ABSTRACT
The cephalopod circulatory system includes an efferent systemic ventricle and two afferent branchial hearts. Branchial hearts lack coronary arteries and thus must be nourished by only venous blood. During routine metabolism in normoxic water, oxygen availability to branchial hearts is only a small percentage of that to the systemic heart; during hypoxic excursions the venous oxygen content may, at least transiently, approach zero (Johansen, Brix & Lykkeboe, 1982; Houlihan, Innes, Wells & Wells, 1982). The extreme difference in oxygen delivery to these tissues suggests that branchial hearts are better able to meet their energetic demands through anaerobic metabolism than are systemic hearts. This hypothesis was tested by following isometric force development of the cardiac tissue of the cuttlefish, Sepia officinalis, during cyanide poisoning. Experiments were also conducted with the systemic heart of Octopus vulgaris, in order to allow a comparison between a decapod and an octapod.
Specimens of Sepia officinalis Lam. (193 + 29g; N = 7) and Octopus vulgaris Cuvier (1550 ± 367 g; N = 6) were maintained in running sea water. Animals were anaesthetized in 3 % ethanol, decapitated and the hearts excised. Sepia systemic ventricles were sectioned along the median line and each half utilized. Octopus ventricles were further sectioned transversely, allowing four preparations per animal. Each Sepia branchial heart was used for a single preparation. Tissues were spread flat to expose the interior surface, one end firmly fixed with a plastic clip, and the other end tied with surgical silk to a Harvard 363 isometric tension transducer interfaced with a Biotronex BL recorder. Hearts were forced to contract via field stimulation from two parallel silver electrodes, one on each side of the tissue. Square wave pulses of 20 ms duration with the minimum voltage necessary to produce the maximal mechanical response were generated from a Grass SD9 stimulator. After positioning, tissues were allowed a 30 min rest period and thereafter paced at 12 beats min−1. Following 5 min of contraction, NaCN (1 mmol 1−1 final concentration) was added to the bathing media in one half of the preparations and the recording of force continued for a further 25 min. Preliminary experiments showed that 2 mmol 1−1 NaCN did not have any greater effect than 1 mmol 1−1 NaCN, a concentration which inhibits respiration of cephalopod tissues (Ghiretti-Magaldi, Guiditta & Ghiretti, 1958). The bathing media was filtered sea water for (37‰) Octopus hearts and sea water plus 10 mmol 1−1 KC1 for Sepia tissues (Robertson, 1953). Media were continuously aerated, maintained at 15 °C, had a pH of 8-0 and contained 1 mmol 1−1 glucose. Control and experimental preparations were run simultaneously. Seven paired preparations were obtained from Sepia hearts and eleven preparations from six Octopus under both control and experimental conditions. In cases of duplicated experiments, values were averaged prior to statistical analysis. Relative force development was calculated as a percentage of that exhibited at time zero. Data are presented as mean±s.E.M.. The effect of the cyanide treatment at the 30-min point was assessed within tissues and between tissues with a paired and independent t-test, respectively.
Sepia systemic ventricles were, on average, able to maintain 98 % of the initial tension development during the experimental period in the absence of cyanide (Fig. 1). Cyanide poisoning led to rapid contractile failure, and after 30 min of treatment, only 11 % of the initial force was developed. The difference in relative performance between paired preparations was 87 ± 14%. Sepia branchial hearts failed both in the presence and absence of cyanide. At the end of the 30-min period, the control and experimental preparations developed only 74 and 37 % of the initial tension, respectively. The relative final performance was not significantly different between the two groups of branchial hearts. Octopus ventricles, even under control conditions, entered into failure and by the end of the experimental period could develop only 82 % of the initial force. Cyanide treatment accelerated the decline in performance and at the 30 min point led to a significant decrease in performance relative to controls of 38 % of the initial tension. Difference in relative performance between paired preparations of Octopus ventricles was 43 ± 12 % The treatment effect was significantly greater on Sepia than Octopus systemic ventricles (P< 0·05).
Both Sepia branchial hearts and Octopus systemic ventricles entered into contractile failure even under control conditions. This suggests that the bathing media were inadequate to support cellular integrity at the imposed level of energy demand. It is possible that cephalopod cardiac tissue has a necessary requirement for amino acids, which are known to be at extremely high levels in plasma (Hochachka et al. 1983). Another feature that may have been inappropriate was the level of CO2 which is higher in Sepia and Octopus blood than in aerated sea water (Johansen et al. 1982; Houlihan et al. 1982) Carbon dioxide tension is known to be extremely critical in the performance of teleost hearts (Gesser, 1977; Farrell, MacLeod, Driedzic & Wood, 1983). The failure of Sepia branchial hearts and Octopus systemic ventricles, however, does not preclude an analysis of the effect of cyanide treatment.
The essentially complete failure of Sepia systemic ventricles under simulated anoxic conditions shows that this tissue is obligately aerobic. This finding is consistent with early manometric studies (Barron, Sights & Wilder, 1953) and the high mitochondria content of squid ventricles (Dykens & Mangum, 1979). As implied by blood gas data, Sepia branchial heart is better able to meet its energetic demands via anaerobic metabolism than the systemic ventricle. Maximal activity levels of enzymes of carbohydrate metabolism do not suggest that better performance of branchial hearts is due to an enhanced ATP generation capability (W. R. Driedzic & B. D. Sidell, unpublished observations). The mechanical work, and presumably the ATP demand, of the cephalopod branchial heart in situ is far lower than that of the systemic ventricle (Bourne, 1982; Wells & Wells, 1983). The present experiments show that anaerobic ATP production can more easily match ATP demands in the former.
The poisoning of oxidative metabolism in Octopus systemic ventricles resulted in an impairment of function but not in a complete collapse over the experimental period. A partial resistance to anoxia has been demonstrated for the ventricle of Octopus dofleini (Pritchard, Huston & Martin, 1963). Octopus ventricle is more resistant to simulated anoxia than is Sepia ventricle. The ventricle of the octopod, Elodone cirrhosa, has similar activity levels of enzymes associated with carbohydrate metabolism but a much lower level of total ATPase activity than Sepia ventricle (W. R. Driedzic & B. D. Sidell, unpublished observations). If Octopus vulgaris is similar to Eledone cirrhosa in this respect, then the difference in performance between Octopus and Sepia ventricles may be related to lower levels of ATP demand and hence a greater capability to match that demand through anaerobic metabolism alone.
In conclusion, the present experiments show that under conditions of impaired oxidative metabolism, both Sepia branchial heart and Octopus systemic ventricle are better able to maintain performance than Sepia systemic ventricle.
ACKNOWLEDGEMENTS
This work was supported by operating grants from N.S.E.R.C. of Canada and the New Brunswick Heart Foundation. Appreciation is extended to the members of the cephalopod group and the staff at the Laboratoire Arago, France for their considerable help.