Expression of G-protein Coupled Receptors in Young and Mature Thrombocytes and Knockdown of Gpr18 in Zebrafish

Expression of G-protein Coupled Receptors in Young and Mature Thrombocytes and Knockdown of Gpr18 in Zebrafish

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Date: May 2013
Creator: Potbhare, Vrinda Nikhil
Description: In this study, a novel method based on biotinylated antibodies and streptavidin coated magnetic beads was used to separate the thrombocyte subpopulations from zebrafish whole blood. DiI-C18, a lipophilic dye, labels only young thrombocytes when used at low concentrations. Commercially available biotinylated anti-Cy3 antibody was used to label the chromophore of DiI-C18 on the young thrombocytes and streptavidin coated magnetic beads were added subsequently, to separate young thrombocytes. The remaining blood cells were probed with custom-made biotinylated anti-GPIIb antibody and streptavidin magnetic beads to separate them from other cells. Further, thrombocytes are equivalents of mammalian platelets. Platelets play a crucial role in thrombus formation. The G-protein coupled receptors (GPCRs) present on the platelet surface are involved during platelet activation and aggregation processes. So, thrombocytes were studied for the presence of GPCRs. The GPCR mRNA transcripts expressed in the young and mature thrombocytes were subjected to densitometry analysis and pixel intensities of the bands were compared using one way ANOVA. This analysis did not show significant differences between the young and mature GPCR mRNA transcripts but identified a novel GPCR, GPR18 that was not reported in platelets earlier. To study the function of this GPCR, it was knocked down using GPR18 ...
Contributing Partner: UNT Libraries
Novel Role of Trypsin in Zebrafish

Novel Role of Trypsin in Zebrafish

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Date: May 2013
Creator: Alsrhani, Abdullah Falleh
Description: It has been shown previously in our laboratory that zebrafish produce trypsin from their gills when they are under stress, and this trypsin is involved in thrombocyte activation via PAR2 during gill bleeding. In this study, I investigated another role of the trypsin that is secreted from zebrafish. This investigation has demonstrated a novel role of trypsin in zebrafish. Not only did this investigation demonstrate the role of trypsin in zebrafish behavior, but also it showed that PAR2 might be the receptor that is involved in trypsin-mediated behavioral response. In addition, we have shown that Gq and ERK inhibitors are able to block the trypsin pathway and prevent the escaping behavior. Finally, the results of this investigation suggest that the cells that respond to trypsin are surface cells, which have an appearance similar to that of neuromast cells.
Contributing Partner: UNT Libraries
Kcnq1-5 (Kv7.1-5) potassium channel expression in the adult zebrafish

Kcnq1-5 (Kv7.1-5) potassium channel expression in the adult zebrafish

Date: February 20, 2014
Creator: Wu, Calvin; Sharma, Kanishk; Laster, Kyle; Hersi, Mohamed; Torres, Christina; Lukas, Thomas J. et al.
Description: In this article, the authors use the zebrafish genome and potassium channels to study cardiac anomalies, epilepsy, convulsions, hearing loss, and tinnitus in humans.
Contributing Partner: UNT College of Arts and Sciences
Zebrafish Thrombocytes: Functions and Origins

Zebrafish Thrombocytes: Functions and Origins

Date: 2012
Creator: Khandekar, Gauri; Kim, Seongcheol & Jagadeeswaran, Pudur
Description: Article integrating the information available on zebrafish thrombocyte function and its formation.
Contributing Partner: UNT College of Arts and Sciences
Zebrafish May Help Solve Ringing in Vets' Ears

Zebrafish May Help Solve Ringing in Vets' Ears

Date: May 6, 2008
Creator: Paul, Marla & Moore, Ernest J.
Description: Article discussing research by Dr. Ernest J. Moore, professor of molecular pharmacology and audiology. This article discusses his research on tinnitus in zebrafish.
Contributing Partner: UNT College of Arts and Sciences
Cardiac and metabolic physiology of early larval zebrafish (Danio rerio) reflects parental swimming stamina

Cardiac and metabolic physiology of early larval zebrafish (Danio rerio) reflects parental swimming stamina

Date: February 24, 2012
Creator: Gore, Matthew & Burggren, Warren W.
Description: Article discussing research on cardiac and metabolic physiology of early larval zebrafish (Danio rerio) reflecting parental swimming stamina.
Contributing Partner: UNT College of Arts and Sciences
Proteomic Responses in the Gill of Zebrafish Following Exposure to Ibuprofen and Naproxen

Proteomic Responses in the Gill of Zebrafish Following Exposure to Ibuprofen and Naproxen

Date: August 2012
Creator: Adhikari, Prem R.
Description: Non-steroidal anti-inflammatory drugs (NSAIDs) are among the most abundant environmental pharmaceutical contaminants. In this study, a proteomic analysis was conducted to identify proteins differentially expressed in gill tissue of zebrafish (Danio rerio) after a 14-day exposure to the NSAIDs ibuprofen or naproxen. A total of 104 proteins with altered expression as indicated by 2-dimensional electrophoresis were analyzed by liquid chromatography with ion trap mass spectrometry (MS/MS). A total of 14 proteins fulfilled our requirements for identification which included consistency among replicate gels as well as successful MS/MS ion searches with the MASCOT database. The most prominent feature of the differential protein expression observed after NSAID exposure was an up-regulation of proteins belonging to the globin family which are involved in the transport of oxygen from gills and availability of heme molecules required for synthesis of cyclooxygenase. Differential expression was observed at exposure concentrations as low as 1-10 µg/L indicating that altered gene expression may occur in fish subjected to environmentally realistic levels of NSAID exposure.
Contributing Partner: UNT Libraries
The Developmental Physiology of the Zebrafish: Influence of Environment and Cardiovascular Attributes

The Developmental Physiology of the Zebrafish: Influence of Environment and Cardiovascular Attributes

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Date: August 2001
Creator: Bagatto, Brian
Description: Temperature effects on the development of the zebrafish embryos and larvae and adults were examined. It was found that the earlier in development a temperature change was performed on an embryo, the more significant the change in survival and/or subsequent development. Thus, viable temperature ranges for zebrafish widened significantly as development proceeded. Adults reared and bred at 25oC produced embryos that were significantly more successful at the lower range of rearing temperatures compared to embryos produced from adults reared at 28oC. The majority of this study focused on the physiological effects of swim training during development in the zebrafish. The earlier in development the zebrafish larvae were trained, the greater the mortality. Trained free swimming larvae had a significantly higher routine oxygen consumption after 11 days of training, and a higher mass specific routine metabolic rate after 8 and 11 days of training. Trained free swimming larvae consumed significantly less oxygen during swimming and were more efficient at locomotion, compared to control larvae. Training enhanced survival during exposure to extreme hypoxia in all age groups. Performance aspects of training were investigated in attempt to quantify training effects and in most cases, trained fish performed significantly better than controls. As blood ...
Contributing Partner: UNT Libraries
Identification of Hox Genes Controlling Thrombopoiesis in Zebrafish

Identification of Hox Genes Controlling Thrombopoiesis in Zebrafish

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Date: December 2015
Creator: Sundaramoorthi, Hemalatha
Description: Thrombocytes are functional equivalents of mammalian platelets and also possess megakaryocyte features. It has been shown earlier that hox genes play a role in megakaryocyte development. Our earlier microarray analysis showed five hox genes, hoxa10b, hoxb2a, hoxc5a, hoxc11b and hoxd3a, were upregulated in zebrafish thrombocytes. However, there is no comprehensive study of genome wide scan of all the hox genes playing a role in megakaryopoiesis. I first measured the expression levels of each of these hox genes in young and mature thrombocytes and observed that all the above hox genes except hoxc11b were expressed equally in both populations of thrombocytes. hoxc11b was expressed only in young thrombocytes and not in mature thrombocytes. The goals of my study were to comprehensively knockdown hox genes and identify the specific hox genes involved in the development of thrombocytes in zebrafish. However, the existing vivo-morpholino knockdown technology was not capable of performing such genome-wide knockdowns. Therefore, I developed a novel cost- effective knockdown method by designing an antisense oligonucleotides against the target mRNA and piggybacking with standard control morpholino to silence the gene of interest. Also, to perform knockdowns of the hox genes and test for the number of thrombocytes, the available techniques were ...
Contributing Partner: UNT Libraries
Role of Gpr17 in Thrombocyte Aggregation in Adult Zebrafish

Role of Gpr17 in Thrombocyte Aggregation in Adult Zebrafish

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Date: December 2015
Creator: Bohassan, Maruah Hejey
Description: GPR17, a uracil nucleotide cysteinyl leukotriene receptor, belongs to the GPCR (G protein coupled receptor) family. It has been shown recently that inhibiting this protein in the nervous system in mice can lead to blockage of oligodendrocyte maturation, which supports myelin repair. Interestingly, our laboratory found GPR17 in thrombocytes. However, we do not know whether it has any function in thrombocyte aggregation or the nature of the ligand. In this paper, we studied the role of GPR17 in hemostasis, which is a fundamental defense mechanism in the event of injury. Using zebrafish as a model system, our laboratory has studied specifically thrombocytes, which play a significant role in hemostasis. The major reasons to use zebrafish as a model system are that their thrombocytes are functionally equivalent to human platelets, the adult fish are amenable to knockdown experiments, and they are readily available in the market. This study was performed by using a piggy back knockdown method where we used a chemical hybrid of control morpholino and an antisense oligonucleotide sequence leads to the degradation the mRNA for GPR17. After knockdown GPR17 in thrombocytes, the percent difference of the thrombocytes aggregation between the control and knockdown blood samples was measured by ...
Contributing Partner: UNT Libraries