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Knockout Rat Models Mimicking Human Atherosclerosis Created By
www.nature.com/scientificreports OPEN Knockout rat models mimicking human atherosclerosis created by Cpf1-mediated gene targeting Received: 23 September 2018 Jong Geol Lee1,2,3, Chang Hoon Ha2,4, Bohyun Yoon2, Seung-A. Cheong1, Globinna Kim1,2,4, Accepted: 8 January 2019 Doo Jae Lee2, Dong-Cheol Woo1,2,4, Young-Hak Kim5, Sang-Yoon Nam3, Sang-wook Lee1,6, Published: xx xx xxxx Young Hoon Sung1,2,4 & In-Jeoung Baek1,2,4 The rat is a time-honored traditional experimental model animal, but its use is limited due to the difculty of genetic modifcation. Although engineered endonucleases enable us to manipulate the rat genome, it is not known whether the newly identifed endonuclease Cpf1 system is applicable to rats. Here we report the frst application of CRISPR-Cpf1 in rats and investigate whether Apoe knockout rat can be used as an atherosclerosis model. We generated Apoe- and/or Ldlr-defcient rats via CRISPR-Cpf1 system, characterized by high efciency, successful germline transmission, multiple gene targeting capacity, and minimal of-target efect. The resulting Apoe knockout rats displayed hyperlipidemia and aortic lesions. In partially ligated carotid arteries of rats and mice fed with high-fat diet, in contrast to Apoe knockout mice showing atherosclerotic lesions, Apoe knockout rats showed only adventitial immune infltrates comprising T lymphocytes and mainly macrophages with no plaque. In addition, adventitial macrophage progenitor cells (AMPCs) were more abundant in Apoe knockout rats than in mice. Our data suggest that the Cpf1 system can target single or multiple genes efciently and specifcally in rats with genetic heritability and that Apoe knockout rats may help understand initial- stage atherosclerosis. -
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Revista de Biología Tropical, ISSN: 2215-2075, Vol. 69(2): 545-556, April-June 2021 (Published Apr. 19, 2021) 545 Rostelato-Ferreira, S., Vettorazzo, O.B., Tribuiani, N., Leal, A.P., Dal Belo, C.A., Rodrigues-Simioni, L., Floriano, R.S., Oshima- Franco, Y. (2021). Action of heparin and acetylcholine modulators on the neurotoxicity of the toad Rhinella schneideri (Anura: Bufonidae) in Brazil. Revista de Biología Tropical, 69(2), 545- 556. DOI 10.15517/rbt.v69i2.44539 DOI 10.15517/rbt.v69i2.44539 Action of heparin and acetylcholine modulators on the neurotoxicity of the toad Rhinella schneideri (Anura: Bufonidae) in Brazil Sandro Rostelato-Ferreira1,2; Orcid: 0000-0002-8987-434X Orlando B. Vettorazzo2; Orcid: 0000-0003-4731-4771 Natália Tribuiani2; Orcid: 0000-0002-7661-475X Allan P. Leal3; Orcid: 0000-0002-4689-4615 Cháriston A. Dal Belo3,4; Orcid: 0000-0001-7010-650 Léa Rodrigues-Simioni5; Orcid: 0000-0002-8712-6639 Rafael S. Floriano6; Orcid: 0000-0003-0759-5863 Yoko Oshima-Franco2*; Orcid: 0000-0002-4972-8444 1. Institute of Health Sciences, Paulista University (UNIP), Sorocaba, São Paulo, Brazil; [email protected] 2. Graduate Program in Pharmaceutical Sciences, University of Sorocaba (UNISO), Sorocaba, São Paulo, Brazil; [email protected], [email protected], [email protected] (*Correspondence) 3. Graduate Program in Toxicological Biochemistry, Federal University of Santa Maria (UFSM), Santa Maria, Rio Grande do Sul, Brazil; [email protected] 4. Laboratory of Neurobiology and Toxinology (Lanetox), Federal University of Pampa (UNIPAMPA), São Gabriel, Rio Grande do Sul, Brazil; [email protected] 5. Department of Pharmacology, Faculty of Medical Sciences, State University of Campinas (UNICAMP), Campinas 13083-970, São Paulo, Brazil; [email protected] 6. -
The Impact of Multiple Climatic and Geographic Factors on the Chemical Defences of Asian Toads (Bufo Gargarizans Cantor)
www.nature.com/scientificreports OPEN The impact of multiple climatic and geographic factors on the chemical defences of Asian toads (Bufo gargarizans Cantor) Yueting Cao, Keke Cui, Hongye Pan, Jiheng Wu & Longhu Wang* Chemical defences are widespread in nature, yet we know little about whether and how climatic and geographic factors afect their evolution. In this study, we investigated the natural variation in the concentration and composition of the main bufogenin toxin in adult Asian toads (Bufo gargarizans Cantor) captured in twenty-two regions. Moreover, we explored the relative importance of eight climatic factors (average temperature, maximum temperature, minimum temperature, average relative humidity, 20–20 time precipitation, maximum continuous precipitation, maximum ground temperature, and minimum ground temperature) in regulating toxin production. We found that compared to toads captured from central and southwestern China, toads from eastern China secreted higher concentrations of cinobufagin (CBG) and resibufogenin (RBG) but lower concentrations of telocinobufagin (TBG) and cinobufotalin (CFL). All 8 climatic variables had signifcant efects on bufogenin production (ri>0.5), while the plastic response of bufogenin toxin to various climate factors was highly variable. The most important climatic driver of total bufogenin production was precipitation: the bufogenin concentration increased with increasing precipitation. This study indicated that the evolution of phenotypic plasticity in chemical defences may depend at least partly on the geographic variation of defensive toxins and their climatic context. Chemical defences is ubiquitous in nature and can serve for deterring predators, competitors, parasites, and path- ogens1–3. Many species can secrete a large diversity of toxic defensive compounds for self-protection. -
UW-Madison Undergraduate Symposium || Abstracts 2014
ABSTRACTS 2014 Undergraduate Symposium Celebrating research, creative endeavor and service-learning We would like to thank our major sponsors and partners: Brittingham Trust General Library System Institute for Biology Education McNair Scholars Program Morgridge Center for Public Service Office of the Provost Undergraduate Academic Awards Office Undergraduate Research Scholars Program University Marketing Wisconsin Union The Writing Center 2014 Undergraduate Symposium Organizing Committee: Jane Harris Cramer, Maya Holtzman, Svetlana Karpe, Kelli Hughes, Linda Kietzer, Laurie Mayberry, Aaron Miller, Christopher Olsen, Janice Rice, Amy Sloane, Julie Stubbs, Malika Taalbi, Beth Tryon, and Berit Ness (coordinator). A special thanks is also extended to Stephanie Diaz de Leon of The Wisconsin Union; Rosemary Bodolay, Patricia Iaccarino, Carrie (Carolyn) Kruse, and Pamela O’Donnell at College Library; and Jeff Crucius of the Division of Information Technology. Cover photos courtesy Office of University Communications and the Undergraduate Symposium Committee ii Abstracts and Art Statements University of Wisconsin–Madison April 10, 2014 • Union South THE SHORT AND DIFFICULT LIFE OF WALES’ DEVOLVED GOVERNMENT: WAS IT ALWAYS A LEGITIMATE INSTITUTION? Lucille Abrams, Yoshiko Herrera (Mentor), Political Science The construction of the regional, or devolved, government of Wales, as well as its corresponding governing body, the National Assembly for Wales, was a protracted process. Through library research, I sought to find the reasons for this dif- ficulty and ask if Welsh citizens believed the Assembly to be a valid, or legitimate, political institution. The political history of the United Kingdom, Wales’ individual history, and analysis of voter turnout at the first Assembly election, contribute to answering this question. The Assembly did not suffer from a lack of legitimacy but from an apathetic and alienated constitu- ency. -
Introducing Gene Deletions by Mouse Zygote Electroporation of Cas12a/Cpf1
Zurich Open Repository and Archive University of Zurich Main Library Strickhofstrasse 39 CH-8057 Zurich www.zora.uzh.ch Year: 2019 Introducing gene deletions by mouse zygote electroporation of Cas12a/Cpf1 Dumeau, Charles-Etienne ; Monfort, Asun ; Kissling, Lucas ; Swarts, Daan C ; Jinek, Martin ; Wutz, Anton Abstract: CRISPR-associated (Cas) nucleases are established tools for engineering of animal genomes. These programmable RNA-guided nucleases have been introduced into zygotes using expression vectors, mRNA, or directly as ribonucleoprotein (RNP) complexes by different delivery methods. Whereas mi- croinjection techniques are well established, more recently developed electroporation methods simplify RNP delivery but can provide less consistent efficiency. Previously, we have designed Cas12a-crRNA pairs to introduce large genomic deletions in the Ubn1, Ubn2, and Rbm12 genes in mouse embryonic stem cells (ESC). Here, we have optimized the conditions for electroporation of the same Cas12a RNP pairs into mouse zygotes. Using our protocol, large genomic deletions can be generated efficiently by electroporation of zygotes with or without an intact zona pellucida. Electroporation of as few as ten zygotes is sufficient to obtain a gene deletion in mice suggesting potential applicability of thismethod for species with limited availability of zygotes. DOI: https://doi.org/10.1007/s11248-019-00168-9 Posted at the Zurich Open Repository and Archive, University of Zurich ZORA URL: https://doi.org/10.5167/uzh-181145 Journal Article Published Version The following work is licensed under a Creative Commons: Attribution 4.0 International (CC BY 4.0) License. Originally published at: Dumeau, Charles-Etienne; Monfort, Asun; Kissling, Lucas; Swarts, Daan C; Jinek, Martin; Wutz, Anton (2019). -
Athe Characteristics of Glucose Metabolism in the Sulfonylurea
Zhou et al. Molecular Medicine (2019) 25:2 Molecular Medicine https://doi.org/10.1186/s10020-018-0067-9 RESEARCHARTICLE Open Access aThe characteristics of glucose metabolism in the sulfonylurea receptor 1 knockout rat model Xiaojun Zhou1†, Chunmei Xu1†, Zhiwei Zou2, Xue Shen3, Tianyue Xie3, Rui Zhang1, Lin Liao1* and Jianjun Dong2* Abstract Background: Sulfonylurea receptor 1 (SUR1) is primarily responsible for glucose regulation in normal conditions. Here, we sought to investigate the glucose metabolism characteristics of SUR1−/− rats. Methods: The TALEN technique was used to construct a SUR1 gene deficiency rat model. Rats were grouped by SUR1 gene knockout or not and sex difference. Body weight; glucose metabolism indicators, including IPGTT, IPITT, glycogen contents and so on; and other molecule changes were examined. Results: Insulin secretion was significantly inhibited by knocking out the SUR1 gene. SUR1−/− rats showed lower body weights compared to wild-type rats, and even SUR1−/− males weighed less than wild-type females. Upon SUR1 gene knockout, the rats showed a peculiar plasma glucose profile. During IPGTT, plasma glucose levels were significantly elevated in SUR1−/− rats at 15 min, which could be explained by SUR1 mainly working in the first phase of insulin secretion. Moreover, SUR1−/− male rats showed obviously impaired glucose tolerance than before and a better insulin sensitivity in the 12th week compared with females, which might be related with excess androgen secretion in adulthood. Increased glycogen content and GLUT4 expression and the inactivation of GSK3 were also observed in SUR1−/− rats, which suggested an enhancement of insulin sensitivity. Conclusions: These results reconfirm the role of SUR1 in systemic glucose metabolism. -
Preeclampsia: Cardiotonic Steroids, Fibrosis, Fli1 and Hint to Carcinogenesis
International Journal of Molecular Sciences Review Preeclampsia: Cardiotonic Steroids, Fibrosis, Fli1 and Hint to Carcinogenesis Natalia I. Agalakova 1, Nikolai I. Kolodkin 2, C. David Adair 3 , Alexander P. Trashkov 4 and Alexei Y. Bagrov 1,* 1 Sechenov Institute of Evolutionary Physiology and Biochemistry, 44 Torez Prospect, 194223 St. Petersburg, Russia; [email protected] 2 State Institute of Highly Pure Biopreparations and Sechenov Institute of Evolutionary Physiology and Biochemistry, 44 Torez Prospect, 194223 St. Petersburg, Russia; [email protected] or [email protected] 3 Department of Obstetrics and Gynecology, University of Tennessee, Chattanooga, TN 37402, USA; [email protected] or [email protected] 4 Konstantinov St. Petersburg Nuclear Physics Institute, National Research Centre Kurchatov Institute, 1 Orlova Roshcha, 188300 Gatchina, Russia; [email protected] * Correspondence: [email protected] Abstract: Despite prophylaxis and attempts to select a therapy, the frequency of preeclampsia does not decrease and it still takes the leading position in the structure of maternal mortality and morbidity worldwide. In this review, we present a new theory of the etiology and pathogenesis of preeclampsia that is based on the interaction of Na/K-ATPase and its endogenous ligands in- cluding marinobufagenin. The signaling pathway of marinobufagenin involves an inhibition of transcriptional factor Fli1, a negative regulator of collagen synthesis, followed by the deposition of collagen in the vascular tissues and altered vascular functions. Moreover, in vitro and in vivo neutralization of marinobufagenin is associated with the restoration of Fli1. The inverse relationship Citation: Agalakova, N.I.; Kolodkin, between marinobufagenin and Fli1 opens new possibilities in the treatment of cancer; as Fli1 is N.I.; Adair, C.D.; Trashkov, A.P.; a proto-oncogene, a hypothesis on the suppression of Fli1 by cardiotonic steroids as a potential Bagrov, A.Y. -
Reproductive Technologies for the Generation and Maintenance Of
Journal of Reproduction and Development, Vol. 64, No 3, 2018 —SRD Innovative Technology Award 2017— Reproductive technologies for the generation and maintenance of valuable animal strains Takehito KANEKO1–3) 1)Division of Science and Engineering, Graduate School of Arts and Science, Iwate University, Iwate 020-8551, Japan 2)Department of Chemistry and Biological Sciences, Faculty of Science and Engineering, Iwate University, Iwate 020-8551, Japan 3)Soft-Path Science and Engineering Research Center (SPERC), Iwate University, Iwate 020-8551, Japan Abstract. Many types of mutant and genetically engineered strains have been produced in various animal species. Their numbers have dramatically increased in recent years, with new strains being rapidly produced using genome editing techniques. In the rat, it has been difficult to produce knockout and knock-in strains because the establishment of stem cells has been insufficient. However, a large number of knockout and knock-in strains can currently be produced using genome editing techniques, including zinc-finger nuclease (ZFN), transcription activator-like effector nuclease (TALEN), and the clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated protein 9 (Cas9) system. Microinjection technique has also contributed widely to the production of various kinds of genome edited animal strains. A novel electroporation method, the “Technique for Animal Knockout system by Electroporation (TAKE)” method, is a simple and highly efficient tool that has accelerated the production of new strains. Gamete preservation is extremely useful for maintaining large numbers of these valuable strains as genetic resources in the long term. These reproductive technologies, including microinjection, TAKE method, and gamete preservation, strongly support biomedical research and the bio-resource banking of animal models. -
Generation of Hprt-Disrupted Rat Through Mouse←Rat ES Chimeras Ayako Isotani1,2, Kazuo Yamagata2,†, Masaru Okabe2 & Masahito Ikawa1,2
www.nature.com/scientificreports OPEN Generation of Hprt-disrupted rat through mouse←rat ES chimeras Ayako Isotani1,2, Kazuo Yamagata2,†, Masaru Okabe2 & Masahito Ikawa1,2 We established rat embryonic stem (ES) cell lines from a double transgenic rat line which harbours Received: 03 February 2016 CAG-GFP for ubiquitous expression of GFP in somatic cells and Acr3-EGFP for expression in sperm Accepted: 22 March 2016 (green body and green sperm: GBGS rat). By injecting the GBGS rat ES cells into mouse blastocysts Published: 11 April 2016 and transplanting them into pseudopregnant mice, rat spermatozoa were produced in mouse←rat ES chimeras. Rat spermatozoa from the chimeric testis were able to fertilize eggs by testicular sperm extraction combined with intracytoplasmic sperm injection (TESE-ICSI). In the present paper, we disrupted rat hypoxanthine-guanine phosphoribosyl transferase (Hprt) gene in ES cells and produced a Hprt-disrupted rat line using the mouse←rat ES chimera system. The mouse←rat ES chimera system demonstrated the dual advantages of space conservation and a clear indication of germ line transmission in knockout rat production. The investigation of gene functions has intensified following completion of genome projects in many species. A significant number of gene disrupted mouse lines have been produced using ES cells and the homologous recom- bination technique, for example, helping clarify the basic biology while serving as animal models for human diseases1. As a result, mice have replaced rats as the most popularly used experimental animal. However, rats retain advantages over mice as experimental animals2,3. Their bigger body size facilitates exper- imental operations and repeated collection of blood samples. -
A Critical Role for the Type I Interferon Receptor in Virus-Induced Autoimmune Diabetes in Rats
Diabetes Volume 66, January 2017 145 Natasha Qaisar,1 Suvana Lin,1 Glennice Ryan,1 Chaoxing Yang,2 Sarah R. Oikemus,3 Michael H. Brodsky,3 Rita Bortell,2 John P. Mordes,1 and Jennifer P. Wang1 A Critical Role for the Type I Interferon Receptor in Virus-Induced Autoimmune Diabetes in Rats Diabetes 2017;66:145–157 | DOI: 10.2337/db16-0462 The pathogenesis of human type 1 diabetes, character- is heritable but non-Mendelian, and genetic susceptibility loci ized by immune-mediated damage of insulin-producing are insufficient for predicting diabetes onset; most people b-cells of pancreatic islets, may involve viral infection. with risk alleles never become diabetic (2). Interaction of Essential components of the innate immune antiviral genes with environmental factors has been invoked as a response, including type I interferon (IFN) and IFN determinant of disease (3,4). Viral infection, particularly receptor–mediated signaling pathways, are candidates with enterovirus, is believed to be a key environmental mod- for determining susceptibility to human type 1 diabetes. Nu- ulator of T1D, and its possible role in pathogenesis has been PATHOPHYSIOLOGY merous aspects of human type 1 diabetes pathogenesis reviewed in detail (5,6). The mechanisms that underlie viral are recapitulated in the LEW.1WR1 rat model. Diabetes triggering of T1D remain unclear; b-cell infection, by- can be induced in LEW.1WR1 weanling rats challenged stander activation, antigenic spreading, and molecular with virus or with the viral mimetic polyinosinic:polycyti- mimicry have been proposed. Alternatively, viruses could dylic acid (poly I:C). We hypothesized that disrupting prevent T1D through immunoregulation or induction of thecognatetypeIIFNreceptor(typeIIFNa/b receptor [IFNAR]) to interrupt IFN signaling would prevent or delay protective immunity (7). -
World Journal of Pharmaceutical Research Pradhan Et Al
World Journal of Pharmaceutical Research Pradhan et al. World Journal of Pharmaceutical SJIF ImpactResearch Factor 6.805 Volume 5, Issue 8, 426-443. Review Article ISSN 2277– 7105 ` BUFO SKIN-SECRETIONS ARE SOURCES OF PHARMACOLOGICALLY AND THERAPEUTICALLY SIGNIFICANT COMPOUNDS Dr. Bishnu Charan Pradhan*1 and Shakti Prasad Pradhan2 1Dept. of Zoology Angul Mahila Mahavidyalaya, Angul, Odisha, India. 759122. 2Dept. of Pharmacy. Utkal University, Vanivihar, Bhubaneswar, Odisha, India. ABSTRACT Article Received on 07 June 2016, Amphibians have been occupying a wide range of habitats since they Revised on 28 June 2016, evolved around 363 million-years-ago. Along with legs and lungs, skin Accepted on 18 July 2016 DOI: 10.20959/wjpr20168-6752 played an important role in survival of amphibians and made it possible for them to exploit diverse ecological conditions. Amphibian skin not only helps in avoiding desiccation but also helps in imposing *Corresponding Author Dr. Bishnu Charan defense against predators as well as pathogens. Amphibian skin Pradhan possesses wide variety of chemical compounds, which have potential Dept. of Zoology Angul significance in pharmacology and therapeutics. Toads especially those Mahila Mahavidyalaya, belonging to genus Bufo, are outstanding source of useful granular- Angul, Odisha, India. 759122. gland secretions. Compounds derived from toad skin-secretions can be used as analgesics, painkillers and as medicine against cardiac- problems, multi-drug resistant bacteria, HIV and Cancer. KEYWORDS: Bufadienolides, pharmacology, Bufo skin-secretions, toxins. INTRODUCTION Amphibians started trolling the landmasses of earth about 363 million-years-ago, with Acanthostega and Ichthyostega probably being the earliest of known amphibians (Evans[20] et al 1998). Fossil records elucidate that ancestors of modern amphibians like Frogs, Toads, Caecilians and Salamanders probably evolved about 200 million years ago during the Triassic period. -
2-Pg. Curr. Vitae 11-3
CURRICULUM VITAE NANCY HUGHES ING May 11, 2015 Departments of Animal Science and Veterinary Integrative Biosciences Texas A&M University 2471 TAMU College Station, TX 77843-2471 Office Phone: (979) 862-2790 Home Telephone:(979) 690-6566 Fax: (979) 862-3399 e-mail: [email protected] EDUCATION: 1976-1979 B.S. University of Florida Zoology (with Honors) 1980-1984 D.V.M. University of Florida Veterinary Medicine 1980-1988 Ph.D. University of Florida Biochemistry & Molecular Biology 1988-1992 Post-Doc Baylor College of Medicine Cell Biology PROFESSIONAL APPOINTMENTS: 1986-1988 Research Assistant Department of Animal Science University of Missouri 1988-1992 Post-Doctoral Fellow Department of Cell Biology Baylor College of Medicine 1992- Assistant Professor Department of Animal Science Texas A&M University 1992- Joint Appointment Department of Veterinary Integrative Biosciences Texas A&M University 1998 Associate Professor Department of Animal Science Texas A&M University HONORS and AWARDS: 1975 Scholarship to attend the Summer Science Research Program of the Florida Foundation for Future Scientists at the University of Florida, American Cancer Society. 1979 Rita McTigue O'Connell Award Gainesville Women's Club 1979 Phi Beta Kappa University of Florida 1980 ERF Award American Medical Association 1980 Graduate Fellowship for Women Entering Non-Traditional Careers University of Florida 2000 Gamma Sigma Delta (Agricultural Honor Society) Texas A&M University 2004 Phi Beta Kappa (Founding Member) Texas A&M University 2005 Phi Zeta (Veterinary Honor