The Role Of Human Sperm Mannose Receptor(hsMR)- Related Molecules In Sperm Egg Fusion

Objective: To investigate the role of relative molecules of the human sperm mannose receptor(hs MR) :Serum amyloid P-component(SAP) and HSP70 family molecules(HSP70/HSPA2/HSC70) in the interaction(fusion)of sperm-oocyte,so as to further understand the relationship of hs MR with SAP and HSP70 s.Methods: Using interspecific in vitro fertilization experiment model, namely human sperm penetration assay(sperm penetration assay(SPA), Preincubated zona-free hamster oocytes with different concentrations of Hsp A2 recombinant antigen about 30 min, and together with human capacitated sperm in vitro fertilization 3h.With different concentrations of HSPA2 antibody preincubated the human sperm 30 min, and then incubate with zona-free hamster oocyte for fertilization in vitro 3h.According to the effective concentration from that result above, with the same concentration of human sperm mannose receptor(hs MR) related molecules preincubating zona-free hamster oocytes about 30 min, and together with human capacitated sperm in vitro fertilization 3h.Or the same concentration of human sperm mannose receptor(hs MR) related molecules antibodies preincubating human capacitated sperm about 30 min, and incubate with zona-free hamster oocyte for fertilization in vitro 3h.To make the fertilized egg tablets, the number of fertilized eggs, the number of the sperm heads penetrated and the number of the sperm binding were observed by the phase contrast microscope. The results of several experiments were statistically analyzed by statistical methods.Results:1. The fertilization rate of Hsp A2 recombinant protein group and BSA control group has significant difference(P < 0.05), indicating a role in inhibition of fertilization, the penetration index and binding index with BSA control group have no significant difference.2.the fertilization rate of the anti HSPA2 antibody test group and BSA control group is significantly different(p<0.05), which showes that it has the effect of inhibiting fertilization, the Fertilization Index and the combination index with BSA control group are not significantly different.3.The fertilization rates are significantly different between BSA group and HSP70 group and HSPA2 group,but not in penetration index and binding index. There are no significant differences in fertilization rate, penetration index, and binding index between SAP and HSC70 group and BSA group. 4 the fertilization rates between the HSP70 antibody and anti HSPA2 antibody group and BSA control group are different, penetration indexes, and binding indexes are not significant different.There are no significant differences in fertilization rate, penetration index, and binding index between SAP antibody and HSC70 antibody group and BSA group.Conclusion:1. SAP and HSC70 have no inhibitory effect on the fusion of human sperm and ha mster oocyte; 2. A certain concentration of HSP70 / HSPA2 protein preincubated zona-free hams ter oocytes and a certain concentration of HSP70 / HSPA2 antibody pretreated s perm can inhibit fertilization; 3. The role of recombinant heat shock protein Hsp A2 and hs MR in human sperm a nd oocyte recognition- Fusion is highly similar, further supporting the hypothes is that the head of human sperm mannose receptor(hs MR) is Hsp A2,one of HS P70 family molecules.