Sel’skokhozyaistvennaya Biologiya [Agricultural Biology], 2017, Vol. 52, № 6, p. 1157-1165
(how to cite this article)

doi: 10.15389/agrobiology.2017.6.1157eng

UDC 636.32/.38:636.082.12:577.21

Acknowledgements:
The equipment of the Center for Bioresources and Bioengineering of Agricultural Animals was used in the study. Supported financially by Russian Science Foundation, project № 14-36-00039, and Federal Agency of Scientific Organizations, theme № 0600-2014-0004.3

 

CHARACTERISTICS OF ALLELE POOL OF THE ROMANOV SHEEP
BREED FOR THE PRION PROTEIN GENE ASSOCIATED
WITH GENETIC SUSTAINABILITY TO SCRAPIE

E.A. Gladyr1, T.E. Deniskova1, V.A. Bagirov1, O.V. Kostyunina1,
N.N. Makarova2, G. Brem1, 3, N.A. Zinovieva1

1L.K. Ernst Federal Science Center for Animal Husbandry,Federal Agency of Scientific Organizations, 60, pos. Dubrovitsy, Podolsk District, Moscow Province, 142132 Russia,
e-mail elenagladyr@mail.ru (corresponding author), tandeniss@rambler.ru, vugarbagirov@mail.ru, kostolan@yandex.ru, n_zinovieva@mail.ru (corresponding author);
2LLC «Agrivolga», 11, ul. Spasskaya, Uglich, Yaroslavl Province,152615 Russia,
e-mail N.Makarova@agrivolga.ru;
3Institut für Tierzucht und Genetik, University of Veterinary Medicine (VMU), Veterinärplatz, A-1210, Vienna, Austria, e-mail gottfried.brem@vetmeduni.ac.at

ORCID:
Gladyr E.A. orcid.org/0000-0002-5210-8932
Bagirov V.A. orcid.org/0000-0001-8385-2433
Makarova N.N. orcid.org/0000-0002-4742-9985
Zinovieva N.A. orcid.org/0000-0003-4017-6863
Deniskova T.E. orcid.org/0000-0002-5809-1262
Kostyunina O.V. orcid.org/0000 0001 8206 3221
Brem G. orcid.org/0000-0002-7522-0708

Received October 4, 2017

 

The Romanov is a unique indigenous sheep breed of Russia, belonging to the group of Northern short-tailed sheep. The breed is known all over the world, due to out-of-season breeding ability, phenomenal fecundity (up to 10 lambs) and unsurpassed quality of sheepskins. Presently the gene pool of the breed is actively involved in creation of new types of modern prolific sheep and it is considered as an important genetic reserve for the sheep breeding of the future. Diseases resistance is the most important selection trait in sheep. One of the diseases which can cause serious economic loses is spongiform encephalopathy of sheep, also known as scrapie. Scrapie is a fatal neurodegenerative disease of sheep and goats, belonging to the class of transmissible spongiform encephalopathies (TSE), which also includes bovine spongiform encephalopathy (BSE). Three polymorphisms in amino acid codons 136 (A/V), 154 (R/H), and 171 (R/Q/H) of the PRPN gene are associated with resistance or susceptibility of sheep to classical scrapie. Depending on the PRNP genotype, there are five classes of genetic sustainability to Scrapie (G1-G5). The ARR allele is desirable regarding the resistance to scrapie. However, discovery of atypical scrapie (Nor98) showed a possibility of transmitting BSE to animals of different sustainability classes, including G1 (ARR/ARR genotype). It is shown, that L/F amino acid substitution at position 141 provides resistance to atypical scrapie. The aim of our work was to study the allele pool of the Romanov sheep by the PRNP gene, associated with sustainability to both classical and atypical forms of scrapie. The material for the work was tissue samples of 364 clinically healthy Romanov animals including three modern populations of the Yaroslavl region and one population introduced for breeding in the Kamchatka. Genomic DNA was isolated using the Nexttec columns (Nexttec Biotechnologie GmbH, Germany). Identification of the alleles in the codons 136 (A/T/V), 141 (L/F), 154 (R/H) and 171 (Q/R/H/K) was performed by pyrosequencing on the PSQ96MA device (Quiagen, USA). We found four alleles, 136/154/171 — ARR, ARQ, AHQ and VRQ, and nine haplotypes of PRNP as ARR/ARR, ARR/ARQ, ARR/AHQ, ARQ/ARQ, AHQ/ARQ, AHQ/AHQ, ARR/VRQ, VRQ/AHQ and ARQ/VRQ, relating to all five classes of genetic sustainability to the classical Scrapie. The allele of wild type ARQ (the frequency from 0.704 to 0.933) and the genotype ARQ/ARQ (sustainability class G3) were the prevalent. In all the studied groups, a desirable ARR allele was identified with frequencies varied from 0.022 to 0.089 and averaged 0.066. The undesirable VRQ allele was found in three of the four groups, while its frequency was relatively low — from 0.011 to 0.022. The study of the PRNP polymorphism by four codons 136/141/154/171 revealed the presence of five different alleles — ALRR, ALRQ, ALHQ, VLRQ, AFRQ and ten genotypes. We detected an animal carrying a sensitive to the atypical scrapie allele F at position 141 of PRNP (genotype VLRQ/AFRQ) with the allele frequency of 0.001. The results will be applied in the development of breeding programs for Romanovs, as well as in strategic planning of conservation of the genetic diversity of this unique Russian Northern short-tailed sheep.

Keywords: prion protein gene (PRNP), allele pool, the Romanov sheep, genetic sustainability, scrapie.

 

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REFERENCES

  1. Ernst L.K., Zinov'eva N.A. Biologicheskie problemy zhivotnovodstva v XXI veke [Challenges in biology of XXI century]. Moscow, 2008 (in Russ.).
  2. Stolpovskii Yu.A. Concept and principles of genetic monitoring for the purpose of preservation in situ of domestical animals kinds. Sel’skokhozyaistvennaya Biologiya [Agricultural Biology], 2010, 6: 3-8 (in Russ.).  
  3. Deniskova T.E., Selionova M.I., Gladyr’ E.A., Dotsev A.V., Bobrysho-va G.T., Kostyunina O.V., Brem G., Zinovieva N.A. Variability of microsatellites in sheep breeds raced in Russia. Agricultural Biology, 2016, 51(6): 801-810 CrossRef (in Engl.).
  4. Ivanov M.F. Ovtsevodstvo [Sheep farming]. Moscow, 1933 (in Russ.).
  5. Kuleshov P.N. Ovtsevodstvo Rossii [Sheep breeding in Russia]. Petrograd, 1916 (in Russ.).
  6. Ernst L.K., Dmitriev N.G., Paronyan I.A.Geneticheskie resursy sel'skokhozyai-stvennykh zhivotnykh v Rossii i sopredel'nykh stranakh [Gene pool of farm animals in Russia and neighboring countries]. St. Petersburg, 1994 (in Russ.). 
  7. Dankvert A.G., Dankvert S.A. Istoriya plemennogo zhivotnovodstva Rossii [History of pedigree animal breeding in Russia]. Moscow, 2004 (in Russ.). 
  8. Prusiner S.B.Prions. PNAS, 1998, 95(23): 13363-13383.
  9. Hunter N., Goldmann W., Foster J. D., Cairns D., Smith G. Natural scrapie and PrP genotype: case-control studies in British sheep. Vet. Rec., 1997, 141: 137-140 CrossRef
  10. Smits M.A., Barillet F., Harders F., Boscher M.Y., Vellema P., Aguer-
    re X., Hellinga M., McLean A.R., Baylis M., Elsen J.M. Genetics of scrapie susceptibility and selection for resistance. Proc. 51st Annual Meeting of the European Association for Animal Production. The Hague, Netherlands, 2000: Paper S.4.4.
  11. Bossers A., Belt P.B.G.M., Raymond G.J., Caughey B., de Vries R., Smits M.A. Scrapie susceptibility-linked polymorphisms modulate the in vitro conversion of sheep prion protein to protease-resistant forms. PNAS, 1997, 94(10): 4931-4936 CrossRef
  12. Ishiguro N., Shinagawa M., Onoe S., Yamanouchi K., Saito T. Rapid analysis of allelic variants of the sheep PrP gene by oligonucleotide probes. Microbiol. Immunol., 1998, 42: 579-582 CrossRef
  13. Buschmann A., Biacabe A.G., Ziegler U., Bencsik A., Madec J.Y., Erhardt G. Atypical scrapie cases in Germany and France are identified by discrepant reaction patterns in BSE rapid tests. J. Virol. Methods, 2004, 117: 27-36 CrossRef
  14. DEFRA. National scrapie plan for great britain. London, 2001: 1-28.
  15. Groschup M.H., Lacroux C., Buschmann F., Luhken G., Mathey J., Eiden M., Lugan S., Hoffmann C., Espinosa J.C., Baron T., Torres J.M., Erhardt G., Andreoletti O. Classic scrapie in sheep with the ARR/ARR prion genotype in Germany and France. Emerg. Infect. Dis., 2007, 13(8): 1201-1207 CrossRef
  16. Renaville R. Résistance génétique a la «Tremblante» (aussi appelée scrapie) chez le mouton. FIOW Gembloux, Belgique, 2002.
  17. Vaccari G., Di Bari M.A., Morelli L., Nonno R. Chiappini B., Antonucci G., Marcon S., Esposito E., Fazzi, P., Palazzini N., Troiano P., Petrella A., Guardo G.-D., Agrimi U. Identification of an allelic variant of the goat PrP gene associated with resistance to scrapie. J. Gen. Virol., 2006, 87: 1395-1402 CrossRef
  18. Houston F., Goldmann W., Chong A., Jeffrey M., Gonzalez L., Foster J. Prion diseases: BSE in sheep bred for resistance to infection. Nature, 2003, 423: 498 CrossRef
  19. Acín C., Martín-Burriel I., Goldmann W., Lyahyai J., Monzón M., Bolea R., Smith A., Rodellar C., Badiola J.J., Zaragoza P. Prion protein gene polymorphisms in healthy and scrapie-affected Spanish sheep. J. Gen. Virol., 2004, 85: 2103-2110 CrossRef
  20. Goldmann W., Chong A., Foster J., Hope J., Hunter N. The shortest known prion protein gene allele occurs in goats, has only three octapeptide repeats and is non-pathogenic. J. Gen. Virol., 1998, 79(12): 3173-3176 CrossRef
  21. Schläpfer J., Saitbekova N., Gaillard C., Dolf G. A new allelic variant in the bovine prion protein gene (PRNP) coding region. Anim. Genet., 1999, 30(5): 386-387 CrossRef
  22. O'Neill G.T., Donnelly K., Marshall E., Cairns D., Goldmann W., Hunter N. Characterization of ovine PrP gene promoter activity in N2a neuroblastoma and ovine foetal brain cell lines. J. Anim. Breed. Genet., 2003, 120(2): 114-123 CrossRef
  23. O'Neill G.T., Cairns D., Toovey L., Goldmann W., Hunter N. New ovine PrP gene haplotypes as a result of single nucleotide polymorphisms in the PrP gene promoter. J. Anim. Breed. Genet., 2005, 122(2): 86-94 CrossRef
  24. Benestad S.L., Sarradin P., Thu B., Schönheit J., Tranulis M.A., Bratberg B. Cases of scrapie with unusual features in Norway and designation of a new type, Nor98. Vet. Rec., 2003, 153: 202-208 CrossRef
  25. Buschmann A., Biacabe A.G., Ziegler U., Bencsik A., Madec J.Y., Erhardt G., Luhken G., Baron T., Groschup M.H. Atypical scrapie cases in Germany and France are identified by discrepant reaction patterns in BSE rapid tests. J. Virol. Methods, 2004, 117: 27-36 CrossRef
  26. Everest S.J., Thorne L., Barnicle D.A., Edwards J.C., Elliott H., Jackman R., Hope J. Atypical prion protein in sheep brain collected during the British scrapie-surveillance programme. J. Gen. Virol., 2006, 87(2): 471-477 CrossRef
  27. Luhken G., Buschmann A., Brandt H., Eiden M., Groschup M.H., Erhardt G. Epidemiological and genetical differences between classical and atypical scrapie cases. Vet. Res., 2007, 38: 65-80 CrossRef
  28. Le Dur A., Béringue V., Andréoletti O., Reine F., Lan Laï T., Baron T., Bratberg B., Vilotte J.-L., Sarradin P., Benestad S.L., Laude H. A newly identified type of scrapie agent can naturally infect sheep with resistant PrP genotypes. PNAS, 2005, 102(44): 16031-16036 CrossRef
  29. Langeveld J.P.M., Jacobs J.G., Hunter N.,  van Keulen L.J.M., Lantier F., van Zijderveld F.G., Bossers A. Prion type-dependent deposition of PRNP allelic products in heterozygous sheep. J. Virol., 2016, 90(2): 805-812 CrossRef
  30. Gonzalez L., Jeffrey M., Dagleish M.P., Goldmann W., Siso S., Eat-
    on S.L., Martin S., Finlayson J., Stewart P., Steele P., Pang Y., Hamilton S., Reid H.W., Chianini F. Susceptibility to scrapie and disease phenotype in sheep: cross-PRNP genotype experimental transmissions with natural sources. Vet. Res., 2012, 43: 55 CrossRef
  31. Gladyr' E.A., Zinov'eva N.A., Burylova S.S., Selionova M.I., Moiseikina L.G., Ernst L.K., Brem G. Dostizheniya nauki i tekhniki APK, 2012, 11: 34-37 (in Russ.).   
  32. Zinov'eva N.A., Popov A.N., Ernst L.K., Marzanov N.S., Bochka-
    rev V.V., Strekozov N.I., Brem G. Metodicheskie rekomendatsii po ispol'zovaniyu metoda polimeraznoi tsepnoi reaktsii v zhivotnovodstve [PCR in livestock farming. Manual guid].Dubrovitsy, 1998 (in Russ.).   
  33. Gladyr' E.A., Zinov'eva N.A., Brem G. Sposob genodiagnostiki ustoichivosti ovets k skrepi. A.S. 2303067 (RF) MPK C12N 15/00, C12N 15/12. FGBNU FNTS VIZH im. L.K. Ernsta (Vserossiiskii gosudarstvennyi nauchno-issledovatel'skii institut zhivotnovodstva (VIZH) (RF). № 2005128754/13. Zayavl. 16.05.2005. Opubl. 20.07.2007. Byul. № 20 [Gene typing of sheep resistance to scrapie. Patent RF № 2005128754/13. Appl. 16.05.2005. Publ. 20.07.2007. Bul. № 20] (in Russ.).  
  34. Belt P.B.G.M., Muileman I.H., Schreuder B.E.C., Bos-de Ruijter J., Giel-
    kens A.L., Smits M.A. Identification of five allelic variants of the sheep PrP gene and their association with natural scrapie. J. Gen. Virol., 1995, 76: 509-517 CrossRef
  35. Goldmann W., Hunte N., Benson G., Foster J.D., Hope J. Different scrapie-associated fibril proteins (PrP) are encoded by lines of sheep selected for different alleles of the Sip gene. J. Gen. Virol., 1991, 72: 2411-2417 CrossRef
  36. Hunter N., Goldmann W., Smith G., Hope J. The association of a codon 136 PrP gene variant with the occurrence of natural scrapie. Arch. Virol., 1994, 137: 171-177 CrossRef
  37. Cameron S., Bell-Rogers P., McDowall R., Rebelo A.R., Cai H.Y. Prion protein genotypes of sheep as determined from 3343 samples submitted from Ontario and other provinces of Canada from 2005 to 2012. Can. J. Vet. Res., 2014, 78: 260-266.
  38. Otelea M.R., Zaulet M.M., Dudu A., Otelea F., Baraitareanu S., Dan-
    es D. The scrapie genetic susceptibility of some sheep breeds in southeast Romanian area and genotype profiles of sheep scrapie infected. Rom. Biotech. Lett., 2011, 16(4): 6419-6429.
  39. Garcia-Crespo D., Juste RA, Hurtado A. Selection of ovine housekeeping genes for normalisation by real-time RT-PCR; analysis of PrP gene expression and genetic susceptibility to scrapie. BMC Vet. Res., 2005, 1: 3 CrossRef

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