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Molekularna struktura, poreklo i funkcija B hromozoma kod žutogrlog šumskog miša Apodemus flavicollis (Mammalia, Rodentia)

Molecular structure, origin and function of B chromosomes in yellow-necked wood mouse Apodemus flavicollis (Mammalia, Rodentia)

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Disertacija.pdf (2.022Mb)
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Author
Rajičić, Marija
Mentor
Adnađević, Tanja
Committee members
Blagojević, Jelena
Jelić, Mihailo
Adnađević, Tanja
Blagojević, Jelena
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Abstract
B hromozomi predstavljaju prekobrojne hromozome koji nisu neophodni za normalan rast i razviće. Ovi prekobrojni hromozomi se javljaju kod svih višećelijskih eukariotskih grupa, osim kod ptica, i različiti su po poreklu, morfologiji, veličini, broju, genetičkoj strukturi i efektima na fenotip. Njihova prisutnost u živom svetu procenjuje se na 3% kariotipski analiziranih vrsta. Karakteriše ih nasleđivanje koje ne prati Mendelova pravila, kao i izostanak rekombinacije sa hromozomima osnovnog A seta. Glavna pitanja koja se postavljaju u savremenim istraživanjima B hromozoma se odnose na njihovo poreklo, mehanizme nastanka i održavanja. Poznavanje genetičkog sadržaja, transkripcionog statusa i efekata prekobrojnih hromozoma, omogućuje rasvetljavanje njihovog biološkog značaja. Glodari su najbrojniji nosioci B hromozoma među sisarima. Rod Apodemus broji ukupno 20 vrsta, od kojih je kod šest zabeleženo prisustvo B hromozoma. U ovoj studiji izučavani su B hromozomi na jedinkama vrste žutogrli ...miš, Apodemus flavicollis. Nosioci imaju najčešće jedan ili dva B, dok je maksimalan zabeležen broj osam B hromozoma. Prekobrojni hromozomi se kod ove vrste javljaju u svim do sada proučavanim populacijama na teritoriji Republike Srbije, sa različitom učestalošću. Brojne populacione studije na ovoj vrsti pokazuju da jedinke sa B hromozomima imaju višu adaptivnu vrednost u poređenju sa jedinkama bez prekobrojnih hromozoma. Cilj ove studije bio je utvrđivanje sekvence, proučavanje porekla, kao i analiza transkripcione aktivnosti pojedinih sekvenci B hromozoma vrste A. flavicollis. U istraživanje su uključene životinje sa lokaliteta u Republici Srbiji, Ruskoj federaciji i Republici Belorusiji. Metodom mikrodisekcije iz kariotipa tri jedinke su izolovani B hromozomi koji su potom umnoženi u lančanoj reakciji polimeraze (PCR). Proizvedene su fluorescentne B-specifične probe. Procena homologije između B-specifičnih proba i hromozoma osnovnog kariotipa iste vrste izvršena je putem fluorescentne in situ hibridizacije (FISH). Utvrđena je homologija između sekvence B hromozoma i: a) pericentromernog regiona (PR) polnih hromozoma, b) subtelomernih regiona dva para malih autozoma, kao i c) slabija homologija sa distalnim regionom Y hromozoma

B chromosomes (Bs) are supernumerary chromosomes in a standard karyotype, unnecessary for normal growth and development. Bs are present in all main groups of multicellular eukaryotes, except birds, and extremely variable considering origin, morphology, size, number, molecular structure, and phenotypic effects. 3% of all analyzed species have B chromosomes. The Bs do not follow rules of Mendelian segregation law and do not recombine with members of the basic A chromosome complement. The main questions considered in contemporary studies of Bs are surrounding their origin, mechanisms of formation and maintenance. Knowledge of their molecular structure, transcriptional activity, and phenotypic effects, will allow a deeper understanding of their biological importance. Within mammals, the most frequent carriers of Bs are rodents. The genus Apodemus includes 20 species, six of which carry Bs, in addition to A chromosome set. In this study, B chromosomes were analyzed in specimens of the yello...w-necked mouse, Apodemus flavicollis. Carriers usually have one or two Bs, and the highest recorded number in this species is eight B chromosomes. In different frequencies, supernumeraries are present in all studied populations of A. flavicollis in the territory of the Republic of Serbia. Numerous population studies of this species are indicating that B carriers have a higher fitness compared to individuals without B chromosomes. The aim of this study was to define sequence, origin and transcriptional activity of selected B regions in species A. flavicollis. Specimens were collected from different localities in the Republic of Serbia, Russian Federation and the Republic of Belarus. B chromosomes have been isolated by microdissection from the karyotype of three individuals and amplified in the polymerase chain reaction (PCR). B-specific fluorescent probes were synthesized. Estimation of homology between B-specific probes and chromosomes of A set had been obtained by fluorescent in situ hybridization (FISH). Homology was shown between the B chromosome and: a) pericentromeric region (PR) of sex chromosomes b) subtelomeric region of the two pairs of small chromosomes and c) lower homology with the distal region of the Y chromosome.

Faculty:
Универзитет у Београду, Биолошки факултет
Date:
28-06-2019
Projects:
  • Science in Motion for Friday Night Commotion 2013 (EU-609724)
  • Science in Motion for Friday Night Commotion 2014-15 (EU-633376)
  • The Road to Friday of Science (EU-722341)
  • Genetic and phenetic diversity in natural populations across different environments - contribution of B chromosome polymorphism (RS-173003)
  • ReFocuS 2.0 - Road to Friday of Science 2.0 (EU-818325)
Keywords:
B hromozomi / B chromosomes / genetika / FISH / mikrodisekcija / sekvenciranje / RT-PCR / Apodemus flavicollis / genetics FISH / microdissection / sequencing / RT-PCR / Apodemus flavicollis
[ Google Scholar ]
Handle
https://hdl.handle.net/21.15107/rcub_nardus_11604
URI
https://nardus.mpn.gov.rs/handle/123456789/11604
http://eteze.bg.ac.rs/application/showtheses?thesesId=6965
https://fedorabg.bg.ac.rs/fedora/get/o:20427/bdef:Content/download
http://vbs.rs/scripts/cobiss?command=DISPLAY&base=70036&RID=1025226418

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