Experimental Physiology

Physiological Society Symposium - Vagal Control: From Axolotl to Man The crocodilian heart; more controlled than we thought?
Michael Axelsson *
Department of Zoology, Goteborg University, PO Box 463, SE-40530 Goteborg, Sweden
CONTENTS
PAGE

Introduction The subpulmonary conus The foramen of Panizza The aortic anastomosis Future directions References Experimental Physiology (200 1) 86.6,785-789.

785 786
787

788

788 788

Introduction There are large differences in the morphology of the vertebrate heart; from the fish heart with its single atrium and single ventricle to the crocodilian, bird and mammalian hearts with two fully separated atria and ventricles (van Mierop & Kutsche, 1985). It is only in crocodilians, birds and mammals where the heart has a complete interventricular septum, that a full intracardiac separation of blood pressure and flow in the systemic and pulmonary circulations can occur. In birds and mammals the left ventricle gives rise to the aorta supplying the body with oxygenated blood and the pulmonary arteries arise from the right ventricle carrying deoxygenated blood to the lungs. In these two animal groups no intra- or extra-cardiac mixing of blood (shunting) is possible, and no shunting of blood between the pulmonary and systemic circulations occurs in healthy adults. The crocodilians are unique in comparison to other reptiles and also to birds and mammals. In comparison to other reptiles (snakes, lizards and turtles) the crocodilian heart is unique in that the ventricle is fully divided into a left and a right ventricle whereas the non-crocodilian reptile heart is subdivided into three intraventricular compartments that are interconnected (no morphological subdivision of the ventricle) allowing intracardiac mixing of oxygenated and deoxygenated blood. The crocodilian heart is also unique compared to the bird and mammalian heart in that shunting of blood away from the pulmonary circulation is still possible as the right ventricle gives rise not only to the pulmonary

arteries but also to the left aorta (Fig. 1). This allows deoxygenated blood from the right ventricle to bypass the lungs and to be recirculated into the systemic circulation (pulmonary-to-systemic shunt). Apart from the extra left aorta from the right ventricle, three other morphological features of the crocodilian cardiovascular system have been the focus of discussion over recent years. (1) The foramen of Panizza; an opening between the right and left aorta situated in the common wall of the left and right aorta (Fig. 1A). (2) The subpulmonary conus situated in the pulmonary outflow tract of the right ventricle (Fig. 1B). (3) The aortic anastomosis that connects the two aortic arches just posterior to the heart (Fig. 1C; van Mierop & Kutsche, 1985). The subpulmonary conus contains connective tissue nodules protruding into the outflow tract that acts as an extra and unique valve mechanism (Fig. 1B). These three areas have received much attention from comparative physiologists interested in the function of the crocodiliadreptile circulation, and the evolution of the cardiovascular system and its regulation. There is increasing evidence for a close regulation of the three unique structures in the crocodilian cardiovascular system and their importance for the normal function of the crocodilian circulation. This short overview will give a summary of the latest findings. Due to the anatomical arrangement of the heart and major vessels, the mean blood pressure in the right and left aortas are equal and usually higher than the pulmonary pressure (Axelsson et al. 1989; Jones & Shelton, 1993). For a

Presented at the Oxford Meeting of the Physiological Society in March 2001. Publication of The Physiological Society
2293

* Email: m.axelsson@zool.gu.se

Downloaded from Exp Physiol (ep.physoc.org) by guest on April 14, 2010

786

M. Axelsson

Exp. Physiol. 86.6

pulmonary-to-systemic shunt to develop the pressure in the right ventricle must exceed the pressure in the left aorta. Numerous mechanisms for the initiation and maintenance of the pulmonary-to-systemic shunt have been described: (I) increasing the end-diastolic volume of the right ventricle (Starling effect; Franklin & Axelsson, 1994; (2) increasing the pulmonary circuit resistance thus increasing right intraventricular pressure (White, 1969; Jones & Shelton, 1993); (3) decreasing the systemic vascular resistance (Jones & Shelton, 1993).

The subpulmonary conus The physiological significance of the subpulmonary conus in the right ventricle of the crocodilian heart has been debated for many years (for discussion see Jones, 1995, 1996; Axelsson & Franklin, 1997). It is electrically separated from the rest of the right ventricle and contains connective tissue nodules that have been called cog-teeth (Grigg, 1989; Fig. 1). These nodules project from the subpulmonary conus

wall into the pulmonary outflow tract just proximal to the pulmonary leaflet-like valves. Evidence for a role of the subpulmonary conus in initiating and regulating pulmonaryto-systemic shunts in crocodilians comes from pressure recordings from the right ventricle and pulmonary artery both in anaesthetised (Jones & Shelton, 1993) and unanaesthetised animals (Axelsson et al. 1996). In both the alligator (Alligator mississippiensis) and saltwater crocodile (Crocodylus porosus) a biphasic pressure development in the right ventricle has been found (Grigg & Johansen, 1987; Jones & Shelton, 1993; Axelsson et al. 1996). The secondary pressure peak in the right ventricle is only possible if there is an increase in resistance in the pulmonary outflow tract, and this cannot be attributed to the normal leaflet-likevalves but has to be a function of the subpulmonary conus with the extracog-teeth valves. It is interesting that the same type of pressure pattern is seen in the left ventricle and aorta of humans diagnosed with left ventricular hypertrophic cardiomyopathy (Murgo et al. 1980). The most accepted

Figure 1 Schematic representation of the crocodilian heart, outflow tract and major arteries; arrows indicate blood flow pattern during non-shunting conditions. Blood is ejected from the left ventricle (LV) into the right aorta (RAo), right subclavian artery (RS) and the common carotid artery (CCA). The right ventricle (RV) ejects blood into the common pulmonary trunk that divides into the left and right pulmonary artery (LPA and RPA). During a pulmonary-to-systemic shunt, blood is also ejected from the right ventricle into the left aorta (LAO)that continues to the gut as the coeliac artery. The right and left aortas communicate through the foramen of Panizza located at the base of the aortas just outside the bicuspid semilunar valves ( A ) and posterior to the heart via the aortic anastomosis (B). The cogteeth valves are found in the subpulmonary conus just proximal to the bicuspid semilunar valves (C).

Downloaded from Exp Physiol (ep.physoc.org) by guest on April 14, 2010

Exp. Pliysiol. 86.6

Control of the crocodilian heart

787

explanation for the ventricular-aortic pressure gradients in these patients is a mechanical obstruction to left ventricular outflow. In humans this is a pathological situation, but in crocodilians it is an effect of the cog-teeth valves in the right ventricular wall and represents a unique possibility for regulating the resistance of the pulmonary outflow tract. Further support for this was found in a study using an intracardiac angioscope to look at the various valves in the beating heart of the Cuban crocodile (Cvocodylus rhomhfer). It was shown that the individual tissue nodules (cog-teeth valves) fitted snugly together and, during systole, reduced the diameter of the pulmonary outflow tract (Axelsson et al. 1996). This could explain the increase in the resistance in the subpulmonary conus and the development of the secondary pressure peak in the right ventricle. In a more recent study it was shown that the subpulmonary conus is also regulated by p-adrenoceptors in such a way that when the p-adrenergic tone on the heart is low the cogteeth valves close more during systole thereby increasing the resistance between the right ventricle and the pulmonary circuit, initiating a pulmonary-to-systemic shunt. When the p-adrenergic tone on the heart is high the cog-teeth valves stay fully open and the crocodilian circulation works in the same way as aviadmammalian hearts without any shunting of blood between the pulmonary and systemic circuits (Fig. 2 4 Franklin & Axelsson, 2000). This fits with the results from long term recordings of flow in unanaesthetised animals where during stress or exercise no shunts can be observed, while at rest the pulmonary-to-systemic shunts are operational up to 85% of the time (Jones, 1996). Of the three mechanisms listed above that can initiate and maintain a pulmonary-to-systemic shunt, the subpulmonary conus fulfils the criteria for mechanisms 1 and 2. By increasing the pulmonary circuit resistance the shunt is initiated (mechanism 2), when a shunt is initiated more blood returns to the right side of the heart leading to the Starling effect that in itself can initiate/maintain a shunt (mechanism 1). The subpulinonary conus is obviously an important site for the control of pulmonary blood flow in crocodilians, but are the functions of the foramen of Panizza and the aortic anastomosis linked to that of the subpulmonaryconus and the capacity of the crocodilian heart to shunt blood away from the pulmonary circulation or do they have other functions?
The foramen of Panizza The foramen of Panizza was first described by the Italian anatomist B. Panizza in 1833 (Panizza, 1833). It is found deep within the pockets formed by the aortic valves and has been described as an insignificant aperture in the common wall of the right and left aortic arch. In an angioscopic study of the beating heart of the Cuban crocodile (Crocodylus rhombfer) it was shown that at physiological pressures the foramen of Panizza is a substantial opening (around 3WO% of the diameter of the right aorta) between the two aortic arches (Fig. 1.4; Axelsson et al. 1996). The mean blood pressure in the two aortic arches is equal, but the phasic pressures and blood flow profiles are more complicated. Under nonshunting conditions there is a multiphasic profile of pressure

and flow in the left aortic arch, which is due to the fact that during systole the medial cusp of the right aorta covers the foramen completely while in diastole when the valves are closed the foramen is uncovered and there is a connection between the two arches (Axelsson et al. 1989; Shelton & Jones, 1991; Malvin et al. 1995). This is further complicated by the fact that there is another connection between the two aortic arches further down in the abdomen called the aortic anastomosis (Fig. 1). During non-shunting conditions there is a small net blood flow from the right aorta into the left via the foramen of Panizza (Axelsson et al. 1989; Shelton & Jones, 1991). In a few studies it was noted that the pressure profile of the right and left aorta was superimposable during the entire

Adr, 0.2 ml

1 Adr, 10-7M Adr, 0

A
M VIP, lo-*M

i2

"

1 min

Adr 10-6M L-NAME 1 e 4 M

Figure 2 A , the effects of a bolus injections of adrenaline on the pulmonary outflow resistance (Rp,,,)is shown. Recordings are taken from a double perfused crocodile heart. Adrenaline decreases the pulmonary outflow tract resistance leading to an avianlmammalian type of circulation with no pulmonary-to-systemic shunting of blood. B, the effects of adrenaline and vasoactive intestinal polypeptide (VIP) on the foramen of Panizza. Note that VIP relaxes the adrenaline pre-contracted preparations. C, the effects of adrenaline and the nitric oxide inhibitor L-NAME on the isolated anastomosis ring preparations. Note the antagonistic action of adrenaline and nitric oxide as indicated by the oscillating tone in the preparation after addition of adrenaline.

Downloaded from Exp Physiol (ep.physoc.org) by guest on April 14, 2010

788

M. Axelsson

Exp. Physiol. 86.6

cardiac cycle, indicating that the foramen of Panizza was open during the entire cardiac cycle (White, 1956,1969;Grigg & Johanssen, 1987). Grigg & Johansen (1987) suggested these variable pressure and flow patterns seen in the left aorta, even in the same animals at different times, could be a consequence of a change in diameter of the foramen of Panizza. This may be important during pulmonary-to-systemicshunts when less blood is ejected from the left ventricle; they also suggested that during shunting a reversed foramen flow could occur with blood flowing from the left into the right aorta. Morphologically the left aortic valves do not reach the foramen of Panizza as is the case with the medial cusp of the right aortic valves, and therefore there is no obstruction to flow from the left to the right aorta during any part of the cardiac cycle (Axelsson et al. 1996). The variable foramen hypothesis was debated for many years but more recently two studies have presented evidence in favour of this hypotheis. In the study by Karila and coworkers it was shown that the foramen of Panizza and the surrounding tissue contained smooth muscle cells and a number of potential neurotransmitters were also identified including adrenaline and vasoactive intestinal polypeptide (VIP) (Karila et a . 1995). In f a more recent study by Axelsson & Franklin (2001) the effects of the identified substances were tested and it was shown that adrenaline produced a reduction in the diameter of the foramen of Panizza while VIP caused a relaxation of the adrenaline-induced contraction of the preparations (Fig. 2B). This is the first evidence in support of the variable foramen hypothesis proposed by Grigg & Johansen (1987).
The aortic anastomosis The last of the three unique structures in the crocodilian cardiovascular system to be discussed in this short overview is the aortic anastomosis, a short muscular connection between the right and left aorta just posterior to the heart (Fig. 1C). In contrast to the view of Webb (1979), Shelton & Jones(l991) pointed out that the aortic anastomosis is a substantial connection between the two aortas and that it might be of importance for the control of circulatory function in crocodilians. In the non-shunting condition, blood flow in the aortic anastomosis is from the right to the left aorta, and from there to the gastrointestinal canal since after the anastomosis the left aorta becomes the coeliac artery (Fig. I). Direct measurements of blood flow in the anastomosis have shown that it varies spontaneously over time indicating a possible regulatory function for the anastomosis (Axelsson et af. 1997). In an immunohistological study by Karila and coworkers (Karila et al. 1995) a number of potential regulatory substances were identified and it was shown that both Substance P (SP) and Neuropeptide Y (NPY) increase the flow through the anastomosis. In a recent study by Axelsson and coworkers it was shown that adrenaline and nitric oxide had profound effects on the isolated anastomosis (Fig. 2C; Axelsson et al. 2001). Adrenaline induced contraction of the anastomosis but this effect was counteracted by the release of nitric oxide resulting in a cyclic variation in the wall tension during exposure to adrenaline; a basal nitric oxide-induced tone on the anastomosis was also found. The aortic anastomosis

with its unusually thick media and adventitia resembles a sphincter similar to the sphincters found in amphibians arid lungfish (Saint Aubain & Wingstrand, 1976; Fishman et al. 1985). The significance of this muscular connection is still unclear but experimentally it has been shown that if the anastomosis is closed, a pulmonary-to-systemic shunt develops (Axelsson et al. 1997). When the blood flow through the anastomosis ceases or is reduced the small net blood flow into the left aorta via the foramen of Panizza is not enough to supply the gastrointestinal canal and therefore the pressure in the left aorta falls below the level of the right intraventricular pressure and this initiates a pulmonary-to-systemic shunt (mechanism 3 above). Another function of the aortic anastomosis might be to prevent back-flow of blood into the right aorta during periods of shunting, to maintain pressure in the left aortic arch for a reversed foramen flow (Axelsson & Franklin, 1997).
Future directions The functional significance of the actively regulated intracardiac cog-teeth valves, the foramen of Panizza with its variable diameter, and the thick sphincter-like aortic anastomosis is still unclear but together with other specialisations, such as a modified haemoglobin (Bauer & Jelkman, 1977; Grigg & Gruca, 1979; Bauer et a . 1981) these f three structures may increase the capacity for prolonged diving in resting crocodilians. It is clear from the published data that the answers to the final questions about the function of this unique circulatory system are not to be found in the laboratory environment but in the field, and that studies should be carried out in a multidisciplinary way since there might not be a single answer.

AXELSSON, & FRANKLIN, E. (1997). From anatomy to M. C. angioscopy: 164 years of crocodilian cardiovascular research, recent advances and speculations. Comparative Biochemistry and Physiology A 118, 51-62. AXELSSON, & FRANKLIN, E. (2001). The calibre of the M. C. foramen of Panizza in Crocodylus porosus is variable and under f adrenergic control. Journal o Comparative Physiology B 171, 341-346. AXELSSON, FRANKLIN, FRITSCHE, GRIGG, & NILSSON, M., C., R., G. S. (1997). The sub-pulmonary conus and the arterial anastomosis as important sites of cardiovascular regulation in the crocodile Crocodylus porosus. Journal of Experimental Biology 200,807-8 14. AXELSSON, FRANKLIN, E., LOFMAN, O., NILSSON, & M., C. C. S. GRIGG,G. C. (1996). Dynamic anatomical study of cardiac shunting in crocodiles using high resolution angioscopy. Journal o j Experimental Biology 199, 359-365. AXELSSON, HOLM, & NILSSON, (1989). Flow dynamics of M., S. S. the crocodile heart. American Journal of Physiology 256, R875-879. AXELSSON, M., OLSSON,C., GIBBINS, HOLMGREN, & I., S. FRANKLIN, E. (2001). Nitric oxide, a potent vasodilator of the C. aortic anastomosis in the estuarine crocodile, Crocodylus porosus. General and Comparative Endocrinology 122, 198-204.

Downloaded from Exp Physiol (ep.physoc.org) by guest on April 14, 2010

Exp. Physiol. 86.6

Control o j the crocodilian heart

789

W. BAUER,C. & JELKMAN, (1977). Carbon dioxide governs the oxygen affinity of crocodile blood. Nature 269, 825-827. BAUER, FORSTER, GROS,G., MOSCA,A., PERRELLA, C., M., M., ROLEMA, S. & VOGEL,D. (1981). Analysis of bicarbonate H. binding to crocodilian hemoglobin. Journal of Biological Chemistry 256, 8429-8435. FISHMAN, F., DELANEY, G., LAURENT, & SZIDON, P. A. R. P. J. (1985). Blood shunting in lungfish and humans. In Cardiovascular Shunts, Phylogenetic, Ontogenetic and Clinical Aspects. ed. W., JOHANNES, & BURGGREN, Alfred Benzon Symposium, vol. K. 21, pp. 88-99. Munksgaard, Copenhagen. FRANKLIN, E. & AXELSSON, (1994). The intrinsic properties of C. M. an in situ perfused crocodile heart. Journal ofExperimenta1 Biology 186, 269-288. FRANKLIN, E. & AXELSSON, (2000). An actively controlled C. M. heart valve. Nature 406, 847-848. GRTGG, C. (1989). The heart patterns of cardiac outflow in G. crocodilia. Proceedings of the Austvalian Physiological and Pharmacological Society 20,43-57. GRIGG, C. & GRUCA, (1979). Possible adaptive significance of G. M. low red cell organic phosphates in crocodiles. Journal of Experimental Zoology 209, 161-169. GRIGG, C. & JOHANSEN, (1987). Cardiovascular dynamics in G. K. Crocodylus porosus breathing air during voluntary aerobic dives. Compurative Physiology 157, 381-392. JONES, R. (1995). Crocodilian cardiac dynamics: A half-hearted D. attempt. Physiological Zoology 68, 9-1 5. JONES, R. (1996). The crocodilian central circulation: reptilian or D. avian? Verhandlungen der Deutschen Zoologischen 82,209-21 8. JONES, R. & SHELTON, (1993). The physiology of the alligator D. G. heart-left aortic flow patterns and right-to-left shunts. Journal of Experimental Biology 176,247-269. KARILA, AXELSSON, P., M., FRANKLIN, E., FRITSCHE, C. R., GIBBINS, L., GRIGG, C., NILSSON, & HOLMGREN, (1995). I. G. S. S. Neuropeptide immunoreactivity and co-existence in cardiovascular nerves and autonomic ganglia of the estuarine crocodile, Crocodylus porosus, and cardiovascular effects of neuropeptides. Regulatory Peptides 58, 25-39. MALVIN, M., HICKS,J. W. & GREENE, R. (1995). Central G. E. vascular flow patterns in the alligator Alligator niississipiensis. American Journal of Physiology 38, R1133-1139. MURGO, P., ALTER, R., DORETHY, F., ALTOBELLI, A. & J. B. J. S. MCGRANAHAN, M. (1980). Dynamics of left ventriular ejection G. in obstructive and nonobstructive hypertrophic cardiomyopathy. Journal qfClinica1 Investigations 66, 1369-1 382. PANIZZA, (1833). Sulla struttura del cuore e sulla circolazione del B. sangue del Crocodilus lucius. Bililioteca italiana 70, 87-9 1. SAINT AUBAIN, L. & WINGSTRAND, G. (1976). A sphincter in M. K. the pulmonary artery of the frog, Rana temporaria, and its influence on blood flow in skin and lungs. Acta Zoologica 60, 163-172. SHELTON, & JONES, R. (1991). The physiology of the alligator G. D. heart: the cardiac cycle. Journal of Experimental Biology 158, 539-564.
VAN

WEBB, J. W. (1979). Comparative cardiac anatomy of the reptilia. G. Journal of Morphology 161,221-240. WHITE, F. N. (1956). Circulation in the reptilian heart (Caiman sclerops). Anatatomicul Records 125, 417-432. WHITE,F. (1969). Redistribution of cardiac output in the diving alligator. Copeia 3, 567-570.

MIEROP, H. S. & KUTSCHE, M. (1985). In Cardiovascular L. L. Shunts: Phylogenetic, Ontogenetic and Clinical Aspects. ed. K. W. JOHANSEN, & BURGGREN W., Alfred Benzon Symposium, vol. 21, pp. 38-55. Munksgaard, Copenhagen.

Downloaded from Exp Physiol (ep.physoc.org) by guest on April 14, 2010