme activities, along with an increase in the lipid peroxidation. MZ also inhibited the expression of steroidogenic genes and proliferating markers in testes. The administration of NAR with MZ partially restored oxidative homeostasis and the reproductive biomarkers. Otherwise, NARNPs were more effective than NAR, which substantially improved the testicular morphology and function, as well as partially returned the gene expression back to normal with higher immunoexpression of proliferating markers in testicular tissues. The study highlights the potential of nano-formulated NAR as a promising strategy to counteract MZ-induced reproductive damage and improve testicular health via enhancing steroidogenic gene expression.</p>","PeriodicalId":9084,"journal":{"name":"Biological Research","volume":" ","pages":""},"PeriodicalIF":5.4,"publicationDate":"2026-07-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13340092/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148395834","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Telomerase as a therapeutic Bullseye: advances in cancer treatment and clinical trials : A comprehensive review of small molecule inhibitors, antisense oligonucleotides, and immunotherapeutic strategies targeting the telomerase catalytic subunit (hTERT).","authors":"Ansar Hussain, Musavir Abbas, Ming Long, Ghulam Mustafa, Hafiz Muhammad Yasir, Tanveer Abbas, Yousaf Raza, Muhammad Lateef, Muhammad Shoaib, Zain Ul Abideen, Wasim Shah","doi":"10.1186/s40659-026-00715-9","DOIUrl":"https://doi.org/10.1186/s40659-026-00715-9","url":null,"abstract":"<p><p>Telomeres play a crucial role in maintaining genomic stability in healthy cells. However, they gradually shorten during the cell cycle, leading to chromosomal instability. Telomere length and telomerase activity are vital factors that counteract cellular degradation in cancer development and tumor persistence. Telomerase, which is activated in most cancer cells due to telomerase catalytic subunit (hTERT) overexpression, serves as a universal biomarker that is essential for cancer cell growth and survival. The upregulation of hTERT, often associated with G > A mutations in its promoter region, is frequently implicated in cancer progression. Consequently, anti-telomerase therapy has been proposed as a potentially more efficacious alternative to conventional treatment. Small-molecule inhibitors have garnered significant attention owing to their selectivity or ability to modulate multiple proteins. However, challenges, such as low response rates, brief response durations, toxicity, and resistance persist. Several strategies have been proposed to target telomerase activity and the telomere structure. These include the utilization of G-quadruplex-stabilizing compounds and telomere-specific oligonucleotide inhibitors of telomerase such as GRN163L and T-oligos. Another therapeutic approach involves the use of biological antisense oligonucleotides that specifically inhibit hTERT and human telomerase RNA component genes, potentially reducing telomerase activity and generating robust DNA signals in cancer cells. Immunotherapy targeting hTERT represents a recent advancement in cancer treatments. This approach leverages the immune system to target cancer cells with high hTERT expression, thereby offering a potentially more reliable treatment strategy. This review provides an overview of current research on telomerase-targeting small-molecule inhibitors, antisense oligonucleotides, and immunotherapy, discussing their mechanisms, clinical applications, and prospects in cancer treatment.</p>","PeriodicalId":9084,"journal":{"name":"Biological Research","volume":" ","pages":""},"PeriodicalIF":5.4,"publicationDate":"2026-07-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148381669","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Tissue-specific reference gene stability for qPCR in thermally conditioned quail embryos.","authors":"Mohd Matin Ansari, K Martina, Gautham Kolluri, Jagbir Singh Tyagi, Jaydip Rokade, Prashant Bhutekar, Adnan Naim, Munish Gangwar, Sheikh Firdous Ahmad","doi":"10.1186/s40659-026-00714-w","DOIUrl":"https://doi.org/10.1186/s40659-026-00714-w","url":null,"abstract":"<p><p>The reliability and accuracy of relative quantification of transcripts through quantitative real-time PCR (qPCR) is dependent on the use of stable reference genes for normalization. However, the expression stability of various reference genes may differ across the tissues and patho-physiological conditions. The present study investigated the stability of multiple candidate reference genes in Japanese quail (Coturnix japonica) embryos, subjected to embryonic thermal conditioning (ETC). Fertilized quail eggs (n = 200) were incubated under standard conditions and heat exposure (ETC) of incubated eggs was undertaken at 39.5 °C for 10 h daily in the embryonic days 6-8 and 12-14, maintaining constant humidity (55%). Four embryos were randomly selected from each group for experimentation. The expression levels of eight candidate reference genes i.e., 18 S rRNA, β-ACTIN, 28 S rRNA, HPRT1, GAPDH, PGK1, RPS8, and TBP were assessed across four tissues (liver, intestine, brain, and heart) under ETC. Gene expression stability was assessed using five different algorithms: geNorm, NormFinder, BestKeeper, Delta Ct, and RefFinder (Comprehensive gene stability). Variations in gene stability were evident across different tissues and algorithms. The HPRT1, GAPDH and RPS8 emerged as the most stable reference genes for qPCR analyses in liver tissue, making them suitable internal controls. Similarly, PGK1, RPS8, and HPRT1 genes exhibited higher expression stability in intestine. On the other hand, TBP and HPRT1 were identified as the most reliable reference genes for normalization in heart tissue, whereas HPRT1 and GAPDH were found to be the most reliable reference genes in brain tissue. On considering all tissues combined (liver, intestine, brain, and heart), TBP and RPS8 emerged as the most suitable gene set for normalization. This present study provides crucial insights into selecting appropriate reference genes for relative expression quantification studies in quail embryos under heat treatment (ETC), thereby improving the reliability and accuracy of qPCR data in avian developmental and stress physiology research.</p>","PeriodicalId":9084,"journal":{"name":"Biological Research","volume":" ","pages":""},"PeriodicalIF":5.4,"publicationDate":"2026-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148366864","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Phenotyping of post-fertilization sperm mitophagy determinants discovered in a mammalian gamete-based cell-free system.","authors":"Alexis Jones, Natálie Zelenková, Erica Mantle, Chloe Gardner, Barbora Klusáčková, Dalen Zuidema, Miriam Sutovsky, Pavla Postlerová, Michal Zigo, Peter Sutovsky","doi":"10.1186/s40659-026-00713-x","DOIUrl":"https://doi.org/10.1186/s40659-026-00713-x","url":null,"abstract":"<p><p>The targeted, substrate-specific degradation of paternal mitochondria inside the zygote, known as post-fertilization sperm mitophagy, is a crucial and evolutionarily conserved early embryonic event. It ensures the exclusive maternal inheritance of the mitochondrial genome. Post-fertilization sperm mitophagy was initially thought to only be achieved via the ubiquitin-proteasome system. Until pro-autophagic receptor proteins such as SQSTM1, GABARAP, as well as the proteasome-interacting ubiquitinated protein dislocase VCP, were identified as contributors to the degradation of the sperm mitochondria early after mammalian fertilization. This synergy of proteasomal and autophagic pathways ensures a timely degradation of sperm mitochondria shortly after fertilization. The discovery of these autophagic receptors lead researchers to believe there might be other autophagic receptors and determinants necessary for proper post-fertilization sperm mitophagy. Based on the established inventory of proteins from mass spectrometry trials of boar spermatozoa exposed to porcine oocyte extracts in an intra-specific porcine cell-free system (CFS), five candidate mitophagy determinants were further investigated in this study, namely LACTB, PRDX3, PSMA8, TOMM34, and FUNDC1. These proteins of interest were studied and validated by using in vitro fertilization (IVF) protocols, cell imaging of spermatids, spermatozoa, oocytes and zygotes, protein interactome analysis, and the porcine CFS. The proteins PSMA8 and TOMM34 behaved in accordance with our proteomic study predictions. The PSMA8 labeling increased after exposure to CFS; in agreement with the classification PSMA8 was given from the mass spectrometry findings. TOMM34 underwent a visible decrease in labeling after exposure to CFS, which also agreed with its proteomic classification; this labeling persisted in IVF zygotes. Except for LACTB, the examined proteins showed mutual interactions as well as interactions with previously identified sperm mitophagy factors in the STRING interactome analysis. Results from this study validate the novel porcine CFS as a valuable tool for the exploration of early fertilization events at a molecular level. Future phenotyping and functional studies using porcine CFS will advance the understanding of mitochondrial inheritance and zygotic development and potentially shed light on the origins of certain mitochondrial diseases arising from the failure of post-fertilization sperm mitophagy.</p>","PeriodicalId":9084,"journal":{"name":"Biological Research","volume":" ","pages":""},"PeriodicalIF":5.4,"publicationDate":"2026-06-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148343882","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}