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Cited by 46Pregnancy and preeclampsia studiesNeurological Complications and SyndromesReproductive System and Pregnancy

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Full-Length Human Placental sFlt-1-e15a Isoform Induces Distinct Maternal Phenotypes of Preeclampsia in Mice

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Szalai, Gabor, Romero, Roberto, Chaiworapongsa, Tinnakorn, Xu, Yi, Wang, Bing, Ahn, Hyunyoung, Xu, Zhonghui, Chiang, Po Jen, Sundell, Birgitta, Wang, Rona, Jiang, Yang, Plazyo, Olesya, Olive, Mary, Tarca, Adi L., Dong, Zhong, Qureshi, Faisal, Papp, Zoltan, Hassan, Sonia S., Hernandez-Andrade, Edgar, Than, Nandor Gabor·2015·PLoS ONE
DOI 10.1371/journal.pone.0119547PMC PMC4393117
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Abstract

OBJECTIVE: Most anti-angiogenic preeclampsia models in rodents utilized the overexpression of a truncated soluble fms-like tyrosine kinase-1 (sFlt-1) not expressed in any species. Other limitations of mouse preeclampsia models included stressful blood pressure measurements and the lack of postpartum monitoring. We aimed to 1) develop a mouse model of preeclampsia by administering the most abundant human placental sFlt-1 isoform (hsFlt-1-e15a) in preeclampsia; 2) determine blood pressures in non-stressed conditions; and 3) develop a survival surgery that enables the collection of fetuses and placentas and postpartum (PP) monitoring. METHODS: Pregnancy status of CD-1 mice was evaluated with high-frequency ultrasound on gestational days (GD) 6 and 7. Telemetry catheters were implanted in the carotid artery on GD7, and their positions were verified by ultrasound on GD13. Mice were injected through tail-vein with adenoviruses expressing hsFlt-1-e15a (n = 11) or green fluorescent protein (GFP; n = 9) on GD8/GD11. Placentas and pups were delivered by cesarean section on GD18 allowing PP monitoring. Urine samples were collected with cystocentesis on GD6/GD7, GD13, GD18, and PPD8, and albumin/creatinine ratios were determined. GFP and hsFlt-1-e15a expression profiles were determined by qRT-PCR. Aortic ring assays were performed to assess the effect of hsFlt-1-e15a on endothelia. RESULTS: Ultrasound predicted pregnancy on GD7 in 97% of cases. Cesarean section survival rate was 100%. Mean arterial blood pressure was higher in hsFlt-1-e15a-treated than in GFP-treated mice (∆MAP = 13.2 mmHg, p = 0.00107; GD18). Focal glomerular changes were found in hsFlt-1-e15a -treated mice, which had higher urine albumin/creatinine ratios than controls (109.3 ± 51.7 μg/mg vs. 19.3 ± 5.6 μg/mg, p = 4.4 x 10(-2); GD18). Aortic ring assays showed a 46% lesser microvessel outgrowth in hsFlt-1-e15a-treated than in GFP-treated mice (p = 1.2 x 10(-2)). Placental and fetal weights did not differ between the groups. One mouse with liver disease developed early-onset preeclampsia-like symptoms with intrauterine growth restriction (IUGR). CONCLUSIONS: A mouse model of late-onset preeclampsia was developed with the overexpression of hsFlt-1-e15a, verifying the in vivo pathologic effects of this primate-specific, predominant placental sFlt-1 isoform. HsFlt-1-e15a induced early-onset preeclampsia-like symptoms associated with IUGR in a mouse with a liver disease. Our findings support that hsFlt-1-e15a is central to the terminal pathway of preeclampsia, and it can induce the full spectrum of symptoms in this obstetrical syndrome.

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Figures

Fig 1 FLT1 protein isoforms and mRNA transcript variants. (A) Flt-1 contains seven extracellular Ig-like domains and an intracellular tyrosine kinase. The first three extracellular Ig-like domains are essential for ligand-binding, while the 4–7th extracellular Ig-like domains for receptor dimerization. The truncated mouse sFlt-1 mutant [msFlt-1(1–3)] contains only 1–329 amino acids of Flt-1, corresponding to the first three Ig-like domains. Mouse and human sFlt-1-i13 contains the first six Ig-like domains corresponding to 1–657 amino acids of Flt-1, as well as a unique 31-amino-acid tail. This unique C-terminus is evolutionarily highly conserved among mammals; the mouse and human amino acid sequences of this tail are only different in two positions (shown with blue letters). Among the human placental expressed sFlt-1 isoforms, hsFlt-1-i14, hsFlt-1-e15a and hsFlt-1-e15b diverge from Flt-1 after amino acid 706, and contain a 31-, 28- and 13-amino-acid unique tails, respectively. (B) Among the placental expressed FLT1 transcripts, the abundance of the mRNA encoding for the transmembrane receptor is about 2.5% in preeclampsia. FLT1 transcript expression data was retrieved from Jebbink et al. and is shown as transcript level divided by total FLT1 transcript level [154]. HsFlt-1-i13, the second most abundant placental FLT1 transcript in preeclampsia, is generated by skipped splicing and extension of exon 13. Similarly, hsFlt-1-i14 is generated by skipped splicing and extension of exon 14. HsFlt-1-e15a and hsFlt-1-e15b contain alternatively spliced exons derived from intronic sequences (exon 15a and exon 15b, respectively). The most abundant placental transcript, hsFlt-1-e15a contains exon 15a, which is located within a primate-specific AluSeq retrotransposon. The graph was adapted with permission from figures in publications of Heydarian et al. [153] and Shibuya M. [243]. Permissions for reuse of these original figures were obtained from Elsevier Ltd. and from the Proceedings of the Japan Academy, Series B, respectively.
Fig 1 FLT1 protein isoforms and mRNA transcript variants. (A) Flt-1 contains seven extracellular Ig-like domains and an intracellular tyrosine kinase. The first three extracellular Ig-like domains are essential for ligand-binding, while the 4–7th extracellular Ig-like domains for receptor dimerization. The truncated mouse sFlt-1 mutant [msFlt-1(1–3)] contains only 1–329 amino acids of Flt-1, corresponding to the first three Ig-like domains. Mouse and human sFlt-1-i13 contains the first six Ig-like domains corresponding to 1–657 amino acids of Flt-1, as well as a unique 31-amino-acid tail. This unique C-terminus is evolutionarily highly conserved among mammals; the mouse and human amino acid sequences of this tail are only different in two positions (shown with blue letters). Among the human placental expressed sFlt-1 isoforms, hsFlt-1-i14, hsFlt-1-e15a and hsFlt-1-e15b diverge from Flt-1 after amino acid 706, and contain a 31-, 28- and 13-amino-acid unique tails, respectively. (B) Among the placental expressed FLT1 transcripts, the abundance of the mRNA encoding for the transmembrane receptor is about 2.5% in preeclampsia. FLT1 transcript expression data was retrieved from Jebbink et al. and is shown as transcript level divided by total FLT1 transcript level [154]. HsFlt-1-i13, the second most abundant placental FLT1 transcript in preeclampsia, is generated by skipped splicing and extension of exon 13. Similarly, hsFlt-1-i14 is generated by skipped splicing and extension of exon 14. HsFlt-1-e15a and hsFlt-1-e15b contain alternatively spliced exons derived from intronic sequences (exon 15a and exon 15b, respectively). The most abundant placental transcript, hsFlt-1-e15a contains exon 15a, which is located within a primate-specific AluSeq retrotransposon. The graph was adapted with permission from figures in publications of Heydarian et al. [153] and Shibuya M. [243]. Permissions for reuse of these original figures were obtained from Elsevier Ltd. and from the Proceedings of the Japan Academy, Series B, respectively.CC0 · PubMed Central
Fig 2 Experimental procedures. The flow-chart shows the experimental procedures performed at certain time-points during the study. GD, gestational day; PPD, postpartum day; PFU, plaque forming unit.
Fig 2 Experimental procedures. The flow-chart shows the experimental procedures performed at certain time-points during the study. GD, gestational day; PPD, postpartum day; PFU, plaque forming unit.CC0 · PubMed Central
Fig 3 Determination of mouse pregnancy status with a 55MHz ultrasound probe. (A) Pregnant uterus on GD6. Gestational sacs of 1.8mm–2.7mm were observed in the proximity of the abdominal surface without visible signs of an embryo. (B) Pregnant uterus on GD7. Advanced endometrial reaction and the presence of an embryo were visible in the gestational sacs. (C) Non-pregnant uterus of a mouse seven days after mating, equivalent to GD7. (D) The pie chart shows that pregnancy could be diagnosed in 32 (white) of the 35 mice with high-frequency ultrasound on GD7. Of three mice diagnosed as non-pregnant on GD7 (shading), two were non-pregnant (grey shading), while one carried a pregnancy to term (white shading).
Fig 3 Determination of mouse pregnancy status with a 55MHz ultrasound probe. (A) Pregnant uterus on GD6. Gestational sacs of 1.8mm–2.7mm were observed in the proximity of the abdominal surface without visible signs of an embryo. (B) Pregnant uterus on GD7. Advanced endometrial reaction and the presence of an embryo were visible in the gestational sacs. (C) Non-pregnant uterus of a mouse seven days after mating, equivalent to GD7. (D) The pie chart shows that pregnancy could be diagnosed in 32 (white) of the 35 mice with high-frequency ultrasound on GD7. Of three mice diagnosed as non-pregnant on GD7 (shading), two were non-pregnant (grey shading), while one carried a pregnancy to term (white shading).CC0 · PubMed Central
Fig 4 Implantation with a telemetric blood pressure monitoring system. (A) On GD8, after isolation and ligation of the left common carotid artery at the level of bifurcation, a small arteriotomy was prepared with a 25G tip needle, (B) and the blood pressure monitoring catheter was positioned into the aortic arch with the assistance of the vessel cannulation forceps. (C) The transmitter was placed into a subcutaneous pocket in the left flank and preformed with blind dissection. (D) On GD13, the position of the telemetry catheter was determined with a 55MHz ultrasound probe. The catheter tip is situated in the aortic arch, and the intra-aortic part of the catheter reaches the optimal 2mm length. (E) On GD18, a pregnant mouse is shown before a cesarean section. The incision line of the telemetry surgery healed completely. The projected graph illustrates the position of the intra-aortic catheter and the subcutaneous telemetric blood pressure transmitter. (F) On PPD8, the catheter position, aortic arch, and main arterial branches were visualized after autopsy in the mouse mediastinum and chest. The dotted lines show the heart and main arteries of the mediastinum. (A-F) Head orientations are shown with asterisks.
Fig 4 Implantation with a telemetric blood pressure monitoring system. (A) On GD8, after isolation and ligation of the left common carotid artery at the level of bifurcation, a small arteriotomy was prepared with a 25G tip needle, (B) and the blood pressure monitoring catheter was positioned into the aortic arch with the assistance of the vessel cannulation forceps. (C) The transmitter was placed into a subcutaneous pocket in the left flank and preformed with blind dissection. (D) On GD13, the position of the telemetry catheter was determined with a 55MHz ultrasound probe. The catheter tip is situated in the aortic arch, and the intra-aortic part of the catheter reaches the optimal 2mm length. (E) On GD18, a pregnant mouse is shown before a cesarean section. The incision line of the telemetry surgery healed completely. The projected graph illustrates the position of the intra-aortic catheter and the subcutaneous telemetric blood pressure transmitter. (F) On PPD8, the catheter position, aortic arch, and main arterial branches were visualized after autopsy in the mouse mediastinum and chest. The dotted lines show the heart and main arteries of the mediastinum. (A-F) Head orientations are shown with asterisks.CC0 · PubMed Central
Fig 5 Survival cesarean section. (A) After a 1cm–1.5cm midline abdominal incision, a short segment of one uterine horn was exteriorized, and a 3mm–5mm longitudinal hysterectomy was performed on the opposite side of the mesometrial arterial arcade. (B-C) As the uterine wall could be easily dilated, this minimal incision enabled the delivery of two to three fetuses and their placentas using gentle fingertip pressure. An arrowhead depicts a placenta, stars depict umbilical cords, and arrows point to the fetuses. (D) Hysterectomies were closed with a single 4/0 absorbable multifilament suture. (E) After abdominal lavage with 0.9% sterile saline, the abdominal wall was closed with an absorbable multifilament continuous suture, and the skin was closed with 7mm-wide staples. The image shows a surgical field before euthanization on PPD8. (F) One uterus harvested after euthanization on PPD8. Arrowheads show hysterectomy sutures. (G) H&E staining of a uterine cross-section of a control mouse euthanized on PPD8 shows granulation tissue with enlarged vessels, foam cells, myofibroblasts, macrophages, and hemosiderin deposition. The image inside the black box is magnified in Subfigure H. (I) SMA immunostaining of the same uterus. The arrow depicts the cesarean section incision site with the disruption of the two-layered myometrium. (J) H&E staining of a uterine cross-section of a control mouse euthanized on PPD77 shows granulation tissue and the complete healing of the two-layered myometrium. The image inside the black box is magnified in Subfigure K. (L) SMA immunostaining of the same uterus. The arrow depicts a suture site. (G, I, J and L: 40x magnifications, H and K: 100x magnifications).
Fig 5 Survival cesarean section. (A) After a 1cm–1.5cm midline abdominal incision, a short segment of one uterine horn was exteriorized, and a 3mm–5mm longitudinal hysterectomy was performed on the opposite side of the mesometrial arterial arcade. (B-C) As the uterine wall could be easily dilated, this minimal incision enabled the delivery of two to three fetuses and their placentas using gentle fingertip pressure. An arrowhead depicts a placenta, stars depict umbilical cords, and arrows point to the fetuses. (D) Hysterectomies were closed with a single 4/0 absorbable multifilament suture. (E) After abdominal lavage with 0.9% sterile saline, the abdominal wall was closed with an absorbable multifilament continuous suture, and the skin was closed with 7mm-wide staples. The image shows a surgical field before euthanization on PPD8. (F) One uterus harvested after euthanization on PPD8. Arrowheads show hysterectomy sutures. (G) H&E staining of a uterine cross-section of a control mouse euthanized on PPD8 shows granulation tissue with enlarged vessels, foam cells, myofibroblasts, macrophages, and hemosiderin deposition. The image inside the black box is magnified in Subfigure H. (I) SMA immunostaining of the same uterus. The arrow depicts the cesarean section incision site with the disruption of the two-layered myometrium. (J) H&E staining of a uterine cross-section of a control mouse euthanized on PPD77 shows granulation tissue and the complete healing of the two-layered myometrium. The image inside the black box is magnified in Subfigure K. (L) SMA immunostaining of the same uterus. The arrow depicts a suture site. (G, I, J and L: 40x magnifications, H and K: 100x magnifications).CC0 · PubMed Central
Fig 6 Profiling of hsFlt-1-e15a and GFP expression. Boxplots show the expression profile of GFP (A) or hsFlt-1-e15a (B) mRNAs in placentas harvested on GD18 and in five tissues harvested on PPD8. Adenovirus doses (1x109 or 2x109 plaque-forming units) are depicted with different colors. Expression of hsFlt-1-e15a and GFP mRNAs was highest in the liver. Viral dose-effect was not seen in GFP expression; however, there was a dose-effect in hsFlt-1-e15a expression. The number of hsFlt-1-e15a expressing placenta, brain and uterine tissues was low in the hsFlt-1-e15a 1x group.
Fig 6 Profiling of hsFlt-1-e15a and GFP expression. Boxplots show the expression profile of GFP (A) or hsFlt-1-e15a (B) mRNAs in placentas harvested on GD18 and in five tissues harvested on PPD8. Adenovirus doses (1x109 or 2x109 plaque-forming units) are depicted with different colors. Expression of hsFlt-1-e15a and GFP mRNAs was highest in the liver. Viral dose-effect was not seen in GFP expression; however, there was a dose-effect in hsFlt-1-e15a expression. The number of hsFlt-1-e15a expressing placenta, brain and uterine tissues was low in the hsFlt-1-e15a 1x group.CC0 · PubMed Central
Fig 7 Blood pressure monitoring. X-axes show gestational days (GDs) and postpartum days (PPDs). Mean arterial pressure changes (ΔMAP) are depicted on the Y-axes. Blue dots represent ΔMAP values for given time-points, blue error bars show +/-standard errors. Red lines depict the ΔMAP patterns, fitted from the linear mixed effects models. During pregnancy (left sides of the sub-figures), there was no significant blood pressure elevation in control mice (A,C; ΔMAP slope = 0.513 mmHg/day; p = 0.187), whereas hsFlt-1-e15a treatment significantly increased blood pressure (B,D; ΔMAP slope = 2.05 mmHg/day; p = 8.09x10-8). The ΔMAP slope in the hsFlt-1-e15a group was higher compared to the controls (1.53 mmHg/day; p = 0.0043). The ΔMAP at parturition (GD18) was 13.2 mmHg higher in hsFlt-1-e15a-treated mice than in the control mice (p = 0.00107). After cesarean delivery (right sides of sub-figures), a similar quadratic pattern of ΔMAP was found in both control and hsFlt-1-e15a-treated mice, but ΔMAP dropped below the baseline in control mice (-1.96 mmHg; p = 0.560), while it remained above this in hsFlt-1-e15a treated mice (6.88 mmHg; p = 0.0346). There was no dose effect of the number of viral construct injections (1x109 PFU vs. 2x109 PFU) in hsFlt-1-e15a-treated or in control mice either before delivery (dose effect: -1.06 mmHg, p = 0.693) or in the postpartum period (dose effect: 1.30 mmHg, p = 0.763).
Fig 7 Blood pressure monitoring. X-axes show gestational days (GDs) and postpartum days (PPDs). Mean arterial pressure changes (ΔMAP) are depicted on the Y-axes. Blue dots represent ΔMAP values for given time-points, blue error bars show +/-standard errors. Red lines depict the ΔMAP patterns, fitted from the linear mixed effects models. During pregnancy (left sides of the sub-figures), there was no significant blood pressure elevation in control mice (A,C; ΔMAP slope = 0.513 mmHg/day; p = 0.187), whereas hsFlt-1-e15a treatment significantly increased blood pressure (B,D; ΔMAP slope = 2.05 mmHg/day; p = 8.09x10-8). The ΔMAP slope in the hsFlt-1-e15a group was higher compared to the controls (1.53 mmHg/day; p = 0.0043). The ΔMAP at parturition (GD18) was 13.2 mmHg higher in hsFlt-1-e15a-treated mice than in the control mice (p = 0.00107). After cesarean delivery (right sides of sub-figures), a similar quadratic pattern of ΔMAP was found in both control and hsFlt-1-e15a-treated mice, but ΔMAP dropped below the baseline in control mice (-1.96 mmHg; p = 0.560), while it remained above this in hsFlt-1-e15a treated mice (6.88 mmHg; p = 0.0346). There was no dose effect of the number of viral construct injections (1x109 PFU vs. 2x109 PFU) in hsFlt-1-e15a-treated or in control mice either before delivery (dose effect: -1.06 mmHg, p = 0.693) or in the postpartum period (dose effect: 1.30 mmHg, p = 0.763).CC0 · PubMed Central
Fig 8 Aortic ring assays. (A) A light image of an aortic ring from a mouse injected with Ad-CMV-GFP. (B) An image of the same aortic ring, illustrating segmentation and classification, with the aortic ring shown in blue and the red color depicting the microvessel outgrowth area. (C) A light image of an aortic ring from a mouse injected with Ad-CMV-hsFlt-1-e15a. (D) An image of the same aortic ring, illustrating segmentation and classification, with the aortic ring shown in blue and the red color depicting the microvessel outgrowth area. (A-D) Scale bars depict 500μm. (E) The bar chart depicts the differences in mean microvessel outgrowth areas (pixels) between the GFP and hsFlt-1-e15a groups.
Fig 8 Aortic ring assays. (A) A light image of an aortic ring from a mouse injected with Ad-CMV-GFP. (B) An image of the same aortic ring, illustrating segmentation and classification, with the aortic ring shown in blue and the red color depicting the microvessel outgrowth area. (C) A light image of an aortic ring from a mouse injected with Ad-CMV-hsFlt-1-e15a. (D) An image of the same aortic ring, illustrating segmentation and classification, with the aortic ring shown in blue and the red color depicting the microvessel outgrowth area. (A-D) Scale bars depict 500μm. (E) The bar chart depicts the differences in mean microvessel outgrowth areas (pixels) between the GFP and hsFlt-1-e15a groups.CC0 · PubMed Central
Fig 9 Histopathological and functional evaluation of the kidneys. Representative H&E (A) and PAS (B) stained sections show a morphologically normal glomerulus in a control animal. Representative H&E (D) and PAS (E) stained sections show a glomerulus with signs of swollen capillary endothelial cells and occlusion of glomerular capillaries in a mouse overexpressing hsFlt-1-e15a. (C,F) Charts depict albumin/creatinine ratios in urine specimens collected on gestational day (GD)7, GD13, GD18, and PPD8 from mice in the GFP (C) and hsFlt-1-e15a (F) groups. Mean urine albumin/creatinine ratios did not change in control mice; however, these increased by gestational days and then dropped postpartum in hsFlt-1-e15a overexpressing mice. Mean urine albumin/creatinine ratio on GD18 was higher in hsFlt-1-e15a-treated than in control mice (p = 4.4x10-2). The mean urine albumin/creatinine ratio on PPD8 was marginally significantly higher in hsFlt-1-e15a-treated (36.6±9.0μg/mg) than in control mice (18.0±4.9μg/mg; p = 0.06).
Fig 9 Histopathological and functional evaluation of the kidneys. Representative H&E (A) and PAS (B) stained sections show a morphologically normal glomerulus in a control animal. Representative H&E (D) and PAS (E) stained sections show a glomerulus with signs of swollen capillary endothelial cells and occlusion of glomerular capillaries in a mouse overexpressing hsFlt-1-e15a. (C,F) Charts depict albumin/creatinine ratios in urine specimens collected on gestational day (GD)7, GD13, GD18, and PPD8 from mice in the GFP (C) and hsFlt-1-e15a (F) groups. Mean urine albumin/creatinine ratios did not change in control mice; however, these increased by gestational days and then dropped postpartum in hsFlt-1-e15a overexpressing mice. Mean urine albumin/creatinine ratio on GD18 was higher in hsFlt-1-e15a-treated than in control mice (p = 4.4x10-2). The mean urine albumin/creatinine ratio on PPD8 was marginally significantly higher in hsFlt-1-e15a-treated (36.6±9.0μg/mg) than in control mice (18.0±4.9μg/mg; p = 0.06).CC0 · PubMed Central
Fig 10 “EM35” mouse had early-onset preeclampsia-like symptoms associated with fetal growth restriction. (A) “EM35” mouse had a higher and earlier blood pressure peak than other hsFlt-1-e15a-treated mice, followed by the normalization of blood pressures and a second blood pressure peak on PPD1. (B) The urinary albumin/creatinine ratio on GD18 was higher in “EM35” mouse than in other hsFlt-1-e15a-treated mice. (C) HsFlt-1-e15a expression in the liver was lower in “EM35” mouse than in other hsFlt-1-e15a-treated mice. (D-E) Histopathological examination of the liver found a multiple cystic biliary hyperplasia in the liver (* in D), and multiple recent (D) and remote (E) infarcts in the parenchyma of liver. H&E staining, 200x magnifications. (F) The endothelial outgrowth area was the lowest in “EM35” mouse among hsFlt-1-e15a-treated mice. (G-H) Mean fetal (G) weights (0.749±0.029) and placental (H) weights (0.080±0.006) in “EM35” mouse were significantly lower than in other hsFlt-1-e15a-treated mice (fetal weight: 1.06±0.023, p = 2.017x10-15, placental weight: 0.111±0.004, p = 2.05x10-6). (I) Histopathological examination of a placenta from “EM35” mouse showed multiple thrombi in maternal decidual vessels (star and black box). H&E staining, 20x magnification. The image inside the black box is magnified to 800x in Subfigure I. (J) The arrow shows one of the thrombi.
Fig 10 “EM35” mouse had early-onset preeclampsia-like symptoms associated with fetal growth restriction. (A) “EM35” mouse had a higher and earlier blood pressure peak than other hsFlt-1-e15a-treated mice, followed by the normalization of blood pressures and a second blood pressure peak on PPD1. (B) The urinary albumin/creatinine ratio on GD18 was higher in “EM35” mouse than in other hsFlt-1-e15a-treated mice. (C) HsFlt-1-e15a expression in the liver was lower in “EM35” mouse than in other hsFlt-1-e15a-treated mice. (D-E) Histopathological examination of the liver found a multiple cystic biliary hyperplasia in the liver (* in D), and multiple recent (D) and remote (E) infarcts in the parenchyma of liver. H&E staining, 200x magnifications. (F) The endothelial outgrowth area was the lowest in “EM35” mouse among hsFlt-1-e15a-treated mice. (G-H) Mean fetal (G) weights (0.749±0.029) and placental (H) weights (0.080±0.006) in “EM35” mouse were significantly lower than in other hsFlt-1-e15a-treated mice (fetal weight: 1.06±0.023, p = 2.017x10-15, placental weight: 0.111±0.004, p = 2.05x10-6). (I) Histopathological examination of a placenta from “EM35” mouse showed multiple thrombi in maternal decidual vessels (star and black box). H&E staining, 20x magnification. The image inside the black box is magnified to 800x in Subfigure I. (J) The arrow shows one of the thrombi.CC0 · PubMed Central
Fig 11 The possible mechanisms of distinct preeclampsia phenotypes induced by hsFlt-1-e15a. Blood pressure peaks (blue arrows) occurred after a ten-day-period following the first and second Ad-CMV-hsFlt-1-e15a injections in all but one mice that developed late-onset or postpartum preeclampsia without growth restriction. In these cases the growth potential of the fetuses was not compromised because they have already reached the top of their growth curve when hsFlt-1-e15a effect reached its plateau (blue stick crossing the curve). In case of “EM35” mouse, blood pressure elevations already peaked at six days after the first Ad-CMV-hsFlt-1-e15a injection (red arrow). In this case the fetal growth potential was severely compromised because the hsFlt-1-e15a effect was the highest when fetuses had reached only about one third or fourth of their growth curve (red stick crossing the curve). Preeclampsia phenotypes are described below the growth curve of CD-1 mouse embryos, which was adapted from a figure in a previous publication of Mu et al. [240]. Permission for reuse of this original figure was obtained from BioMed Central.
Fig 11 The possible mechanisms of distinct preeclampsia phenotypes induced by hsFlt-1-e15a. Blood pressure peaks (blue arrows) occurred after a ten-day-period following the first and second Ad-CMV-hsFlt-1-e15a injections in all but one mice that developed late-onset or postpartum preeclampsia without growth restriction. In these cases the growth potential of the fetuses was not compromised because they have already reached the top of their growth curve when hsFlt-1-e15a effect reached its plateau (blue stick crossing the curve). In case of “EM35” mouse, blood pressure elevations already peaked at six days after the first Ad-CMV-hsFlt-1-e15a injection (red arrow). In this case the fetal growth potential was severely compromised because the hsFlt-1-e15a effect was the highest when fetuses had reached only about one third or fourth of their growth curve (red stick crossing the curve). Preeclampsia phenotypes are described below the growth curve of CD-1 mouse embryos, which was adapted from a figure in a previous publication of Mu et al. [240]. Permission for reuse of this original figure was obtained from BioMed Central.CC0 · PubMed Central

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Authors
Szalai, G., Romero, R., Chaiworapongsa, T., Xu, Y., Wang, B., Ahn, H., Xu, Z., Chiang, P. J., Sundell, B., Wang, R., Jiang, Y., Plazyo, O., Olive, M., Tarca, A. L., Dong, Z., Qureshi, F., Papp, Z., Hassan, S. S., Hernandez-Andrade, E., & Than, N. G.
Affiliations
  • National Institutes of Health (US)
  • Wayne State University (US)
Corresponding author
Romero, Roberto
ORCID
Venue
PLoS ONE
Publisher
Public Library of Science (PLoS)
Published
2015-04-10
Volume
10
Issue
4
Pages
e0119547
ISSN
1932-6203 (ISSN-L)
DOI
10.1371/journal.pone.0119547
PMC
PMC4393117
OpenAlex
W1967129767
Cited by
46
References
292
Citations by year
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FWCI
4.55
Type
journal article
Language
en
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  • PLoS ONE (publishedVersion, public-domain)
  • PubMed Central (publishedVersion, public-domain)
  • PubMed (publishedVersion)
  • Repository of the Academy's Library (Library of the Hungarian Academy of Sciences) (acceptedVersion)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
  • Figshare (submittedVersion, cc-by)
  • Semmelweis University Repository (Semmelweis University) (submittedVersion)
Funders
  • U.S. Department of Health and Human Services (HHSN275201300006C)
  • Wayne State University (HHSN275201300006C)
  • Barbara Ann Karmanos Cancer Institute
  • National Institutes of Health (Contract No. HHSN275201300006C, HHSN 275201300006C)
  • U.S. Public Health Service (HHSN275201300006C)
  • National Cancer Institute (P30CA 22453, P30CA022453)
  • National Institute of Child Health and Human Development (HHSN275201300006C, Contract No. HHSN275201300006C)
  • Eunice Kennedy Shriver National Institute of Child Health and Human Development (HHSN275201300006C, Contract No. HHSN275201300006C.)
  • NHLBI Division of Intramural Research
Topics
  • Health Sciences › Medicine › Obstetrics and Gynecology › Pregnancy and preeclampsia studies
  • Health Sciences › Medicine › Clinical Neurology › Neurological Complications and Syndromes
  • Life Sciences › Immunology and Microbiology › Immunology › Reproductive System and Pregnancy
Keywords
preeclampsia, soluble fms-like tyrosine kinase-1, anti-angiogenic therapy, maternal hypertension, albuminuria, glomerular changes, postpartum monitoring, endothelial dysfunction, aortic ring assay
MeSH terms
Amino Acid Sequence, Animals, Blood Pressure, Disease Models, Animal, Female, Gestational Age, Humans, Mice, Transgenic, Molecular Sequence Data, Phenotype, Placenta, Pre-Eclampsia, Pregnancy, Recombinant Proteins, RNA, Messenger, Species Specificity, Telemetry, Ultrasonography, Blood Pressure Monitors, Sequence Homology, Amino Acid, Protein Isoforms, Gene Expression Profiling, Vascular Endothelial Growth Factor Receptor-1, Green Fluorescent Proteins, Mice

About these checks

Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.

Statistics and arithmetic are recomputed, not judged.

Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.

What counts as a claim ›How we measure confidence ›How we check this ›

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