MONOCLONAL ANTIBODY-AG7 INHIBITS FERTILIZATION POST SPERM-ZONA BINDING

被引:19
作者
BRUCKER, C
SANDOW, BA
BLACKMORE, PF
LIPFORD, GB
HODGEN, GD
机构
[1] EASTERN VIRGINIA MED SCH,DEPT OBSTET & GYNECOL,JONES INST REPROD MED,601 COLLEY AVE,NORFOLK,VA 23507
[2] EASTERN VIRGINIA SCH MED,DEPT ANAT & NEUROBIOL,NORFOLK,VA 23507
[3] EASTERN VIRGINIA SCH MED,DEPT PHARMACOL,NORFOLK,VA 23507
[4] EASTERN VIRGINIA SCH MED,DEPT IMMUNOL & MICROBIOL,NORFOLK,VA 23507
[5] UNIV MUNICH,WOMENS HOSP,W-8000 MUNICH 2,GERMANY
[6] TECH UNIV MUNICH,DEPT MICROBIOL & HYG,W-8000 MUNICH 2,GERMANY
关键词
SPERM ANTIGENS; ACROSOME REACTION; CALCIUM INFLUX; ZONA-PELLUCIDA;
D O I
10.1002/mrd.1080330412
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Monoclonal antibodies (mAbs) against sperm cells are currently being used in an effort to define spermatozoal antigens involved in the fertilization process. We have produced a number of anti-human sperm mAbs by immunization of female mice with the 100,000 x g supernatant of octylglycoside-solubilized washed human sperm. From a panel of mAbs, 1 antibody, AG7, was selected and characterized due to its fertilization-inhibiting characteristics. MAb AG7 defines a sperm acrosome antigen-1 (SAA-1) located in the acrosomal region of human sperm as evaluated by indirect immunofluorescence. Staining of life sperm cells indicated that the antigen is present on the sperm surface. SAA-1 was also found on sperm of several other mammalian species, implying evolutionary conservation of the antigen. SAA-1 was first observed on testicular sperm and can be followed through epididymal transit, ejaculation, and capacitation. When applied in a mouse in vitro fertilization assay, mAb AG7 inhibits fertilization by greater than 95%, and inhibition is dose dependent, with half-maximal inhibition at 0.8 mug/ml. The block to fertilization could not be attributed to sperm agglutination, inhibition of motility, interference with adhesion to the zona pellucida, or inhibition of fusion with the oocyte membrane. MAb AG7 was demonstrated to inhibit calcium influx in spermatozoa in vitro (measured using the fluorescent indicator fura 2), a prerequisite for the acrosome reaction. Initial biochemical characterization of the antigen suggests it is proteinlike in nature, with a molecular weight of approximately 220 kD. The results suggest that SAA-1, identified by mAb AG7, is a sperm antigen crucially involved in the fertilization process, possibly an atypical steroid receptor or ion channel located within the sperm plasma membrane.
引用
收藏
页码:451 / 462
页数:12
相关论文
共 49 条
[1]  
BIGGERS J. D., 1971, Methods in mammalian embryology, P86
[2]  
BLACKMORE PF, 1990, J BIOL CHEM, V265, P1376
[3]   CELL-SURFACE LOCALIZATION OF A NOVEL NON-GENOMIC PROGESTERONE-RECEPTOR ON THE HEAD OF HUMAN SPERM [J].
BLACKMORE, PF ;
LATTANZIO, FA .
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 1991, 181 (01) :331-336
[4]  
BRUCKER C, 1992, 8TH ANN M EUR SOC HU, P136
[5]  
BURKMAN LJ, 1988, FERTIL STERIL, V49, P688
[6]   INVITRO STUDIES OF THE GOLDEN-HAMSTER SPERM ACROSOME REACTION - COMPLETION ON THE ZONA-PELLUCIDA AND INDUCTION BY HOMOLOGOUS SOLUBLE ZONAE-PELLUCIDAE [J].
CHERR, GN ;
LAMBERT, H ;
MEIZEL, S ;
KATZ, DF .
DEVELOPMENTAL BIOLOGY, 1986, 114 (01) :119-131
[7]   ROLE OF CALCIUM-IONS IN ACROSOME REACTION OF SEA-URCHIN SPERM - REGULATION OF EXOCYTOSIS [J].
COLLINS, F ;
EPEL, D .
EXPERIMENTAL CELL RESEARCH, 1977, 106 (01) :211-222
[8]   STUDIES ON ACROSOME .7. FORMATION OF ACROSOMAL PROCESS IN SEA URCHIN SPERMATOZOA [J].
DAN, J ;
OHORI, Y ;
KUSHIDA, H .
JOURNAL OF ULTRASTRUCTURE RESEARCH, 1964, 11 (5-6) :508-&
[9]  
Dan J.C., 1967, FERTILIZATION, V1, P237
[10]   ENZYMATIC DISSECTION OF THE FUNCTIONS OF THE MOUSE EGGS RECEPTOR FOR SPERM [J].
FLORMAN, HM ;
BECHTOL, KB ;
WASSARMAN, PM .
DEVELOPMENTAL BIOLOGY, 1984, 106 (01) :243-255