Xueyun Huo

Establishment and Characterization of a Newly Established Diabetic Gerbil Line

Objectives We aimed to selectively breed a spontaneous diabetic gerbil when a sub-line of inbred gerbil showed increased blood glucose levels was found recently. Then we investigated the characteristics including the serum insulin, triglyceride, cholesterol, leptin, adiponectin and explored the underlying molecular mechanism for the diabetic phenotype. Methods The spontaneous diabetic line of gerbils was selectively inbreed the sub-line of gerbil by monitoring blood glucose of each animal. The serum insulin, adiponectin, and leptin levels were tested using an ELISA kit. The expression levels of GLUT4, Akt, leptin, adiponectin, and calpain 10 (CAPN10) were tested by western blot and Quantitative Real-time PCR (qPCR) in liver, skeletal muscle, and white adipose. Results Our results show that the percentages of animals with FPG≥5.2 (mmol/l), PG2h≥6.8 (mmol/l) and both FPG≥5.2 and PG2h≥6.8 (mmol/l) were increased with the number of breeding generations from F0 (21.33%) to F6 (38.46%). These diabetic gerbils exhibited insulin resistance and leptin resistance as well as decreased adiponectin level in the serum. We also observed decreased expression of adiponectin and increased expression of leptin in the skeletal muscle, respectively. Conclusions These results indicate that we have primarily established a spontaneous diabetic gerbil line, and the diabetic phenotypes may have been accounted for by altered expression of leptin and adiponectin.

Selection of genes associated with variations in the Circle of Willis in gerbils using suppression subtractive hybridization.

Deformities in the Circle of Willis (CoW) can significantly increase the risk of cerebrovascular disease in humans. However, the molecular mechanisms underlying these deformities have not been understood. Based on our previous studies, variations in the CoW of gerbils are hereditary. A normal CoW is observed in approximately 60% of gerbils, a percentage that also applies to humans. Thus, gerbil is an ideal experimental model for studying variations in the CoW. To study the mechanisms underlying these variations, we selected genes associated with different types of the CoW using suppression subtractive hybridization (SSH). After evaluating the efficiency of SSH using quantitative real-time polymerase chain reaction (qPCR) on subtracted and unsubtracted cDNA and Southern blotting on SSH PCR products, 12 SSH libraries were established. We identified 4 genes (CST3, GNAS, GPx4 and PFN2) associated with variations in the CoW. These genes were identified with qPCR and Western blotting using 70 expressed sequence tags from the SSH libraries. Cloning and sequencing allowed us to demonstrate that the 4 genes were closely related to mouse genes. We may assume that these 4 genes play an important role in the development of variations in the CoW. This study provides a foundation for further research of genes related to development of variations in the CoW and the mechanisms of dysmorphosis of cerebral vessels.

Analysis of the relationship between microsatellite instability and thymic lymphoma induced by N-methyl-N-nitrosourea in C57BL/6J mice.

Microsatellite instability (MSI) has been found to be closely associated with many types of human tumors and often shows strong correlations with specific tumor features. However, the relationship between MSI and tumors are still unclear. The aim of the present study is to explore the relationships between MSI, tumor formation under the mutagenic effects of N-methyl-N-nitrosourea (MNU). Mice were administered with either MNU (90 mg/kg) or PBS and DMSO (control) at the beginning of the 1st week of the experiment. Of the 31 mice that survived the entire experimental time course, 19 (61.3%) mice developed thymic lymphomas. In addition, 52.6% (10/19) of the tumors had metastasized to the liver. We detected MSI in MNU-treated mice using a panel of 42 mutation-sensitive loci. Nineteen loci (45.2%) in six organs showed 70 MSI events. Locus D8Mit14 showed enhanced MSI compared with the other examined loci. MSI frequency in thymus was higher than in other organs. Interestingly, there was no significant difference observed between the metastatic and non-metastatic livers. The MSI frequency (4.6%, 23/(42×12)) in the MNU-treated thymus that had never developed tumor was significantly higher than this in the thymus that had developed lymphoma (0.5%, 4/(42×19)) (p<0.0001). These results indicate that, although thymic tumorigenesis is associated with MSI, it occurs with higher frequency in these that have not developed tumors upon the MNU-treatment. Our study provides additional insights into the relationship between MSI occurrence and tumorigenesis.

Newly breeding an inbred strain of ischemia-prone Mongolian gerbils and its reproduction and genetic characteristics

The Mongolian gerbil has been a useful laboratory animal in many research fields, especially in ischemia studies. However, due to the variation of the circle of Willis (COW), the ischemic model is unstable and various. To solve this problem, we newly established an inbred strain of gerbils, restricting breeding and keeping to F23. The data on the breeding and growth of the animals are described in the present study. The genetic characteristics of F4 to F20 detected by microsatellite DNA and biochemical markers are also shown here. The results demonstrated that the frequency of ischemic model by unilateral carotid occlusion and the frequency of incomplete COW increased, increasing from 50% and 75% in F1 to 88.89% and 100% in F20, respectively. The ratios of consistent patterns of COW in parents were positively related with the number of inbred generations. A reproductive performance analysis indicated that the average size of litters in the inbred gerbils was less than that of outbred gerbils and that adult body weight was also lower in inbred gerbils; also, the pups in the 2nd litter were the best ones chosen to reproduce. The genetic detection results indicated that 26 out of 28 microsatellite loci and all 26 biochemical markers were homozygous in F20, showing comparably identical genetic composition in inbred gerbils. All the data demonstrated that an inbred strain of ischemia-prone gerbil has been established successfully. This strain can be used in stroke research and can largely reduce the number of animals needed in experiments.

The complete mitochondrial genome of the Tamarisk gerbil, Meriones tamariscinus (Rodentia: Muridae)

Abstract The complete mitochondrial genome of the Tamarisk jird, Meriones tamariscinus, was sequenced. The 16,389bp genome contains 37 genes, typical for rodent mitogenomes, including 22 tRNA genes, 2 rRNA genes, and 13 protein-coding genes. The total GC content of the mitochondrial genome is 36.8%, with a base composition of 34.0% A, 24.5% C, 12.3% G, and 29.2% T. The phylogenetic analysis showed that M. tamariscinus was classified in the genus Meriones, Muridae.