2013 Vol. 34, No. 2

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2013, 34(2): 0-0.
Review
Animal models are indispensible in biomedical research and have made tremendous contributions to answer fundamental questions on human biology, disease mechanisms, and to the development of new drugs and diagnostic tools. Due to the limitations of rodent models in translational medicine, tree shrews (Tupaia belangeri chinensis), the closest relative of primates, have attracted increasing attention in modeling human diseases and therapeutic responses. Here we discuss the recent progress in tree shrew biology and the development of tree shrews as human disease models including infectious diseases, metabolic diseases, neurological and psychiatric diseases, and cancers. Meanwhile, the current problems and future perspectives of the tree shrew model are explored.
The tree shrew is currently located in the Order Scandentia and is widely distributed in Southeast Asia, South Asia, and South China. Due to its unique characteristics, such as small body size, high brain-to-body mass ratio, short reproductive cycle and life span, and low-cost of maintenance, the tree shrew has been proposed as an alternative experimental animal to primates in biomedical research. However, there is unresolved debate regarding the phylogenetic affinity of tree shrews to primates and their phylogenetic position in Euarchontoglires. To help settle this debate, we summarized the available molecular evidence on the phylogenetic position of the tree shrew. Most nuclear DNA data, including recent genome data, suggested that the tree shrew belongs to the Euarchonta clade harboring primates and flying lemurs (colugos). However, analyses of mitochondrial DNA (mtDNA) data suggested a close relationship to lagomorphs and rodents. These different clustering patterns could be explained by nuclear gene data and mtDNA data discrepancies, as well as the different phylogenetic approaches used in previous studies. Taking all available conclusions together, the robust data from whole genome of this species supports tree shrews being genetically closely related to primates.
Nonhuman primate animal models play an important role in studying HIV-1 pathogenesis, developing antiviral drugs and vaccines. Due to the lack of animals that can be directly infected with HIV-1, SIV/SHIV-infected macaques have been widely used in AIDS research. Although these models are somewhat similar to human AIDS, there are many limitations due to genetic differences between SIV/SHIV and HIV-1. Developing a suitable nonhuman primate animal model is still an important topic in HIV/AIDS research. The pigtailed macaque is the only primate in Old World monkeys that can be infected with HIV-1 and offer many benefits as HIV-1 intravenous and sexual transmission models. Here we reviewed the characteristics of pigtailed macaque models infected by SIV, HIV, SHIV, and HSIV via intravenous and mucosal routes. In addition, we briefly introduced the molecular mechanisms of viral replication in pigtailed macaque cells, and discussed the limitations and prospects of pigtailed macaque models in AIDS research.
Articles
The pig-tailed macaque is an important non-human primate experimental animal model that has been widely used in the research of AIDS and other diseases. Pig-tailed macaques include Mentawai macaques (Macaca pagensis), Sunda pig-tailed macaques (M. nemestrina) and northern pig-tailed macaques (M. leonina). Northern pig-tailed macaques inhabit China and surrounding Southeast Asia countries. To our knowledge, no reports have been published regarding the hematology and blood chemistry parameters of northern pig-tailed macaques, which are important for the objective evaluation of experimental results. We measured and analyzed 18 hematology parameters and 13 blood chemistry parameters in juvenile (aged 2?4 years) and adult (aged 5?10 years) northern pig-tailed macaques. We found that red blood cells, hemoglobin and alkaline phosphatase values were lower in female macaques than male macaques in both juvenile and adult groups. White blood cells, lymphocyte, monocytes, platelet distribution width, cholesterol, aspartate aminotransferase and alkaline phosphatase values were higher in juvenile macaques than adult macaques, while creatinine and triglycerides values were lower in juvenile macaques. Mean corpuscular hemoglobin and creatinine values were positively correlated with weight in juvenile groups. In adult groups, mean corpuscular hemoglobin, percentage of granulocyte, hemoglobin and creatinine were also positively correlated with weight, and lymphocyte, percentage of lymphocyte, red cell distribution width, aspartate aminotransferase and cholesterol values were negatively correlated with weight. The results suggest that age, gender and weight of northern pig-tailed macaques affected their hematology and blood chemistry parameters. This hematological and blood chemistry study has great significance in biomedical research and animal models using northern pig-tailed macaque as an experimental animal.
The aim of this study was to induce type 2 diabetes in tree shrew (Tupaia belangeri chinensis). Streptozotocin (STZ), a glucosamine derivative of nitrosourea and preferentially toxic to pancreatic beta cells, has been commonly used to induce type 1 and 2 diabetes in experimental animals. Tree shrews were treated with different low doses of STZ (60, 70, and 80 mg/kg), with six control tree shrews receiving citrate buffer. After STZ injection, tree shrews displayed increased fasting blood and urine glucose, impaired oral glucose tolerance test, and disturbed lipids metabolism and renal function. However, STZ induced tree shrews showed no diabetic complications such as diabetic lactic acidosis and hyperglycemic hyperosmolar. Animals with the above type 2 diabetic-like symptoms were variable across the three groups from 66.7% to 100%, and mortality ranged from 16.7% to 33.3%. Thus, two 80 mg/kg STZ dose injections were appeared more appropriate than other doses to induce tree shrew model of type 2 diabetes. Our results demonstrated that type 2 diabetes could be induced with favorable STZ application in tree shrew.
Note
In most Old world monkey species, TRIM5α plays a role in combating retroviruses and restricting HIV-1. Alongside TRIM5α, the TRIMCyp fusion gene formed by the retrotransposition of a CypA pseudogene cDNA to 3' terminal or 3'-UTR of TRIM5 gene in these monkeys has become a key research area in anti HIV-1 factors. The regional differences, gene frequencies, genotypes, and retrovirus restrictive activities of TRIMCyp vary among different primate species. While the frequencies of cynomolgus TRIMCyp have been studied in several areas of Southeast Asia, the frequency and prevalence of cynomolgus TRIMCyp in China remains unclear. In this study, we screened 1, 594 cynomolgus samples from 11 monkey manufacturers located across 5 provinces in China. Our results showed that the frequencies of TRIMCyp range from 7.65% to 19.79%, markedly lower than the frequencies found in monkey species in the Philippines, Malaysia and Indonesia (ranging from 34.85% to 100%). We speculate that potentially the latter were isolated groups established since 1978. The NE haplotype frequencies of cynomolgus TRIMCyp were 4.93% in China, also significantly lower than those found in species in the Philippines, Malaysia and Indonesia (from 11.1% to 14.3%). Our research provides interesting findings that contribute towards a more firm basis of further studies of HIV-1 animal models and relevant pathogenesis.
Recent studies found that a loss of motor function following corticospinal tract (CST) damage can, to some extent, be restored. Few studies, however, examine how space-occupying lesions in the brain motor area may affect the contralateral CTS structure and function. We performed a simulation of intracranial space-occupying lesions in the brain motor area by implanting of balloons into the brains of the two healthy macaques. Diffusion tensor imaging (DTI) was performed on the macaques’ brains four times to measure the FA values of the contralateral CST operative area. The results showed that on the day of balloon implantation, the FA values had no obvious effect, but with time the effect increased, becoming increasingly apparent one week after removing the balloons. Experimental results demonstrated that this model was both feasible and reliable. After the simulated space-occupying lesions occurred in the brain motor area, DTI showed a compensatory response of the contralateral CTS, which remained for a short period of time even after the lesions were removed. This result suggests that the contralateral CST may then also contribute to recovery of limb function.
Pathogenic viruses can harm acutely the life and health of laboratory tree shrews acutely; however, few papers exist regarding natural pathogenic virus infection in this species. Six fecal samples obtained from dead tree shrews were collected. The fecal supernatant infected Vero cell line resulted in cytopathic effects (CPE) after 72 h. The CPE included granulating, shrinking, rounding, seining and falling off. Electron microscopy showed the isolation was spherical, double-layered capsid, and about 75 nm in diameter. The purified isolation genome was 10 segments in a typical 3:3:4 arrangements, as shown by polyacrylamide gel electrophoresis (PAGE). The isolation was confirmed by RT-PCR assays targeting the conserved region of the L1 gene, sequence analysis and reconstruction of a phylogenetic tree. The isolation was a Tupaia Orthoreovirus (TRV), belonging to Mammalian Orthoreovirus (MRV). The obtained strain had the closest phylogenetic relationship to the MRV strain T3/Bat/Germany/342/08. As a zoonotic virus, the novel TRV strain was first isolated from wild tree shrews, which is significant for promoting tree shrew standardization and providing scientific data for preventing zoonotic tree shrew-to-human transmission.
While the tree shrew (Tupaia belangeri chinensis) is an excellent animal model for studying the mechanisms of human diseases, but few studies examine interleukin-2 (IL-2), an important immune factor in disease model evaluation. In this study, a 465 bp of the full-length IL-2 cDNA encoding sequence was cloned from the RNA of tree shrew spleen lymphocytes, which were then cultivated and stimulated with ConA (concanavalin). Clustal W 2.0 was used to compare and analyze the sequence and molecular characteristics, and establish the similarity of the overall structure of IL-2 between tree shrews and other mammals. The homology of the IL-2 nucleotide sequence between tree shrews and humans was 93%, and the amino acid homology was 80%. The phylogenetic tree results, derived through the Neighbour-Joining method using MEGA5.0, indicated a close genetic relationship between tree shrews, Homo sapiens, and Macaca mulatta. The three-dimensional structure analysis showed that the surface charges in most regions of tree shrew IL-2 were similar to between tree shrews and humans; however, the N-glycosylation sites and local structures were different, which may affect antibody binding. These results provide a fundamental basis for the future study of IL-2 monoclonal antibody in tree shrews, thereby improving their utility as a model.
In this paper, partial sequences of the tree shrew (Tupaia belangeri) Klf4, Sox2, and c-Myc genes were cloned and sequenced, which were 382, 612, and 485 bp in length and encoded 127, 204, and 161 amino acids, respectively. Whereas, their cDNA sequence identities with those of human were 89%, 98%, and 89%, respectively. Their phylogenetic tree results indicated different topologies and suggested individual evolutional pathways. These results can facilitate further functional studies.
Anatomical parameter values in tree shrews are major biological characteristic indicators in laboratory animals. Body size, bones and mammilla, organ weights, coefficient intestinal canal and other anatomical data were measured and analyzed in laboratory domesticated tree shrews (7 to 9 months of age). Measurement of 31 anatomical parameters showed that body height, width of the right ear, ileum and colon had significant differences between males and females (P<0.05). Highly significant differences were also found in body slanting length, chest depth, torso length, left and right forelimb length, right hind limb length, left and right ear length, left ear width, keel bone length, left and right tibia length, duodenum and jejunum (P<0.01). With body length as the dependent variable, and tail length, torso length, right and left forelimb length, and left and right hind limb length as independent variables for stepwise regression analysis, the regression equation for body length = 13.90 + tail length ? 0.16. The results of 37 organs weights between female and male tree shrews showed very significant differences (P<0.01) for weight of heart, lungs, spleen, left and right kidney, bladder, left and right hippocampus, left submandibular gland, and left and right thyroid gland, as well as significant (P<0.05) differences in the small intestine, right submandibular gland, and left adrenal gland. The coefficient of heart, lung, stomach, bladder, small and large intestine, brain, right hippocampus, and left adrenal gland showed highly significant differences (P<0.01), while differences in the right kidney, left hippocampus, left submandibular gland, right adrenal gland, and left and right thyroid gland were significant (P<0.05). With animal weight as the dependent variable and indicators of heart, lung, liver, spleen, left and right kidney and brain as independent variables for stepwise regression analysis, the regression equation showed that weight = 62.73 + left kidney ? 79.21 + heart ? 24.09. Female and male laboratory domesticated tree shrews showed certain influences in body size, organ weight and coefficient, and intestinal canal regarding anatomical parameters. This experiment provides basic data for studies on laboratory tree shrews and animal models.