Embryonic and Larval Development in the Caecilian Ichthyophis Kohtaoensis (Amphibia, Gymnophiona): a Staging Table
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JOURNAL OF MORPHOLOGY 243:3–34 (2000) Embryonic and Larval Development in the Caecilian Ichthyophis kohtaoensis (Amphibia, Gymnophiona): A Staging Table Nicole Du¨ nker,1 Marvalee H. Wake,2* and Wendy M. Olson2 1 Department of Zoology, Technical University of Darmstadt, Darmstadt, Germany 2 Department of Integrative Biology and Museum of Vertebrate Zoology, University of California, Berkeley, California ABSTRACT Little is known about the developmental of the lateral line organs, formation of three pairs of ex- biology of caecilians—tropical, elongate, limbless, mostly ternal gills, development of the eyes, changes in yolk fossorial amphibians that are members of the Order Gym- structure, changes in the structure of the cloacal aperture nophiona. Ichthyophis kohtaoensis (Family Ichthyophi- and growth of the tail, including the formation and regres- idae; southeast Asia) is an oviparous species in which sion of the tail fin. This study provides a comparison with maternal care of the clutch is provided. The clutch is laid descriptions of embryonic stages of I. glutinosus and Hy- in a burrow on land, and the embryos develop in their egg pogeophis rostratus and with a recent staging table for the membranes, curved around a large yolk mass. Larvae are aquatic, viviparous caecilian Typhlonectes compressi- aquatic and exhibit characteristic features that are not cauda, the only other caecilians for which reasonably com- present in the terrestrial adults. Because accurate de- plete ontogenetic information exists in the literature. scriptions of ontogenies and the establishment of stan- Comparisons with established staging tables for selected dardized stages of embryonic and larval development are useful for both experimental and comparative embryology, frogs and salamanders are also presented. J. Morphol. a staging table for I. kohtaoensis was developed based on 243:3–34, 2000. © 2000 Wiley-Liss, Inc. external morphological features. Development from the end of neurulation to metamorphosis was divided into 20 KEY WORDS: Ichthyophis kohtaoensis; amphibians; cae- stages. Principal diagnostic features include development cilians; ontogeny; development; staging Caecilians, members of the Order Gymnophiona, the yolk. A number of specific features of develop- are tropical, limbless, elongate, tailless or nearly so, ment are evaluated below. amphibians that occupy subterranean, leaf-litter, or There are very few descriptions of caecilian devel- semi-aquatic to aquatic habitats. They are among opment from early stages to metamorphosis or birth the least studied of vertebrates. Rather little is (Sarasin and Sarasin, 1887–1890; Brauer, 1897, known of the development of caecilians, especially 1899; Sammouri et al., 1990); some authors have compared to the relative wealth of knowledge about described only one or a few specimens (not staged) selected frogs and salamanders. available for study. Sarasin and Sarasin (1887– The development of Ichthyophis glutinosus (Fam- 1890) and Brauer (1899) presented detailed descrip- ily Ichthyophiidae) was described and beautifully tions of the changes in the course of development, illustrated by Sarasin and Sarasin (1887–1890). referring variously to the formation of gills, eyes, the Cleavage in amphibian eggs is typically holoblastic tentacle, the lateral line organs, changes in yolk and unequal. In I. glutinosus, the only caecilian for structure, and general external morphology. Brauer which cleavage has been described (Sarasin and (1899) provided a rather complete series of drawings Sarasin, 1887–1890), cleavage is nearly meroblastic, of early embryos of Hypogeophis rostratus, with dividing the egg into numerous separate blas- tomeres and a residual multinucleate mass of cyto- plasm. At the tail-bud stage, caecilians are similar Contract grant sponsor: National Science Foundation; Contract to salamanders and have a relatively long head and grant number: IBN95-27681. pharyngeal region, including the gill plates, that projects above the yolk mass, but they have only a Current address for Nicole Du¨ nker, Anatomy, Medical Faculty 3.1, short tail bud. In contrast, in anurans the gill plate University of Saarland, 66421 Homburg/Saar, Germany. region lies dorsal to the yolk mass with only the *Correspondence to: Marvalee H. Wake, M.D., Department of Integra- anterior most part of the head projecting beyond the tive Biology, University of California, Berkeley, CA 94720-3140, USA. yolk, but a relatively long tail bud extends beyond E-mail: [email protected] © 2000 WILEY-LISS, INC. 4N.DU¨ NKER ET AL. some later stages of that species and a few for Gran- disonia alternans; Marcus described the develop- ment of specific elements of Hypogeophis rostratus, but mostly internal features so only his work rele- vant to external morphology is cited herein; Sarasin and Sarasin (1887–1890) described various develop- mental stages of Ichthyophis glutinosus, including those of very early development. However, these studies did not present their results in the form of a staging table. The continuously changing appearance of em- bryos and larvae during ontogenesis necessitates a method of quantifying the progress of development. Tables of normal stages of development have been established for a number of amphibian species, nearly all anuran and urodelan (see Duellman and Trueb, 1986). Each normal stage is designated by a number and represents a specific (but highly subjec- tive) interval in ontogeny, defined by the presence or absence of ontogenetic character states, such as body size or morphology (Bartsch et al., 1997). By subdividing development into discrete stages, it is possible to group and compare individuals at ap- proximately the same point in ontogeny. Staging tables are useful tools and have long been used in the study of amphibian development. Only one such table exists for caecilians; it is for a highly derived taxon. Sammouri et al. (1990) presented a detailed description of the embryonic development of Typhlonectes compressicauda, a viviparous aquatic caecilian of the family Typhlonectidae that has many developmental features unique to the family. They included a formal staging table and Fig. 1. Ichthyophis kohtaoensis. An adult specimen with a compared development in T. compressicauda with total body length of approximately 300 mm is shown in A. Larvae, that of the oviparous frog Alytes obstetricans. approximately 100–150 mm in length, are shown in B. Photos by We describe the development of Ichthyophis W. Himstedt. kohtaoensis, an oviparous species with free-living larvae from Thailand, and compared it to that re- ported for Ichthyophis glutinosus, Hypogeophis ros- second oviparous ichthyophiid caecilian, I. kohtaoen- tratus, and Typhlonectes compressicauda. This com- sis (Fig. 1A), thus facilitating comparison within a F1 parison affords an examination of development in genus and family of caecilians. caecilians representing both basal and derived taxa, Ichthyophis kohtaoensis typifies the biology of and provides a basis for evaluation of evolutionary oviparous caecilians, so far as is known (see Himst- trends in development in caecilians. The two species edt, 1996). Copulation in I. kohtaoensis has never of Ichthyophis are members of the Family Ich- been observed, but internal fertilization is pre- thyophiidae. The families Rhinatrematidae (the sumed, as for all caecilians, via the male’s inserting most basal caecilian family) and Ichthyophiidae con- his phallodaeum into the vent of the female to effect stitute the sister-group to all other caecilians. H. sperm transport (see Wake, 1972, for summary). In rostratus is oviparous and direct-developing (i.e., de- the course of the rainy season from May to October, velopment through metamorphosis occurs before during which 90% of the annual rainfall in northeast hatching in terrestrially laid clutches), thus obviat- Thailand occurs and floods the rice fields, the fe- ing the free-living aquatic larval stage, and is a males lay their fertilized, developing eggs in moist member of the derived family Caeciliidae. The mo- cavities in the ground. I. kohtaoensis practices ma- notypic Hypogeophis occurs in the Seychelles Is- ternal care of the laid clutch, as has been reported lands. T. compressicauda, with its obligate vivipar- for several egg-laying species (e.g., Sarasin and ity, and with other features of its biology that are Sarasin, 1887–1890; Sanderson, 1936). The eggs of a construed as correlates of its aquatic habitus, repre- clutch are bound together with strands that extend sents a highly derived clade of caecilians; it occurs in from the jelly coats, forming a cluster; the parental South America. The present study provides develop- female curls around the clutch and turns the eggs mental information, including a staging table, for a at regular intervals. Similar to embryos of DEVELOPMENT OF ICHTHYOPHIS KOHTAOENSIS 5 To our knowledge, there is not yet available a table of normal stages of development for oviparous caecilians. Consequently we evaluate our data on the development of Ichthyophis kohtaoensis and present a staging table based on external morpho- logical characters (Table 1). Because the present T1 study is intended to provide a staging table for a very poorly known species, a detailed description of the embryonic and larval development is given and as many characters are described as possible. Those features facilitate a comparison to other caecilians, and to frogs and salamanders, allowing a much broader comparison of development among amphib-