Abstract
Objectives
An approach is presented for high-field MRI studies of the cardiovascular system (CVS) of a marine crustacean, the edible crab Cancer pagurus, submerged in highly conductive seawater.
Materials and methods
Structure and function of the CVS were investigated at 9.4 T. Cardiac motion was studied using self-gated CINE MRI. Imaging protocols and radio-frequency coil arrangements were tested for anatomical imaging. Haemolymph flow was quantified using phase-contrast angiography. Signal-to-noise-ratios and flow velocities in afferent and efferent branchial veins were compared with Student’s t test (n = 5).
Results
Seawater induced signal losses were dependent on imaging protocols and RF coil setup. Internal cardiac structures could be visualized with high spatial resolution within 8 min using a gradient-echo technique. Variations in haemolymph flow in different vessels could be determined over time. Maximum flow was similar within individual vessels and corresponded to literature values from Doppler measurements. Heart contractions were more pronounced in lateral and dorso-ventral directions than in the anterior–posterior direction.
Discussion
Choosing adequate imaging protocols in combination with a specific RF coil arrangement allows to monitor various parts of the crustacean CVS with exceptionally high spatial resolution despite the adverse effects of seawater at 9.4 T.
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Abbreviations
- AA:
-
Anterior aorta
- ALA:
-
Antero-lateral arteries
- BPV:
-
Branchiopericardial veins
- BV:
-
Branchial veins
- CVS:
-
Cardiovascular system
- (Fc)FLASH:
-
(Flow-compensated) fast low-angle shot
- HA:
-
Hepatic arteries
- MIP:
-
Maximum intensity projection
- MRI:
-
Magnetic resonance imaging
- NMR:
-
Nuclear magnetic resonance
- PA:
-
Posterior artery
- RARE:
-
Rapid acquisition with relaxation enhancement
- RF:
-
Radio frequency
- ROI:
-
Region of interest
- SA:
-
Sternal artery, arteria sternalis
- SD:
-
Standard deviation
- SNR:
-
Signal-to-noise ratio
- SUC:
-
(receive-only) surface RF coil
- TE:
-
Echo time
- TOF:
-
Time-of-flight
- TR:
-
Repetition time
References
Benveniste H, Blackband S (2002) MR microscopy and high resolution small animal MRI: applications in neuroscience research. Prog Neurobiol 67:393–420
Ahrens ET, Narasimhan PT, Nakada T, Jacobs RE (2002) Small animal neuroimaging using magnetic resonance microscopy. Prog Nucl Magn Reson Spectrosc. https://doi.org/10.1016/S0079-6565(02)00003-1
Uǧurbil K, Adriany G, Andersen P, Chen W, Garwood M, Gruetter R, Henry PG, Kim SG, Lieu H, Tkac I, Vaughan T, Van De Moortele PF, Yacoub E, Zhu XH (2003) Ultrahigh field magnetic resonance imaging and spectroscopy. Magn Reson Imaging 21:1263–1281
Vallée JP, Ivancevic MK, Nguyen D, Morel DR, Jaconi M (2004) Current status of cardiac MRI in small animals. Magn Reson Mater Phy 17:149–156
Tsui BMW, Kraitchman DL (2009) Recent advances in small-animal cardiovascular imaging. J Nucl Med 50:667–670
Herberholz J (2004) Anatomy of a live invertebrate revealed by manganese-enhanced magnetic resonance imaging. J Exp Biol 207:4543–4550
Brinkley CK, Kolodny NH, Kohler SJ, Sandeman DC, Beltz BS (2005) Magnetic resonance imaging at 9.4 T as a tool for studying neural anatomy in non-vertebrates. J Neurosci Methods 146:124–132
Kabli S, Alia A, Spaink HP, Verbeek FJ, De Groot HJM (2006) Magnetic resonance microscopy of the adult zebrafish. Zebrafish 3:431–439
Bock C, Wermter FC, Mintenbeck K (2017) MRI and MRS on preserved samples as a tool in fish ecology. Magn Reson Imaging 38:39–46
Köhnk S, Baudewig J, Brandis D, Boretius S (2017) What’s in this crab? MRI providing high-resolution three-dimensional insights into recent finds and historical collections of Brachyura. Zoology 121:1–9
Ziegler A, Kunth M, Mueller S, Bock C, Pohmann R, Schröder L, Faber C, Giribet G (2011) Application of magnetic resonance imaging in zoology. Zoomorphology 130:227–254
Van Der Linden A, Verhoye M, Pörtner H-O, Bock C (2004) The strengths of in vivo magnetic resonance imaging (MRI) to study environmental adaptational physiology in fish. Magn Reson Mater Phy 17:236–248
Wermter FC, Maus B, Pörtner H-O, Dreher W, Bock C (2018) CO2 induced pHi changes in the brain of polar fish: a TauCEST application. NMR Biomed. https://doi.org/10.1002/nbm.3955
Melzner F, Bock C, Pörtner H-O (2007) Coordination between ventilatory pressure oscillations and venous return in the cephalopod Sepia officinalis under control conditions, spontaneous exercise and recovery. J Comp Physiol B Biochem Syst Environ Physiol 177:1–17
Lannig G, Cherkasov AS, Pörtner H-O, Bock C, Sokolova IM (2008) Cadmium-dependent oxygen limitation affects temperature tolerance in eastern oysters (Crassostrea virginica Gmelin). AJP Regul Integr Comp Physiol 294:R1338–R1346
Sukhotin A, Fokina N, Ruokolainen T, Bock C, Pörtner H-O, Lannig G (2017) Does the membrane pacemaker theory of metabolism explain the size dependence of metabolic rate in marine mussels? J Exp Biol 220:1423–1434
Koth J, Maguire ML, McClymont D, Diffley L, Thornton VL, Beech J, Patient RK, Riley PR, Schneider JE (2017) High-resolution magnetic resonance imaging of the regenerating adult zebrafish heart. Sci Rep 7:1–12
Seo E, Ohishi K, Maruyama T, Imaizumi-Ohashi Y, Murakami M, Seo Y (2014) Testing the constant-volume hypothesis by magnetic resonance imaging of Mytilus galloprovincialis heart. J Exp Biol 217:964–973
Seo E, Sazi T, Togawa M, Nagata O, Murakami M, Kojima S, Seo Y (2016) A portable infrared photoplethysmograph: heartbeat of Mytilus galloprovincialis analyzed by MRI and application to Bathymodiolus septemdierum. Biol Open 5:1752–1757
Bock C, Frederich M, Wittig R-M, Pörtner H-O (2001) Simultaneous observations of haemolymph flow and ventilation in marine spider crabs at different temperatures: a flow weighted MRI study. Magn Reson Imaging 19:1113–1124
Maus B, Bock C, Pörtner H-O (2018) Water bicarbonate modulates the response of the shore crab Carcinus maenas to ocean acidification. J Comp Physiol B Biochem Syst Environ Physiol 188:749–764
McMahon BR, Wilkens JL (1977) Periodic respiratory and circulatory performance in the red rock crab Cancer productus. J Exp Zool 202:363–374
Bock C, Dogan F, Pörtner H-O (2012) Coping with a changing ocean: real-time cardiac MRI on an animal model with a natural cardiovascular disorder. Proc Int Soc Magn Reson Med Sci Meet Exhib 20:5021
McMahon BR, Burnett LE (1990) The crustacean open circulatory system: a reexamination. Physiol Zool 63:35–71
Frederich M, DeWachter B, Sartoris F-J, Pörtner H-O (2000) Cold tolerance and the regulation of cardiac performance and hemolymph distribution in maja squinado (Crustacea: Decapoda). Physiol Biochem Zool 73:406–415
Frederich M, Pörtner H-O (2000) Oxygen limitation of thermal tolerance defined by cardiac and ventilatory performance in spider crab, Maja squinado. Am J Physiol Regul Integr Comp Physiol 279:R1531–R1538
Giomi F, Pörtner H-O (2013) A role for haemolymph oxygen capacity in heat tolerance of eurythermal crabs. Front Physiol 4:1–12
De Wachter B, McMahon BR (1996) Temperature effects on heart performance and regional hemolymph flow in the crab Cancer magister. Comp Biochem Physiol A Physiol 114:27–33
Bradford SM, Taylor AC (1982) The respiration of Cancer pagurus under normoxic and hypoxic conditions. J Exp Biol 97:273–288
Walther K, Sartoris F-J, Bock C, Pörtner H-O (2009) Impact of anthropogenic ocean acidification on thermal tolerance of the spider crab Hyas araneus. Biogeosciences 6:2207–2215
Taylor AC (1977) The respiratory responses of Carcinus maenas (L.) to changes in environmental salinity. J Exp Mar Bio Ecol 29:197–210
McGaw IJ, Reiber CL (2002) Cardiovascular system of the blue crab Callinectes sapidus. J Morphol 251:1–21
Airriess CN, McMahon BR, McGaw IJ, Bourne GB (1994) Application and in situ calibration of a pulsed-doppler flowmeter for blood flow measurement in crustaceans. J Mar Biol Assoc UK 74:455–458
Reiber CL, McMahon BR, Burggren WW (1997) Cardiovascular functions in two macruran decapod crustaceans (Procambarus clarkii and Homarus americanus) during periods of inactivity, tail flexion and cardiorespiratory pauses. J Exp Biol 200:1103–1113
Bock C, Sartoris F-J, Wittig R-M, Pörtner H-O (2001) Temperature-dependent pH regulation in stenothermal Antarctic and eurythermal temperate eelpout (Zoarcidae): an in-vivo NMR study. Polar Biol 24:869–874
Pope JM, Yao S (1993) Quantitative NMR imaging of flow. Concepts Magn Reson 5:281–302
Roberts L, Cheesman S, Elliott M, Breithaupt T (2016) Sensitivity of Pagurus bernhardus (L.) to substrate-borne vibration and anthropogenic noise. J Exp Mar Bio Ecol 474:185–194
Florey E, Kriebel ME (1974) The effects of temperature, anoxia and sensory stimulation on the heart rate of unrestrained crabs. Comp Biochem Physiol A 48:285–300
McDonald DG, McMahon BR, Wood CM (1977) Patterns of heart and scaphognathite activity in the crab Cancer magister. J Exp Zool 202:33–43
McMahon BR, McDonald DG, Wood CM (1979) Ventilation, oxygen uptake and haemolymph oxygen transport, following enforced exhausting activity in the Dungeness crab Cancer magister. J Exp Biol 80:271–285
Wilkens JL, Wilkens LA, McMahon BR (1974) Central control of cardiac and scaphognathite pacemakers in the crab, Cancer magister. J Comp Physiol 90:89–104
Brouwer M, Engel DW, Bonaventura J, Johnson GA (1992) In vivo magnetic resonance imaging of the blue crab, Callinectes sapidus: effect of cadmium accumulation in tissues on proton relaxation properties. J Exp Zool 263:32–40
Libove JM, Singer JR (1980) Resolution and signal-to-noise relationships in NMR imaging in the human body. J Phys E Sci Instrum 13:38–44
Beck BL, Jenkins K, Caserta J, Padgett K, Fitzsimmons J, Blackband SJ (2004) Observation of significant signal voids in images of large biological samples at 11.1 T. Magn Reson Med 51:1103–1107
Hartung MP, Grist TM, François CJ (2011) Magnetic resonance angiography: current status and future directions. J Cardiovasc Magn Reson 13:1–11
Wheaton AJ, Miyazaki M (2012) Non-contrast enhanced MR angiography: physical principles. J Magn Reson Imaging 36:286–304
McGaw IJ, Airriess CN, McMahon BR (1994) Patterns of haemolymph-flow variation in decapod crustaceans. Mar Biol 121:53–60
Bock C, Sartoris F-J, Pörtner H-O (2002) In vivo MR spectroscopy and MR imaging on non-anaesthetized marine fish: techniques and first results. Magn Reson Imaging 20:165–172
Airriess CN, McMahon BR (1994) Cardiovascular adaptations enhance tolerance of environmental hypoxia in the crab cancer magister. J Exp Biol 190:23–41
Taylor EW (1982) Control and co-ordination of ventilation and circulation in crustaceans: responses to hypoxia and exercise. J Exp Biol 100:289–319
Maceira AM, Prasad SK, Khan M, Pennell DJ (2006) Normalized left ventricular systolic and diastolic function by steady state free precession cardiovascular magnetic resonance. J Cardiovasc Magn Reson 8:417–426
Chuang ML, Hibberd MG, Salton CJ, Beaudin RA, Riley MF, Parker RA, Douglas PS, Manning WJ (2000) Importance of imaging method over imaging modality in noninvasive determination of left ventricular volumes and ejection fraction: assessment by two- and three-dimensional echocardiography and magnetic resonance imaging. J Am Coll Cardiol 35:477–484
Walsh TF, Hundley WG (2007) Assessment of ventricular function with cardiovascular magnetic resonance. Cardiol Clin 25:15–33
Acknowledgements
The authors thank Fredy Veliz Moraleda for assistance in animal care, Rolf Wittig, Felizitas Wermter and Sebastian Gutsfeld for assistance during in vivo experiments and data evaluation. Further, thanks to the AWI workshop, in particular Erich Dunker, as well as Sven Junge and Martin Tabbert from the Bruker BioSpin coil development group (Ettlingen, Germany).
Funding
The study is a contribution to the PACES II research program (WP 1.6) of the Alfred-Wegener-Institute, funded by the Helmholtz Association.
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Authors and Affiliations
Contributions
BM and CB conceived the study design and experimental setup. BM performed the experiments with assistance from CB. BM analyzed the data. BM and CB interpreted the data. BM, HOP and CB drafted and approved of the final version of the manuscript.
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The authors declare that they have no conflict of interest.
Ethical approval
All applicable international, national and institutional guidelines for the care and use of animals were followed. All procedures performed in studies involving animals were in accordance with the ethical standards of the institution at which the studies were conducted.
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10334_2019_752_MOESM1_ESM.pdf
Spin-echo MRI of the heart region of C. pagurus. Image was recorded with a RARE (rapid acquisition with relaxation enhancement) technique, using the resonator in transmit-receive mode. TE = 85 ms; TR = 4500 ms; 4 averages, RARE factor = 8; bandwidth = 50000 Hz, 512×384 pixels; FOV = 100×75 mm²; slice thickness = 1.0 mm. Compared to the gradient-echo scans, signal losses in the center of the image reduce the visibility of the inner structures. Scale bar = 2 cm (PDF 533 kb)
10334_2019_752_MOESM2_ESM.mp4
3D MIP reconstruction of the cardiovascular system of Cancer pagurus based on coronal multi-slice TOF angiography scans. The scans and their parameters are identical to those in fig. 4a-c. Total volume = 100×100×49 mm³ (l×w×h) (MP4 29288 kb)
10334_2019_752_MOESM3_ESM.mp4
3D MIP reconstruction of the cardiovascular system of Cancer pagurus based on axial multi-slice TOF angiography scans. The scans and their parameters are identical to those in fig. 4d-f. Total volume = 100×99×50 mm³ (l×w×h) (MP4 29783 kb)
10334_2019_752_MOESM4_ESM.mp4
3D MIP reconstruction of the cardiovascular system of Cancer pagurus based on sagittal multi-slice TOF angiography scans. The scans and their parameters are identical to those in fig. 4g-h. Total volume = 80×103.2×60 mm³ (l×w×h) (MP4 571 kb)
10334_2019_752_MOESM5_ESM.mp4
IntraGate© movie for the contraction of the heart in coronal orientation. For scan parameters, see tab. 1. One cardiac cycle was split into 10 frames, each lasting 100 ms in the movie. The real duration of the reconstructed cardiac cycle is 666 ms at a heart rate of 90 bpm. End-systolic and end-diastolic frames can be seen in fig. 7a-b (MP4 526 kb)
10334_2019_752_MOESM6_ESM.mp4
IntraGate© movie for the contraction of the heart in axial orientation. For scan parameters, see tab. 1. One cardiac cycle was split into 10 frames, each lasting 100 ms in the movie. The real duration of the reconstructed cardiac cycle is 666 ms at a heart rate of 90 bpm. End-systolic and end-diastolic frames can be seen in fig. 7c-d (MP4 617 kb)
10334_2019_752_MOESM7_ESM.mp4
IntraGate© movie for the contraction of the heart in sagittal orientation. For scan parameters, see tab. 1. One cardiac cycle was split into 10 frames, each lasting 100 ms in the movie. The real duration of the reconstructed cardiac cycle is 750 ms at a heart rate of 80 bpm. End-systolic and end-diastolic frames can be seen in fig. 7e-f (MP4 29808 kb)
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Maus, B., Pörtner, HO. & Bock, C. Studying the cardiovascular system of a marine crustacean with magnetic resonance imaging at 9.4 T. Magn Reson Mater Phy 32, 567–579 (2019). https://doi.org/10.1007/s10334-019-00752-4
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DOI: https://doi.org/10.1007/s10334-019-00752-4