Altered protein profile and possible hypoxia in the placenta of 2,3,7,8-tetrachlorodibenzo-p-dioxin-exposed rats.
Exposure to 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) during pregnancy causes fetal death in many animal species. In an earlier study we observed alteration of placental glucose kinetics at the same TCDD exposure level that resulted in fetal death (Ishimura et al., Toxicol. Appl. Pharmacol. 178, 161-171, 2002). In the present study, in order to identify the molecules that might explain the alterations of placental function and the mechanism of fetal death, we used two-dimensional gel electrophoresis (2D/E) to detect and identify placental proteins whose amounts changed after exposure to TCDD and we examined the expression properties of these proteins in the placenta during hypoxia by using the uterine artery ligation model. Pregnant Holtzman rats were given a single oral dose of 1600 ng TCDD/kg body wt or an equivalent volume of vehicle (control) on gestational day (GD) 15 and placental tissue was collected on GD16 and GD20. The 15,000 g supernatant fractions of placental homogenates from the control group and TCDD-exposed group were subjected to the 2D/E analysis, and the protein spots whose amounts had changed after exposure to TCDD were characterized by amino acid sequence analysis. The amounts of heat shock protein 27 (Hsp27) and beta-tropomyosin (beta-TM) in TCDD-exposed placentas tended to have increased on GD16 and had increased significantly on GD20, and these changes were followed by an approximately twofold increase in glyceraldehyde 3-phosphate dehydrogenase (GAPDH) on GD20. Next, the uterine-artery ligation model was prepared on GD15, and the hypoxic placentas were collected on GD20. Two-D/E analysis of the 15,000 g supernatant proteins of the placentas revealed an increased level of GAPDH but not of other proteins, including Hsp27 and beta-TM. The results of this study showed that the increase in GAPDH level during hypoxia previously observed in endothelial cells occurs in the placenta and indicated that the TCDD-exposed placentas were in a hypoxic state at the end of pregnancy. Finally, the results of this study suggested the possibility that the increased incidence of fetal death after exposure to TCDD was due to the placental hypoxia.
Ishimura
Ryuta
R
Environmental Health Sciences Division, National Institute for Environmental Studies, 16-2 Onogawa, Tsukuba 305-8506, Japan.
Ohsako
Seiichiroh
S
Kawakami
Takashige
T
Sakaue
Motoharu
M
Aoki
Yasunobu
Y
Tohyama
Chiharu
C
D000595:Amino Acid Sequence; D000818:Animals; D015180:Electrophoresis, Gel, Two-Dimensional; D005260:Female; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000096:biosynthesis; D055551:HSP27 Heat-Shock Proteins; D006360:Heat-Shock Proteins; D000860:Hypoxia / Q000378:metabolism*; D007150:Immunohistochemistry; D008297:Male; D008969:Molecular Sequence Data; D009363:Neoplasm Proteins / Q000096:biosynthesis; D010920:Placenta / Q000187:drug effects / Q000378:metabolism*; D000072317:Polychlorinated Dibenzodioxins / Q000633:toxicity*; D011247:Pregnancy; D011506:Proteins / Q000378:metabolism*; D051381:Rats; D017207:Rats, Sprague-Dawley; D012039:Regional Blood Flow / Q000502:physiology; D014335:Tropomyosin / Q000096:biosynthesis; D014599:Uterus / Q000098:blood supply
D055551:HSP27 Heat-Shock Proteins; D006360:Heat-Shock Proteins; C486114:Hspb1 protein, rat; D009363:Neoplasm Proteins; D000072317:Polychlorinated Dibenzodioxins; D011506:Proteins; D014335:Tropomyosin; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases