Reproduction in South American camelids is poorly studied. To extend our knowledge of the development and cellular physiology of the placenta in the alpaca Lama pacos, we have examined specimens from day 150 of pregnancy to term. Morphological investigations using light, transmission and scanning electron microscopy, the histochemical localization of iron, alkaline and acid phosphatase activity, and the immunodetection of placental lactogen hormone were performed. Throughout pregnancy there was a progressive increase in the depths of folds on the uterine mucosa surface together with a thickening of the endometrium. Glandular cells exhibited PAS and acid phosphatase (AcP) positive secretion granules. In the chorion, giant trophoblast polyploid cells gradually became more numerous and larger. Non-giant cells exhibited positive granules for PAS, alkaline phosphatase (AkP) reaction and immunostaining for bovine placental lactogen hormone (PLH). SDS -PAGE electrophoresis and Western blotting procedures also confirmed the presence of a bovine PLH-like glycoprotein in the fetal alpaca placenta. Over the glandular openings, the chorion formed typical areolae, where the trophoblast exhibited AcP and PAS positive reactions. At these sites, the fetal endothelial cells contained iron-storage granules in their cytoplasm. The trophoblast-epithelial interface exhibited a complex microvillous interdigitation, in which an AkP reaction was very prominent. The chorionic capillaries progressively indented adjacent trophoblast cells. These data suggest that although the epitheliochorial alpaca placenta is diffuse, various trophoblast cell types and specialized areas of the maternofetal interface give the placenta micro-regional functions where histiotrophic nutrition, hormone production and molecular exchange are prevalent.

This study analyses the manner in which trophoblast cells adhere to uterine epithelium and the subsequent interactions that contribute to the establishment of epitheliochorial placentation in the alpaca Lama pacos. Specimens at the luteal and follicular phases and at 22, 26, 30 and 45 days of pregnancy (op) were processed for morphological studies. On day 15 op, the blastocysts are completely free within the uterine lumen, with implantation starting around day 20. On days 22 and 26 of gestation, the trophoblast is apposed to the epithelial surface of the uterus, with areas of contact and adhesion by means of complex interdigitation. Implantation sites occur prevalently in the left uterine horn, but an expanded trophoblast also occupies large extensions of the right horn, where the maternofetal interaction shows peculiar areas of apposition. As development continues, attachment areas become more extensive. On days 30 and 45, many secretory granules can be seen in the uterine epithelium, while giant multinucleate cells appear interposed between the remaining trophoblast cells, showing intense alkaline phosphatase activity, deposits containing iron and PAS-positive granules. Placental lactogen hormone is not present within the cytoplasm of the binucleate or multinucleate trophoblast cells. By day 30 of gestation, the trophoblast layer is lined by an extraembryonic connective tissue that by day 45 is well vascularized, thus indicating the starting point of placental formation. Fetal and maternal capillaries indent the epithelium and the trophoblast, narrowing the specialized areas of exchange, which occur along the entire maternofetal interface, characterizing the diffuse nature of this placenta.

No abstract

The pattern of expression of a variety of placental nitric oxide synthase isoforms has contributed to elucidating the regulatory mechanisms of nitric oxide (NO) synthesis during gestation. The maintenance of vascular tone, attenuation of vasoconstriction, prevention of platelet and leukocyte adhesion to the trophoblast surface, and possible participation in uterine blood flow seem to be the main functions of NO generated at the fetal‐maternal interface in humans and mice. Extending this knowledge to other rodent species commonly used as laboratory animals, in this study we focus on NADPH‐diaphorase activity and the distribution of nitric oxide synthase isoforms (NOS) in the trophoblast cells of Calomys callosus during different phases of pregnancy. NADPH‐diaphorase activity was evaluated cytochemically and the presence of NOS isoforms detected by immunohistochemistry. These techniques were performed on pre‐ and postimplantation embryos in situ and in vitro, as well as in placentae on d 14 and 18 of pregnancy. Neither NADPH‐diaphorase activity nor inducible or endothelial NOS isoforms were found in pre‐implanting embryos except after culturing for at least 48 h, when some of the embryonic cells were positive for the diaphorase reaction. On d 6·5 of pregnancy, trophoblast cells showed intense diaphorase activity both in situ and under in vitro conditions. A positive reaction was also found in the different placental trophoblast cells on d 14 and 18 of pregnancy. The inducible NOS (iNOS) isoform, but not the endothelial isoform, was immunodetected in trophoblast cells from the placenta and from postimplantation embryos in situ and under in vitro conditions. These results strongly suggest the production of NO by the iNOS isoform in the trophoblast of Calomys callosus after embryo implantation. The data also emphasise a possible role for the trophoblast in producing and releasing cytotoxic molecules at the fetal‐maternal interface.