doctoral thesis
Human cumulus cell expression profile of AMH, AMHR2, FSHR and AR genes in predicting oocyte fertilization potential

Sanja Dević Pavlić (2017)
University of Rijeka
Department of Biotechnology
Metadata
TitleEKSPRESIJSKI PROFIL AMH, AMHR2, FSHR I AR GENA HUMANIH STANICA KUMULUSA U PREDVIĐANJU FERTILIZACIJSKOG POTENCIJALA JAJNE STANICE
AuthorSanja Dević Pavlić
Mentor(s)Anđelka Radojčić Badovinac
Abstract
Temelj uspješnosti medicinski pomognute oplodnje je pravilno sazrijevanje jajnih stanica. Na kvalitetu jajne stanice velik utjecaj ima okruženje u kojem se ona nalazi, tzv. mikrookoliš koji uključuje stanice kumulusa i folikularnu tekućinu. Određivanje pojedinih komponenti tog mikrookoliša koje se jednostavno i brzo mogu detektirati, a koje omogućavaju razlikovanje kvalitete jajnih stanica, doprinijelo bi povećanju uspješnosti metoda pomognute oplodnje. Anti-Müllerov hormon (AMH) je glikoproteinski faktor rasta, koji luče stanice kumulusa. S obzirom na njegovu ulogu u folikulogenezi, ovim je doktorskim radom istražen ekspresijski profil ovog gena, i s njim povezanih receptora (AMHR2, FSHR i AR), te njihova korelacija s kvalitetom jajnih stanica u postupku medicinski pomognute oplodnje, odnosno s morfološkim karakteristikama jajnih stanica, zigota i embrija te njihovim fertilizacijskim potencijalom. U istraživanje je uključeno 129 uzoraka stanica kumulusa i 35 uzoraka folikularne tekućine koji su sakupljeni od ukupno 52 pacijentice u postupku medicinski pomognute oplodnje. Primijenjena je metoda relativne kvantifikacije genskih ekspresija istraživanih gena u stanicama kumulusa, enzimatski pojačani dvostruki imunološki test (ELISA) za određivanje koncentracije AMH u folikularnoj tekućini istog folikula te morfološka analiza jajnih stanica, zigota i embrija invertnom mikroskopijom. Rezultati dobiveni istraživanjem po prvi put ukazuju na negativnu povezanost ekspresije AMH i AR gena u stanicama kumulusa s morfološkom kvalitetom pripadajuće jajne stanice, odnosno s njenom zrelošću. Osim navedenog, istraživanjem je utvrđeno i postojanje korelacija među ekspresijama ispitivanih gena: između FSHR gena i svih ostalih istraživanih gena (AMH, AMHR2 i AR), između AMHR2 i AR gena, te između AMH i AR gena. Dobiveni rezultati pridonose razjašnjenju potencijalne veze AMH (i receptora koji su na direktni ili manje direktni način povezani s ovim hormonom) i morfološke kvalitete jajne stanice. Istraživanjem je potvrđena važnost kompleksa kumulusa i oocite, odnosno komunikacije između jajne stanice i njenog mikrookoliša, za razvoj kvalitetne oocite koja je preduvjet za uspješnu oplodnju i razvoj kvalitetnog embrija.
KeywordsAnti-Müllerian hormone (AMH) cumulus cells follicular fluid oocyte morphology oocyte quality
Parallel title (English)Human cumulus cell expression profile of AMH, AMHR2, FSHR and AR genes in predicting oocyte fertilization potential
Committee MembersKrešimir Pavelić (committee chairperson)
Miljenko Kapović (committee member)
Neda Smiljan Severinski (committee member)
GranterUniversity of Rijeka
Lower level organizational unitsDepartment of Biotechnology
PlaceRijeka
StateCroatia
Scientific field, discipline, subdisciplineBIOTECHNICAL SCIENCES
Biotechnology
UDK612
APPLIED SCIENCES. MEDICINE. TECHNOLOGY
Physiology
Study programme typeuniversity
Study levelpostgraduate
Study programmeMedicinal chemistry
Academic title abbreviationdr. sc.
Genredoctoral thesis
Language Croatian
Defense date2017-02-10
Promoted date2017
Parallel abstract (English)
The core notion determining the success rate of the medically assisted reproduction is the right growth of the oocytes. The environment, or the so called microenvironment, greatly influences the quality of the oocyte and it includes the cumulus cells and the follicular fluid. Determining the specific components of this microenvironment, which can be easily and swiftly detected, and which enable us to distinguish between good and bad quality oocytes, would contribute to the success rate of the medically assisted reproduction. Anti-Müllerian hormone is a glycoprotein growth factor emitted by the cumulus cells. Considering its role in the folliculogenesis, this doctoral thesis looked into the expression profile of the stated gene and its respective receptors. Their correlation with the oocyte quality in the process of medically assisted reproduction was examined, as well as morphological characteristics of the oocytes, zygotes and embryos together with their fertilization potential. The research included 129 cumulus cells samples and 35 follicular fluid samples which had been harvested from 52 patients during the process of medically assisted reproduction. The method of relative quantification of gene expressions in the cumulus cells for the researched genes was employed, the enzyme - linked immunosorbent assay (ELISA) was used to determine the concentration of AMH in the follicular fluid of the respective follicle, and the morphological analysis of the oocytes, zygotes and embryos was conducted by inverted microscopy. The results obtained in the research for the first time show the negative correlation between the AMH and AR genes expression in the cumulus cells and the morphological quality, i.e. maturity, of the respective egg cell. Furthermore, the research determined the existence of correlations between the expressions of the examined genes as follows: between FSHR genes and all the other researched genes (AMH, AMHR2 and AR), between AMHR2 and AR genes, as well as between AMH and AR genes. The obtained results also contribute to the clarification of the potential connection between AMH (and the receptors directly or less directly linked to this hormone) and morphological quality of the egg cell. The research confirmed the importance of the cumulus - oocyte complex, i.e. of the communication between the egg cell and its microenvironment, for the growth of a high - quality oocyte, which is a prerequisite for a successful fertilization and growth of a high - quality embryo.
Parallel keywords (Croatian)Anti-Müllerov hormon (AMH) folikularna tekućina kvaliteta jajne stanice morfologija jajne stanice stanice kumulusa
Versionaccepted version
Resource typetext
Access conditionOpen access
Terms of usehttp://creativecommons.org/licenses/by-nc-nd/4.0/
Noteaccepted version
URN:NBNhttps://urn.nsk.hr/urn:nbn:hr:193:794933
CommitterLea Lazzarich