Astyanax Mexicanus) Atukorallaya Devi Sewvandini Atukorala1*, Christine Hammer1, Megan Dufton2 and Tamara Anne Franz-Odendaal1
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Atukorala et al. EvoDevo 2013, 4:28 http://www.evodevojournal.com/content/4/1/28 RESEARCH Open Access Adaptive evolution of the lower jaw dentition in Mexican tetra (Astyanax mexicanus) Atukorallaya Devi Sewvandini Atukorala1*, Christine Hammer1, Megan Dufton2 and Tamara Anne Franz-Odendaal1 Abstract Background: The Mexican tetra (Astyanax mexicanus) has emerged as a good animal model to study the constructive and regressive changes associated with living in cave environments, as both the ancestral sighted morph and the cave dwelling morph are extant. The cave dwelling morphs lack eyes and body pigmentation, but have well developed oral and sensory systems that are essential for survival in dark environments. The cave forms and surface forms are interfertile and give rise to F1 hybrids progeny known as intermediates. In cavefish, degeneration of the lens is one of the key events leading to eye regression. We have previously shown that surgical lens removal in surface fish embryos has an effect on the craniofacial skeleton. Surprisingly, lens removal was also found to have an effect on the caudal teeth in the lower jaw. In order to understand this result, we analyzed the lower jaw and upper jaw dentitions of surface, cavefish and F1 hybrids of surface and cavefish and compared our findings with surface fish that underwent lens removal. We also investigated the upper jaw (premaxillae and maxillae) dentition in these fish. Results: Our tooth analyses shows that cavefish have the highest numbers of teeth in the mandible and maxillae, surface forms have the lowest numbers and F1 hybrids are between these groups. These differences are not observed in the premaxillae. A wide diversity of cuspal morphology can also be found in these fish. Jaw size also differs amongst the groups, with the mandible exhibiting the greatest differences. Interestingly, tooth number in surgery fish is different only in the caudal region of the mandible; this is the region that is constrained in size in all morphs. Conclusion: Our data provides the first detailed description of the jaw dentitions of two morphs of Astyanax mexicanus, as well as in F1 hybrids. Tooth number, patterning and cuspal morphology are enhanced in cavefish in all jaws. This is in contrast to the increase in tooth number previously observed on the lens ablated side of the surgery fish. These findings indicate that the mechanisms which govern the constructive traits in cavefish are different to the mechanisms causing an increase tooth number in surgery fish. Keywords: Cavefish, Oral jaws, Mandible, Premaxilla, Maxilla, Tooth, Cusps Background In recent years, the teleost fish Mexican tetra, Over the course of vertebrate evolution organisms have Astyanax mexicanus, has been identified as a useful adopted constructive and regressive changes favorable model for studying the evolutionary biology of eye for their habitats. The creatures living in the extreme en- development. This is a single species consisting of two vironments, for instance cave dwelling animals, are morphological types, an eyed and pigmented surface- popular organisms to study because of their wide variety dwelling form (surface fish) and an eyeless unpigmented of adaptive features. Thus great insight can be gained cave-dwelling (cavefish) form [1-5]. These cavefish di- when comparing cave-adapted animals to their close rel- verged from their surface fish ancestors, probably about atives who do not live in these environments. 1 million years ago, and approximately 29 different cave- fish populations have been identified [1-3,6]. The two * Correspondence: [email protected] morphs can easily interbreed giving rise to intermediates. 1 Department of Biology, Mount Saint Vincent University, 166 Bedford The most highly investigated feature of the cavefish is Highway, Halifax, Nova Scotia B3M3J8, Canada Full list of author information is available at the end of the article their lack of eyes in adulthood. Despite the fact that © 2013 Atukorala et al.; licensee BioMed Central Ltd. This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. Atukorala et al. EvoDevo 2013, 4:28 Page 2 of 11 http://www.evodevojournal.com/content/4/1/28 cavefish do not have eyes as adults, eye development be- gins in the same manner in both the surface and cavefish morphs up until 24 hours postfertilization (hpf) [7,8]. At this age, the eye starts to degenerate. In addition to eye degeneration, cavefish have other regressive changes in- Pre maxilla cluding loss of pigmentation, reduction in the size of the E optic tectum, and a reduction in aggressive and school- ing behavior [1,2,5,9]. Less commonly studied are the constructive changes, which include changes in body Mandible position while feeding, increased size and number of cra- nial neuromasts, larger fat stores, increased number of taste buds, increased number of teeth and increased jaw Maxilla size [2,5,9]. Figure 1 Lateral view of the surgery side of the 3 days Although the increased tooth number in cave forms is postfertilization surgery fish. Alizarin red stained and cleared adult generally accepted, only one study has investigated tooth Astyanax mexicanus surface fish which has undergone surgical differences between cavefish and surface fish and it removal of the lens at 3 days postfertilization. The surgery eye is largely focused on teeth of only one oral jaw, namely the indicated by E. White arrowhead indicates the caudal region of the mandible. maxillary bone [10]. In addition, there are no data on tooth patterning in F1 hybrids of cavefish and surface fish crosses, which are commonly known as intermedi- Furthermore, we determined that the surgery fish are ates. The Mexican tetra bears teeth on the premaxilla, a useful group in which to be begin to unravel the tran- maxillary, and dentary bones of the oral jaws and on the sition from surface to cavefish [12]. gill rakers, upper pharyngeal tooth plates and the fifth To date, a detailed comparison of the characteristics of ceratobranchials of the pharyngeal skeleton [11]. The the lower jaw dentition in the Astyanax system has not unicuspid larval dentition is gradually replaced by the bi- been conducted. In this present study, we analyze and cuspid then tricuspid teeth resulting in the oral adult describe the tooth morphology, patterning, cuspal dentition. Generally the dentary bone of the adult sur- morphology and jaw dimensions in surface, cavefish and face morphs bears eight large multicuspid teeth rostrally their F1 hybrids. For a more comprehensive analysis, and several small multicuspid and unicuspid teeth caud- and since the mandible occludes with the upper jaws, we ally [11]. The diverse cuspal morphology can be found include both premaxillae and maxillae in our analysis. in the rostral teeth, which varies from five to eight cusps. Finally we compared the above results with the surgery- During food acquisition the lower jaws occlude with the induced jaw phenotypes, which we described in our pre- upper jaws (the premaxillae and maxillae). Premaxillary vious study [12] in order to fully understand the gross teeth directly contact the mandibular teeth while the morphological diversity of the oral jaws associated with maxillary teeth are positioned 180° to the functional pos- eye regression in Astyanax mexicanus. ition. The premaxilla has two rows of teeth with seven to nine teeth per side of the jaw while each maxilla bears Methods none, one or two teeth on each side [11]. Cuspal morph- Material ology in the upper jaw teeth are similar as described Surface Mexican tetra (Astyanax mexicanus) adults were above for the mandible. These teeth help the fish to maintained at 21°C on a 12-hour light, 12-hour dark grasp and chew food; nearly 40 tooth replacement cycles cycle. To induce spawning, tank temperature was in- occur throughout the life of the surface morph [11]. creased to 26°C and two males were added to a tank We previously demonstrated that, by removing the containing one female. Eggs were collected the next day. lens of one eye at several time points in surface tetra Fish were housed at Mount Saint Vincent University, embryos, lens removal has an effect on tooth develop- Halifax Nova Scotia, Canada. Animals were raised ment of the caudal region of the mandible (Figure 1) according to the guidelines of Canadian Council of Ani- [12]. An increased number of caudal teeth were found mal Care. All protocols were approved annually. on the surgery side compared to the non-surgery side. Tinaja cavefish and F1 hybrids of Tinaja cavefish and Generally, caudal teeth are susceptible to natural vari- surface fish were received in 10% neutral buffered for- ation making them more plastic in nature [11]. Their malin from Dr R Borowsky (New York University, New small size, the timing and manner of their development, York City, USA). These Tinaja cavefish were second gen- and their location close to the optic cup might make eration from wild populations. these teeth more susceptible to influences from the de- We examined a total of 27 adult fish: Tinaja cavefish veloping eye than the central larger multicuspid teeth. (n=8, 4.08 ± 0.29 cm Standard Length, age 6 years old), Atukorala et al. EvoDevo 2013, 4:28 Page 3 of 11 http://www.evodevojournal.com/content/4/1/28 F1 hybrid fish (n=6, 5.1 ± 0.17 cm Standard Length, age 7 years old), surface fish (n=8, 4.4 ± 0.3 cm Standard A Length , age 3.5± 1.4), and surgery fish (unilateral lens ablation of surface fish embryo at three days postfertilization (dpf) as described in [12]; n=5, 3.8 ± 0.3 cm Standard Length, age 1.0± 2.1).