TY - JOUR
T1 - Recovery of penetration ability in protease‐treated zona‐free mouse eggs occurs coincident with recovery of a cell surface 94 kD protein
AU - Kellom, Theresa
AU - Vick, Angela
AU - Boldt, Jeffrey
PY - 1992/9
Y1 - 1992/9
N2 - Previous studies have demonstrated that protease treatment of zona‐free mouse eggs impairs sperm‐egg interaction (Boldt et al.: Biol Reprod 39:19–27. 1988) and causes modification of a 94 kD egg plasma membrane protein (Boldt et al., Gamete Res 23:91–101, 1989), In this report, the ability of eggs to recover penetration ability following protease treatment was examined. Zona‐free mouse eggs were isolated and treated with either trypsin or chymotrypsin (1 mg/ml, 20 min), then cultured for 0, 3, or 6 hr before insemination. Eggs cultured for 3 or 6 hr displayed significantly higher penetration levels than eggs inseminated immediately after protease treatment, indicating a recovery of penetration ability during the 3 or 6 hr incubation period. The recovery of penetration ability was not blocked by inclusion of cyclohexamide (50 μg/ml) during the 3 or 6 hr culture period, indicating that protein synthesis was not required for recovery of fusion ability. Cell surface radiolabeling studies with 125I revealed that a 94 kD cell surface protein was lost immediately following trypsin or chymotrypsin treatment but was found on the egg surface after the 3 or 6 hr recovery period. Recovery of the 94 kD egg surface protein occurred in the presence of cyclohexamide, and metabolic radiolabeling studies with 35S‐methionine confirmed that synthesis of a 94 kD protein was blocked by cyclohexamide. These results suggest that the recovery of penetration ability after protease treatment of zona‐free eggs is due to recovery of the 94 kD cell surface protein, providing further evidence for the involvement of the 94 kD protein in sperm‐egg interaction. © 1992 Wiley‐Liss, Inc.
AB - Previous studies have demonstrated that protease treatment of zona‐free mouse eggs impairs sperm‐egg interaction (Boldt et al.: Biol Reprod 39:19–27. 1988) and causes modification of a 94 kD egg plasma membrane protein (Boldt et al., Gamete Res 23:91–101, 1989), In this report, the ability of eggs to recover penetration ability following protease treatment was examined. Zona‐free mouse eggs were isolated and treated with either trypsin or chymotrypsin (1 mg/ml, 20 min), then cultured for 0, 3, or 6 hr before insemination. Eggs cultured for 3 or 6 hr displayed significantly higher penetration levels than eggs inseminated immediately after protease treatment, indicating a recovery of penetration ability during the 3 or 6 hr incubation period. The recovery of penetration ability was not blocked by inclusion of cyclohexamide (50 μg/ml) during the 3 or 6 hr culture period, indicating that protein synthesis was not required for recovery of fusion ability. Cell surface radiolabeling studies with 125I revealed that a 94 kD cell surface protein was lost immediately following trypsin or chymotrypsin treatment but was found on the egg surface after the 3 or 6 hr recovery period. Recovery of the 94 kD egg surface protein occurred in the presence of cyclohexamide, and metabolic radiolabeling studies with 35S‐methionine confirmed that synthesis of a 94 kD protein was blocked by cyclohexamide. These results suggest that the recovery of penetration ability after protease treatment of zona‐free eggs is due to recovery of the 94 kD cell surface protein, providing further evidence for the involvement of the 94 kD protein in sperm‐egg interaction. © 1992 Wiley‐Liss, Inc.
KW - Cyclohexamide
KW - Fusion ability
KW - Sperm‐egg interaction
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U2 - 10.1002/mrd.1080330107
DO - 10.1002/mrd.1080330107
M3 - Article
C2 - 1510843
AN - SCOPUS:0026687270
SN - 1040-452X
VL - 33
SP - 46
EP - 52
JO - Molecular Reproduction and Development
JF - Molecular Reproduction and Development
IS - 1
ER -