Device-mediated MODification of the Effects of Laser and led systems on sperm cells of local animals and an evaluation in terms of artificial insemination application: milk analysis and valorization in female generations

This project aims to evaluate individual photostimulation procedures in the process of developing a

practical device that uses laser radiation in common artificial insemination applications in farm animals. In

this respect, laboratory analysis of spermatological parameters, monitoring of pregnancy rates as a result of

artificial insemination with sperm cells exposed to photostimulation, and milk quality and valorization of

the female generation will be evaluated. For this purpose, ejaculates will be collected from each local

mature, fertile animal types [ Bos taurus – Eastern Anatolian Red, Bos taurus –Zavot, Norduz Sheep; at

least 5 animals of each species] via an artificial vagina (as an alternative method with electroejaculator).

Spermatological parameters will be evaluated in terms of viability, acrosome structure, motility,

cytochrome C level, lipid peroxidation, comet test and MTT. Besides, it was aimed to evaluate the effect of

photo-stimulation on frozen thawed sperm cells subjected to in vitro capacitation in terms of changes in

mitochondrial membrane potential. In the presented project, in order to investigate the effect of

photostimulation on DNA damage mechanism, PARP-1 mRNA transcript level will be determined by RTPCR method. ProAKAP4, ENO1, VDAC2 and UQCRC2 proteins are a biomarker for sperm quality. In

determining sperm quality, expression levels of ProAKAP4, ENO1, VDAC and UQCRC2 proteins will be

determined by immunofluorescence, western blot and real time PCR methods. In order to examine the

thawing efficiency of the device, first, the sperm common lipid profile (ColE, FFA, TAG, Col, PL) will be

examined by HPTLC, then, the fatty acid contents (eg. saturated vs unsaturated) of phospholipids will be

analyzed by GC-MS. In addition, it is planned to evaluate the results of pregnancy test (ultrasonography

method on 30th and 60th days) in fertile and healthy females. Six experimental replicates will run.