Junjie Xu, Lei Zhang, Peng Zhang, Yanhong Su, Yuxia Gao
{"title":"The protective effects of retinoic acid-induced protein 14 on ischemia/reperfusion-induced myocardial apoptosis involves over-autophagy repression.","authors":"Junjie Xu, Lei Zhang, Peng Zhang, Yanhong Su, Yuxia Gao","doi":"10.1538/expanim.25-0006","DOIUrl":"https://doi.org/10.1538/expanim.25-0006","url":null,"abstract":"<p><p>Uncontrolled activation of autophagy following ischemia/reperfusion (I/R) injury leads to cell death. The superfamily of ankyrin repeat proteins (N-Ank protein) was reported to be involved in autophagy regulation and cardiac protection. Bioinformatics analysis was performed (GSE61592 and GSE160516) and ten N-Ank proteins were differentially expressed in I/R models. Retinoic acid-induced protein 14 (RAI14), a member of N-Ank protein family, was upregulated in I/R-injured cardiac tissue and was first selected for research. A mouse I/R model was established by ligating the left anterior descending coronary artery to induce 90 min of ischemia, followed by 72 h of reperfusion. RAI14 was found upregulated in ischemic penumbra. RAI14 overexpression in cardiac tissue by injecting adeno-associated virus-9-RAI14 plasmid system via tail vein improved cardiac function and reduced infarct and apoptosis. Furthermore, the activated autophagy in ischemic penumbra of I/R mice was reversed by RAI14 overexpression along with decreased LC3-II and increased p62 expressions. RAI14 silence showed an opposite effect. A cell model was established by using mouse cardiomyocytes HL-1 underwent hypoxia/reoxygenation (H/R) treatment. Similarly, H/R also enhanced RAI14 expression and RAI14 overexpression inhibited H/R-induced apoptosis and autophagy in HL-1 cells. Mechanistically, autophagy inhibitor, the AKT/mTOR pathway, was found to be suppressed in mouse and cell models whereas RAI14 overexpression activated this pathway. Collectively, we demonstrated that compensatory increase of RAI14 inhibited I/R-induced myocardial injury by preventing excessive autophagy through activating the AKT/mTOR pathway, which providing an idea to explore strategies for preventing I/R injury.</p>","PeriodicalId":12102,"journal":{"name":"Experimental Animals","volume":" ","pages":""},"PeriodicalIF":2.2,"publicationDate":"2025-06-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144247101","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Evaluation of a mixture of medetomidine, alfaxalone and butorphanol as an alternative drug for euthanasia in mice.","authors":"Megumi Kiyoto, Kenta Nakano, Yukiyoshi Watai, Yukiko Shimizu, Mayu Uchihashi, Tadashi Okamura","doi":"10.1538/expanim.25-0052","DOIUrl":"https://doi.org/10.1538/expanim.25-0052","url":null,"abstract":"<p><p>Euthanasia agents should induce a rapid and painless loss of consciousness, followed by cardiopulmonary arrest and subsequent brain death. Injectable drugs such as pentobarbital sodium are commonly used for laboratory rodents due to their quick and smooth action. However, the discontinuation of pharmaceutical-grade pentobarbital sodium and secobarbital sodium in Japan, along with a global shortage of pentobarbital in late 2020, has increased the demand for new injectable euthanasia drugs. In Japan, the combination of medetomidine, midazolam, and butorphanol (MMB), as well as a newer formulation in which midazolam is replaced with alfaxalone (MAB), have been widely used as balanced anesthesia for rodents. To evaluate their potential as alternative euthanasia agents in mice, we compared mortality rates and the time intervals to the loss of the righting reflex, respiratory arrest, and cardiac arrest following anesthetic administration. An intraperitoneal injection of MAB at five times the anesthetic dose induced death within 10 min with the loss of the righting reflex, respiratory arrest, and cardiac arrest occurring at 1.5 min, 4 min, and 9 min respectively, in all mice, which was comparable to those observed with 300 mg/kg of secobarbital. In contrast, none of the mice administered MMB at five times the anesthetic dose experienced cardiopulmonary arrest within 30 min. Intraperitoneal overdose of MAB induces rapid and irreversible death, supporting its potential use as an effective euthanasia agent in mice.</p>","PeriodicalId":12102,"journal":{"name":"Experimental Animals","volume":" ","pages":""},"PeriodicalIF":2.2,"publicationDate":"2025-06-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144247100","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Effect of gamma-ray exposure on the genome-editing efficiency of improved genome-editing via oviductal nucleic acids delivery (i-GONAD).","authors":"Anarkhuu Bold-Erdene, Kento Miura, Norimasa Yamasaki, Shuka Miura, Sawako Ogata, Osamu Kaminuma","doi":"10.1538/expanim.25-0036","DOIUrl":"https://doi.org/10.1538/expanim.25-0036","url":null,"abstract":"<p><p>DNA double-strand breaks (DSBs) are among the most hazardous cellular damages, potentially leading to cell death or oncogenesis if unrepaired. Genome editing methods, such as the CRISPR/Cas9 system, induce DSBs and utilize these repair pathways for gene knockout and knock-in. Although ionizing radiation also induces DSBs, it is not clear whether the efficiency of genome editing is affected by ionizing radiation. This study investigated the impact of gamma-ray exposure on the genome editing efficiency of the improved genome editing via oviductal nucleic acid delivery (i-GONAD) method. Gamma-rays were exposed to pregnant mice receiving i-GONAD targeting the Hr gene, whose mutation causes hair loss in mice. The exposure on the fertilization day (Day 0) decreased natural delivery rates and litter sizes, with notable effects at 0.3 Gy or higher. Although the proportions of hairless offspring obtained by i-GONAD differed greatly between single-guide RNAs (sgRNAs) used, total mutation rates, including hairless, mosaic, and indel, were equivalent. Gamma-ray exposure on Day 0 and the day after fertilization (Day 1) similarly and almost dose-dependently enhanced the genome editing efficiency evaluated by the total mutation rate. This study suggests the improvement of genome editing efficiency by gamma-ray exposure, at least in i-GONAD method, potentially facilitating the creation of diverse experimental animal models.</p>","PeriodicalId":12102,"journal":{"name":"Experimental Animals","volume":" ","pages":""},"PeriodicalIF":2.2,"publicationDate":"2025-05-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144172812","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Seungwoo Lee, Jae-Hong Min, Myoung Jun Kim, Somi Yun, Min Kyoung Seo, Jong Kwon Lee
{"title":"Comparison of the characteristics of non-alcoholic fatty liver disease in Lepr<sup>em1hwl</sup> and Lepr<sup>db/db</sup> mice.","authors":"Seungwoo Lee, Jae-Hong Min, Myoung Jun Kim, Somi Yun, Min Kyoung Seo, Jong Kwon Lee","doi":"10.1538/expanim.24-0100","DOIUrl":"https://doi.org/10.1538/expanim.24-0100","url":null,"abstract":"<p><p>The Lepr gene encodes a receptor for leptin, a hormone instrumental in the regulation of appetite and metabolism. Mutations in the Lepr gene impair leptin signaling, leading to metabolic dysfunctions and facilitating the development of non-alcoholic fatty liver disease (NAFLD). In this study, we compared the NAFLD-associated phenotypes of two mutant strains of mice, C57BL/6J-Lepr<sup>em1hwl</sup>/Korl (Lepr<sup>em1hwl</sup>) and C57BLKS/J-Lepr<sup>db</sup>/J (Lepr<sup>db/db</sup>), carrying different alleles of the Lepr gene. Although both Lepr<sup>em1hwl</sup> and Lepr<sup>db/db</sup>mice were characterized by similar obesity phenotypes, leptin resistance, insulin resistance, and glucose intolerance, comparatively, Lepr<sup>em1hwl</sup> mice were found to have relatively more severe hepatic steatosis, along with the upregulated expression of enzymes associated with lipogenesis and triglyceride synthesis, and, notably, the histological characteristics of steatohepatitis were observed only in these mice. In addition, compared with the Lepr<sup>db/db</sup>mice, Lepr<sup>em1hwl</sup> mice developed hepatic fibrosis characterized by elevated levels of collagen deposition and expression of profibrotic factors. Moreover, we detected elevated levels of pro-inflammatory mediators and increases in M1 macrophage markers in the serum and liver, respectively, of Lepr<sup>em1hwl</sup> mice. These findings highlight the distinct NAFLD-associated phenotypic differences between Lepr<sup>em1hwl</sup> and Lepr<sup>db/db</sup>mice, and thereby indicate that Lepr<sup>em1hwl</sup> mice could serve as a valuable model for studying NAFLD, including steatohepatitis and fibrosis.</p>","PeriodicalId":12102,"journal":{"name":"Experimental Animals","volume":" ","pages":""},"PeriodicalIF":2.2,"publicationDate":"2025-05-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144076517","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Sexual activity is predicted by digit ratio in rats.","authors":"Himeka Hayashi, Hirotaka Sakamoto","doi":"10.1538/expanim.24-0159","DOIUrl":"https://doi.org/10.1538/expanim.24-0159","url":null,"abstract":"<p><p>The ratio of the second and fourth digits (2D:4D) is a morphological marker reflecting fetal exposure to sex steroid hormones. This ratio exhibits sexual dimorphism, with males typically showing a significantly lower ratio than females, which results from higher androgen exposure during the fetal period. While studies in humans have suggested a relationship between sexual orientation and the 2D:4D ratio, this relationship in rodents remains elusive. Here, we investigated this relationship using rats as an experimental model. We found that male rats exhibited significantly shorter 2D length than females, resulting in a lower 2D:4D ratio in males, similar to humans. Observations of sexual behavior revealed that males that ejaculated during the first mating test exhibited shorter 2D length compared to males those that did not ejaculate. When males were classified into two groups based on 2D length (long-2D and short-2D groups), short-2D males were more sexually active than long-2D males. Additionally, only short-2D males showed a preference for female odors. These findings suggest that, in rats, 2D length is a useful morphological marker reflecting sexual activity and preference. Furthermore, they provide evidence supporting the potential use of the 2D:4D ratio as a tool for studying the relationship between sexual orientation and the 2D:4D ratio in humans.</p>","PeriodicalId":12102,"journal":{"name":"Experimental Animals","volume":" ","pages":""},"PeriodicalIF":2.2,"publicationDate":"2025-05-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144076631","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Loss of non-canonical translation initiation factors impairs perinatal cardiac function in mice.","authors":"Takehiro Asai, Ryota Tochinai, Yoshiharu Tsuru, Marie Sekiguchi, Atsushi Minami, Wataru Fujii, Shigeru Kyuwa, Tetsuhiro Ogawa, Shigeru Kakuta","doi":"10.1538/expanim.25-0021","DOIUrl":"https://doi.org/10.1538/expanim.25-0021","url":null,"abstract":"<p><p>Translation regulation is crucial for cellular homeostasis. Recent studies have demonstrated that, in addition to the conventional AUG start codon, eukaryotic mRNA can also possess non-canonical start codons. These non-canonical start codons, including non-AUG codons, can be found both upstream and downstream of the conventional AUG start codon. Translation of these non-canonical open reading frames (ORFs) has been implicated in the development of diseases, such as cardiac diseases, neurodegeneration and cancer development. Non-AUG translation initiation is regulated by eukaryotic initiation factor (eIF) 2A and eIF2D; however, their target non-canonical ORFs, roles in disease development, and the underlying precise mechanisms of translation regulation remain poorly understood. To address these gaps, we generated mice lacking either or both of Eif2a and Eif2d genes on an ICR background and investigated their cardiac function using echocardiography. The results indicated that simultaneous disruption of both Eif2a and Eif2d led to perinatal cardiac impairment, as evidenced by a significant reduction in cardiac contractility as measured by ejection fraction. Furthermore, the absence of phenotypic changes in single knockouts of either Eif2a or Eif2d suggests that eIF2A and eIF2D function redundantly in their molecular roles. These findings underscore the importance of non-AUG translation initiation in maintaining cardiac function and suggest its broader implications in other physiological and pathological processes. We propose the Eif2a and Eif2d double-knockout mice as a novel stress-sensitive animal model to investigate the molecular mechanisms of translation regulation and their contribution to disease pathogenesis.</p>","PeriodicalId":12102,"journal":{"name":"Experimental Animals","volume":" ","pages":""},"PeriodicalIF":2.2,"publicationDate":"2025-05-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143974467","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Luminescent mouse model of endometriosis: three-dimensional morphology of lesions and cytokine profiles.","authors":"Nanda Yuli Rahmawati, Tra Thi Huong Dinh, Tomona Oikawa, Akiko Shinogi, Kyoko Ikeda, Masayo Kadota, Masaru Tamura, Takanori Amano, Atsushi Yoshiki","doi":"10.1538/expanim.25-0044","DOIUrl":"https://doi.org/10.1538/expanim.25-0044","url":null,"abstract":"<p><p>The pathophysiology of endometriosis remains incompletely understood, necessitating the development of effective animal models for research. We generated and characterized a luminescent endometriosis mouse model utilizing luminescent B6-CAG-ELuc transgenic mice as uterine tissue donors and B6.Cg-c/c-hr/hr mice as recipients, enabling non-invasive in vivo imaging. Following transplantation of minced uterine tissue fragments into the peritoneal cavity of recipients, we monitored lesion growth via in vivo imaging system on 0, 14, 28, 42 days post transplantation. Morphology of the lesion was observed by dissecting microscopy, X-ray micro-computed tomography, and conventional histology. Inflammation-related serum cytokines were quantified using multiplex immunobeads assay. The growth of endometriotic lesions was efficiently observed by bioluminescence from day 0 through 42 days post transplantation. Comprehensive morphological observations revealed typical endometriotic lesions consisted of multiple fluid-filled cysts lined with single-layered epithelium, associated with glandular epithelial tissues and interstitial stroma. The level of IL-1β, IL-2, IL-6, IL-10, IL-12p70, IFN-γ, and TNF-α was quantified simultaneously in each serum sample to evaluate the temporal changes of each cytokine, showing four distinct patterns: IFN-γ and TNF-α showed continuous increase, IL-12p70 and IL-1β demonstrated gradual increase followed by marked elevation, IL-6 and IL-2 exhibited dramatic increase in later stages, while IL-10 showed transient increase followed by gradual decrease. In conclusion, this luminescent endometriosis mouse model using B6 luminescent transgenic mice as uterine tissue donor and B6.Cg-c/c-hr/hr recipient could be used to investigate comprehensive cytokine profiling in the development of endometriosis.</p>","PeriodicalId":12102,"journal":{"name":"Experimental Animals","volume":" ","pages":""},"PeriodicalIF":2.2,"publicationDate":"2025-05-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144005130","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Soyasaponin I alleviates inflammation and oxidative stress in chronic obstructive pulmonary disease through inhibiting the MAPK signaling pathway.","authors":"Ruoqi Zhang, Jiabo Yuan, Congyao Wang, Ruiqi Zhao, Fengli Gao, Zhuying Li","doi":"10.1538/expanim.25-0007","DOIUrl":"https://doi.org/10.1538/expanim.25-0007","url":null,"abstract":"<p><p>Chronic obstructive pulmonary disease (COPD) is a prevalent lung disease that mainly induced by cigarette smoking (CS). Soyasaponin I is an amphiphilic oleanane triterpenoid glycoside extracted form Astragali Radix. In order to investigate treatment strategies of COPD, this study focused on the effect of soyasaponin I on the lung tissue of COPD model. The mouse model of COPD was induced by CS exposure for 12 weeks, and was administrated with different doses of soyasaponin I. Subsequently, the morphology and histopathology of lung tissue, the proportion of inflammatory cell, the levels of inflammatory cytokines, and indicators of oxidative stress were assessed and analyzed. The signaling pathway potentially regulated by soyasaponin I in the pathogenesis of COPD were predicted by network pharmacology analysis and validated by western blot. Our results demonstrated that soyasaponin I mitigated the lung injury and bronchial lesions induced by COPD through reducing the lung coefficient, wall area of the bronchioles and Periodic Acid Schiff (PAS)-positive cells in the lung tissue. The CS-induced inflammation and oxidative stress was alleviated by soyasaponin I through reversing the levels of inflammatory cytokines and oxidative stress indicators. In addition, the phosphorylation of p38, JNK and ERK1/2 was activated in COPD model, and was reverted by soyasaponin I in the lung tissue. Collectively, the present study confirmed that soyasaponin I is an effective compound that attenuates the lung injury through inhibiting inflammatory response and oxidative stress via the MAPK signaling pathway.</p>","PeriodicalId":12102,"journal":{"name":"Experimental Animals","volume":" ","pages":""},"PeriodicalIF":2.2,"publicationDate":"2025-05-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143994469","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Experimental AnimalsPub Date : 2025-04-20Epub Date: 2024-11-29DOI: 10.1538/expanim.24-0107
Haruhisa Tsuji, Rei Maeyama, Yoko Kato
{"title":"Optimization of culture-preservation methods to maintain developmental competence in porcine metaphase II (MII) oocytes post-in vitro maturation (IVM).","authors":"Haruhisa Tsuji, Rei Maeyama, Yoko Kato","doi":"10.1538/expanim.24-0107","DOIUrl":"10.1538/expanim.24-0107","url":null,"abstract":"<p><p>After in vitro maturation (IVM) of porcine germinal vesicle (GV) oocytes, those that matured to the metaphase II (MII) stage were selected for further culture over a period of 24-48 h. Subsequently, these oocytes were either parthenogenetically activated or used for somatic cell nuclear transfer (SCNT) to evaluate their in vitro developmental competence. Parthenogenetically activated MII oocytes developed to the blastocyst stage after 42 h of continuous culture, whereas SCNT oocytes reached the blastocyst stage within 30 h of culture. These findings suggest that porcine MII oocytes retain their developmental competence after extended in vitro culture exceeding 30 h. This study highlights the potential of prolonged culture in enhancing the utility of MII-stage oocytes for livestock applications and possibly for future advancements in human infertility treatments.</p>","PeriodicalId":12102,"journal":{"name":"Experimental Animals","volume":" ","pages":"189-196"},"PeriodicalIF":2.2,"publicationDate":"2025-04-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12044358/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142767466","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Experimental AnimalsPub Date : 2025-04-20Epub Date: 2024-11-23DOI: 10.1538/expanim.24-0132
Risa Iwanaga, Kanako Sumi, Chizuko Kodama, Munekatsu Ita, Mohammad Ibrahim Qasimi, Jun Tamura, Ko Nakanishi, Yasuhiro Yoshida, Masami Morimatsu, Kayoko Matsumura, Teppei Nakamura
{"title":"Dual-route administration of balanced anesthesia using medetomidine, midazolam, and butorphanol provides both suitable anesthetic depth and reduced tissue injury in rabbits.","authors":"Risa Iwanaga, Kanako Sumi, Chizuko Kodama, Munekatsu Ita, Mohammad Ibrahim Qasimi, Jun Tamura, Ko Nakanishi, Yasuhiro Yoshida, Masami Morimatsu, Kayoko Matsumura, Teppei Nakamura","doi":"10.1538/expanim.24-0132","DOIUrl":"10.1538/expanim.24-0132","url":null,"abstract":"<p><p>Medetomidine, midazolam, and butorphanol (MMB) anesthesia is the preferred choice for rodents but requires excess volume of intramuscular injection in rabbits, which can lead to muscular damage. This study aimed to evaluate a dual-route MMB administration via the intravenous and subcutaneous routes in rabbits. MMB was administered to male Kbs:JW rabbits with an intravenous injection of 0.2 ml/kg followed by a subcutaneous injection of 0.8 ml/kg, totaling 0.2 mg/kg medetomidine, 2.0 mg/kg midazolam, and 2.0 mg/kg butorphanol. We compared the anesthetic effects of this dual-route method with those of intramuscular administration. The dual-route method resulted in a shorter induction time and similar anesthetic duration compared with those of the intramuscular route. While it induced a temporary decrease in body temperature within 30 min post-injection, other vital signs, such as respiration rate, heart rate, and O<sub>2</sub> saturation, remained similar. Notably, unlike intramuscular administration, dual-route administration did not increase tissue injury marker levels. This dual-route MMB administration provided sufficient anesthetic depth during surgery, eliminating pain reflexes. Double-dose administration extended anesthetic duration but resulted in rare fatalities, indicating room for protocol improvement. In conclusion, the novel anesthetic method is preferable for injectable anesthesia in rabbits, providing rapid induction and sufficient anesthetic duration, while potentially minimizing muscle injury. This technique may be beneficial for both laboratory and companion animals and significantly enhance animal welfare in anesthesia by reducing the pain associated with injectable anesthesia.</p>","PeriodicalId":12102,"journal":{"name":"Experimental Animals","volume":" ","pages":"181-188"},"PeriodicalIF":2.2,"publicationDate":"2025-04-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12044349/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142709669","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}