Fenotipske promjene vrste Asellus aquaticus (Isopoda, Crustacea) uslijed izloženosti mraku i farmakološkim modulatorima

master's thesis

• Title (english): Phenotypic changes in Asellus aquaticus (Isopoda, Crustacea) as a result of exposure to darkness and to pharmacological modulators
• Author: Manuela Žugaj
• Mentor: Helena Bilandžija (mentor)
• Mentor: Sofia Ana Blažević (komentor)
• Committee member: Sofia Ana Blažević (predsjednik povjerenstva)
• Committee member: Tvrtko Dražina (član povjerenstva)
• Committee member: Romana Gračan (član povjerenstva)
• Committee member: Anamaria Štambuk (član povjerenstva)
• Granter: University of Zagreb; Faculty of Science; (Department of Biology); Zagreb
• Defense date and country: 2023-09-20, Croatia
• Scientific / art field, discipline and subdiscipline: NATURAL SCIENCES; Biology

Abstract

Slatkovodni jednakonožni rakovi vrste Asellus aquaticus naseljavaju površinska i podzemna staništa u kojima poprimaju različite fenotipove. Fenotipska plastičnost, sposobnost genoma da proizvede različite fenotipove uslijed djelovanja okoliša, je potencijalni mehanizam kojim se površinske populacije prilagođavaju podzemlju. Poznato je da epigenetski mehanizmi imaju važnu ulogu u odgovoru genoma na okoliš. Stoga sam u ovom radu istražila kako mrak, najbitnija karakteristika podzemlja, te farmakološki modulatori, djeluju na preživljenje i fenotip vrste Asellus aquaticus. Koristila sam bisfenol A (BPA; endokrini i epigenetički disruptor), 5-azacitidin (inhibitor metilacije DNA), radicicol (inhibitor proteina HSP90) i natrij butirat (inhibitor histon deacetilaza). Jedinke su izložene otopinama farmakoloških modulatora u periodu od tjedan dana, dio u potpunom mraku, a dio na fotoperiodu svjetlo-tama uz pripadajuće kontrolne skupine u vodi. Kontrolne skupine u mraku su bile manje pigmentirane te imale dulje antene od kontrolnih skupina na svjetlu. Jedinke u eksperimentalnim grupama su imale dulje antene uslijed djelovanja 5-azacitidina i natrij butirata, pigmentiranije su kada ih se izloži 5-azacitidinu, a manje pigmentirane u otopini BPA. Inhibicija proteina Hsp90 radicicolom dala je varijabilne fenotipove, ali ne statistički značajne promjene. Zaključno, endokrino i/ili epigenetičko djelovanje BPA, metilacija DNA, promjene strukture kromatina uslijed ostalih farmakoloških modulatora uključeni su u promjene fenotipova kod ovih rakova.

Abstract (english)

Asellus aquaticus is a freshwater isopod, which inhabits surface and cave habitats expressing different phenotypes. Phenotypic plasticity, the genome's ability to produce different phenotypes in response to differing environments, is a potential mechanism of surface-dwellers’ adaptation to caves and epigenetic mechanisms often play a crucial role in the genome's response to the environment. Here, I investigated the effects of darkness, a key characteristic of caves, and pharmacological modulators on survival and phenotype of Asellus aquaticus. I used bisphenol A (BPA, endocrine and epigenetic disruptor), 5-azacytidine (DNA methylation inhibitor), radicicol (Hsp90 protein inhibitor), and sodium butyrate (histone deacetylase inhibitor). Individuals were exposed to pharmacological modulators for a week, some in complete darkness and others in light-dark photoperiod, alongside respective control groups in water. Dark-exposed group exhibited reduced pigmentation and longer antennae compared to light-dark controls. Individuals in the experimental groups exhibited longer antennae when exposed to 5-azacytidine and sodium butyrate; increased pigmentation with 5-azacytidine; and decreased pigmentation in the BPA solution. Inhibition of Hsp90 protein using radicicol resulted in statistically insignificant phenotypic changes. In conclusion, the endocrine and/or epigenetic effects of BPA, DNA methylation and changes in chromatin structure due to other chemicals, are involved in the phenotypic changes observed in these isopods.

• Keywords: Fenotipska plastičnost; epigenetika; podzemne životinje; prilagodbe na špilje; isopoda
• Keywords (english): phenotypic plasticity; epigenetics; subterranean animals; adaptations to caves; isopods
• Language: croatian
• URN:NBN: urn:nbn:hr:217:283576
• Study programme: Title: Experimental Biology-University graduate programme; Study programme type: university; Study level: graduate; Academic / professional title: sveučilišni/a magistar/magistra eksperimentalne biologije (sveučilišni/a magistar/magistra eksperimentalne biologije)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Embargoed access; Embargo expiration date: 2025-09-20
• Created on: 2023-11-29 13:12:22