Signalizacija kalcija v človeških lečnih epitelnih celicah po lokalnem mehanskem dražljaju

Gojić, Dajana (author), Andjelić, Sofija (mentor), Gošak, Marko (comentor)

Porušena signalizacija kalcija (Ca2+) v lečnih epitelnih celicah (LECs) je eden izmed ključnih dejavnikov za razvoj sive mrene. Da bi bolje razumeli vlogo medcelične komunikacije povezane s sivo mreno in nastajanjem zamotnitve zadnje lečne ovojnice (PCO) ter regeneracijo leče, smo raziskali in opredelili medcelično Ca2+ signalizacijo v človeških LECs ob lokalni mehanski stimulaciji. Uporabili smo sveže postoperativne lečne ovojnice (LCs), pridobljene pri operaciji sive mrene. Sprednje LECs smo lokalno mehansko stimulirali s stekleno mikropipeto, kar je povzročilo dvig znotrajcelične koncentracije kalcija ([Ca2+]i). Ob natančnem sledenju prostorsko-časovnim spremembam [Ca2+]i smo ugotovili, da se Ca2+ signal širi radialno od točke stimulacije in da se amplitude povečanja [Ca2+]i zmanjšujejo z večanjem razdalje od mesta stimulacije. Pri analizi značilnosti Ca2+ signalov glede na stopnjo katarakte smo ugotovili, da se v LCs močneje razvitih katarakt Ca2+ signal širi hitreje, s tem da so amplitude Ca2+ signala manjše, a njegovo trajanje daljše. Primerjali smo tudi signalizacijo Ca2+ med različnimi vrstami katarakte, pri čimer nismo ugotovili nobenih signifikantnih razlik. Lahko zaključimo, da kataraktogeneza vključuje spremembe tako v znotrajceličnem vidiku Ca2+ signalizacije kakor tudi na ravni medcelične komunikacije. Za spremljanje funkcionalnih lastnosti LECs, ki so pomembne za PCO in regeneracijo leče, smo iz postoperativnih LCs vzgojili primarne lečne kulture. Z lokalno mehansko stimulacijo smo pokazali odzivanje zraslih LECs, ki se v prihodnosti lahko uporabi za testiranje različnih fizičnih in farmakoloških učinkovin.

• Language: Slovenian
• Keywords: lečne epitelne celice, signalizacija kalcija, medcelična komunikacija, katarakta, očesna leča, kulture lečnih epitelnih celic, mehanska stimulacija
• Work type: Master's thesis/paper
• Organization: BF - Biotechnical Faculty
• Year: 2018
• PID: 20.500.12556/RUL-101843
• COBISS.SI-ID: 4774991
• Publication date in RUL: 08.07.2018

GOJIĆ, Dajana, 2018, Signalizacija kalcija v človeških lečnih epitelnih celicah po lokalnem mehanskem dražljaju [online]. Master’s thesis. [Accessed 11 April 2025]. Retrieved from: https://repozitorij.uni-lj.si/IzpisGradiva.php?lang=eng&id=101843

Secondary language

• Language: Unknown
• Title: Calcium signaling in human lens epithelial cells upon local mechanical stimulation

Abstract

Calcium (Ca2+) signalling disturbances in lens epithelial cells (LECs) are one of the key factors involved in cataract formation. We decided to explore and identify intercellular Ca2+ signalization in human LECs upon local mechanic stimulation, to understand better the role of intercellular communication and its connection to cataractogenesis and posterior capsule opacification (PCO) development as well as lens regeneration. We used fresh postoperative lens capsules (LCs) obtained during cataract surgery. We mechanically stimulated anterior LECs with glass micropipette, which induced an increase in intracellular Ca2+ concentration ([Ca2+]i). By following the spatio-temporal changes in [Ca2+]i we discovered that Ca2+ signal spreads radially from stimulation point and that the amplitude of Ca2+ transients decreases with increasing distance from stimulation. When we analysed signalling characteristics with respect to degree of cataract progression, we discovered that in LCs from more developed cataracts the Ca2+ wave propagates faster, the amplitudes of Ca2+ signals are lower, whereas their durations are longer. On contrary, no significant differences were observed when comparing LCs with regard to the type of the cataract. From this we can conclude that cataractogenesis is associated with changes in intracellular signalization of Ca2+, as well as with changes in intercellular communication. Moreover, we grew ex vivo cultures of the LC explants to follow functional properties of LECs, which are important for studying PCO and lens regeneration. With local mechanical stimulation we showed responding of grown LECs, which can be in future used for testing of different physical and pharmacological agents.

• Keywords: lens epithelial cells, calcium signaling, intercellular communication, cataract, eye lens, cultures of lens epithelial cells, mechanical stimulation