Experimental Animals最新文献

The Cynomolgus monkey is the most widely used models in non-clinical studies. As factors like age, gender, and breeding province may affect hematologic and serum biochemical parameters, it is important to establish base values of these parameters by these three factors and to determine the effects of these factors on the parameters. In total, 1794 Cynomolgus monkeys (Male: 901, Female: 893) were selected. A total of 24 hematologic and 21 serum biochemical parameters were measured, and the effects of age, gender, and breeding province were analyzed. Base values for hematologic and serum biochemical parameters were established by age, gender, and breeding province. A significant neutrophil percent, alkaline phosphatase, and creatinine differences were observed between different ages; a significant alkaline phosphatase, gamma glutamyl transpeptidase, and creatinine differences were observed between males and females; a significant lymphocyte percent, neutrophil percent, reticulocyte count, alkaline phosphatase, gamma glutamyl transpeptidase, and creatinine differences were observed between different breeding provinces. The results emphasize the importance of improving base values by age, gender, and breeding provinces. There was no statistically significant difference in most of the above parameters, and Cynomolgus monkeys from different breeding provinces can be used in the same study.

Spinocerebellar ataxia 13 (SCA13) is an autosomal dominant neurological disorder caused by mutations in KCNC3. Our previous studies revealed that KCNC3 (Potassium Voltage-Gated Channel Subfamily C Member 3) mutation R423H results in an early-onset form of SCA13. Previous biological models of SCA13 include zebrafish and Drosophila but no mammalian systems. More recently, mouse models with Kcnc3 mutations presented behavioral abnormalities but without obvious pathological changes in the cerebellum, a hallmark of patients with SCA13. Here, we present a novel transgenic mouse model by bacterial artificial chromosome (BAC) recombineering to express the full-length mouse Kcnc3 expressing the R424H mutation. This BAC-R424H mice exhibited behavioral and pathological changes mimicking the clinical phenotype of the disease. The BAC-R424H mice (homologous to R423H in human) developed early onset clinical symptoms with aberrant gait, tremor, and cerebellar atrophy. Histopathological analysis of the cerebellum in BAC-R424H mice showed progressive Purkinje cell loss and thinning of the molecular cell layer. Additionally, Purkinje cells of BAC-R424H mice showed significantly lower spontaneous firing frequency with a corresponding increase in inter-spike interval compared to that of wild-type mice. Our SCA13 transgenic mice recapitulate both neuropathological and behavioral changes manifested in human SCA13 R423H patients and provide an advantageous approach to understanding the role of voltage-gated potassium channel in cerebellar morphogenesis and function. This mammalian in vivo model will lead to further understanding of the R423H allelic form of SCA13 from the molecular to the behavioral level and serve as a platform for testing potential therapeutic compounds.

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.

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 O₂ 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.

There are few ultrasonographic studies on the spontaneous type 2 diabetes mellitus db/db mouse. Our objective was to dynamically investigate and assess renal morphological and hemodynamic changes in spontaneous T2DM db/db mice through high-frequency ultrasound. Eighteen male db/db mice (the model group) and twelve male db/+ mice (the control group) were included. Body weight and fasting blood glucose were measured at the ages of 8, 16 and 32 weeks. High-frequency ultrasound examinations were conducted at the same ages. Compared with those in the control group, hematoxylin-eosin and Masson staining revealed pathological changes in the renal tissue of the db/db mice at 16 weeks of age, and the lesions were significantly aggravated at 32 weeks of age. The body mass of the mice in the model group increased significantly at 8, 16 and 32 weeks of age, and the kidney volume measured by ultrasound also increased with age. Compared with those of the control group, the blood flow scores determined via power Doppler were significantly different. The peak systolic velocity (PSV), end diastolic velocity (EDV), and resistive index (RI) of the renal artery and the PSV, EDV, and RI of the segmental artery were significantly different at the sixteenth week compared with those that at the eighth week. The results of high-frequency ultrasound revealed that the renal hemodynamics of db/db mice changed at the sixteenth weeks.

Beige adipocytes arise from white adipocytes in response to cold or other stimuli, known as browning of white adipose. Beige adipocytes play a role similar to that of brown adipocytes, express high levels of uncoupling protein 1 (UCP1), and are responsible for energy consumption via heat production, thus aiding in fat loss. Although histidine (His) and soy isoflavones (Iso) co-ingestion reportedly reduces food intake, body weight, and fat accumulation in female rats, the underlying mechanism remains unclear. Therefore, this study aimed to elucidate the mechanisms whereby histidine and soy isoflavones (His-Iso) co-ingestion suppresses fat accumulation. Female rats were fed a control diet or diet containing Iso, His, or His-Iso for 2 weeks, followed by sampling of periovarian white adipose tissue (poWAT) and retroperitoneal white adipose tissue (rWAT) and adipocyte morphology analysis. Additionally, the expression of browning- and lipid metabolism-related genes was examined. Histochemical analysis revealed the presence of multilocular lipid droplets, representative of beige adipocytes, in the poWAT and rWAT of rats in the His-Iso co-ingestion group. Quantitative PCR analysis showed that His-Iso co-ingestion upregulated brown adipocyte and beige adipocyte markers, including UCP1, indicating that His-Iso intake induces beige adipocytes. Moreover, His-Iso co-ingestion upregulated genes related to fatty acid oxidation (carnitine palmitoyl transferase 1A) and lipolysis (adipose triglyceride lipase) in WATs. In conclusion, His-Iso co-ingestion increases UCP1 expression and morphological changes to beige adipocytes, and suppresses fat accumulation by promotion of lipolysis and fatty acid oxidation in WAT.

Atopic dermatitis (AD) is a chronic skin disease that causes itching and is characterized by recurrent flares and remissions. The interactions among type 2 inflammation, skin barrier dysfunction, and pruritus play important roles in the pathogenesis of AD. AD symptoms persist for a long period; thus, it is desirable to have disease models that reproduce a prolonged AD-like phenotype. Although MC903-induced AD model mice reportedly exhibit type 2 inflammation, skin barrier dysfunction, and pruritus, the effects of long-term application of MC903 on the changes in these symptoms over time are not fully understood. To clarify this point, we conducted a long-term time course analysis of these symptoms by applying MC903 to the ears of mice every other day for four weeks. Increased ear thickness, transepidermal water loss, number of scratching events, and serum IgE levels were observed in the MC903 model. Histological analysis revealed the infiltration of granulocytes and CD3-positive T cells and an increase in mast cells in the dermis. Furthermore, analyses of mRNA and protein expression in ear tissue revealed increased expression of thymic stromal lymphopoietin, IL-4, IL-13, and IL-33, which are involved in type 2 inflammation. All these changes were observed within two weeks after the initial application of MC903 and thereafter persisted throughout the experimental period. In conclusion, our data indicate that the long-term application of MC903 prolongs the duration of the three major symptoms of AD.

The Adriamycin-induced nephropathy (AN) model plays a crucial role in advancing our understanding of and research on chronic kidney disease (CKD). This review outlines methodologies for generating AN models in mice and rats, discusses their pathophysiologic and molecular characteristics, highlights their advantages and limitations, describes therapeutic interventions that have been evaluated in these models, and presents future research perspectives. The AN model replicates key features observed in human CKD, such as proteinuria, podocyte injury, glomerulosclerosis, and tubulointerstitial fibrosis. Notably, genetic factors significantly influence the onset and severity of AN, with mutations in the Prkdc gene linked to nephrotoxicity and systemic toxicity. To evaluate therapeutic interventions for CKD, agents such as ACE inhibitors, corticosteroids, and SGLT2 inhibitors have been tested in the AN model, demonstrating promising renoprotective effects. However, the systemic toxicity of Adriamycin and variability across models pose limitations, highlighting the need for caution when translating findings to human CKD. Future advancements in genetic engineering and the application of CRISPR-Cas9 technology are expected to improve the fidelity of AN models to human disease. Additionally, discovery of biomarkers by using the AN model enables us to improve early diagnosis. These efforts are anticipated to deepen our understanding of CKD pathophysiology and contribute to developing more effective diagnostic tools and targeted therapies.

At present, there lacks a definitive pharmaceutical intervention or therapeutic approach for diabetes-associated cognitive impairment. Herein, we delved into the impact of electroacupuncture on cognitive function in high-fat diet/streptozocin (HFD/STZ)-induced type 2 diabetes mellitus (T2DM) mice and underlying mechanisms. Hippocampal insulin resistance was determined by western blot analysis. Cognitive function was evaluated by Morris water maze test. The morphology of the hippocampal neurons was observed through hematoxylin & eosin staining and Nissl staining. Synaptic plasticity was assessed by western blot analysis. Immunofluorescence, immunohistochemistry, western blot and real-time PCR were employed to detect the levels of ferroptosis markers, autophagy markers, and netrin-1. Electroacupuncture treatment exhibited ameliorative outcomes on hippocampal insulin resistance, spatial learning, memory function, neuronal damage, and synaptic plasticity in T2DM mice. Furthermore, it effectively suppressed neuronal ferroptosis in the hippocampus by upregulating GPX4 and SLC7A11 expression, and reducing 4-HNE expression. Meanwhile, electroacupuncture intervention increased the levels of Beclin1 and LC3II/LC3I, as well as decreased the levels of p62 and phosphorylated-mTOR in the hippocampus of T2DM mice, suggesting that electroacupuncture facilitated autophagy activation by inhibiting mTOR activity. 3-MA-mediated autophagy inhibition undermined the beneficial effects of electroacupuncture on neuronal ferroptosis and cognitive deficits in T2DM mice. Additionally, the beneficial effects of electroacupuncture on autophagy and ferroptosis was achieved by upregulation of netrin-1 in the hippocampus. Our study revealed that electroacupuncture therapy inhibited neuronal ferroptosis via the activation of autophagy, thereby ameliorating cognitive deficits in T2DM mice.

Gracile axonal dystrophy (gad) mutant mice present with autosomal recessive inherited sensory ataxia in the early stages, followed by age-dependent motor ataxia. This phenotype is caused by a mutation in the ubiquitin carboxyl-terminal hydrolase isozyme L1 (UCH-L1) gene and leads to a lack of expression of UCH-L1 protein, ubiquitin-proteasome which is related to the autophagy pathway and the ubiquitin-proteasome system (UPS). To elucidate the pathophysiology of abnormal protein accumulation in gad mice, we focused on macroautophagy. Using electron microscopy, we detected a double-membrane structure, which was characteristic of autophagosomes, in gad mice. In addition, in immunohistochemistry to investigate the expression levels of autophagy-related proteins in the gracile nuclei of the gad mouse, we found upregulation of LC3 and p62 but not LAMP-2A. These results suggested that a lack of UCH-L1 expression might induce the formation of autophagosomes, but the resulting autophagy flux might be disturbed.