Ekspresija i kolokalizacija razvojnog markera Nkx2.1 s proteinima parvalbumin i nNOS u subtalamičkoj jezgri miša i štakora

master's thesis

• Title (english): Expression and colocalization of developmental marker Nkx2.1 and proteins parvalbumin and nNOS the mouse and rat subthalamic nucleus
• Author: Tila Medenica
• Mentor: Goran Sedmak (mentor)
• Mentor: Sofia Ana Blažević (sumentor)
• Committee member: Sofia Ana Blažević (predsjednik povjerenstva)
• Committee member: Rosa Karlić (član povjerenstva)
• Committee member: Damjan Franjević (član povjerenstva)
• Committee member: Ana Previšić (član povjerenstva)
• Granter: University of Zagreb; Faculty of Science; (Department of Biology); Zagreb
• Defense date and country: 2020-02-28, Croatia
• Scientific / art field, discipline and subdiscipline: NATURAL SCIENCES; Biology

Abstract

Subtalamička jezgra (STh) je struktura diencefalona koja funkcionalno pripada bazalnim ganglijima, a klinički je važna u liječenju Parkinsonove bolesti kao mjesto dubokomozgovne stimulacije. Untoč tome, podaci o sastavu podtipova neurona STh, njihovom molekularnom profilu, te razvojnom porijeklu nedostaju. Cilj ovog istraživanja bio je analizirati prostornu ekspresiju razvojnog markera Nkx2.1 u STh odraslog mozga miša i štakora te njegovu kolokalizaciju s proteinima parvalbumin i nNOS kako bi se utvrdio molekularni profil neurona STh te kako bi se utvrdile potencijalne evolucijske razlike u razvoju STh između dvije vrste. Korišteni su uzorci mozga miša i štakora muških jedinki starosti 3,5 mjeseci. Ekspresija proteina u STh analizirana je pomoću DAB imunohistokemije, a kolokalizacija proteina u pojedinačnim neuronima analizirana je imunohistokemijskom metodom dvostrukog obilježavanja s fluorescentnim protutijelima. Utvrđeni su obrasci ekspresije navedenih proteina u STh odraslog miša i štakora, no zbog nespecifičnosti sekundarnih protutijela nije utvrđena njihova kolokalizacija. U ovom je istraživanju uočena razlika u ekspresiji markera Nkx2.1 između miša i štakora, što do sada nije opisano u literaturi. Mogući razlozi za ovakve rezultate su da se s dobi smanjuje translacija proteina Nkx2.1, ili da je miš tijekom evolucije izgubio neurone koji eksprimiraju Nkx2.1. Za razrješenje ovih dvojbi potrebno je provesti dodatna istraživanja.

Abstract (english)

Subthalamic nucleus (STh) is a diencephalic basal ganglia structure, clinically important as a target in the treatment for Parkinson's disease. Data describing its neuronal composition, molecular profile and developmental origin is still missing. The aim of this research was to analyze spatial expression of the developmental marker Nkx2.1 in the adult mouse and rat STh and colocalization of Nkx2.1 with proteins parvalbumin and nNOS. Another aim was to determine the potential evolutionary difference in STh development between the two species. Brain samples were obtained from male mouse and rat individiuals aged 3,5 months. Protein expression was analyzed using DAB immunohistochemistry, and protein colocalization was analyzed using immunofluorescence with double-labeling. Expression patterns of these proteins were obtained, but their colocalization remains unknown due to the unspecificity of secondary antibodies used in the experiment. Difference in the expression of Nkx2.1 between mouse and rat was noticed, which has not yet been described in literature. Possible cause of this result could be the age-dependent downregulation of the protein translation, or that the mouse has lost Nkx2.1-expressing neuron population in the STh during evolutionary time. For definite conclusions further experiments need to be done.

• Keywords: bazalni gangliji; imunohistokemija; diencefalon
• Keywords (english): basal ganglia; diencephalon; immunohistochemistry
• Language: croatian
• URN:NBN: urn:nbn:hr:217:732803
• Study programme: Title: Molecular Biology; Study programme type: university; Study level: graduate; Academic / professional title: magistar/magistra molekularne biologije (magistar/magistra molekularne biologije)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Open access; Embargo expiration date: 2021-02-28
• Created on: 2020-07-13 06:58:09