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Perinatal Center, Departments of Physiology and Pharmacology (N.J., T.L.P., T.J.) and Anatomy and Cell Biology (B.R.J.), Göteborg University, 405 30 Göteborg, Sweden; and Academic Unit of Child Health (S.L.G.), University of Manchester, Manchester M13 0JH, United Kingdom
Address all correspondence and requests for reprints to: Nina Jansson, Perinatal Center, Department of Physiology and Pharmacology, Göteborg University, P.O. Box 432, 405 30 Göteborg, Sweden. E-mail: nina.jansson{at}fysiologi.gu.se.
The activity and expression of placental nutrient transporters are primary determinants for the supply of nutrients to the fetus, and these nutrients in turn regulate fetal growth. We developed an experimental system to assess amino acid uptake in single primary villous fragments to study hormonal regulation of the amino acid transporter system A in term human placenta. Validation of the method, using electron microscopy and studies of hormone production, indicated that fragments maintained ultrastructural and functional integrity for at least 3 h. The activity of system A was measured as the Na+-dependent uptake of methylaminoisobutyric acid (MeAIB), and the effect of 1 h incubation in various hormones was investigated. Uptake of MeAIB into villous fragments in the presence of Na+ was linear up to at least 30 min. Insulin (300 ng/ml, n = 14) increased system A activity by 56% (P < 0.05). This effect was also present at insulin concentrations in the physiological range (+47% at 0.6 ng/ml, n = 10, P < 0.05). Leptin (500 ng/ml, n = 14) increased Na+-dependent MeAIB uptake by 37% (P < 0.05). System A activity increased in a concentration-dependent fashion in response to leptin (n = 10). However, neither epidermal GF (600 ng/ml), cortisol (340 ng/ml), nor GH (500 ng/ml) altered system A activity significantly (n = 14). We conclude that primary single isolated villous fragments can be used in studies of hormonal regulation of nutrient uptake into the syncytiotrophoblast. These data suggest that leptin regulates system A, a key amino acid transporter.
This work was supported by grants from the Swedish Medical Research Council (Grant 10838), the Swedish Diabetes Association, the Torsten and Ragnar Söderbergs Foundation, the Åke Wiberg Foundation, The Magnus Bergvall Foundation, Frimurare-Barnhus-direktionen, the Åhlens Foundation, and the Willhelm and Martina Lundgrens Foundation.
Abbreviations: BM, Basal membrane; EGF, epidermal growth factor; hPL, human placental lactogen; IUGR, intrauterine growth restriction; LDH, lactate dehydrogenase; MeAIB, methylaminoisobutyric acid; MVM, microvillous membrane; SEM, scanning-electron microscopy; TEM, transmission-electron microscopy.
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