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Developmental Biology

High Frequency Ultrasound for the Analysis of Fetal and Placental Development In Vivo

Published: November 8th, 2018



1Experimental Obstetrics and Gynecology, Medical Faculty, Otto-von-Guericke University, 2Institute of Molecular and Clinical Immunology, Medical Faculty, Otto-von-Guericke University

Here we describe the technique of high frequency ultrasound for in vivo analysis of fetuses in mice. This method allows the follow-up of fetuses and the analysis of placental parameters as well as maternal and fetal blood flow throughout pregnancy.

Ultrasound imaging is a widespread method used to detect organ anomalies and tumors in human and animal tissues. The method is non-invasive, harmless, and painless, and the application is easy, fast, and can be done anywhere, even with mobile devices. During pregnancy, ultrasound imaging is standardly used to closely monitor fetal development. The technique is important to assess intrauterine growth restriction (IUGR), a pregnancy complication with short- and long-term health consequences for both the mother and fetus. Understanding the process of IUGR is indispensable for developing effective therapeutic strategies.

The ultrasound system used in this manuscript is an ultrasound device produced for the analysis of small animals and can be used in various research fields, including pregnancy research. Here we describe the usage of the system for in vivo analysis of fetuses from natural killer (NK) cell/mast cell (MC)-deficient mothers that give birth to growth-restricted pups. The protocol includes preparation of the system, handling of the mice before and during measurements, and the usage of the B-mode, color doppler mode, and pulse-wave doppler mode. Fetal size, placental size, and blood supply to the fetus were analyzed. We found reduced implantation sizes and smaller placentas in NK/MC-deficient mice from mid-gestation onwards. In addition, MC/NK-deficiency was associated with absent and reversed end diastolic flow in the fetal Arteria umbilicalis(UmA) and an elevated resistance index. The methods described in the protocol can easily be used for related and non-related research topics.

Ultrasound is sound waves with frequencies above the audible range of the human ear, higher than about 20 kHz1. Animals like bats, wales, dolphins2,3, mice4, rats5, and mouse lemurs6 all use ultrasound for orientation or communication. Humans take advantage of ultrasound for several technical and medical applications. An ultrasound device is able to create the sound wave and distribute and represent the signal. If ultrasound encounters an obstacle, the sound is reflected, absorbed or can go through it. The applicati....

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All methods described here have been approved by the “Landesverwaltungsamt Sachsen Anhalt: 42502-2-1296UniMD.”

1. Experimental Procedure

  1. Mate 6 to 8-week-old female MC-deficient C57BL/6J-Cpa3Cre/+ (Cpa3Cre/+) mice and MC-sufficient C57BL/6J-Cpa3+/+ (colony controls; Cpa3+/+) with BALB/c males.
  2. Define the gestation day (gd) 0 after confirmation of the v.......

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Individual components of the ultrasound system used in this manuscript are shown in Figure 1. Figure 2 shows representative ultrasound images acquired in B-mode at gd5, 8, 10, and 12 (B) and corresponding implantation area measurement results (A), demonstrating a significant reduced implantation area of anti-CD122-treated Cpa3Cre/+ mice from gd10 onwards.

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Using our ultrasound system, we demonstrated fetal growth restriction in MC/NK-deficient mothers from gd10 on. Furthermore, at gd10 and 12, we observed reduced placental dimensions, and at gd14 the absence or reversion of end diastolic flow in the UmAs of some fetuses of uMC/uNK-deficient mice. This sign of poor vascularization was associated with a significant resistance index of the arteries indicating IUGR. Results confirm the important role of uMCs and uNKs in pregnancy and fetal well-being and in understanding the c.......

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Many thanks to the Imaging Instrument company (especially to Magdalena Steiner, Katrin Suppelt, and Sandra Meyer) for their pleasant and fast support and for answering all our questions concerning the Imaging System and its usage promptly and completely. We are grateful to Prof. Hans-Reimer Rodewald and Dr. Thorsten Feyerabend (DKFZ Heidelberg, Germany) for providing the Cpa3 colony. Additionally, we thank Stefanie Langwisch, who was in charge of the mouse colonies and who generated the pictures in Figure 1.

The work and the Imaging System were funded by grants from the Deutsche Forschungsgemeinschaft (DFG) to A.C.Z. (ZE526/6-1 and AZ526/6-....

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Name Company Catalog Number Comments
LEAF anti-Maus CD122 (IL-2Rb) BioLegend 123204 Klon TM-β1; 500 µg
Vevo 2100 System  FujiFilm VisualSonics Inc. Transducer MS550D-0421
Vevo LAB Software  FujiFilm VisualSonics Inc.
Isoflurane Baxter PZN: 6497131
Electrode gel Parker 12_8
Surgical tape 3M Transpore 1527-1
Eye cream Bayer PZN: 1578675
Cotton tipped applicators Raucotupf 11969 100 pieces
Depilatory cream Reckitt Benckiser 2077626
Compresses Nobamed Paul Danz AG 856110 10 x 10 cm
Ultrasound gel Gello GmbH 246000

  1. Abramowicz, J. S., Kremkau, F. W., Merz, E. Ultraschall in der Geburtshilfe: Kann der Fötus die Ultraschallwelle hören und die Hitze spüren?. Ultraschall in der Medizin. 33 (3), 215-217 (1980).
  2. Jones, G. Echolocation. Current Biology. 15 (13), R484-R488 (2005).
  3. Simmons, J. A. The sonar receiver of the bat. Annals of the New York Academy of Sciences. 188, 161-174 (1971).
  4. Zala, S. M., Reitschmidt, D., Noll, A., Balazs, P., Penn, D. J. Sex-dependent modulation of ultrasonic vocalizations in house mice (Mus musculus musculus). Public Library of Science ONE. 12 (12), e0188647 (2017).
  5. Wöhr, M., Seffer, D., Schwarting, R. K. W. Studying Socio-Affective Communication in Rats through Playback of Ultrasonic Vocalizations. Current Protocols in Neuroscience. 75, 1-8 (2016).
  6. Hasiniaina, A. F., et al. High frequency/ultrasonic communication in a critically endangered nocturnal primate, Claire's mouse lemur (Microcebus mamiratra). American Journal of Primatology. , e22866 (2018).
  7. Yeo, L., Romero, R. Color and power Doppler combined with Fetal Intelligent Navigation Echocardiography (FINE) to evaluate the fetal heart. Ultrasound in Obstetrics & Gynecology. 50 (4), 476-491 (2017).
  8. Teichholz, L. E. Echocardiography in valvular heart disease. Progress in Cardiovascular Diseases. 17 (4), 283-302 (1975).
  9. Zechner, P. M., et al. Lungensonographie in der Akut- und Intensivmedizin. Der Anaesthesist. 61 (7), 608-617 (2012).
  10. Blank, W., Schuler, A. Sonografie der Schilddrüse - Update 2017. Praxis. 106 (12), 631-640 (2017).
  11. Hansen, K. L., Nielsen, M. B., Ewertsen, C. Ultrasonography of the Kidney: A Pictorial Review. Diagnostics. 6 (1), (2015).
  12. Older, R. A., Watson, L. R. Ultrasound anatomy of the normal male reproductive tract. Journal of Clinical Ultrasound. 24 (8), 389-404 (1996).
  13. Reeves, J. J., Rantanen, N. W., Hauser, M. Transrectal real-time ultrasound scanning of the cow reproductive tract. Theriogenology. 21 (3), 485-494 (1984).
  14. Sharma, M., Somani, P., Sunkara, T. Imaging of gall bladder by endoscopic ultrasound. World Journal of Gastrointestinal Endoscopy. 10 (1), 10-15 (2018).
  15. Weskott, H. -. P. Ultraschall in der Diagnostik maligner Lymphome. Der Radiologe. 52 (4), 347-359 (2012).
  16. Shirinifard, A., Thiagarajan, S., Johnson, M. D., Calabrese, C., Sablauer, A. Measuring Absolute Blood Perfusion in Mice Using Dynamic Contrast-Enhanced Ultrasound. Ultrasound in Medicine & Biology. 43 (8), 1628-1638 (2017).
  17. Quaia, E. Assessment of tissue perfusion by contrast-enhanced ultrasound. European Radiology. 21 (3), 604-615 (2011).
  18. Saw, S. N., Poh, Y. W., Chia, D., Biswas, A., Zaini Mattar, C. N., Yap, C. H. Characterization of the hemodynamic wall shear stresses in human umbilical vessels from normal and intrauterine growth restricted pregnancies. Biomechanics and Modeling in Mechanobiology. , (2018).
  19. Kessler, J., Rasmussen, S., Godfrey, K., Hanson, M., Kiserud, T. Fetal growth restriction is associated with prioritization of umbilical blood flow to the left hepatic lobe at the expense of the right lobe. Pediatric Research. 66 (1), 113-117 (2009).
  20. Laurin, J., Lingman, G., Marsál, K., Persson, P. H. Fetal blood flow in pregnancies complicated by intrauterine growth retardation. Obstetrics and Gynecology. 69 (6), 895-902 (1987).
  21. Arduini, D., Rizzo, G., Romanini, C., Mancuso, S. Fetal blood flow velocity waveforms as predictors of growth retardation. Obstetrics and Gynecology. 70 (1), 7-10 (1987).
  22. Meyer, N., et al. Chymase-producing cells of the innate immune system are required for decidual vascular remodeling and fetal growth. Scientific Reports. 7, 45106 (2017).
  23. Meyer, N., Schüler, T., Zenclussen, A. C. Simultaneous Ablation of Uterine Natural Killer Cells and Uterine Mast Cells in Mice Leads to Poor Vascularization and Abnormal Doppler Measurements That Compromise Fetal Well-being. Frontiers in Immunology. 8, 1913 (2017).
  24. Evans, D. H., Jensen, J. A., Nielsen, M. B. Ultrasonic color Doppler imaging. Interface Focus. 1 (4), 490-502 (2011).

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