1. Figure 4. In-vitro sperm binding to ovulated eggs
Figure 4. In-vitro sperm binding to ovulated eggs. Eggs were incubated with 1.5×10⁶ mouse spermatozoa/ml for 30 min and washed (to remove non-adherent spermatozoa) until 2–6 spermatozoa were bound to control 2-cell mouse embryos (insets). (A) Eggs from huZP3 rescue females (moZP1, moZP2, huZP3) injected with mAb to human ZP3. (B) Same as (A) but injected with mAb to mouse ZP3. (C) Eggs from normal (wild-type) females injected with mAb to human ZP3. (D) Same as (C) but injected with mAb to mouse ZP3. Scale bar = 50 μm.
From: Antibodies to human ZP3 induce reversible contraception in transgenic mice with `humanized' zonae pellucidae
Hum Reprod. 1999;14(3): doi: /humrep/
Hum Reprod | © European Society of Human Reproduction and Embryology

2. Figure 5. Western blot of ovulated eggs of HuZP3 rescue and normal mice injected with antibodies. (A) Proteolytic cleavage of mouse ZP2 associated with the post-fertilization block to polyspermy can be detected with a mAb to the N-terminal portion of ZP2 (amino acids 114–129). Top: ZP2 in the zona matrix of ovulated eggs is intact. Bottom: ZP2 in the zona matrix of 2-cell embryos is cleaved but the two subunits (20–30 kDa and 90 kDa) remain attached unless linking disulphide bond(s) are reduced (Moller and Wassarman, 1989). (B) 13 eggs or 2-cell embryos were analysed by Western blot analysis using the mAb to mouse ZP2: ovulated eggs (lanes 1,3) and 2-cell embryos (lanes 2,4) from normal (wild-type) females were incubated without (lanes 1,2) and with (lanes 3,4) 50 mM β-mercaptoethanol (β-SH), respectively, prior to SDS–PAGE. (C) Lane 1, ovulated eggs from huZP3 rescue mice injected with mAb to human ZP3; lane 2, same as lane 1 but injected with mAb to mouse ZP3; lane 3, ovulated eggs from normal mice injected with mAb to human ZP3; and, lane 4, same as lane 3 but injected with mAb to mouse ZP3. All samples in (C) contained 13 eggs and were incubated with 50 mM β-mercaptoethanol. Molecular weight markers (kDa) are indicated at the left.
From: Antibodies to human ZP3 induce reversible contraception in transgenic mice with `humanized' zonae pellucidae
Hum Reprod. 1999;14(3): doi: /humrep/
Hum Reprod | © European Society of Human Reproduction and Embryology

3. Figure 6. Antibodies prevent fertilization
Figure 6. Antibodies prevent fertilization. MAb to human or mouse ZP3 were administered to huZP3 rescue and normal (wild-type) female mice, respectively. At 20 h after superovulation and mating with males, eggs/zygotes were recovered from the oviduct. (A) Total number of eggs/zygotes (± SEM) recovered from: bar 1, huZP3 rescue females treated with mAb to human ZP3; bar 3, same as bar 1 but treated with mAb to mouse ZP3; bar 5, normal females treated with mAb to human ZP3; bar 7, same as bar 5 but treated with mAb to mouse ZP3. Number of 1-cell zygotes (▪) recovered from: bar 2, huZP3 rescue females treated with mAb to human ZP3; bar 4, same as bar 2 but treated with mAb to mouse ZP3; bar 6, normal females treated with mAb to human ZP3; bar 8, same as bar 6 but treated with mAb to mouse ZP3. Standard errors and percentage of cells recovered as 1-cell zygotes are indicated. Photomicrographs of: (B) unfertilized eggs recovered from huZP3 rescue female mice treated with mAb to human ZP3; (C) 1-cell zygotes recovered from huZP3 rescue female mice treated with mAb to mouse ZP3; (D) 1-cell zygotes recovered from normal female mice treated with mAb to human ZP3; (E) unfertilized eggs recovered from normal female mice treated with mAb to mouse ZP3. Pronuclei in 1-cell zygotes are indicated with arrows. Scale bar = 50 μm.
From: Antibodies to human ZP3 induce reversible contraception in transgenic mice with `humanized' zonae pellucidae
Hum Reprod. 1999;14(3): doi: /humrep/
Hum Reprod | © European Society of Human Reproduction and Embryology

4. Figure 1. Reversible contraception in mice with `humanized' zonae pellucidae. (A) Fertility of 10 huZP3 rescue mice injected with mAb to human (•) or mouse (▪) ZP3. At 2 days after antibody administration, females were continuously mated with males until just prior to delivery. After giving birth, eight of the female mice injected with mAb to human ZP3 were re-mated (○). (B) Fertility of 10 normal (wild-type) mice injected with mAb to human (•) or mouse (▪) ZP3. After giving birth, eight of the female mice injected with mAb to mouse ZP3 were re-mated (□).
From: Antibodies to human ZP3 induce reversible contraception in transgenic mice with `humanized' zonae pellucidae
Hum Reprod. 1999;14(3): doi: /humrep/
Hum Reprod | © European Society of Human Reproduction and Embryology