Cryobiology最新文献

{"title":"Effect of temperature on acute kidney injury in a rat model of renal ischemia‒reperfusion","authors":"Jing Mei ,&nbsp;Minhui Xie,&nbsp;Congcong Dai,&nbsp;Yuhong Tang,&nbsp;Na Huang,&nbsp;Jun Ou","doi":"10.1016/j.cryobiol.2025.105276","DOIUrl":"10.1016/j.cryobiol.2025.105276","url":null,"abstract":"<div><div>Renal ischemia-reperfusion (I/R) injury is the main cause of acute kidney injury (AKI). A reliable and stable animal model is crucial for experimental research. Various factors affect the stability of animal models. In the present study, the effect of intraoperative temperature on the establishment of AKI model in rats, induced by I/R through bilateral renal pedicles clamping via the dorsal approach surgery, was investigated. Forty-eight male Sprague Dawley rats of 8–10 weeks were randomly divided into 6 groups (8 per group). The renal ischemia-reperfusion injury model was induced using a retroperitoneal double pedicle clamp procedure, using a clamping time of 50 min. The modeling procedure and sham operation were performed at different ambient air temperature, i.e., 15–20 °C, 20–25 °C and 25–30 °C. The general state of the rats, the ascites volume and mortality were observed after the operation. The serum creatinine (Scr) and urea nitrogen (BUN) levels of rats in each group were measured by an automatic biochemical analyzer 24 h post-operation. HE staining was used to observe pathological changes in renal tissues, and NF-κB p65 and interleukin-6 (IL-6) expression was measured by immunohistochemistry (IHC). TUNEL staining was used to assess apoptosis in renal tissues, and the expression of inflammatory markers, such as NF-κB and IL-6, and the apoptosis-related protein Bax was measured by Western blotting. There were no significant differences among the different sham operation groups. However, the abovementioned parameters were greater in the hypothermia, normothermia and hyperthermia model groups than in the corresponding sham operation groups, indicating successful model establishment. Scr and BUN levels were greater in the hyperthermia model group than in the other model groups, and there was obvious pathological damage to renal tissue. Notably, in the hypothermia model group, the general state of rats was good, Scr and BUN levels were not significantly increased 24 h post-operation, pathological damage to renal tissue and renal tubular epithelial cell apoptosis were mild, NF-κB and IL-6 protein expression was increased, and Bax protein expression was decreased. In conclusion, the induction of AKI by I/R at 20–25 °C has a high successful modeling rate and a low mortality rate. It may affect the stability of animal models by down-regulating the NF-κB/IL-6 signaling pathway and the NF-κB/Bax pathway through its anti-apoptotic and anti-inflammatory effects.</div></div>","PeriodicalId":10897,"journal":{"name":"Cryobiology","volume":"120 ","pages":"Article 105276"},"PeriodicalIF":2.3,"publicationDate":"2025-06-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144279590","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Supplemental effect of metallic nanoparticles on cryopreserved semen quality, lipid peroxidation, sperm head ultrastructure and field fertility levels in crossbred (Hampshire x Niang Megha) boars","authors":"Manoj Kumar Kalita ,&nbsp;Prithviraj Mazumdar Baruah ,&nbsp;Kutubuddin Ahmed ,&nbsp;Sudip Sinha ,&nbsp;Shantanu Tamuly ,&nbsp;Sourabh Deori ,&nbsp;Rahul Katiyar ,&nbsp;Sayed Nabil Abedin ,&nbsp;Prerona Patowary ,&nbsp;Raju Dewry ,&nbsp;Jitumoni Das ,&nbsp;Biplab Sarkar ,&nbsp;Gautam Khargharia ,&nbsp;Govindasamy Kadirvel","doi":"10.1016/j.cryobiol.2025.105277","DOIUrl":"10.1016/j.cryobiol.2025.105277","url":null,"abstract":"<div><div>Reactive oxygen species (ROS) produced by sperm contributes to oxidative stress (OS), leading to a decline in sperm quality. Different antioxidants have shown potential in preventing such damage. The present study explored the effect of metallic nanoparticle (NPs) supplementation on sperm quality in cryopreserved boar spermatozoa. Forty-two (42) ejaculates (7 ejaculates per animal) were collected from six (6) physically and sexually healthy and fertile Lumsniang (Hampshire x Niang Megha) crossbred boars, aged 18–42 months and then diluted in Beltsville thawing solution (BTS) extender containing the different treatments (CON: no NP addition; 10 μM ZnO NPs, 1 μg Se NPs and 0.192 mg Fe₃O₄ NPs per mL of semen). Semen was diluted, packed into 0.5 mL French medium straws, and subsequently frozen in liquid nitrogen (LN₂). Sperm quality, lipid peroxidation (LPO), antioxidant enzyme levels, ultrastructural sperm head morphology and field fertility levels were determined. Significantly (p &lt; 0.05) higher sperm <em>in vitro</em> quality attributes were recorded in the zinc oxide (ZnO) and selenium (Se) NP treatments in comparison to CON whereas iron-oxide (Fe₃O₄) NP treatment significantly (p &lt; 0.05) lowered sperm quality. Post-thaw LPO levels significantly (p &lt; 0.05) decreased in the ZnO and Se NP groups when compared to CON. LPO levels were significantly (p &lt; 0.05) higher in the Fe₃O₄ NP group. The ZnO and Se NP treatments showed significantly (p &lt; 0.05) higher post-thaw antioxidant enzyme levels. Ultrastructurally, Group 1 spermatozoa were significantly (p &lt; 0.05) higher and Group 2 were significantly (p &lt; 0.05) lower in the ZnO and Se NP supplemented group in comparison to CON. No significant (p &gt; 0.05) difference was observed in the <em>in vivo</em> pregnancy rate; however, the ZnO and Se NP treatments significantly (p &lt; 0.05) enhanced the average litter sizes at birth and weaning compared to CON. In conclusion, 10 μM ZnO and 1 μg Se NPs improved post-thaw quality of boar spermatozoa and safeguarded the ultrastructure of their membranes. On the other hand, 0.192 mg Fe₃O₄ NPs had deleterious effects on sperm <em>in vitro</em> quality.</div></div>","PeriodicalId":10897,"journal":{"name":"Cryobiology","volume":"120 ","pages":"Article 105277"},"PeriodicalIF":2.3,"publicationDate":"2025-06-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144254748","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Rat one-cell embryo vitrification in F344, Long-Evans, and SD strain via small-volume vitrification and rapid warming in cryotubes","authors":"Shinsuke Seki ,&nbsp;Toshiaki Kawabe ,&nbsp;Kazuaki Matsumura ,&nbsp;Misako Higashiya ,&nbsp;Takanori Oikawa ,&nbsp;Yuriko Fujii ,&nbsp;Megumi Yano ,&nbsp;Wataru Yamazaki ,&nbsp;Tomoo Eto","doi":"10.1016/j.cryobiol.2025.105260","DOIUrl":"10.1016/j.cryobiol.2025.105260","url":null,"abstract":"<div><div>Genome-edited animals can be created by introducing CRISPR/Cas9 systems into one-cell stage embryos, even in non-mice embryos. We developed a vitrification method for rat one-cell embryos of the F344 inbred, Long-Evans, and SD strains. Successful cryopreservation requires the avoidance of intracellular ice formation (IIF). We hypothesized that a new cryopreservation method could be developed by rapid warming to avoid IIF during warming, via cryopreservation in conventional cryotubes using small-volume vitrification and rapid warming. When cryopreserved one-cell embryos in a cryotube with 15 μL cryopreservation solution (mixture of 5 μL 5 % propylene glycol and 10 μL PEPeS) were warmed by adding 1 ml of 0.3 M sucrose solution at 50 °C, they developed to term at 67.2 % (F344), 56.3 % (Long-Evans), and 65.0 % (SD), which were comparable with those (60.3 %, 58.3 %, 67.0 %, respectively) of non-cryopreserved control embryos. Thus, rat one-cell embryos from several strains can be cryopreserved even with cryotubes via rapid warming.</div></div>","PeriodicalId":10897,"journal":{"name":"Cryobiology","volume":"120 ","pages":"Article 105260"},"PeriodicalIF":2.3,"publicationDate":"2025-06-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144243323","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Development of an automated device for the optimization of oocyte and embryo vitrification","authors":"Haoyang Lin ,&nbsp;Chenxi Liu ,&nbsp;Yukun Cao ,&nbsp;Xinli Zhou","doi":"10.1016/j.cryobiol.2025.105275","DOIUrl":"10.1016/j.cryobiol.2025.105275","url":null,"abstract":"<div><div>Oocyte/embryo vitrification is one of the basic techniques employed in assisted reproduction and fertility preservation. However, the current process typically requires manual operation by experienced embryologists, which is both time-consuming and exhibits inconsistent outcomes. To resolve this issue, we herein developed an automated vitrification device for oocytes/embryos. The device consists of a microfluidic mixing unit, a microgrid capillary, and a mechanical sliding unit. The microfluidic mixing unit was adopted to determine continuous changes in cryoprotective agent (CPA) concentration, reducing osmotic damage during CPA loading/removal; and the microgrid capillary was used to load/remove the CPA and vitrify the oocytes/embryos in the same carrier so as to reduce cellular loss due to cell transfer. The medical absorbent dressing was placed under the carrier, and the excess liquid was absorbed to minimize the remaining CPA solution after CPA loading. Eight different loading/removal curves were developed for CPA loading and removal protocols, and we conducted oocyte vitrification with automated vitrification equipment. Our results revealed that a quadratic function-based, loading-removal curve achieved the highest oocyte survival rate within an 8-min loading and removal duration. In addition, there was no significant difference in oocyte survival rates between the automated vitrification device and the Cryotop multi-step equilibration method (90.33 % and 94.33 %, respectively); nor did the two methods differ in terms of survival or hatching rates of 8-cell embryos. The current system automates and standardizes the oocyte/embryo vitrification process while achieving survival and developmental potential.</div></div>","PeriodicalId":10897,"journal":{"name":"Cryobiology","volume":"120 ","pages":"Article 105275"},"PeriodicalIF":2.3,"publicationDate":"2025-06-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144243321","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Viewing heat through ice: An infrared camera monitors hydrogel freezing and thawing during cryoapplication","authors":"Gennadiy O. Kovalov ,&nbsp;Mykola O. Chyzh ,&nbsp;Vyacheslav Yu Globa ,&nbsp;Oleksandr F. Todrin ,&nbsp;Galyna V. Shustakova ,&nbsp;Eduard Yu Gordiyenko ,&nbsp;Yuliya V. Fomenko ,&nbsp;Oleh V. Ivakhnenko ,&nbsp;Polina O. Kofman ,&nbsp;Sergey N. Shevchenko","doi":"10.1016/j.cryobiol.2025.105259","DOIUrl":"10.1016/j.cryobiol.2025.105259","url":null,"abstract":"<div><div>Cryosurgery employs a safe and relatively simple technique of exposure and is an advantageous and highly rated method. For its effective application, it is necessary to control both the volume of the expanding freezing zone and volumetric thermal field dynamics. The aim of this study was to perform a thermal imaging study of freezing and thawing in a model system (gel phantom) to predict the dynamics of the freezing zone during cryodestruction of biological tissues in vivo. Here, the thermal imager is an effective tool for demonstrating the surface temperature distribution. We have studied how the observed infrared image relates to the distribution and change of the thermal field in depth. For this purpose, we created test measuring equipment for simultaneous analysis of the dynamics of thermal fields on the surface, video recording of freezing and thawing on the surface as well as in the depth of the gel phantom, measuring the temperature at any given point in the depth and modeling in the zone of low temperature exposure of vessels with different blood flow parameters. It was revealed that with a modelled vessel in the low-temperature exposure zone, the surface thermal fields deformed and they gained the shape of butterfly wings. Our experimental study in a gel phantom is supported by numerical calculations, demonstrating how the freezing zone and thermal isotherms on the surface and in depth evolve under real conditions, thereby providing a basis for assessing the cryoeffect time and intensity in practice.</div></div>","PeriodicalId":10897,"journal":{"name":"Cryobiology","volume":"120 ","pages":"Article 105259"},"PeriodicalIF":2.3,"publicationDate":"2025-06-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144243322","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"The antioxidant capacity and protective ability of astaxanthin in cryopreservation of mouse spermatogonial stem cells","authors":"Shiva Fathi ,&nbsp;Hassan Nazari ,&nbsp;Mehran Arabi ,&nbsp;Azita Afzali ,&nbsp;Ebrahim Ahmadi","doi":"10.1016/j.cryobiol.2025.105261","DOIUrl":"10.1016/j.cryobiol.2025.105261","url":null,"abstract":"<div><div>Cryopreservation of spermatogonial stem cells (SSCs) offers several benefits, but it can also cause various forms of damage that may reduce the functionality of these cells. Incorporating antioxidants into the cryopreservation medium can provide protection against the detrimental effects of cryopreservation by reducing levels of reactive oxygen species (ROS). In this study, the protective effect of astaxanthin (AST) was evaluated to establish an optimal cryopreservation method for SSCs obtained from the testes of neonatal male mice. AST was added to the freezing base medium at 1, 10, and 100 μM concentrations, and then compared with the control (freezing medium without any additives) and 100 μM vitamin E as a conventional antioxidant. Viability, oxidative stress status, intracellular ROS generation levels, and the expression of <em>Bax</em> and <em>Bcl2</em> were measured in frozen-thawed SSCs 3 weeks after culture and purification. The data showed that the presence of antioxidants, especially 10 μM AST, in the freezing medium significantly increases the viability and total antioxidant capacity (TAC) (P &lt; 0.05), and reduces the levels of lipid peroxidation and intracellular ROS accumulation in the frozen–thawed SSCs. Vitamin E, as well as 10 and 100 μM AST reduced apoptosis in mouse SSCs by downregulating <em>Bax</em> and upregulating <em>Bcl2</em>. The results of this study suggest that adding 10 μM of AST to the freezing medium provides protection to SSCs after thawing. Therefore, the inclusion of 10 μM AST in the cryopreservation medium significantly improves the post-thaw viability and antioxidant capacity of SSCs. These findings indicate that AST could serve as a valuable additive for improving the cryopreservation process of SSCs, thereby offering potential benefits for the preservation of male fertility.</div></div>","PeriodicalId":10897,"journal":{"name":"Cryobiology","volume":"120 ","pages":"Article 105261"},"PeriodicalIF":2.3,"publicationDate":"2025-06-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144220881","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Protein carbonylation reveals the molecular basis of cryoinjury in buffalo spermatozoa","authors":"S.K. Bhure ,&nbsp;D.S. Kumara Wodeyar ,&nbsp;Atul Kumar Sharma ,&nbsp;Abhishek Kumar ,&nbsp;Ajay Kumar ,&nbsp;Manish Mahawar ,&nbsp;S.K. Ghosh ,&nbsp;G. Taru Sharma","doi":"10.1016/j.cryobiol.2025.105271","DOIUrl":"10.1016/j.cryobiol.2025.105271","url":null,"abstract":"<div><div>During cryopreservation, oxidative damage to sperm biomolecules occurs, ultimately compromising its fertilizing ability. Simple proteome analysis may not accurately reflect the cellular functions that are affected, as most carbonylated proteins lose their functions. The carbonylated proteins act as a good marker of oxidative stress. Choosing buffalo spermatozoa as a model, carbonylated proteins from fresh-extended (FESCL) and frozen-thawed (FTSCL) spermatozoa were enriched by affinity chromatography using biotin hydrazide and a monomeric avidin agarose matrix followed by mass spectrometry. Data were analyzed with Proteome Discoverer (v2.2), post-translational modification analysis was performed using SEQUEST, and affected functional activities were predicted using the FunRich tool (v3.0). The mass spectrometric analysis of sperm proteins led to the identification of 415 carbonylated proteins, 151 in FESCL and 405 in FTSCL. Twelve highly abundant carbonylated proteins identified only in FTSCL can be the major contributors to the cryoinjury. The cryopreservation induces the carbonylation of selective sperm proteins. A total of 98 peptides have been precisely annotated that have oxidative modifications. In comparison to FESCL, more proteins in various cellular components, pathways, and processes in FTSCL were carbonylated. The findings indicate the molecular injury to spermatozoa through carbonylated proteins affecting energy metabolism, free-radical scavenging, cytoskeleton, plasma membrane, capacitation, zona-pellucida binding, and sperm-oocyte interaction and are well correlated with the functional parameters of the spermatozoa. The findings provide strong evidence that protein carbonylation is one of the major factors and molecular basis of cryodamage, which compromises the functions of FTSCL. With cryopreservation becoming an inevitable tool, the study becomes more relevant, and this experimental design would give better-quality frozen-thawed spermatozoa with good molecular profiles.</div></div>","PeriodicalId":10897,"journal":{"name":"Cryobiology","volume":"120 ","pages":"Article 105271"},"PeriodicalIF":2.3,"publicationDate":"2025-06-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144205258","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Ice ball size and temperature change during cryoablation in a lard and an ethiodized-oil tissue phantom","authors":"Nai-Wen Chang ,&nbsp;Shu-Huei Shen ,&nbsp;Chien-An Liu ,&nbsp;Felix Antony Halim ,&nbsp;Jia-An Hong ,&nbsp;Chih-Chien Li ,&nbsp;Ping-Lang Yen","doi":"10.1016/j.cryobiol.2025.105274","DOIUrl":"10.1016/j.cryobiol.2025.105274","url":null,"abstract":"<div><div>This in vitro study evaluates the effects of varying concentrations of lard and ethiodized-oil (Lipiodol) on temperature changes and ice ball diameters during cryotherapy. A phantom was constructed using six glass bottles, one filled with 0.9 % normal saline (NS) and the others containing agar phantoms mixed with different proportions of lard and NS (0 %, 10 %, 40 %, 70 %, and 100 % lard). Similarly, a phantom was prepared with ethiodized-oil. Six cryoprobes were inserted into the bottles, and freezing was initiated simultaneously, with temperature readings recorded every 10 s over a 9-min freezing period. CT scans were performed before freezing and at 3, 6, and 9 min post-freezing to measure ice ball diameters, with each phantom undergoing a single freezing cycle. Phantoms with lard or ethiodized-oil showed increased freezing rates with higher concentrations, stabilizing at approximately −140 to −150 °C. The largest ice ball diameters were observed in the 0 % and 10 % lard and ethiodized-oil phantoms. Negative correlations were identified between ice ball diameter and lard concentration at 3, 6, and 9 min (R² = 0.910, 0.838, 0.778; p = 0.008, 0.019, 0.030), as well as between ice ball diameter and ethiodized-oil concentration at 6 min (R² = 0.881; p = 0.040). These results suggest that distinct concentrations of lard and ethiodized-oil significantly influence the temporal temperature changes and ice ball dimensions during cryotherapy.</div></div>","PeriodicalId":10897,"journal":{"name":"Cryobiology","volume":"120 ","pages":"Article 105274"},"PeriodicalIF":2.3,"publicationDate":"2025-06-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144205257","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Corrigendum to \"Ex vivo use of a Rho-kinase inhibitor during renal preservation improves graft function upon reperfusion\" [Cryobiology 70 (2015) 71-75].","authors":"Carmen Kirchner, Bastian Lüer, Patrik Efferz, Jeremias Wohlschlaeger, Andreas Paul, Thomas Minor","doi":"10.1016/j.cryobiol.2025.105255","DOIUrl":"https://doi.org/10.1016/j.cryobiol.2025.105255","url":null,"abstract":"","PeriodicalId":10897,"journal":{"name":"Cryobiology","volume":" ","pages":"105255"},"PeriodicalIF":2.3,"publicationDate":"2025-05-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144181368","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Identification of the hub genes involved in ATF5 mediated stress response in vitrified mouse oocytes based on WGCNA","authors":"Guizhen Zhou ,&nbsp;Aiju Liu ,&nbsp;Jiachen Bai ,&nbsp;Yixiao Zhu ,&nbsp;Xinhua Zou ,&nbsp;Yuwen Luo ,&nbsp;Yizaitiguli Reyimjan ,&nbsp;Yan Ma ,&nbsp;Shuaixiang Huang ,&nbsp;Yunpeng Hou ,&nbsp;Jun Li ,&nbsp;Xiangwei Fu","doi":"10.1016/j.cryobiol.2025.105258","DOIUrl":"10.1016/j.cryobiol.2025.105258","url":null,"abstract":"<div><div>Activating transcription factor 5 (ATF5) is a key regulator in the stress response and plays a crucial role in cellular adaptation to the nonphysical environment. Our previous study indicated that a homeostatic state of ATF5 level could improve mitochondrial function in vitrified oocytes. However, the molecular mechanisms underlying the ATF5-mediated gene network in response to oocyte vitrification remain largely unknown. In this study, specific ATF5 siRNA was microinjected into oocytes to suppress ATF5 expression, and oocytes with ATF5 deficiency under normal and vitrification stress conditions were collected for Smart RNA-seq analysis. Weighted gene co-expression network analysis (WGCNA) was used to characterize oocyte co-expression modules involved in the ATF5 mediated response. Our results identified three key gene modules related to ATF5 knockdownRed, Green and, Yellow-using the screening criteria of |R| ≥0.5 and <em>P</em> ≤ 0.05 with WGCNA. Functional enrichment analysis of key gene modules showed that genes in the modules were mainly enriched in apoptosis, ubiquitin mediated proteolysis, the PI3K-Akt signaling pathway, and protein digestion and absorption. STEM and KEGG functional enrichment analysis showed that most genes were involved in protein digestion and absorption, the AMPK signaling pathway, the JAK-STAT signaling pathway, and the apoptosis signaling pathway. Importantly, <em>Diablo</em>, <em>Map3k1</em>, <em>Spta1,</em> and <em>Ubqln4</em>, obtained by sequential scanning of WGCNA and combined functional enrichment analysis, were identified as candidate genes under ATF5 regulation in response to vitrification. These findings demonstrate that the ATF5 mediated gene network exerts regulatory roles in various cellular events and provide novel insights for a deeper understanding of stress response associated impairments in vitrified oocytes.</div></div>","PeriodicalId":10897,"journal":{"name":"Cryobiology","volume":"119 ","pages":"Article 105258"},"PeriodicalIF":2.3,"publicationDate":"2025-05-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144107214","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}