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Telomerase is highly expressed in oocyte cumulus cells and plays a significant role in follicular development and oocyte maturation. In this study, we hypothesized that in vitro culture conditions may affect telomerase activity during in vitro embryo production (IVP) and that its activation may improve embryo quality. We first examined telomerase protein levels and localization in bovine cumulus-oocyte complexes via immunofluorescence assays. The results showed that healthy cumulus-oocyte complexes have the nuclear localization of the telomerase while the degraded cumulus-oocyte complex had reduced telomerase levels and that telomerase was localized in the cytoplasm. We activated telomerase via Epitalon, a tetrapeptide with the amino acid sequence Ala-Glut-Asp-Gly. We observed a significant improvement in the oocyte maturation rate compared with the control group (p < 0.05). Furthermore, telomerase activity was significantly compromised in post-thawed embryos, and Epitalon treatment significantly improved blastocyst hatching rate and implantation potential (p < 0.05). Moreover, we performed qPCR, reactive oxygen species, and JC-1 (ΔΨm) assays to evaluate the effect of Epitalon on the health of in vitro mature oocytes, cumulus cells, and post-thawed blastocysts, and the result showed that Epitalon highly enhances the quality and health of the oocyte, cumulus cell, and post-thawed blastocyst. Our results suggest that telomerase activation via Epitalon improves bovine in vitro embryo production., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Kong Il Keun reports financial support was provided by National Research Foundation of Korea. Il Keun Kong reports a relationship with Gyeongsang National University that includes: BK21 four and school of Agriculture and Life Science. No conflict of interest between the co-authors. If there are other authors, they declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2025 Elsevier Inc. All rights reserved.)

Extensive mitochondrial replication during oogenesis and its role in oocyte maturation and fertilization indicate that the amount of mitochondrial DNA (mtDNA) may play a significant role in early embryonic development. Early embryos express peroxisome proliferator-activated receptor gamma co-activator alpha (PPARGC1A/PGC-1a), a protein essential for mitochondrial biogenesis. This study investigated the role of PGC-1α from a single-cell zygotic stage to day-8 bovine blastocyst and the effect of PGC-1a knockdown (KD) on embryo mitochondria and development. PGC-1α KD via siRNA injection into single-cell zygotes does not substantially affect embryonic cleavage up to the morula stage but considerably reduces blastocyst development (18.42%) and hatching than the control (32.81%). PGC-1α regulates transcription of the gene encoding mitochondrial transcription factor A (TFAM), and immunofluorescence analysis indicated significantly lower TFAM expression in the 16-cell KD embryos and day-8 KD blastocysts. Reduction in NRF1 protein's nuclear localization in bovine blastomeres was also observed in PGC-1α-KD embryos. Furthermore, to understand the effect of PGC-1α-KD on the mitochondrial genome, we found a low mtDNA copy number in PGC-1α-KD day-8 bovine blastocysts. Several genes related to mitochondrial functioning, like ND1, ND3, ND5, ATPase8, COI, COII, and CYTB, were significantly downregulated in PGC-1α-KD embryos. Moreover, high mitochondrial depolarization (ΔΨm) and abnormal lipid depositions were observed in the PGC-1α KD blastocysts. SIRT1 is the upstream regulator of PGC-1α, but SIRT1 activation via Hesperetin does not affect PGC-1α-KD embryonic development considerably. In conclusion, PGC-1α plays a critical role in early embryo mitochondrial functioning, and any perturbation in its expression significantly disrupts early embryonic development., Competing Interests: Author I-KK was employed by The King Kong Corp. Ltd. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. The author(s) declared that they were an editorial board member of Frontiers, at the time of submission. This had no impact on the peer review process and the final decision., (Copyright © 2025 Idrees, Haider, Perera, Ullah, Lee, Lee, Kang, Kim and Kong.)

Heavy metals (HMs) have emerged as a critical global concern, not only limiting crop productivity but also posing risks to public health. Among them, nickel (Ni) is an essential micronutrient for plant growth; however, it becomes toxic at higher concentrations. Nano-enabled approaches, on the other hand, have emerged as promising eco-friendly alternatives for mitigating the negative impact associated with HMs. Here, we investigated the potential of titanium dioxide nanoparticles (TiO 2 NPs) against Ni-induced stress in maize. Our results showed that Ni stress caused negative changes in maize by the excessive production of reactive oxygen species (ROS), inhibiting photosynthetic attributes, and damaging cellular ultrastructure. In contrast, TiO 2 NPs priming significantly enhanced the antioxidant mechanism, photosynthetic efficacy, and nutrient uptake while reducing ultrastructural damage caused by Ni stress. Furthermore, TiO 2 NPs efficiently reduced Ni accumulation, MDA (28%/32%), H 2 O 2 (23%/26%), and O 2 •‒ (31%/34%) levels in shoot/root tissues, respectively, compared to Ni treatment. Moreover, TiO 2 NPs priming has modulated the expression of antioxidant and defense-related genes, thereby restoring cellular redox homeostasis. Collectively, this is the first piece of evidence demonstrating the potential of TiO 2 NPs as an efficient and sustainable alternative for enhancing crop tolerance in Ni-contaminated areas., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)