Ahmad K, Naz R K
Department of Obstetrics and Gynecology, Albert Einstein College of Medicine, Bronx, New York 10461.
Biol Reprod. 1991 Dec;45(6):841-50. doi: 10.1095/biolreprod45.6.841.
The effects of antibodies against sperm antigens and the c-myc proto-oncogene product on early embryonic development were investigated in mice. Affinity-purified Fab' antibodies against lithium diiodosalicylate (LIS)-solubilized murine sperm extract and fertilization antigen (FA-1) reduced (p less than 0.01 to p less than 0.001) blastulation rates of in vitro cultured 2-cell murine embryos primarily because of an arrest of development at the morula stage. Similarly, the c-myc monoclonal antibody (mAb) affected early embryonic development in a dose-dependent manner. These effects were specific, since immunoabsorption, with its respective peptide, completely blocked the inhibitory effect of the c-myc mAb. Anti-LIS sperm Fab' identified four protein bands (approx. 36, 29, 24.6, and 17.6 kDa) on Western blots of extracts from unfertilized and fertilized ova, one band (approx. 68 kDa) each on 4-8-cell embryo and morula extracts, and one band (approx. 53 kDa) on blastocyst extracts. Anti-FA-1 Fab' did not react with unfertilized or fertilized ova, but specifically identified two protein bands (approx. 53 and 25.7 kDa) on blots of 2-cell-embryo extract, one band (approx. 25.7 kDa) on morula extract, and one band (approx. 53 kDa) on blastocyst extract. The c-myc mAb did not react with any band corresponding to the c-myc protein on blots of extracts from unfertilized or fertilized ova, 2-cell embryos, 4-8-cell embryos, morulae, or blastocysts. These results suggest that some of the cross-reacting sperm antigens that are expressed during early cleavages, and the product of the c-myc proto-oncogene may have a role in normal early embryonic development.
在小鼠中研究了抗精子抗原抗体和c-myc原癌基因产物对早期胚胎发育的影响。针对二碘水杨酸锂(LIS)溶解的小鼠精子提取物和受精抗原(FA-1)的亲和纯化Fab'抗体降低了(p小于0.01至p小于0.001)体外培养的2细胞小鼠胚胎的囊胚形成率,主要是因为在桑椹胚阶段发育停滞。同样,c-myc单克隆抗体(mAb)以剂量依赖的方式影响早期胚胎发育。这些作用是特异性的,因为用其各自的肽进行免疫吸附完全阻断了c-myc mAb的抑制作用。抗LIS精子Fab'在未受精和受精卵提取物的蛋白质印迹上鉴定出四条蛋白带(约36、29、24.6和17.6 kDa),在4-8细胞胚胎和桑椹胚提取物上各有一条带(约68 kDa),在囊胚提取物上有一条带(约53 kDa)。抗FA-1 Fab'与未受精或受精卵不反应,但在2细胞胚胎提取物的印迹上特异性鉴定出两条蛋白带(约53和25.7 kDa),在桑椹胚提取物上有一条带(约25.7 kDa),在囊胚提取物上有一条带(约53 kDa)。c-myc mAb在未受精或受精卵、2细胞胚胎、4-8细胞胚胎、桑椹胚或囊胚提取物的印迹上未与任何对应于c-myc蛋白的条带反应。这些结果表明,一些在早期卵裂过程中表达的交叉反应性精子抗原以及c-myc原癌基因的产物可能在正常早期胚胎发育中起作用。