Study On Yellow Cattle Frozen Semen In Vitro Capacitation And The Expression Of Prostaglandin (PGF2α) Receptor Gene In Ovary

The study intended to increase the rate of frozen semen in vitro capacitation andinvestigate the function mechanism of prostaglandin on mammals theriogenology.weselected BO, Percoll, heparin,caffeine,that is related to frozen semen in vitrocapacitation;and PGF2αreceptors that is relevant to reproduction of animal asresearch objects. making use of different methods to study on frozen semen in vitrocapacitationaction. and using PCR clone Partial PGF2αreceptor gene sequence;immunohistochemistry locate the receptor’s distribution in ovary and corpus luteum.bioinfomatics software predict the physico-chemical property, hydrophobicity,secondary structure，domain and tertiary structure and so on. The purpose of theexperiment is to set up the best condition for frozen semen in vitro capacitation. andoffer the basic datas for the research of the effect of prostaglandin on mammalstheriogenology. The main results were as follow:1.with BO, Percoll, two different methods deal with semen, the results show thatthey BO is better than Percoll.2. The time for caffeine add to insemination medium is thawed semen incubationin it10min~15min, The ratio is1%.3. The best condition of frozen semen in vitro capacitation is the concentration ofheparin is80μg/ml，incubation time is20min; the best condition for acrosomereaction is the concentration of heparin is20μg/ml,incubation time is40min. the restof group have no significant difference.4. Sequence analysis showed the PGF2αcontained an ORF of633bp, coded210amino acids. The product was hydrophobicity and mainly composed of random coilandα-helix.5. Phylogenetic tree analysis of the predicted animo sequence showed a highhomology between cattle and sheep(91.80%).6. Immunohistochemictry result showed that the PGF2α recepter geneexpressed in both granular luteal cell and theca luteal cell of luteum, but higher in thegranular luteal cell type. While in ovary, expression level from high to low is granular cell(high), intimal cell of follicle (middle) and the stroma cell(low) in turn, but alllocated in cytoplasm.