project - Innovative project
Development of new technologies for bull sperm sex selection and fertility management
Veise seemnerakkude suguselektsiooni- ja viljakustehnoloogiate arendus
Objectives
The aim of the current study was to develop new transglutaminase-1 (TGM1)- biomarker based technology for bull sperm sex selection. In addition, the project aimed to pinpoint and validate bovine sperm fertility related biomarkers. With the help of these new biomarkers it would be possible to develop bull fertility test and artificial supplement for the improvement of bull semen quality
Objectives
Projekti eesmärgiks oli veise spermide transglutaminaas-1 (TGM1)-biomarkeri põhise sooselektsioonitehnoloogia väljaarendamine. Lisaeesmärgiks pulli spermide viljakusmarkerite väljaselgitamine, mis võimaldaks välja töötada veise viljakuse hindamise metoodika (genoomikatesti) ning luua sperma kvaliteeti ja hilisemat viljastamise määra tõstva seemendusdooside sünteetilise lisandi
Activities
During the project TGM1-biomarker and other monoclonal antisperm antibodies were tested for their sex chromosome specificity using semen from bulls of different breeds. As a result, TGM1-marker resulted to be not sufficiently effective for the use in sex selection technology development due to large biological variation between the bulls. As an alternative, colloid centrifugation method was tested to provide cheaper method for the sperm sex ratio alteration. It resulted in small shift in sperm sex ratio, but not economically sufficient to be used for commercial production of sex selected semen.
To identify sperm fertility markers, semen from high and low fertility bulls was investigated. In transcriptome analysis, 32 genes were revealed, which showed at least four-fold expression difference between the bull groups. Two genes showing relation to bull semen quality, were further validated. These results have a very good perspective for the further research on bovine fertility markers.
Additionally, 2 original FISH (fluorescence in situ hybridisation) methods were developed, which can be used for a) simultaneous analysis of Y- ja X-chromosomes in the sperm sample; b) simultaneous detection of antibodies and presence of X-chromosome in the sperm sample. These methods are important in quality control of sexed semen samples and in the development of novel sex selection technologies.
Activities
Projekti jooksul analüüsiti TGM1-biomarkeri jt. spermivastaste monoklonaalsete antikehade soospetsiifilisust erinevat tõugu pullide spermas. Töö tulemusena selgus et tingituna pullide bioloogilisest varieeruvusest, ei ole TGM1-biomarker piisavalt efektiivne sooselektsiooni tehnoloogia väljaarendamiseks. Projekti vältel toimus ka kõrgtehnoloogilisele pulli spermide sooselektsiooni meetodile alternatiivse, odavama meetodi arendus. Kolloidtsentrifuugimise meetod kallutas spermide osakaalu X-kromosoomi kandvate rakkude kasuks, kuid mitte piisavalt, et ületada majanduslikult otstarbekas lävend.
Sperma viljakusmarkerite määramiseks võrreldi andmeid viljakate ja vähem viljakate pullide vahel. Spermide transkriptoomide võrdlusel tuvastati 32 geeni, mille ekspressiooni muutus nende kahe pulligrupi vahel oli neljakordne või suurem. Valideerimise käigus leidis kinnitust, et 2 geeni ekspressioon on seotud pulli sperma kvaliteediga. Saadud tulemused veise sperma viljakusmarkerite määramisel omavad perspektiivi uuringute jätkamiseks ja on tõuaretuse seisukohast aktuaalsed.
Lisaks eeltoodule loodi kaks originaalset meetodit, mille abil saab a) analüüsida samaaegselt Y- ja X-kromosoomide esinemissagedust sperma preparaadis; b) uurida ühes preparaadis samu sperme nii antikehade immuunreaktsiooni kui ka rakkude X-kromosoomi kandvuse suhtes. Vastavad metoodikad on vajalikud suguselekteeritud sperma kvaliteedi hindamisel ja sooselektsiooni tehnoloogiate edasiarendamisel.
Project details
- Main funding source
- Rural development 2014-2020
- Agricultural sectors
- Dairy cattle
€ 392611
Total budget
Total contributions including EU funding.
€ 349613
EU contribution
Any type of EU funding.
Project keyword
Contacts
Triin Hallap
Project coordinator
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Animal Breeders Association of Estonia
Project coordinator
Project partners
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Estonian University of Life Sciences
Project partner
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The Competence Centre on Health Technologies
Project partner