{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,5]],"date-time":"2026-06-05T05:01:10Z","timestamp":1780635670108,"version":"3.54.1"},"reference-count":34,"publisher":"MDPI AG","issue":"6","license":[{"start":{"date-parts":[[2026,6,1]],"date-time":"2026-06-01T00:00:00Z","timestamp":1780272000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["CIMB"],"abstract":"<jats:p>The brown frog (Rana dybowskii) is an important cold-temperate amphibian species in northeastern China, with considerable ecological and resource value. Because its survival, metabolism, and reproduction are highly sensitive to environmental temperature, elucidating the molecular mechanism\u2019s underlying temperature response is of great significance for understanding environmental adaptation and reproductive regulation in amphibians. In this study, R. dybowskii was used as the experimental model, and a multi-tissue transcriptomic analysis was conducted on the brain, liver, spleen, ovary, oviduct, and testis from both females and males under 2 \u00b0C and 12 \u00b0C conditions to characterize tissue- and sex-specific responses to temperature variation. The results showed that the global transcriptomic landscape of R. dybowskii was primarily driven by tissue type, whereas temperature effects were mainly manifested within individual tissues. The liver, brain, and spleen exhibited pronounced temperature responsiveness in both sexes, with the liver mainly showing metabolic reprogramming-related functional changes. Intersection analysis further revealed that temperature-responsive genes included not only conserved modules shared across tissues but also a large number of tissue-specific genes. Reproductive tissues displayed more pronounced functional divergence: the ovary showed relatively limited transcriptional changes and remained comparatively stable; the oviduct underwent marked transcriptional remodeling, with upregulated genes mainly involved in glycosylation and macromolecule processing, whereas downregulated genes were primarily associated with ribosomes, translation, and RNA splicing; the testis was likewise highly sensitive to temperature changes, with upregulated genes mainly enriched in amino acid metabolism, mitochondrial function, and DNA repair, while downregulated genes were mainly related to cellular structure and intercellular junctions. Collectively, these findings indicate that the molecular adaptation of R. dybowskii to temperature variation is characterized by strong tissue specificity and sex-biased responses in reproductive tissues. This study provides a transcriptomic basis for elucidating the molecular responses to short-term temperature variation is important for understanding environmental sensitivity and reproductive regulation in amphibians.<\/jats:p>","DOI":"10.3390\/cimb48060583","type":"journal-article","created":{"date-parts":[[2026,6,3]],"date-time":"2026-06-03T02:37:26Z","timestamp":1780454246000},"page":"583","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Multi-Tissue Transcriptomic Analysis Reveals Tissue-Specific Thermal Responses and Sex-Biased Functional Differentiation in Reproductive Tissues of the Brown Frog (Rana dybowskii)"],"prefix":"10.3390","volume":"48","author":[{"given":"Shifang","family":"Zhang","sequence":"first","affiliation":[{"name":"Institute of Natural Resources and Ecology, Heilongjiang Academy of Sciences, Harbin 150040, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4437-0797","authenticated-orcid":false,"given":"Guo","family":"Hu","sequence":"additional","affiliation":[{"name":"Heilongjiang River Fisheries Research Institute, Chinese Academy of Fishery Sciences, Harbin 150070, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yuan","family":"Guo","sequence":"additional","affiliation":[{"name":"Institute of Natural Resources and Ecology, Heilongjiang Academy of Sciences, Harbin 150040, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xudong","family":"Chang","sequence":"additional","affiliation":[{"name":"Institute of Natural Resources and Ecology, Heilongjiang Academy of Sciences, Harbin 150040, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Haijun","family":"Li","sequence":"additional","affiliation":[{"name":"Institute of Natural Resources and Ecology, Heilongjiang Academy of Sciences, Harbin 150040, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yingzhu","family":"Liu","sequence":"additional","affiliation":[{"name":"Institute of Natural Resources and Ecology, Heilongjiang Academy of Sciences, Harbin 150040, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,6,1]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"13","DOI":"10.1002\/jez.2396","article-title":"The thermal ecology and physiology of reptiles and amphibians: A user\u2019s guide","volume":"335","author":"Taylor","year":"2021","journal-title":"J. Exp. Zool. Part A Ecol. Integr. Physiol."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"2373","DOI":"10.1111\/gcb.12240","article-title":"Temperature variation makes ectotherms more sensitive to climate change","volume":"19","author":"Paaijmans","year":"2013","journal-title":"Glob. Change Biol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"20220132","DOI":"10.1098\/rstb.2022.0132","article-title":"Heat stress and amphibian immunity in a time of climate change","volume":"378","year":"2023","journal-title":"Philos. Trans. R. Soc. Lond. B Biol. Sci."},{"key":"ref_4","doi-asserted-by":"crossref","unstructured":"Nitzan, T., Kokou, F., Doron-Faigenboim, A., Slosman, T., Biran, J., Mizrahi, I., Zak, T., Benet, A., and Cnaani, A. (2019). Transcriptome analysis reveals common and differential response to low temperature exposure between tolerant and sensitive blue tilapia (Oreochromis aureus). Front. Genet., 10.","DOI":"10.3389\/fgene.2019.00100"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"24069","DOI":"10.1038\/srep24069","article-title":"Comparative transcriptome analyses of seven anurans reveal functions and adaptations of amphibian skin","volume":"6","author":"Huang","year":"2016","journal-title":"Sci. Rep."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"281","DOI":"10.3390\/d2020281","article-title":"Direct and indirect effects of climate change on amphibian populations","volume":"2","author":"Blaustein","year":"2010","journal-title":"Diversity"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"3","DOI":"10.1186\/s40851-023-00201-0","article-title":"A review of the reproductive system in anuran amphibians","volume":"9","year":"2023","journal-title":"Zool. Lett."},{"key":"ref_8","unstructured":"Exbrayat, J.-M. (2009). Spermatogenesis and Male Reproductive System in Amphibia\u2014Gymnophiona. Reproduction of Amphibians, CRC Press."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"2632","DOI":"10.1016\/j.bbagen.2018.07.032","article-title":"Transcriptome analyses reveal molecular mechanisms that regulate endochondral ossification in amphibian Bufo gargarizans during metamorphosis","volume":"1862","author":"Bo","year":"2018","journal-title":"Biochim. Biophys. Acta Gen. Subj."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"13","DOI":"10.1186\/s12977-025-00669-y","article-title":"Transcriptome mining reveals diversity and evolution of circulating and endogenous amphibian retroviruses","volume":"22","author":"Harding","year":"2025","journal-title":"Retrovirology"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"78","DOI":"10.1016\/j.theriogenology.2022.03.006","article-title":"Expression of glycogenic genes in the oviduct of Chinese brown frog (Rana dybowskii) during pre-brumation","volume":"185","author":"Fan","year":"2022","journal-title":"Theriogenology"},{"key":"ref_12","first-page":"1859","article-title":"Cloning and tissue expression analysis of the LepROT gene of Rana dybowskii","volume":"38","author":"Wu","year":"2022","journal-title":"Chin. J. Biotechnol."},{"key":"ref_13","doi-asserted-by":"crossref","unstructured":"Xu, Y., Cui, S., Li, T., Du, Z., Ning, F., Jiang, H., Bai, X., Wang, X., and Bao, J. (2022). Testicular Transcriptome of Males and Pseudo-Males Provides Important New Insight into Sex Reversal of Rana dybowskii. Animals, 12.","DOI":"10.3390\/ani12202887"},{"key":"ref_14","doi-asserted-by":"crossref","unstructured":"Xu, Y., Liu, H., Jiang, X., Zhang, X., Liu, J., Tian, Y., Bai, X., Cui, S., and Di, S. (2024). Genome Survey of Male Rana dybowskii to Further Understand the Sex Determination Mechanism. Animals, 14.","DOI":"10.3390\/ani14202968"},{"key":"ref_15","doi-asserted-by":"crossref","unstructured":"Su, H., Zhang, H., Wei, X., Pan, D., Jing, L., Zhao, D., Zhao, Y., and Qi, B. (2018). Comparative Proteomic Analysis of Rana chensinensis Oviduct. Molecules, 23.","DOI":"10.3390\/molecules23061384"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"2114","DOI":"10.1093\/bioinformatics\/btu170","article-title":"Trimmomatic: A flexible trimmer for Illumina sequence data","volume":"30","author":"Bolger","year":"2014","journal-title":"Bioinformatics"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"907","DOI":"10.1038\/s41587-019-0201-4","article-title":"Graph-based genome alignment and genotyping with HISAT2 and HISAT-genotype","volume":"37","author":"Kim","year":"2019","journal-title":"Nat. Biotechnol."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"923","DOI":"10.1093\/bioinformatics\/btt656","article-title":"featureCounts: An efficient general purpose program for assigning sequence reads to genomic features","volume":"30","author":"Liao","year":"2014","journal-title":"Bioinformatics"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"550","DOI":"10.1186\/s13059-014-0550-8","article-title":"Moderated estimation of fold change and dispersion for RNA-seq data with DESeq2","volume":"15","author":"Love","year":"2014","journal-title":"Genome Biol."},{"key":"ref_20","first-page":"e62528","article-title":"Three Differential Expression Analysis Methods for RNA Sequencing: Limma, EdgeR, DESeq2","volume":"175","author":"Liu","year":"2021","journal-title":"J. Vis. Exp."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"284","DOI":"10.1089\/omi.2011.0118","article-title":"clusterProfiler: An R package for comparing biological themes among gene clusters","volume":"16","author":"Yu","year":"2012","journal-title":"Omics"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"3844","DOI":"10.1093\/bioinformatics\/btac409","article-title":"ggtranscript: An R package for the visualization and interpretation of transcript isoforms using ggplot2","volume":"38","author":"Gustavsson","year":"2022","journal-title":"Bioinformatics"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"103355","DOI":"10.1016\/j.jtherbio.2022.103355","article-title":"Effects of acute heat exposure on oxidative stress and antioxidant defenses in overwintering frogs, Nanorana parkeri","volume":"110","author":"Zhang","year":"2022","journal-title":"J. Therm. Biol."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"2937","DOI":"10.3390\/cimb45040192","article-title":"Potential effects of high temperature and heat wave on Nanorana pleskei Based on transcriptomic analysis","volume":"45","author":"Zhang","year":"2023","journal-title":"Curr. Issues Mol. Biol."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"93","DOI":"10.1086\/282976","article-title":"Temperature adaptations in amphibians","volume":"109","author":"Snyder","year":"1975","journal-title":"Am. Nat."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"9","DOI":"10.1093\/icb\/icy005","article-title":"High temperature, oxygen, and performance: Insights from reptiles and amphibians","volume":"58","author":"Gangloff","year":"2018","journal-title":"Integr. Comp. Biol."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"60","DOI":"10.1046\/j.1365-2435.2001.00495.x","article-title":"Allometric and ecological relationships of ventricle and liver mass in anuran amphibians","volume":"15","author":"Withers","year":"2001","journal-title":"Funct. Ecol."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"jeb186379","DOI":"10.1242\/jeb.186379","article-title":"Effects of both cold and heat stress on the liver of the giant spiny frog (Quasipaa spinosa): Stress response and histological changes","volume":"221","author":"Liu","year":"2018","journal-title":"J. Exp. Biol."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"499","DOI":"10.1002\/(SICI)1096-9861(19970616)382:4<499::AID-CNE6>3.0.CO;2-Y","article-title":"Distribution of choline acetyltransferase immunoreactivity in the brain of anuran (Rana perezi, Xenopus laevis) and urodele (Pleurodeles waltl) amphibians","volume":"382","author":"Smeets","year":"1997","journal-title":"J. Comp. Neurol."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"1205","DOI":"10.1007\/s12192-018-0926-x","article-title":"Stress-induced antioxidant defense and protein chaperone response in the freeze-tolerant wood frog Rana sylvatica","volume":"23","author":"Wu","year":"2018","journal-title":"Cell Stress Chaperones"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"R545","DOI":"10.1152\/ajpregu.1996.271.3.R545","article-title":"Oxidative damage and antioxidants in Rana sylvatica, the freeze-tolerant wood frog","volume":"271","author":"Joanisse","year":"1996","journal-title":"Am. J. Physiol. Regul. Integr. Comp. Physiol."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"6350","DOI":"10.1111\/gcb.15321","article-title":"Future winters present a complex energetic landscape of decreased costs and reduced risk for a freeze-tolerant amphibian, the Wood Frog (Lithobates sylvaticus)","volume":"26","author":"Fitzpatrick","year":"2020","journal-title":"Glob. Change Biol."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"e02788","DOI":"10.1002\/ecs2.2788","article-title":"Modeling the distribution of niche space and risk for a freeze-tolerant ectotherm, Lithobates sylvaticus","volume":"10","author":"Fitzpatrick","year":"2019","journal-title":"Ecosphere"},{"key":"ref_34","doi-asserted-by":"crossref","unstructured":"Forz\u00e1n, M.J., and Horney, B.S. (2020). Amphibians. Exotic Animal Laboratory Diagnosis, John Wiley & Sons, Inc.","DOI":"10.1002\/9781119108610.ch19"}],"container-title":["Current Issues in Molecular Biology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1467-3045\/48\/6\/583\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,6,5]],"date-time":"2026-06-05T04:21:45Z","timestamp":1780633305000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1467-3045\/48\/6\/583"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,6,1]]},"references-count":34,"journal-issue":{"issue":"6","published-online":{"date-parts":[[2026,6]]}},"alternative-id":["cimb48060583"],"URL":"https:\/\/doi.org\/10.3390\/cimb48060583","relation":{},"ISSN":["1467-3045"],"issn-type":[{"value":"1467-3045","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,6,1]]}}}