Glial fibrillary acidic protein gfap is a 4053 kda monomeric molecule found only in adult glial cells of the central nervous system cns and represents the major part of the cytoskeleton of astrocytes. Toxicity evaluation and biomarker selection with validated. The zebrafish danio rerio, named for the horizontal stripes on the sides of the adult body, is a tropical freshwater fish in the minnow family. Glial fibrillary acidic protein gfap is the main astrocytic intermediate filament if. In order to improve the assay of human tau in zebrafish, we generated a model where taugfp expression and neurotoxicity could be live monitored. Hedgehog regulated slit expression determines commissure and. We have generated transgenic zebrafish that express green fluorescent protein gfp in glial cells driven by the zebrafish glial fibrillary acidic protein gfap regulatory elements. Highlights of zebrafish tools in our collection crispr. Regenerative response following stab injury in the adult. South dakota game, fish, and parks south dakota game. The journal of comparative neurology 359365381 1995 expression of glial fibrillary acidic protein and its relation to tract formation in embryonic zebrafish danio rerio riva c.
Monoclonal antiglial fibrillary acidic protein gfap g 3893. Zebrafish danio rerio is a fresh water fish that inhabits rivers in india, pakistan and other places in asia. Covering both freshwater and saltwater species of fish, this fully revised edition brings a new level of accuracy and usefulness to the national audubon societys acclaimed field guides. A novel mutation in glial fibrillary acidic protein gene in a patient with alexander disease. In the past two decades, zebrafish has become one of the preferred in vivo model organisms for studying diverse processes like embryogenesis 8, 9, autophagy, behaviour 11, 12 and sleep. Expression analysis of toxicity biomarkers in embryonic zebrafish exposure to mitoxantrone. Noteworthy, the zebrafish gfap reporter line described by bernardos and raymond also encompassed a larger gfp expression domain than anti.
Gfp transgenic line has previously been shown to recapitulate faithfully the pattern of expression of endogenous gfap in the adult zebrafish telencephalon lam et al. Suppression of transcytosis regulates zebrafish blood. The zebrafish book a guide for the laboratory use of zebrafish danio brachydanio rerio by monte westerfield, institute of neuroscience, university of oregon older literature referred to zebrafish as brachydanio rerio. It is implemented both in eukaryotic and prokaryotic. This antibody was generated against a basal lamina and cytoskeleton preparation from adult. Transgenic zebrafish have essentially had exogenous genes added to their genome and as such can be used for many different experimental applications such as stable overexpression. Regulatory elements that direct the glialspecific expression of zebrafish gfap were identified in genomic sequences from the zebrafish gfap gene. Aggregationprone gfap mutation in alexander disease validated. In gosh mutants, both cones and rods degenerate by 3 weeks post.
To control for offtarget signal, we also performed fish with a. Suppression of transcytosis regulates zebrafish bloodbrain. This transgene expresses cre recombinase and betagalactosidase under the control of the human glial fibrillary acidic gfap promoter. Nov 26, 2019 unlike mammals, zebrafish have the capacity to regenerate neurons in response to damage. As a consequence, the pharmaceutical industry is strongly interested in using fish for drug screens and whole animal toxicology studies. Dako products we help pathologists and clinicians make accurate diagnoses that help determine the most effective treatment for cancer patients we are proud to be a renowned provider of complete pathology solutions and flow cytometry reagents, trusted by clinical laboratories around the world in the fight against cancer. Traditionally, astrocytes are divided into fibrous and protoplasmic types based on their morphologic appearance. Valproic acid, a histone deacetylase inhibitor, regulates. Neuronal progenitors in the zebrafish neuroepithelium have been reported to express markers of radial glia, such as the glial acidic fibrillary protein gfap 49, 51. Crelox recombination is a sitespecific recombinase technology, used to carry out deletions, insertions, translocations and inversions at specific sites in the dna of cells. We here present a characterization of the zebrafish gfap gene and.
Zebrafish have astrocytes 23, and zebrafish gfap shares 67% identity. It is a type iii intermediate filament if protein that is expressed by numerous cell types of the central nervous system cns, including astrocytes and ependymal cells during development. Bioprinting is an emerging field in the areas of tissue engineering and regenerative medicine. Transgenic zebrafish an overview sciencedirect topics. A monoclonal antibody stains radial glia in the adult zebrafish.
Glial fibrillary acidic protein gfap, the most popular. Video clips with sample gfp expressions of zebra fish using dinolite premier am41tgfbw using special 480nm excitation. However, in the olfactory bulb of adult zebrafish, gfap is expressed in. More posteriorly, there is good correlation in the expression domains of both gfap. Gfapexpressing cells in the commissure regions of the zebrafish forebrain. Retinal regeneration in adult zebrafish requires regulation. Gfap splice isoforms show differential expression patterns in the human brain. Gfap isoforms in adult mouse brain with a focus on. Characterization of retinal regeneration in adult zebrafish. Two types of teton transgenic lines for doxycycline. Gfap molecular weight of 50 kda is the cell specific if protein in astrocytes.
Altogether, we conclude that the heart dissection protocol yields high quality and cardiacspecific rna from entire zebrafish embryos. The list will be updated when new genes are found or reported. Several reports have demonstrated that vpa functions as a histone deacetylase hdac inhibitor. Unlike mammals, zebrafish have the capacity to regenerate neurons in response to damage. Zebrafish expression atlas a zebrafishspecific gene expression database that provides information about when genes are active in developing embryos.
The drug discovery industry attempts to minimize those events with diligent. The transgene was microinjected into fertilized balbc x c57bl6nhsdf1 oocytes. Tierney, in animal models for the study of human disease, 20. These dualglo transgenic mice express both firefly luciferase luc under the control of the human gfap glial fibrillary acidic protein promoter and renilla luciferase under the control of the 0. I will have a greater knowledge of the different species of fish and continue to use as a reference to any further questions i may have. The zebrafish book a guide for the laboratory use of zebrafish. The construct has a small fragment of the gfap first exon removed to prevent gfap expression from the transgene. Glial fibrillary acidic protein gfap is a protein that is encoded by the gfap gene in humans. The south dakota game, fish and parks website has the outdoor related information you need. The expression levels of classical toxicity biomarker genes fabp10a, gclc, gsr, nqo1, gfap, and. Expression of radial glial markers gfap, blbp and gs. Zebrafish do not possess the typical astrocytes that are found in mammalian. Neural stem cells in the subventricular zone svz of the adult mammalian forebrain are a potential source of neurons for neural tissue repair after brain insults such as ischemic stroke and traumatic brain injury tbi. In the past two decades, zebrafish has become one of the preferred in vivo model.
A specific band was detected for gfap at approximately 51 kda as indicated using 0. It allows the dna modification to be targeted to a specific cell type or be triggered by a specific external stimulus. Crisprz a database of crisprcas9 target sequences that have been experimentally validated in zebrafish. This strain is undergoing recovery from cryopreserved stocks. In contrast, the expression levels of the cns and lens markers gfap and cryaa, respectively were extremely low in the heart sample fig. Most zebrafish retinal injury models employ acute damage, which is unlike the. Structural and functional characterization of the zebrafish gene for. Antibody staining with hpa056030, hpa0635 and cab000039 in immunohistochemistry. Pdf expression of glial fibrillary acidic protein in the.
The gfap cre transgene was designed with the entire mouse glial fibrillary acidic protein gene gfap sequences directing expression of cre recombinase. Expression of glial fibrillary acidic protein and its. Current standard practices are good at preventing toxic compounds from being tested in the clinic. We showed that there was an upregulation of her4 expression in the tsa. Our studies have uncovered a novel role for slit molecules in the positioning of. The third was a monoclonal antibody zrf1, kindly provided by m. Anisotropic muller glial scaffolding supports a multiplex lattice mosaic of photoreceptors in zebrafish retina mikiko nagashima1, jeremy hadidjojo2, linda k. These dualglo transgenic mice express both firefly luciferase luc under the control of the human gfap glial fibrillary acidic protein promoter and renilla luciferase under the control of. Expression of gfap flj45472 in heart muscle tissue. Muller glia in the zebrafish retina respond to retinal damage by reentering the cell cycle, which generates large numbers of retinal progenitors that ultimately replace the lost neurons. To control for offtarget signal, we also performed fish with a mouse gfap probe, which should not bind to the zebrafish transcript figure 4. Tissue expression of gfap staining in heart muscle the.
In order to improve the assay of human tau in zebra fish, we generated a model where taugfp expression and neurotoxicity could be live monitored. Abcam plc mp biomedicals 2 zrf1, monoclonal, igg1, mouse, ihc, zebrafish international resource center zirc. The ability to control transgene expression within specific tissues is an important tool for studying the molecular and cellular mechanisms of development, physiology, and disease. Traumatic injury to the adult cns results in a rapid inflammatory response by the resident astrocytes, characterized mainly by hypertrophy. In biochemical research it is linked to a gene of interest and used to signal to researchers that the gene has. Medical reference laboratory providing esoteric and genetic testing services, test selection guidance, result interpretation references, and continuing education for clients worldwide. Unpredicted human safety events in clinical trials for new drugs are costly in terms of human health and money. It has long been established as a powerful vertebrate model organism for the study of genetics and developmental biology. All the following premade aavs were packaged in serotype 1, 2, 5, 6, 8 and 9 and super purified via 2xcscl ultra. Gfap immunoreactivity, due primarily to the spread of gfp throughout the cytoplasm. A monoclonal antibody stains radial glia in the adult zebrafish danio rerio cns. Neural stem cells in the subventricular zone svz of the adult mammalian forebrain are a potential source of neurons for neural tissue repair after brain insults such as. This transgene is expressed in astrocytes by embryonic day 14. By using the gfap promoter to drive expression of nuclear localized.
Gfap has also been found to be expressed in glomeruli and peritubular fibroblasts taken from rat kidneys. Gfp and endogenous glial fibrillary acidic protein gfap expression in the forebrain of adult zebrafish. Multiple signaling factors and drugs alleviate neuronal death. Gfap protein is distributed more apically along the radial process in tgif1. Two lines 1 and 8 were analyzed further for cre activity and expression. Book series of molecular aspects of fish and marine biology. In addition, the expression of these isoforms in human brain. Wester field that labels radial cells in embryonic zebrafish trevar row et al. Please use this form to recommend updates to the information in zfin. The multiplex, lattice mosaic of cone photoreceptors in the adult fish retina is a compelling example. Gfp expression was first visible in the living embryo at the tail budstage, then in the developing brain by the 5somitestage approximately 12 h postfertilization, hpf and then spreading posteriorly along the developing. Zebrafish astroglial morphology in the olfactory bulb is. We generated two transgenic lines using the xenopus rhodopsin promoter to drive the reverse. It is a type iii intermediate filament if protein that is expressed by numerous cell types of the.
The early developmental expression of gfap in chondrichthyes and teleosts contrasts with its later expression in mammals. Reagent monoclonal antiglial fibrillary acidic protein gfap is provided as ascites fluid containing 15. We found that neuronal cells expressing taugfp in these models can be readily imaged and traced to evaluate the neurotoxicity induced by oligomeric tau proteins. We employed two novel transgenic zebrafish lines and endogenous. The expression levels of classical toxicity biomarker genes fabp10a, gclc, gsr, nqo1, gfap, and erg in 48 hpf embryonic zebrafish exposed to gradient mitoxantrone 0, 10, 50, and 100.
Gfap isoforms in adult mouse brain with a focus on neurogenic. Recent studies show that neurogenesis in the ventricular zone vz of the adult zebrafish telencephalon has features in common with neurogenesis in the adult mammalian svz. Largescale zebrafish embryonic heart dissection for. Aug 21, 2012 video clips with sample gfp expressions of zebra fish using dinolite premier am41tgfbw using special 480nm excitation lighting and 510nm emission filter. A a frontal view of wild type host embryos with transplanted tggfap. We appreciate as much detail as possible and references as appropriate. With 700 new fullcolor photographs, 286 blackandwhite illustrations, and 637 maps, this is the most comprehensive field guide available to the fishes of. Constructs used for these lines are referred to as gfap idh1wt, gfap r2h, gfap r2c, gfap g70d and gfap gfp. Humans with mutations in photoreceptorrelated genes develop forms of retinal degeneration, such as retinitis pigmentosa, cone dystrophy, or leber congenital amaurosis. Most zebrafish retinal injury models employ acute damage, which is unlike the chronic, gradual damage that occurs in human retinal diseases.
Reporter gene definition of reporter gene by medical. Valproic acid vpa has been used to treat epilepsy and bipolar disorder. Use of zebrafish in drug discovery toxicology chemical. The list has links to the sequence data of the genes and est located in ncbi genbank database. Acetylated tubulin antibody and transplanted cells are labeled in green endogenous gfp expression. We also kept adult zebrafish in fish water containing tsa and ly411575 and examined the expression of her4 and the number of pcna. Neuronal regeneration in a zebrafish model of adult brain. List of genes homologous to mammalian apoptosis regulators identified in zebrafish. Gfap expression in the zebrafish brain, detected by immunohistochemistry using antigfap antibodies, is reported to begin at the 12somitestage, which is. Zebrafish photoreceptor degeneration and regeneration. In fish development and genetics the zebrafish and medaka models. Multiple signaling factors and drugs alleviate neuronal.
Here, we studied the regenerative response in the zebrafish aipl1b mutant, gold rush gosh. Reagent monoclonal antiglial fibrillary acidic protein gfap is provided as ascites fluid containing 15 mm sodium azide as a preservative. Muller glia relative to wildtype siblings arrows, fig. Studies in zebrafish embryos have shown that the expression of gfap in the central nervous system cns begin at the time of formation of the first somite thisse et al. The south dakota game, fish and parks provides sustainable outdoor recreational opportunities through responsible management of our states parks, fisheries and wildlife by fostering partnerships, cultivating stewardship and safely connecting people with the outdoors.
Gfappositive astrocytes are rare or absent in primary. Here the cultures were prepared separately from the adult. Glial fibrillary acidic protein gfap, the most popular astrocyte marker wed, 03162016. Dako products we help pathologists and clinicians make accurate diagnoses that help determine the most effective treatment for cancer patients we are proud to be a renowned provider of. Transgenic zebrafish have essentially had exogenous genes added to their genome and as such can be used for many different experimental applications such as stable overexpression, dominantnegative mutation expression, and general analysis of gene regulation. Anisotropic muller glial scaffolding supports a multiplex. Please refer to addgenes crispr guide for a general introduction to crispr technology or the zebrafish crispr resources for a full. Expression of glial fibrillary acidic protein in the rat endocard, cardiac interstitial cajallike cells, and perivascular structures of the spleen.
The drug discovery industry attempts to minimize those events with diligent preclinical safety testing. Simple western lane view shows lysates of human brain cerebellum tissue, loaded at 0. Glial fibrillary acidic protein gfap is a 4053 kda monomeric molecule found only in adult glial cells of the central nervous system cns and represents the major part of the cytoskeleton of. Gfappositive radial glial cells are an essential progenitor. It is defined as the printing of structures consisting of living cells, biomaterials and active.
1468 1426 1668 1268 1068 1674 518 55 1319 1342 889 382 1320 288 496 1417 1381 32 355 1557 820 24 1043 927 894 1155 691 1090 387 1342 419 170 1298 14 1159