The influence of hyproxia an GATA-1 and Epo expression levels in developing zebrafish (PDF)

Inhaltsangabe:Abstract:
The transcription factor GATA-1 is essential for the development of the erythroid cell lineage in vertebrates. In this article we introduce a method to easily determine the approximately development status of red blood cells and the progression of blood formation by intensity of fluorescence in GATA-1/Ds-Red marked zebrafish. We classified the blood cells on the basis of their fluorescence intensity in 5 intensity stages (IS) with the brightest in IS 1. The comparison with our erythropoietin (Epo) data showed a noticeable correlation between GATA-1, Epo mRNA and EPO protein level. Between 2 and 3 dpf we observed a major increase in blood cell concentration to circa 1200 cells*nl-1, until 15 dpf this value decreased to about the half.
The appearance of IS 1 cells correspond approximately to the peaks in Epo cRNA copies and the highest values in EPO protein emerged about 1 day later. Our data show that the blood cell concentration, Epo and Gata-1 expression in zebrafish larvae is subjected to large fluctuations in the first few days of development.
The zebrafish Danio rerio, also known as Brachidanio rerio, Cyprinus rerio and others, is a omnivorous, tropical fish of the family Cyprinidae and was at fist described by Hamilton in 1822. Its natural range within Asia are slow moving or stagnant water bodies in India, Bangladesh, Nepal and Pakistan.
In the past few decades the zebrafish became an important model organism for genetical, developmental and physiological studies. Fish are vertebrates and thus the genetic program is more similar to that of mammals than invertebrate models like the fruity (Drosophila melanogaster). Because of this relationship most of the zebrafish genes have human orthologs.
Due to its short generation time of approximately 3 month, the rapid development and the transparency up to adulthood this tropical teleost is predestined for in vivo hematologic studies and digital imaging techniques. Especially digital imaging is a gentle, non-invasive and thereby seminal method for researcher who are working with transparent animals.
The combination with fluorescent reporter genes allows real time imaging of gene expression and cell migration studies over their whole lifespan. Inhaltsverzeichnis:Table of Contents:
Abbreviations4
Abstract5
1.Introduction6
1.1Zebra_sh hematopoiesis7
1.2GATA-1 and Co-factors8
1.2.1The role of GATA-18
1.2.2GATA-1 related factors10
1.3Epo and EpoR11
1.4Oxygen [...]