JOURNAL OF MORPHOLOGY 218:195-201 (1993)

Heterodonty and Patterns of Tooth Replacement in Crocodylus niloticus

J.A. KIESER, C . KLAPSIDIS, L. LAW, AND M. MARION Departments of Oral Pathology (J.A.K.)and General Anatomy (C.K., L.L., M.M.), School of Dentistry, Witwatersrand University, Johannesburg 2050, South Africa

ABSTRACT Patterns of tooth replacement in the Nile crocodile Crocodylus niloticus were evaluated in the lateral radiographs of six dried skulls of 1-year-old individuals. Additionally, the ontogeny of the succession of teeth in this species was investigated in four embryos and a hatchling, which were serially sectioned. Three-dimensional, computer-generated reconstructions of these individuals suggested that there was a regular pattern of odontogenic progression. Three morphogenetic zones could be identified in each of the age classes: an incisor, a canine, and a molar region. It was thus concluded that the Nile crocodile is hetero- rather than homodont. Zahnreihen, or tooth rows, with a periodicity of one were easily identified. Q 1993 Wiley-Liss, Inc. The regional specialization of tooth shape in reptiles had its beginnings before the Triassic, with the development of canine, postcanine, and precanine regions within the dental arcades of some cynodont therapsids (Brown, 83). The extensive literature on reptilian dentitions has evoked the image of a simple dichotomy between undifferentiated homodont dentitions and differentiated heterodont dentitions (for review, see Keene, 9 1 ) . What exactly is meant by heterodont is, however, a bone of contention. Simpson (36) suggested that the term incipient heterodonty might be applied to those Permian therapsids (gorgonopsians)in which the first maxillary tooth and a corresponding tooth in the lower jaw was the only tooth in an otherwise homodont arch to be enlarged. This canine, it was argued, divided the dentition into incisors mesial to it and cheek teeth posterior to it. In contrast to incipient heterodonty stood the advanced heterodonty that Simpson (36)had originally observed in the Triassic mammallike reptile Cynognathus and has more recently been documented in cynodonts (Crompton, 63; Fourie, 63; Hopson, 641, which showed a distinct separation of the cheek teeth into premolars and molars. Osborn (75) pointed out that in mammallike reptiles such as Thrinaxodon and Diademodon successive postcanine replacement teeth had different shapes, a condition he termed intrafamilial heterodonty. More recently, Ferguson (81)introduced yet an1993 WILEY-LISS, INC.

other label, the term pseudoheterodont, to describe the arrangement of teeth in the modern alligator. Few empirical stuhes have been applied to the problems of tooth replacement and heterodonty in extant reptiles. To date, studies have been limited to the lizards Lacertu uiuiparu (Osborn, 71) and Zguana iguana (Kline and Cullum, 851, and crocodilians Crocodylus niloticus (Poole, 61) and Aligator mississippiensis (Westergaard and Ferguson, 86, 87). None of these studies have, however, attempted to evaluate the temporal relationships among teeth in a series of different ages of a given species. As a consequence, our empirical knowledge of the developmental origin of molars, canines, and incisors remains scant. Three major hypotheses provide theoretical explanations for aspects of heterodonty such as variation in tooth size, number, shape, and position. The first of these is the field theory of Butler (39, 78, 83), which states that tooth size and shape are determined by the type and intensity of the morphogenetic field into which ectomesenchymal cells migrate during odontogenesis. The disadvantage of this model is that it relies on an as yet unidentified chemical substance, which has the same effect over a large temporal span. The clonal model of Osborn (71, 75, 78)
Address reprint requests to J.A. Kieser, Dept. Oral Pathology, School of Dentistry, Witwatersrand University, Private Bag 3, Johannesburg 2050, South Africa.

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avoids morphogenetic substances and may be applied to molecular biological phenomena. Osborn suggested that two ancestral clones (incisor and molar) were phylogenetically the oldest, with a canine clone that was the last to evolve. Recently, Westergaard ('80, '86) proposed a tooth position theory; in this the number and position of functional tooth positions are controlled by differential jaw growth. I n this study, we address the problem of heterodonty from a n empirical perspective by investigating the patterns of tooth replacement in the large modern reptile, Crocodylus niloticus.
MATERIALS AND METHODS

after which they were traced with the aid of a camera lucida a t 250x magnification. Tracings were stacked and aligned and then retraced onto a Kontron Image Analyser. Reconstructed images were then printed in hard copy with a vidioplan matrix printer.
RESULTS

Six dried skulls of 1-year-old Nile crocodiles Crocodylus niloticus were obtained from the Cango Cave Crocodile Ranch near Oudtshoorn, South Africa. Each individual was sagittally hemisected and radiographed on a n At-gostat equipped with regular intensifying screens. True lateral views were taken on each specimen, with a tube-object distance set at 100 cm. Radiographs were produced on 3 M Medical film for 0.06 seconds a t 100 M.A. and 52 K.U. To evaluate tooth eruption patterns in embryonic crocodiles, we serially sectioned the paraffin-embedded heads of five embryos: at 14 days, 28 days, 42 days, 60 days, and fullterm. Sections were cut in the coronal plane and stained with haematoxylin and eosin,

Figure 1 shows a lateral radiograph of the skull of a 1-year-old crocodile, together with its attendant tracing. It is immediately obvious that there is a gap between tooth 5 and tooth 6 and that this defines the location of the maxillary-premaxillary suture. Posterior to the suture is a newly erupted canine, followed by five other teeth and then another gap, which in turn is followed by six teeth that all share a coronal indentation, as does the tooth immediately preceeding the gap. This pattern was consistent in all six dried skulls examined. Examination of the reconstruction of a crocodile hatchling (Fig. 2) shows that there are five premaxillary teeth, followed by a group of five teeth that are at roughly the same stage of replacement. There follows a gap and then five teeth that are similarly sized. In the mandible, there are three incisors followed by the large canine and a group of four smaller teeth. Then follows a n arrangement of seven distal teeth. The reconstruction of the 60-day-old embryo provides a similar pattern (Fig. 3). In

Fig. 1. Crocodylus niloticus. Lateral radiograph and tracing of the hemiseded skull of a 1-year-oldcrocodile. Functional teeth, white; replacement teeth, black.

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Maxilla

Mandible
Fig. 2. Crocodylus niloticus. Diagram of the reconstruction of a crocodile hatchling head, showing incisor, canine, and molar areas. Large ovals are fully formed teeth, smaller ovals are replacement teeth. In some cases there are second replacement teeth (small ovals), which are at early bell/cap stages of development.

the upper jaw, five teeth precede a group of five teeth, which are roughly in ascending order of size and are followed by six similar size teeth. The mandible showed three incisors and a large canine followed by a group of four subequal teeth and then seven larger teeth. Comparison of the reconstructions for 42 and 28-day embryos (Fig 4) discloses a break in the dental lamina of the upper jaw. In the 28-day embryo, this is preceded by two bud or early cap stages and an anterior caruncle. With maturity, the caruncle is lost and the two early bud/cap areas progress to late cap/ early bell stages. Behind the lamina1 break are two clones that ultimately mature at 42 days to four late cap/early bell regions, respectively. A similar arrangement is found in the lower jaw. However, instead of a clear break in the dental lamina in the 28-day embryo, there is a marked thinning behind the first two buds. The 14-day-oldembryo provides no evidence of tooth bud formation. I t appears, however, that tooth initiation starts distally in the form of thickenings of the oral ectoderm.

Comparison of the tracings of upper and lower teeth of the juvenile crocodile (Fig. 5) discloses patterning of morphological types. As in the embryonic reconstructions of the maxilla, there are five anterior incisors, followed posteriorly by five teeth of ascending size, and in turn by teeth with distal coronal constrictions. The lower jaw has three incisors, five canines, and seven teeth with distal coronal constrictions.
DISCUSSION

De Mar ('74) has stated that the most important contribution of Edmund's work on reptilian tooth replacement was his demonstration that a single cause underlay the organization of most, if not all, dentitions. Edmund's theory of Zahnreihen ('60, '62, '69) is an empirical generalization that asserts that there is a region at the front of the jaw from whence impulses are generated at regular time intervals. These impulses sweep caudally along the jaws at regular speed, initiating tooth formation a t regular intervals. Because pulses were evenly spaced, waves of immature teeth were initiated at

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Fig. 3. Crocodylus niloticus. Diagram of the reconstruction of the head of a 60-day-old crocodile embryo, showing incisor (I), canine (C), and molar (M) areas. Wavy lines indicate borders between tooth types.

each tooth position where they move occlusad to replace functioning teeth. Hence the dentition will develop in sequence from the front to the back with teeth forming posteriorly sloping arrays called Zahnreihen. In the lifetime of the reptile, numerous Zahnreihen pass down the jaw. Despite lengthy discussion (De Mar, '72, '73, '74; ThuIborn, '71; Bolt and De Mar, '75, '83; Hopson, '80; Kline and Cullum, '85)and general acceptance, the theory has been criticized by Osborn ('71, '75, '78) on the grounds that it assumes horizontal control of tooth replacement instead of vertical control within each tooth family. Osborn postulates that teeth develop from growing clones of ectomesenchymal cells that have already acquired morphogenetic properties prior to their migration into the presumptive tooth bearing regions of the upper and lower jaws. Osborn ('78) further suggested that temporal changes in the embryonic competence is responsible for producing intrafamilial similarities of teeth. In contrast, Westergaard ('80, '86) and Westergaard and Ferguson ('86, '87) feel that the position and number of teeth are dependent on the genetically controlled differential growth of the embryonic jaw anlage and the

dental lamina, which would create more or less space around each existing tooth for new teeth t o form. The focal question that remains is: what is the bearing of the present evidence on Edmund's theory of tooth rows or Zahnreihen? The addition of tooth succession lines to Figure 5 appears to provide a very clear indication that Zahnreihen exist in the crocodile and that these are one tooth position apart as they sweep anteriorly through both upper and lower jaws. Berkowitz and Shellis ('78) were the first to call attention to the simultaneous replacement of teeth in the piranha Serrasalmus rhombicus. Although they noted a complex sequence of tooth movements and a n asynchronous eruption between left and right sides of the jaws, these authors were able to show synchronous replacement of teeth within quadrants. A subsequent study by Bolt and De Mar ('83) showed the existence of simultaneous replacement (with a Z-spacing of 1) in the Lower Permian gymnarthrid microsaurs Euryodus and Cardiocephalus. From this study and from their reexamination of data on Necturus, these authors concluded that simultaneous replacement may be more common

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0Dental Lamina
I BudEarly Cap
Late Cap/Early Bell

A Egg Tooth

28 Day

c)

3,

42Day

Fig. 4. Crocodylus niloticus.Diagram of the reconstructionsof the jaws of 28- and 42-day-old crocodile embryos.Wavy lines represent the differentiationinto incisor, canine, and molar areas.

than had been previously supposed. Whereas our study clearly provides additional data in support of the foregoing, there remains the unexplained lengthening of the Z-spacing in the distal segment of the distal segment of

the maxilla (Fig. 5 ) . If one accepts that members of a single Zahnreihe may be recognized by identifying the oldest functional tooth in a series and then progressing to the youngest tooth, then there appears compelling evi-

Fig. 5. Crocodylus niloticus. Lateral cephalometric tracing of a l-year-old crocodile, showing incisor (I), premolar ( C ) ,and molar (M) areas, and Zahnreihen with a periodicity of one.

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J.A. KIESER ET AL. LITERATURE CITED Berkowitz, B.K.B., and R.P. Shellis (1978)A longitudinal study of tooth succession in piranhas with an analysis of the tooth replacement cycle. Z. Zool. Lond. 184.545561. Bolt, J.R., and R.E. De Mar (1975) An explanatory model of the evolution of multiple rows of teeth in Cuptorhinus aguti. J . Paleo. 49:814-832. Bolt, J.R., and R.E. De Mar (1983) Simultaneous tooth replacement in Euryodus and Cardiocephalus. J. Paleo. 57r911-923. Brown, K.S. (1983) Evolution and development of the dentition. Birth Defects 19t29-66. Butler, P.M. (1939) Studies on the mammalian dentition: Differentiation of the postcanine dentition. Proc. Zool. SOC. Lond. 109:l-36. Butler, P.M. (1978) The ontogeny of mammalian heterodonty. J . Biol. Buccale 6:217-227. Butler, P.M. (1983) Evolution and mammalian dental morphology. J. Biol. Buccale 11:285-302. Crompton, A.W. (1963) Tooth replacement in the cynodont Thrinanodon Ziorhznus Seeley. Ann. S. Afr. Mus. 46:479-521. De Mar, R.E. (1972) Evolutionary implications ofZahnreihe. Evolution 26:435450. De Mar, R.E. (1973) The functional implications of geometrical organization of dentitions. J. Paleo. 47:452461. De Mar, R.E. (1974) On the reality ofZahnreihepien and the nature of reality in morphological studies. Evolution 28t328-330. Edmund, A.G. (1960) Tooth replacement phenomena in lower vertebrates. Contr. Life Sci. Div. Roy. Ont. Mus. 52:l-190. Edmund, A.G. (1962) Sequence and rate of tooth replacement in the crocodilia. Contr. Life Sci. Div. Roy. Ont. Mus.56r1-42. Edmund, A.G. (1969) Dentition. In C. Gans, A.A. Bellairs and T.S. Parsons (eds): Biology of the Reptilia I Morphology. London: Academic Press, pp. 115-200. Ferguson, M.W.G. (1981) The value of the American alligator Alligator mississippiepiensis as a model for research in craniofacial development. J. Craniofac. Genet. Dev. Biol. 1t123-144. Fourie, S. (1963) Tooth replacement in the gomphodont cynodont, Diudemodon. S. Afr. J . Sci. 59t211-213. Hopson, J.A. (1964) Tooth replacement in cynodont, dicynodont and therocephalian reptiles. Proc. Zool. SOC. Lond. 142:625-654. Hopson, J.A. (1980) Tooth function and replacement in early Mesozoic ornithischian dinosaurs: Implications for aestivation. Lethaia I13r93-105. Keene, H.J. (1991) On heterochrony in heterodonty: A review of some problems in tooth morphogenesis and evolution. Yearbk. Phys. Anthropol. 34:251-282. Kieser, J.A. (1984)Wave superpositioning and the initiation of tooth morphogenesis: An application of the Bandwidth Theorem. Med. Hypoth. 14:249-252. Kline, L.W., and D.R. Cullum (1985) Tooth replacement and growth in the young green Iguana, Iguana iguana. J. Morph. 186:265-269. Osborn, J.W. (1971) The ontogeny of tooth succession in Lucerta uiuiparu. Proc. Roy. Soe. Lond. B. 179:261289. Osborn. J.W. (1975) Tooth redacement Efficiencv. Datternsand evolution. Evolutibn 29:180-186. Osborn, J.W. (1978) Morphogenetic gradients: fields versus clones. In P.M. Butler and K.A. Joysey (eds):Devel-

dence for alternation in size of the replacing teeth 2-4-6, which are larger than 3-5-7, and hence or a Z-spacing of 2. Another fundamental question is: what are the three groups of teeth observed here? That is, can these teeth be homologized? If one accepts the definition of Wagner ('89), which states that structures from two individuals or from within the same individual are homologous if they share those developmental constraints imposed by locally acting selfregulatory machanisms or organ differentiation, then one may suggest that the three groups of teeth observed in the developing crocodile are developmentally individualized parts of its phenotype. In the presence of a discontinuity of the dental lamina in the maxilla, we suggest that the Nile crocodile has five premaxillary incisors, followed by five canines and six or more postcanines. In the lower jaw there are three incisors, five canines, and up to seven postcanines. Hence, the Nile crocodile appears t o be a truly heterodont animal. Ultimately, development depends on molecular synthesis that results when various chemical precursors exceed critical threshold concentrations. The synthesis of precursors, in turn, depends on exceeding of the threshold concentrations of any number of lower level precursors, and so on (Kieser, '84). Thus development occurs in waves compounded on waves in such complexity that the precise dynamics at any one time is unpredictable from even a complete historical knowledge of the process. What the present study does suggest, however, is that there are four presumptive toothforming loci in each jaw (indicated in black in Fig. 4).The two incisor loci are separated from the other two loci by a distinct break in the maxillary dental lamina and a thinning in the lower jaw. One might hypothesize that from these presumptive loci mature loci are developed, the formation of which is governed by a phase-locking system of feedback involving upper and lower jaw lengths. In the crocodile,Zuhnreihen progress through every tooth locus t o result in a tooth replacement wave with a periodicity of one.
ACKNOWLEDGMENTS

The authors thank the Cango Cave Crocodile Farm for supplying the specimens. Glynny Kieser edited and typed the text. Jeff Osborn provided stimulating and useful insights into his clonal model, and the comments of two anonymous reviewers vastly improved the text.
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opment, Function and Evolution of Teeth. New York: Academic Press, pp. 171-201. Poole, D.F.G. (1961) Notes on tooth replacement in the Nile crocodile. C r o c o d i h niloticus. Proc. Zool. SOC. Lond. 136:131-160. Simpson, G.G. (1936) Studies of the earliest mammalian dentitions. Dent. Cosmos 78t791-800. Thulborn, R.A. (1971) Tooth wear and jaw action in the Triassic ornithischian dinosaur Fabrosaurus. J. 2001. 164t165-179. Wagner, G.P. (1989) The biological homology concept. Ann. Rev. Ecol. Syst. 20:51-69.

Westergaard, B. (1980) Evolution of the mammalian dentition. Mem. SOC. Geol. France 139:191-200. Westergaard, B. (1986) The pattern of embryonic tooth initiation in reptiles. Mem. Mus. Nat. Hist. Paris 53:5563. Westergaard, B., and M.W.J. Ferguson (1986) Development of' the dentition in Alligator mississippiensis: Early development of the lower jaw. J. Zool. 210:575597. Westergaard, B., and M.W.J. Ferguson (1987) Development of the dentition of Alligator mississippiensis: Later development in the lower jaws of hatchlings and youngjuveniles. J. 2001.212:191-222.