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Transkriptomska analiza in celično ozadje nastanka marmoriranega pigmentnega vzorca pri soški postrvi (Salmo marmoratus, Cuvier 1817)
ID Djurdjevič, Ida (Author), ID Sušnik Bajec, Simona (Mentor) More about this mentor... This link opens in a new window

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Abstract
S celovitim pristopom in z uporabo tako celično-bioloških kot genetskih pristopov smo raziskovali mehanizem nastanka vzorca v koži soške in potočne postrvi. Sorodni vrsti se namreč izrazito razlikujeta glede na vzorec, saj v koži soške postrvi najdemo labirinten, pri potočni pa pikast vzorec. Na celični ravni smo s presevnim elektronskim mikroskopom raziskali položaj in ultrastrukturo pigmentnih celic, ki se med vrstama razlikuje. Potočna postrv ima v koži prisotne eritrofore (tipa 1 in tipa 2), ki jih pri soški postrvi ne najdemo in smo jih v naši raziskavi tudi prvič opisali. Potočna postrv ima višjo organiziranost pigmentnih celic v koži v primerjavi s soško postrvjo, nismo pa našli neposredne povezave med položajem različnih tipov pigmentnih celic in različnim vzorcem v koži. Z metodo NGS smo posekvencirali transkriptom celic iz različno obarvanih delov kože obeh vrst, pridobljenih s kriorezinami iz tarčnih območij, kjer so locirane kromatofore. Rezultati so pokazali velik nabor diferenčno izraženih transkriptov med vrstama, pa tudi med posameznimi različno obarvanimi regijami. Med temi transkripti je bilo veliko takih, ki nosijo zapis za membranske proteine, vključene v celično komunikacijo. Rdeča pika je po izraženosti izstopala, saj je edina, ki vsebuje eritrofore tipa 2. Z metodo qPCR smo na večjem številu vzorcev preverili izražanje 22 kandidatnih genov. Med temi smo jih izbrali šest, katerih izražanje smo preverili še v različno obarvanih predelih kože križancev med vrstama. Geni p21, gja5 in tjp1 so pokazali najvišjo povezavo med ravnijo izražanja in vzorcem v koži. Na podlagi pridobljenih rezultatov lahko zaključimo, da je za nastanek vzorca pri postrvih najverjetneje odločilna specifična medcelična komunikacija med pigmentnimi celicami (prek presledkovnih stikov, tesnih stikov in ionskih kanalov).

Language:Slovenian
Keywords:animalna genetika, salmonidi, postrvi, Salmo marmoratus, pigmentni vzorec, RNA, transkriptom, NGS, celična biologija, mikroskopija, TEM
Work type:Doctoral dissertation
Typology:2.08 - Doctoral Dissertation
Organization:BF - Biotechnical Faculty
Year:2018
PID:20.500.12556/RUL-101605 This link opens in a new window
COBISS.SI-ID:4103560 This link opens in a new window
Publication date in RUL:21.06.2018
Views:1923
Downloads:674
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Secondary language

Language:English
Title:Transcriptome analysis and cellular background of the marble pigment pattern formation in marble trout (Salmo marmoratus, Cuvier 1817)
Abstract:
In order to comprehensively address the mechanism responsible for the pigment pattern formation in the skin of marble and brown trout, different cellular and genetic approaches were applied. The related species have a different skin pigment pattern, marble trout expressing a unique labyrinthine, while brown trout has a spot pattern. Pigment cell ultrastructure and their position in the skin were examined with TEM. In the brown trout skin, we found erythrophores (type 1 and type 2), which were not found in the skin of the marble trout and were described here for the first time. The pigment cell organization in the skin of brown trout is more organised, compared to that in marble trout. Based on our findings, we couldn’t find a connection between a position of specific pigment cell type and different pigment pattern in the skin. Transcriptomes of cells from differently pigmented regions, obtained by cryosections from the region where chromatophores are located, were sequenced using NGS. The results have shown a large set of differently expressed transcripts between species, but also between differently pigmented regions. Among these transcripts, there were several coding for membrane proteins, which are involved in cell-cell communication. The red dot in brown trout had a unique expression profile, this region is the only one containing the type 2 erythrophore. Using qPCR, we examined the expression of 22 candidate genes on a larger number of samples. The expression of six of these genes was tested in hybrids between the two species. The genes p21, gja5 and tjp1 were displaying the highest correlation between their expression level and skin pattern. Based on our results, we can conclude that specific cell-cell communication (gap junctions, tight junctions, channels et. al.) between pigment cells is responsible for the pattern formation in trout.

Keywords:animal genetics, salmonids, trouts, Salmo marmoratus, pigment pattern, RNA, transcriptome, NGS, cell biology, microscopy, TEM

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