Purification of heparin binding oviduct specific proteins and its effect on in vitro embryo development in cattle.

Aditya K Sharma, Sushil K Mohapatra, A K Mohanty, S K Das

Dairy Biotechnology Laboratory, Eastern Regional Station, National Dairy Research Institute, Kalyani 741 235, India.

Indian journal of experimental biology 2013 May

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The objective of this study was to see the effect of purified heparin binding oviduct specific proteins (OSP) as media supplement on in vitro embryo developmental competence in cattle. The oviduct specific proteins were isolated from abattoir cattle oviducts and precipitated, dialyzed and at the end purified by high performance liquid chromatography system. The SDS-PAGE profile of eluted heparin binding protein (HBP) fraction showed bands between approximately 66-approximately 97 kDa, while heparin unbinding protein (HUBP) fraction showed two bands at approximately 66 kDa and in total protein (TP) bands were approximately 60-approximately 95 kDa. Collected all three OSP fractions were used as a media supplement in three different concentrations (0, 5 and 20 microg/mL) for in vitro maturation of immature oocytes, in vitro fertilization and culture of presumptive embryos at 38.5 degrees C in 5% CO2 incubator with maximum humidity. The highest cleavage rate (73.40 +/- 2.36%) was observed at 5 microg/mL concentration level and lowest cleavage rate (27.63 +/- 1.89%) was obtained in 20 microg/mL total protein (TP) fraction. The highest blastocyst formation (26.47 +/- 1.47%) also occurred in 5 microg/mL concentration of total protein (TP) fraction and the lowest blastocyst rate (3.60 +/- 1.80%) was achieved at 20 microg/mL HBP fraction. The highest cleavage rate in the control group was 60.45 +/- 2.66% in TP fraction and blastocyst formation was 11.66 +/- 2.54% in HUBP fraction which was not significantly differ from HBP fraction. These results indicate that at 5 microg/mL of total OSP fraction (TP) and HBP used as media supplement increased the cleavage rate significantly as compared to HUBP fraction, and total OSP fraction (TP) increased blastocyst formation significantly (P < 0.05) as compared to HBP & HUBP fraction.

Citation

Aditya K Sharma, Sushil K Mohapatra, A K Mohanty, S K Das. Purification of heparin binding oviduct specific proteins and its effect on in vitro embryo development in cattle. Indian journal of experimental biology. 2013 May;51(5):347-51