ABSTRACT
Spermatogonial stem cells (SSCs) transmit genetic information to the next progeny in males. Thus, SSCs are a potential target for germline modifications to generate transgenic animals. In this study, we report a technique for the generation of transgenic rats by in vivo manipulation of SSCs with a high success rate. SSCs in juvenile rats were transduced in vivo with high titers of lentivirus harboring enhanced green fluorescent protein and mated with wild-type females to create founder rats. These founder rats expressed the transgene and passed on the transgene with an overall success rate of 50.0%. Subsequent generations of progeny from the founder rats both expressed and passed on the transgene. Thus, direct modification of SSCs in juvenile rats is an effective means of generating transgenic rats through the male germline. This technology could be adapted to larger animals, in which existing methods for gene modification are inadequate or inapplicable, resulting in the generation of transgenic animals in a variety of species.
Subject(s)
Animals , Male , Rats , Green Fluorescent Proteins , Lentivirus , Rats, Transgenic , Spermatogonia/metabolismABSTRACT
Spermatogonial stem cells (SSCs) transmit genetic information to the next progeny in males. Thus, SSCs are a potential target for germline modifications to generate transgenic animals. In this study, we report a technique for the generation of transgenic rats by in vivo manipulation of SSCs with a high success rate. SSCs in juvenile rats were transduced in vivo with high titers of lentivirus harboring enhanced green fluorescent protein and mated with wild-type females to create founder rats. These founder rats expressed the transgene and passed on the transgene with an overall success rate of 50.0%. Subsequent generations of progeny from the founder rats both expressed and passed on the transgene. Thus, direct modification of SSCs in juvenile rats is an effective means of generating transgenic rats through the male germline. This technology could be adapted to larger animals, in which existing methods for gene modification are inadequate or inapplicable, resulting in the generation of transgenic animals in a variety of species.
ABSTRACT
Objective To investigate the effects of Epimedium ,Astragalus ,Radix Puerariae on DMT1 expression in the cere‐bral cortex of APPswe/PS1ΔE9 double transgenic mice model of AD .Methods A total of 60 specific‐pathogen‐free male APPswe/PS1ΔE9 double transgenic mice aged 6 months were equally and randomly assigned to model ,Epimedium ,Astragalus ,Radix puerari‐ae ,compound and DFO groups .An additional 10 6‐month‐old C57BL/6J mice served as negative control group .Using immunohisto‐chemistry and molecular biology methods to investigate the effects of a compound combining the effective components of Epimedi‐um ,Astragalus ,Radix puerariae on DMT1 expression in the cerebral cortex of APPswe/PS1ΔE9 double transgenic mice model of AD . Results Immunohistochemical staining results revealed that DM T 1 positive cell did not show in negative control group .DM T1 ex‐pression was higher in model group compared with the negative control group .DMT1 expression was lower in the compound and deferoxamine groups than in the model group .No significant difference was detected in DM T 1 expression between deferoxamine and compound groups .RT‐PCR ,Western blot and immunohistochemical staining results showed no significant difference .Conclusion These compounds can downregulate DMT1 expression and inhibit iron overload in the cerebral cortex of mice with Alzheimer′s dis‐ease ,reduce iron overload induced impairment of the central nervous system .
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Olfactory dysfunction is one of the most debilitating problem in chronic rhinosinusitis (CRS) patients, and exact mechanism underlying sinusitis induced olfactory dysfunction was not fully understood. In vivo manipulation for olfactory epithelium and fresh specimen for histopathological analysis are essential for research, but it is nearly impossible to do in human due to inaccessibility of olfactory epithelium and risk for complication. For this reason, several animal models using toxic materials, such as 3-methylindole or bromomethane, have been suggested for mimicking olfactory epithelial damage in CRS, but none of them could truly imitate the event which happens in real patient. Inducible olfactory inflammation (IOI) mouse is a transgenic mouse model selectively producing tumor necrosis factor-alpha (TNF-alpha) in sustentacular cell of olfactory epithelium. The production of TNF-alpha can be actively initiated by giving food containing doxycycline to IOI mouse, and inflammation is stopped in the absence of doxycycline. Both toxicity model and transgenic model have their own advantages and disadvantages, therefore appropriate model should be selected for optimal results.
Subject(s)
Animals , Humans , Mice , Animals, Genetically Modified , Doxycycline , Inflammation , Mice, Transgenic , Models, Animal , Olfactory Mucosa , Sinusitis , Skatole , Smell , Tumor Necrosis Factor-alphaABSTRACT
Abstract: This article examines whether using Bovine Somatotrophin (bST) and Transgenic Animal is compatible with the norms of animal welfare, environment, and public health. We cannot oppose its usefulness all on a sudden. Despite the usefulness of animal biotechnology, we cannot ignore the different adverse effects of this technology. All of these bring forth different ethical challenges. What is the environmental impact of this technology? Another ethical challenge is related to animal‘s welfare and human‘s health. In order to assessing the ethical challenges, this article has opted for Mepham‘s ethical matrix, which is a practical approach for addressing broader policy issues. I have focused on the application of this ethical matrix upon some contexts of animal biotechnology, such as bST and transgenic animal. Through the analysis, this article came to the conclusion that there are no short curt ways to reach an agreement on the application of animal biotechnology.
ABSTRACT
Skin keratinocytes form a tightly knit and layered epithelium at the surface of the body protecting the body from the outside environment. The formation and maintenance of skin epidermis is governed by dynamic and well-coordinated processes of cell proliferation, differentiation, and self-renewal. Such important cell fate decisions are made possible in part by transcription factors, which activate and repress unique sets of genes in a temporal and spatial pattern. The Tp63 gene encodes for multiple isoforms for one such transcription factor that serves as a key regulator of epidermal development and differentiation. The crucial function of p63 is epitomized by the phenotype of p63 knockout mice – in the absence of p63, there is a profound block in the development of skin epidermis and all related appendages such as hair follicles. Human syndromes resulting from Tp63 gene mutations phenocopy the p63 knockout phenotype, highlighting the evolutionarily conserved function of this factor in epithelial biology. Although the function of p63 as an important hub in transcriptional and signaling networks of keratinocytes is well established, the underlying molecular mechanisms of p63 action is continually redefined with the development of new genetic models and more extensive biochemical analysis. In this review the biological role of Np63, the predominant isoform that is expressed in skin keratinocytes has been described. Results from transgenic animal models that have shed new information on the function of Np63 in the epidermis and hair follicles have been discussed. Further, the molecular mechanisms that maintain the fine-tuned expression of Np63 in skin keratinocytes are also described.
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Uveitis is an important autoimmune disease.Human leukocyte antigen (HLA)-B27-associated anterior uveitis is the most common form of anterior uveitis.HLA-B27-associated anterior uveitis has distinct ocular,systemic,and genetic features.It attacks young subjects and leads to the damage of visual function due to the recurrence and complications.However,the pathologic mechanism of HLA-B27-associated anterior uveitis is still not thoroughly understood,and effective therapy is still unavailable.There are currently many experimental researches which attamp to study the etiology and develop an effective therapy for HLA-B27 acute anterior uveitis.Thus,various animal models of acute anterior uveitis such as endotoxin-induced uveitis and HLA-B27 transgenic animals including rat and mouse are established.Different therapy regimens have been designed both in clinical trial and experimental study and aquired promising achievements.Epidemiology,pathologic mechanism,different animal models and potential new therapies such as anti-TNFα,oral associated peptides tolerance and gene therapy are overviewed.
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A new and effective method to produce transgenic animals was established. Without a surgical incision, the recombinant plasmid containing green fluorescence protein (GFP) cDNA was repeatedly injected into male mouse testis at multi-sites. After few weeks of the final injection, the injected male was mated with normal oestrus female to produce transgenic mice. The presence of the GFP cDNA in F1 transgenic individuals were detected by polymerase chain reaction and Southern blot hybridization, which showed that the transgenic rate of mouse F1 offspring was 41%. The transferred gene was integrated into the host genome and could be transmitted to its offspring. When the positive F1 individuals were mated with the wild type ICR mice, the F2 individuals had a transgenic rate of 37%. The results indicate that the high efficiency of gene transfer and the limited number of manipulations make the method suitable for creating a large number of transgenic animals, especially, for producing domestic animals.
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MicroRNAs (miRNAs) are small non-coding RNAs which are essential for posttranscriptional gene regulation and have important roles in physiology and pathology. The researches on function of miRNA will be a focus in the future. Several animal models have been built by transgenic technology, making important contributions to our understanding of gene function at the whole scale. Recently, the number of transgenic animal models for microRNAs and construction strategies have been increasing and diversifying. Some roles of miRNA in tumor and cardiovascular disease have been revealed by transgenic animals. Transgenic animals are becoming a kind of powerful tool in microRNA researches.
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Objective:To improve the genetic stability of HBV gene in transgenic mice.Methods:HBV transgenic mice were bred by backcross and double cross.The HBV gene expression and replication were studied with real-time PCR,ELISA and chemiluminescence.Results:The HBV transgenic mice have stably bred to 23rd generation.The serum HBsAg level is 4122.31?2044.74IU/ml;The rate of HBV transgenic mice whose serum HBV DNA reach 104~106copies/ml was 93.93%.The HBV replication and expression were improved markedly.There is no difference between male and female mice about serum HBsAg level.Conclusion:After breeding the HBV gene was expressed stably with high-level in transgenic mice.
ABSTRACT
Transgenic animals are those in which recombinant DNA technology is used to remove a gene or a specific part of DNA with exogenous DNA, which is hereditable and can be expressed stably. Transgenic rodent technology, first demonstrated by Gordon in 1980,opened the door to the next step in biotechnology and gene regulation, which has been used to reproduce or mimic a part of a human disease in a complex cellular environment by creating in vivo models with germ line transmission of the introduced genetic regulatory elements. Animal models for drug discovery are developing increasingly, which is promoting the research and development new drugs greatly. The use and development of transgenic animals in pharmacological research is reviewed in this article.