Utjecaj prenatalnog izlaganja etanolu na ekspresiju transkripcijskih faktora u timocitima odraslih štakora

master's thesis

• Title (english): Transcription factor expression in thymocytes of adult rats prenatally exposed to etanol
• Author: Sanja Novak
• Mentor: Mariastefania Antica (mentor)
• Committee member: Mariastefania Antica (predsjednik povjerenstva)
• Committee member: Nada Oršolić (član povjerenstva)
• Committee member: Jasna Lajtner (član povjerenstva)
• Granter: University of Zagreb; Faculty of Science; (Department of Biology); Zagreb
• Defense date and country: 2010-10-29, Croatia
• Scientific / art field, discipline and subdiscipline: NATURAL SCIENCES; Biology

Abstract

Hematopoeza je proces stvaranja novih krvnih stanica. Transkripcijski faktori iz obitelji Ikaros i gen Hes igraju ključnu ulogu u razvoju limfocita. Obitelj transkripcijskih faktora Ikaros čine Ikaros, Aiolos, Helios, Eos i Pegasus. Njihova ekspresija, kao i Notch dojavljivanje važni su faktori u hematopoezi i razvoju limfocita. Visoko su homologni među različitim vrstama zbog svoje važnosti. U ovom radu je po prvi put potvrđene i izmjerena ekspresija transkripcijskih faktora iz obitelji Ikaros, Aiolos, Eos i Pegasus u timusu štakora. Također je utvrđena ekspresija Hes, gena koji je aktiviran nizvodno od puta dojavljivanja Notch. Prethodno je pokazano da etanol može utjecati na razvoj hematopoeze. Analizom ekspresije CD3, CD8 i CD4 molekula na stanicama timusa praćena je raspodjela glavnih timocitnih subpopulacija u štakora prenatalno izloženih etanolu. Majke navedenih štakora su tijekom gestacije dobivale 10% etanola u vodi za piće. Osim protočne citometrije također je korišten semi-kvantitativni RT-PCR u svrhu otkrivanja moguće promjene u razinama timusne mRNA za transkripcijske faktore Aiolos, Eos, Pegasus i Hes. Uspoređene su kontrolne životinje sa štakorima prenatalno izloženim etanolu. Referentni gen, HPRT, služio je kao standard. Ekspresija Aiolosa, Eosa, Pegasusa i Hesa kao i raspodjela subpopulacija timocita u odraslih štakora ukazuje na mogućnost da u timusu štakora prenatalno izloženih etanolu dolazi do poremećaja u razvoju CD3 pozitivnih limfocita T što može biti posljedica jače ekspresije transkripcijskih faktora važnih u diferencijaciji.

Abstract (english)

Hematopoesis is a proces of new blood cell formation. Transcription factors from the Ikaros and Notch families play a crucial role in lymphocyte development. Ikaros family transcription factors are Ikaros, Aiolos, Helios, Eos and Pegasus. The expression of these factors as well as the Notch signaling pathway is limiting for hematopoietic and lymphocyte development. Due to their importance Ikaros and Notch are highly homologous between species. In this work we show for the first time the expression of Ikaros family transcription factors, Aiolos, Eos and Pegasus in the rat thymus. We also analyzed the rat thymus for Hes expression, a protein which is activated downstream of the Notch signaling pathway. It has been previously shown that ethanol can influence hematopoietic development. Therefore we analyzed the thymus of adult rats whose mothers during gestation drunk 10% ethanol in their drinking water. By four colour flow cytometry we studied the distribution of the major lymphocyte subpopulations in the rat thymus according to CD3, CD4 and CD8 expression. We found a higher incidence of CD3 positive cells in the double positive CD4+CD8+ thymic subpopulation of rats prenatally exposed to ethanol when compared to controls. Further, we established a semi-quantitative RT-PCR in order to detect possible changes in thymus mRNA expression levels of Aiolos, Eos, Pegasus and Hes in rats prenatally exposed to ethanol. A housekeeping gene, HPRT was used as a standard. Rats prenatally exposed to ethanol show a slightly higher expression of Aiolos, Eos, Pegasus and Hes in the thymus when compared to control rats. However these differences were not statisticaly significant. Our findings indicate that ethanol exposure of pregnant rats might influence the developmnet of CD3 positive cells in the thymus which is related to an increase in transcription factor expression.

• Keywords: hematopoeza; transkripcijski faktori; semi-kvantitativni RT-PCR; protočna citometrija
• Keywords (english): hemathopoesis; transcription factor; semi-quantitative PCR; flow cythometry
• Language: croatian
• URN:NBN: urn:nbn:hr:217:337920
• Study programme: Title: Experimental Biology; Study programme type: university; Study level: graduate; Academic / professional title: magistar/magistra eksperimentalne biologije (magistar/magistra eksperimentalne biologije)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Access restricted to students and staff of home institution
• Created on: 2017-05-04 07:44:03