Abstract
Studies focusing on molecular mechanisms of cell cycles have been lagging in unicellular eukaryotes compared to other groups. Ciliates, a group of unicellular eukaryotes, have complex cell division cycles characterized by multiple events. During their vegetative cell cycle, ciliates undergo macronuclear amitosis, micronuclear mitosis, stomatogenesis and somatic cortex morphogenesis, and cytokinesis. Herein, we used the hypotrich ciliate Pseudokeronopsis erythrina, whose morphogenesis has been well studied, to examine molecular mechanisms of ciliate vegetative cell cycles. Single-cell transcriptomes of the growth (G) and cell division (D) stages were compared. The results showed that (i) More than 2051 significantly differentially expressed genes (DEGs) were detected, among which 1545 were up-regulated, while 256 were down-regulated at the D stage. Of these, 11 randomly picked DEGs were validated by reverse transcription quantitative polymerase chain reaction (RT-qPCR); (ii) Enriched DEGs during the D stage of the vegetative cell cycle of P. erythrina were involved in development, cortex modifications, and several organelle-related biological processes, showing correspondence of molecular evidence to morphogenetic changes for the first time; (iii) Several individual components of molecular mechanisms of ciliate vegetative division, the sexual cell cycle and cellular regeneration overlap; and (iv) The P. erythrina cell cycle and division have the same essential components as other eukaryotes, including cyclin-dependent kinases (CDKs), cyclins, and genes closely related to cell proliferation, indicating the conserved nature of this biological process. Further studies are needed focusing on detailed inventory and gene interactions that regulate specific ciliated cell-phase events.
Highlights
The cell cycle is a series of processes during the life history of a cell that lead to growth and proliferation [1]
reverse transcription quantitative polymerase chain reaction (RT-qPCR) was performed on an Applied Biosystems® QuantStudio® 5 instrument (Applied Biosystems, Carlsbad, CA, USA), and the reaction was conducted in a 20 μL reaction system containing 10 μL of the QuantiNova SYBR Green PCR kit (Qiagen, Hilden, Germany), 10 μM of each primer, cDNA and nuclease-free water (Qiagen, Hilden, Germany)
5489 unigenes were aligned to the Clusters of Orthologous Groups (COG) database, 7730 unigenes were annotated with Gene Ontology (GO) functions, and 4354 unigenes were mapped using Kyoto Encyclopedia of Genes and Genomes (KEGG)
Summary
The cell cycle is a series of processes during the life history of a cell that lead to growth and proliferation [1]. A cell undergoes multiple rounds of division. The cell cycle of unicellular eukaryotes is of interest because the process is inexoCrailbialytelsi,nakegdrotoupceollfduivniiscieolnluilnartheeuskeaorrygoatneiss,mhsa.ve been used extensively as model organisms to stuCdilyiaatebs,roaagdroruanpgoef oufntioceplilcusladrueeutkoardyuoatel sg, ehnaovme ebeaernchuitseecdtuerxet,enusniivqeulye amsamcroodneulcoleragrangiesnmosmtoe csthuadrayctaerbsr(oFaidgurraen1g)e, aonfdtcoopmicpsledxume otorpdhuoalolggye[n1o4m–2e4]a.rRcheicteencttluyr,em, oulneicquulaermevaecnrotsnuucnldeaerrlygienngotmhee scehxauraacltceerlsl (cFyicglueroef1t)h,eamndodcoeml cpillieaxtemToetrrpahhyomloegnya [t1h4e–rm24o]p. Vegetative proliferation is asexual cell division in ciliates and occurs by binary fission. Ciliate cell division is more complex than that of other eukaryotes, comprising at least 4 types and 15 modes of stomatogenesis, and is of great significance for understanding the evolution of this process in eukaryotes [42]. TThhee llaatttteerr oorriiggiinnaatteeddfrfroommththeesasmame ececlle3ll03m0 imnuinteusteasftearftceerllcdelilvidsiivonis.ioInn.oIrndeorrtdoerretdourceedcuelclehceetellrohgeetenreoitgye, ntheriteye, rtehpreliecarteepsliwcaetreescwolelerectceodllfeocrteedacfhorsteaagceh: sGta1g, eG: 2G, 1G,3Ga2n, dGD3 1a,nDd2D, D1,3D. 2E,aDch3.cEelalcwhacselwl wasahsewd afisvheetdimfievse wtimithesinwacitthivainteadctcivaalctieudmcaalncdiummaganndesimumag-nfreeseiuPmB-SfrbeueffPeBr Susbinugffearnuuscilnegasae-nfruecelepaipsee-tftreeaenpdipweattsethaennd twraansstfherernedtratonsafenrurceldeatosea-fnreuecElepapsee-nfrdeoerfEtpupbeenwdoitrhf atumbeinwimituhma mvoinluimmuemofvlioqluuimd.e of liquid
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