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Placentation in Coendou prehensilis (Rodentia, Erethizontidae)
A12
Abstracts / Placenta 35 (2014) A1eA112
character of endothelial intercellular junctions. It suggests more active angiogenesis, i.e. proliferation and following differentiation of endothelium and pericytes. We tested the hypothesis that cells of the capillary wall in diabetic term placenta have higher ability to proliferate and to differentiate into functionally efficient structure. Methods: Specimens taken from eight normal and sixteen diabetic placentas were fixed and paraffin embedded. In sections cut from five haphazardly chosen blocks per placenta, detection of Ki-67 antigen as a marker of proliferation was performed. In 20 fields of view per section, area of terminal villi was measured and Ki-67 positive nuclei in capillary wall were counted. Subsequently, mean numbers of labeled nuclei per square millimeter of section were calculated. The Ki-67 positive nuclei and total number of nuclei in capillary wall were counted and proliferation index was calculated as well. The same pattern of sections was used for immunocytochemical detection of nestin as a marker of developing vessels. The degree of cell differentiation in capillaries (approx. 50 per placenta) was evaluated as the percentage of nestin-labeled segments of capillary circumference. Results: There were found significant differences in the number of Ki-67 labeled nuclei/mm2 (10.11 ± 4.65 vs. 5.23 ± 4.50), in the proliferative index (0.040 ± 0.015 vs. 0.016 ± 0.011) and in the percentage of nestin-labeled segments of capillary circumference (0.199 ± 0.049 vs. 0.325 ± 0.065) in normal and in diabetic placentas respectively. Conclusions: The results indicate that in diabetic placentas at term, the cells of villous capillaries are less able to respond immediately on fetal demand by enlargement of capillary surface area. Higher proportion of undifferentiated cells in the capillary wall makes evident a delay of functional maturation. Supported by PRVOUK P25/LF1/2.
P1.4-N. NOVEL AND EFFICIENT 3D MICROSCOPIC ANALYSIS OF ISOLATED VILLOUS TREES OF HUMAN PLACENTAS Eva Haeussner a, Christoph Schmitz a, Franz Edler von Koch b, Hans-Georg Frank a a Ludwig-Maximilians-University, Munich, Germany; b Dritter Order Hospital, Munich, Germany The villous trees of human placentas delineate the fetomaternal border and are complex three-dimensional structures. Thus far, they are primarily analysed from thin two-dimensional histological sections. Such sections cannot provide access to key aspects like branching nodes and branch order. As a consequence, histologic nomenclature of the villous tree does not include characterization of villous nodes. Using samples taken from 50 normal human placentas at birth, in the present study, we show that analysis procedures for the three-dimensional reconstruction of neuronal dendrite trees can also analyse villous trees of human placentas. We demonstrate how parameters of dendrite analysis can be adapted to the analysis of villous trees. Nodes and their branches (e.g., branching hierarchy, branching angles, diameters and lengths of branches) can be efficiently measured in wholemount preparations of isolated villous trees using high-end light microscopy. Nodes are classified according to their distance from the terminal end of the villous trees. These novel data differ qualitatively from the data obtainable from histological sections and supplement such data. Since this novel method can be applied conveniently, an effective and technically feasible access to threedimensional morphology of villous trees is opened.
P1.5. PLACENTATION IN COENDOU PREHENSILIS (RODENTIA, ERETHIZONTIDAE) Fabio Sergio Cury a, Phelipe Oliveira Favaron a, Carlos Eduardo ~o sio a, b a Faculty of Veterinary Medicine and Animal Sciences, Sa Ambro ~o Paulo, Brazil; b Faculty of Animal Sciences and Food Engineering, Paulo, Sa ~o Paulo, Brazil Pirassununga, Sa Objectives: The Coendou prehensilis (Rodentia) belongs to the subordem Hystricomorpha. It is closely related to the African porcupines and share
several placental characteristics with hystricognath rodents. Although C. prehensilis is widely distributed in Central and South Americas data about placentation and fetal membranes is this group are very sparse. Herein we investigated the chorioallantoic and yolk sac placentation. Methods: Placental samples from early gestation were investigated by means of gross morphology, histology (H.E and Masson’s tricome), and immunohistochemistry (vimentin, 1:100). Results: The chorioallantoic placenta had a discoidal shape with a lobulated structure. The labyrinth corresponded to the major region for maternal fetal exchanges. In this region, fetal vessels were identified by vimentin-positive endothelium, in contrast to the maternal blood spaces which did not have endotelial cells. Sinusoidal giant cells were identified in the labyrinth. Another layer of trophoblast giant cells were identified near to the decidua. Syncytial and cellular trophoblast were interposed between maternal and fetal circulations. The visceral yolk sac was composed by cubic endoderm cells, a mesenchyme layer rich in vessels and mesothelium. Conclusion: Preliminary data confirm that placental characteristics in Erethizontidae are very similar to other caviomorph rodents. In addition, the next steps will focus in the placental barrier and trophoblast invasion and differentiation.
P1.6. SUBTROPHOBLASTIC DEPOSITS IN DIABETIC PLACENTAS OBSERVED BY ELECTRON MICROSCOPY Roberta– Rossi, Maria Grazia Fiore, Domenico Piscitelli, Antonella degli Studi Aldo Vimercati, Graziana Arborea, Leonardo Resta Universita Moro, Bari, Italy, Italy Objectives: In the last years we performed different studies about immunohistochemical, morphometrical and electronmicroscopy of diabetic placenta at term. In particular we observed a group of consecutive diabetic placenta in which a several amount of electron dense deposits near the basal membrane of villar trophoblast. In this report, we refer about the significance of such deposit. Methods: We exanimate 8 consecutive patients with normal pregnancy and delivery and 8 cases of diabetic well treated patients The mean age was very similar (31-36 year old in normal and 24-39 in diabetic patients). The weeks of gestation were respectively 36-41 and 39-41. Birth weight varied from 2700 to 3050g in control and from 2900 to 4550 g in diabetic women. The placental weight was 430-520 g and 520-850 g in two groups. Small fragments of placental tissue was collected at delivery and submitted to the usual process for TEM examination. Results: In 6 out of the 8 cases of diabetic placenta we observed a different amount of electrondense material near the basal membrane of villar trophoblast. These deposits were present beneath the membrane or between the cytotrophoblast and they had different intensity and form: linear, nodular, figurate. Conclusion: The presence of proteins linked to basal membrane are described in light microscopy and judged as a an effect of thickness of the membrane. In a recent work, a large spectrum of immunoglobulin has been demonstrated in basal membrane of diabetic placenta. In our experience the deposits observed in diabetic placentas had some special aspects similar to immunogenic deposits observed in glomerulopathies. The real thickness of the basal membrane without deposits is equal in normal and diabetic placentas.
P1.7-N. DECIDUAL ACUTE ATHEROSIS IN NORMAL, DIABETIC AND PREECLAMPTIC PREGNANCIES: USING REVISED DIAGNOSTIC CRITERIA Patji Alnæs-Katjavivi a, Fiona Lyall b, Borghild Roald d, Christopher Redman c, Annetine Staff a a Women and Children's Division, Oslo University Hospital (OUS), Oslo, Norway; b Institute of Medical Genetics, University of Glasgow, Glasgow, UK; c Nuffield Department of Obstetrics&Gynaecology, University of Oxford, Oxford, UK; d Department of Pathology, Oslo University Hospital (OUS), Oslo, Norway
Abstracts / Placenta 35 (2014) A1eA112
character of endothelial intercellular junctions. It suggests more active angiogenesis, i.e. proliferation and following differentiation of endothelium and pericytes. We tested the hypothesis that cells of the capillary wall in diabetic term placenta have higher ability to proliferate and to differentiate into functionally efficient structure. Methods: Specimens taken from eight normal and sixteen diabetic placentas were fixed and paraffin embedded. In sections cut from five haphazardly chosen blocks per placenta, detection of Ki-67 antigen as a marker of proliferation was performed. In 20 fields of view per section, area of terminal villi was measured and Ki-67 positive nuclei in capillary wall were counted. Subsequently, mean numbers of labeled nuclei per square millimeter of section were calculated. The Ki-67 positive nuclei and total number of nuclei in capillary wall were counted and proliferation index was calculated as well. The same pattern of sections was used for immunocytochemical detection of nestin as a marker of developing vessels. The degree of cell differentiation in capillaries (approx. 50 per placenta) was evaluated as the percentage of nestin-labeled segments of capillary circumference. Results: There were found significant differences in the number of Ki-67 labeled nuclei/mm2 (10.11 ± 4.65 vs. 5.23 ± 4.50), in the proliferative index (0.040 ± 0.015 vs. 0.016 ± 0.011) and in the percentage of nestin-labeled segments of capillary circumference (0.199 ± 0.049 vs. 0.325 ± 0.065) in normal and in diabetic placentas respectively. Conclusions: The results indicate that in diabetic placentas at term, the cells of villous capillaries are less able to respond immediately on fetal demand by enlargement of capillary surface area. Higher proportion of undifferentiated cells in the capillary wall makes evident a delay of functional maturation. Supported by PRVOUK P25/LF1/2.
P1.4-N. NOVEL AND EFFICIENT 3D MICROSCOPIC ANALYSIS OF ISOLATED VILLOUS TREES OF HUMAN PLACENTAS Eva Haeussner a, Christoph Schmitz a, Franz Edler von Koch b, Hans-Georg Frank a a Ludwig-Maximilians-University, Munich, Germany; b Dritter Order Hospital, Munich, Germany The villous trees of human placentas delineate the fetomaternal border and are complex three-dimensional structures. Thus far, they are primarily analysed from thin two-dimensional histological sections. Such sections cannot provide access to key aspects like branching nodes and branch order. As a consequence, histologic nomenclature of the villous tree does not include characterization of villous nodes. Using samples taken from 50 normal human placentas at birth, in the present study, we show that analysis procedures for the three-dimensional reconstruction of neuronal dendrite trees can also analyse villous trees of human placentas. We demonstrate how parameters of dendrite analysis can be adapted to the analysis of villous trees. Nodes and their branches (e.g., branching hierarchy, branching angles, diameters and lengths of branches) can be efficiently measured in wholemount preparations of isolated villous trees using high-end light microscopy. Nodes are classified according to their distance from the terminal end of the villous trees. These novel data differ qualitatively from the data obtainable from histological sections and supplement such data. Since this novel method can be applied conveniently, an effective and technically feasible access to threedimensional morphology of villous trees is opened.
P1.5. PLACENTATION IN COENDOU PREHENSILIS (RODENTIA, ERETHIZONTIDAE) Fabio Sergio Cury a, Phelipe Oliveira Favaron a, Carlos Eduardo ~o sio a, b a Faculty of Veterinary Medicine and Animal Sciences, Sa Ambro ~o Paulo, Brazil; b Faculty of Animal Sciences and Food Engineering, Paulo, Sa ~o Paulo, Brazil Pirassununga, Sa Objectives: The Coendou prehensilis (Rodentia) belongs to the subordem Hystricomorpha. It is closely related to the African porcupines and share
several placental characteristics with hystricognath rodents. Although C. prehensilis is widely distributed in Central and South Americas data about placentation and fetal membranes is this group are very sparse. Herein we investigated the chorioallantoic and yolk sac placentation. Methods: Placental samples from early gestation were investigated by means of gross morphology, histology (H.E and Masson’s tricome), and immunohistochemistry (vimentin, 1:100). Results: The chorioallantoic placenta had a discoidal shape with a lobulated structure. The labyrinth corresponded to the major region for maternal fetal exchanges. In this region, fetal vessels were identified by vimentin-positive endothelium, in contrast to the maternal blood spaces which did not have endotelial cells. Sinusoidal giant cells were identified in the labyrinth. Another layer of trophoblast giant cells were identified near to the decidua. Syncytial and cellular trophoblast were interposed between maternal and fetal circulations. The visceral yolk sac was composed by cubic endoderm cells, a mesenchyme layer rich in vessels and mesothelium. Conclusion: Preliminary data confirm that placental characteristics in Erethizontidae are very similar to other caviomorph rodents. In addition, the next steps will focus in the placental barrier and trophoblast invasion and differentiation.
P1.6. SUBTROPHOBLASTIC DEPOSITS IN DIABETIC PLACENTAS OBSERVED BY ELECTRON MICROSCOPY Roberta– Rossi, Maria Grazia Fiore, Domenico Piscitelli, Antonella degli Studi Aldo Vimercati, Graziana Arborea, Leonardo Resta Universita Moro, Bari, Italy, Italy Objectives: In the last years we performed different studies about immunohistochemical, morphometrical and electronmicroscopy of diabetic placenta at term. In particular we observed a group of consecutive diabetic placenta in which a several amount of electron dense deposits near the basal membrane of villar trophoblast. In this report, we refer about the significance of such deposit. Methods: We exanimate 8 consecutive patients with normal pregnancy and delivery and 8 cases of diabetic well treated patients The mean age was very similar (31-36 year old in normal and 24-39 in diabetic patients). The weeks of gestation were respectively 36-41 and 39-41. Birth weight varied from 2700 to 3050g in control and from 2900 to 4550 g in diabetic women. The placental weight was 430-520 g and 520-850 g in two groups. Small fragments of placental tissue was collected at delivery and submitted to the usual process for TEM examination. Results: In 6 out of the 8 cases of diabetic placenta we observed a different amount of electrondense material near the basal membrane of villar trophoblast. These deposits were present beneath the membrane or between the cytotrophoblast and they had different intensity and form: linear, nodular, figurate. Conclusion: The presence of proteins linked to basal membrane are described in light microscopy and judged as a an effect of thickness of the membrane. In a recent work, a large spectrum of immunoglobulin has been demonstrated in basal membrane of diabetic placenta. In our experience the deposits observed in diabetic placentas had some special aspects similar to immunogenic deposits observed in glomerulopathies. The real thickness of the basal membrane without deposits is equal in normal and diabetic placentas.
P1.7-N. DECIDUAL ACUTE ATHEROSIS IN NORMAL, DIABETIC AND PREECLAMPTIC PREGNANCIES: USING REVISED DIAGNOSTIC CRITERIA Patji Alnæs-Katjavivi a, Fiona Lyall b, Borghild Roald d, Christopher Redman c, Annetine Staff a a Women and Children's Division, Oslo University Hospital (OUS), Oslo, Norway; b Institute of Medical Genetics, University of Glasgow, Glasgow, UK; c Nuffield Department of Obstetrics&Gynaecology, University of Oxford, Oxford, UK; d Department of Pathology, Oslo University Hospital (OUS), Oslo, Norway