Mechanism of multinucleate myotomal muscle fibre formation in Hymenochirus boettgeri (Anura, Pipidae)

During somitogenesis in Hymenochirus boettgeri, somites separate from non-segmented mesoderm. Somite formation involves changes in position of myotomal cells from perpendicular to parallel relative to axial organs; the changes are asynchronous and show a dorsoventral gradient. After the rotation has been completed, the myotomal cells (primary myoblasts) occupy the whole length of the myotomes. MyoD is present in nuclei of non-segmented mesoderm cells, of myotomal cells during their rotation and of myoblasts occupying the whole length of the myotomes. The effect of MyoD which activates muscle-specific genes is confirmed by the appearance of skeletal α-actin in mononucleate myoblasts in which myofibrils and the sarcotubular system develop. Differentiation of primary myoblasts results in development of mononucleate, morphologically mature myotubes. Differentiating myotubes are initially not accompanied by any other cells. In further developmental stages, mesenchymal cells appear in intermyotomal fissures and then in myotomes. Their role depends on their position: mesenchymal cells remaining in the intermyotomal fissures differentiate into fibroblasts while those that have migrated into the myotomes, between the myotubes, transform into secondary myoblasts. Their myogenic function is evidenced by the presence of MyoD in their nuclei. These cells fuse with the already existing mononucleate myotubes, resulting in an increase in their size and number of nuclei. Accepted: 30 January 2001     

Effects of hind limb denervation on the development of appendicular ossicles in the Dwarf African Clawed Frog,Hymenochirus boettgeri (Anura: Pipidae)

Sesamoids and other appendicular ossicles are common in other classes of vertebrates but comparatively rare in amphibians. The pipid frog Hymenochirus boettgeri (Boulenger, G. A. 1899. On Hymenochirus, a new type of aglossal batrachians. – Annals of the Magazine of Natural History Series 7: 122–125) is unusual among anurans in having seven (or more) appendicular ossicles in each hind limb. Sesamoids are often associated with muscles and tendons, and their development is usually regarded as mediated by or correlated with function. This study investigated the effects of paralysis (loss of function) on development of ossicles in the hind limb of Hymenochirus. Complete denervation of the right sciatic nerve was performed at developmental stages 63 and 66, and the animals maintained for a further 6–7 or 12–13 weeks. Specimens were cleared and double stained for cartilage and bone. There were no gross morphological differences between control and sham operated groups. The lunulae were not affected by paralysis, whereas the fabella arose later and/or regressed in some specimens. The distal os sesamoides tarsalia (OST) was shorter in paralysed individuals, and both the distal OST and cartilagines plantares showed delayed maturation. Denervation of the hind limb thus affected the timing of appearance, maintenance and rate of maturation of some sesamoid bones in Hymenochirus, but had no effect on others.     

The hymenochirins: a family of host-defense peptides from the Congo dwarf clawed frog Hymenochirus boettgeri (Pipidae)

Skin secretions of frogs from the subfamily Xenopodinae (Xenopus+Silurana) within the family Pipidae are a rich source of antimicrobial peptides with therapeutic potential but species from the sister taxon Hymenochirus in the subfamily Pipinae (Hymenochirus+Pseudhymenochirus+Pipa) have not been investigated. Peptidomic analysis of norepinephrine-stimulated skin secretions from two distinct populations of the Congo dwarf clawed frog Hymenochirus boettgeri (Tornier, 1896) has led to identification of five structurally related peptides with broad-spectrum antimicrobial activity. Hymenochirin-1B (IKLSPETKDNLKKVLKGAIKGAIAVAKMV.NH(2)) is C-terminally α-amidated whereas hymenochirins-2B-5B have the general structure XKIPX(2)VKDTLKKVAKGX(2)SX(2)AGAX(3).COOH. Hymenochirin-3B (IKIPAVVKDTLKKVAKGVLSAVAGALTQ) was the most abundant peptide in the secretions. The hymenochirins show very low structural similarity with the antimicrobial peptides isolated from skin secretions of Silurana tropicalis and Xenopus laevis consistent with the proposed ancient divergence of the Pipinae and Xenopodinae. Synthetic replicates of hymenochirin-1B-4B inhibit the growth of Escherichia coli, Pseudomonas aeruginosa, Klebsiella pneumoniae, and Staphylococcus aureus (MIC in the range 10-40 μM) and Candida albicans (MIC=80 μM). The peptides display relatively weak hemolytic activity against human erythrocytes (LC(50) in the range 160 to >300 μM).     

Morphology of skin incubation in Pipa carvalhoi (Anura: Pipidae)

South American Pipidae show a unique reproductive mode, in which the fertilized eggs develop in temporarily formed brood chambers of the dorsal skin after eggs have been deposited on the back of the female. We studied the skin incubation of Pipa carvalhoi using light microscopy and scanning electron microscopy. The skin consists of a stratified epithelium with a one‐layered stratum corneum, and the dermis. The dermis of the dorsal skin of nonreproductive and reproductive females lacks a distinct stratum compactum, which is typical for most anuran skins. The entire dermis shows irregularly arranged collagen bundles like a stratum spongiosum. Before egg laying, the skin swells, primarily by thickening and further by loosening of the middle zone of the dermis. In the epidermis, large furrows develop that are the prospective sites of egg nidation. The epidermis, which forms a brood chamber around the developing egg becomes bi‐layered and very thin and lacks a stratum corneum. Further, the dermis loosens and becomes heavily vascularized. Egg carrying females do not have mature oocytes in their ovaries indicating a slow down or interruption of egg maturation during this period. Similarities with the brood pouch of marsupial frogs are discussed. J. Morphol., 2009. © 2009 Wiley‐Liss, Inc.     

Evolution of globin expression in the genus Xenopus (Anura: Pipidae)

Comparison of electrophoretic globin phenotypes of 18 different Xenopus taxa reveals four different basic types of banding patterns. Each type includes species that also are similar in their morphological, cytogenetical, and biochemical characteristics. Three of these patterns reflect distinct evolutionary lines, while the fourth may be interpreted as the intersection of two of these lines. The composition of the basic pattern of the highly polyploid species is consistent with an allopolyploid origin of most of these species. The number of distinct globin polypeptides--four in the only extant diploid species, X. tropicalis, and five or more in most of the tetraploid species, including X. laevis--suggests that primordial globin genes had undergone duplication either before or after the tetraploidization event. Finally, the individual globin phenotypes are excellent molecular markers that are of great help in identifying the various species but not the subspecies.      

Skeletal development in Xenopus laevis (Anura: Pipidae).

Postembryonic skeletal development of the pipid frog Xenopus laevis is described from cleared-and-stained whole-mount specimens and sectioned material representing Nieuwkoop and Faber developmental Stages 46-65, plus postmetamorphic individuals up to 6 months old. An assessment of variation of skeletogenesis within a single population of larvae and comparison with earlier studies revealed that the timing, but not the sequence, of skeletal development in X. laevis is more variable than previously reported and poorly correlated with the development of external morphology. Examination of chondrocranial development indicates that the rostral cartilages of X. laevis are homologous with the suprarostral cartilages of non-pipoid anurans, and suggests that the peculiar chondrocranium of this taxon is derived from a more generalized pattern typical of non-pipoid frogs. Derived features of skeletal development not previously reported for X. laevis include 1) bipartite formation of the palatoquadrate; 2) precocious formation of the adult mandible; 3) origin of the angulosplenial from two centers of ossification; 4) complete erosion of the orbital cartilage during the later stages of metamorphosis; 5) development of the sphenethmoid as a membrane, rather than an endochondral bone; and 6) a pattern of timing of ossification that more closely coincides with that of the pelobatid frog Spea than that recorded for neobatrachian species.     

The morphology and distribution of Xenopus vestitus (Anura: Pipidae) in Central Africa

Studies on the collections of the Musée Royal de l'Afrique Centrale and the Institut Royal des Sciences Naturelles de Belgique have shown that Xenopus kigesiensis Tinsley, 1973 is synonymous with X. vestitus Laurent, 1972. Previously unrecognized representatives of the species have been found in the museum collections and these considerably enlarge the known geographical distribution of X. vestitus. Samples have been recorded principally from highland areas adjacent to the western Rift in Zaire, Uganda and Ruanda; comprehensive morphological analysis demonstrates relatively limited intraspecific variation throughout the known range.
The distribution of X. vestitus overlaps with that of three other Xenopus taxa: X. laevis victorianus, X. bunyoniensis and a new Xenopus species. There is evidence from a large number of collections from widely separated localities that X. vestitus is commonly sympatric with X. l. victorianus. On the other hand, it is postulated that X. vestitus has recently invaded Lake Mutanda, Uganda, and has replaced the original X. bunyoniensis population during the past 30 years. The museum record provides a potential origin of this invasion: X. vestitus populations are well documented at Rutshuru, only 25 km downstream from Lake Mutanda.
A comparison of the morphological characters of X. vestitus, X. l. victorianus and X. bunyoniensis is presented to provide information on the systematic relationships and possible ecological adaptations of the overlapping species.     

Übertragung von Verhaltensweisen durch Transplantation von Anlagen neuroanatomischer Strukturen bei Amphibienlarven: I. Xenoplastischer Austausch von Nachhirnanlagen zwischen Xenopus laevis und Hymenochirus boettgeri (Amphibia,Anura)1

By transplantation of parts of the neural plate (later medulla oblongata) from Xenopus laevis (Daud.) to Hymenochirus boettgeri (Torn.) larval chimaeras were obtained, which showed donorlike rhythmical movements of the mouth and pharyngeal region, movements which the host species lacks. Nevertheless, some of these tadpoles were still able to catch small prey in a reaction which is typical for the host and lacking in the donor.     

Evolution of pipoid frogs:intergeneric relationships of the aquatic frog family Pipidae (Anura)

The 27 species of the aquatic frog family Pipidae are currently arranged in four genera: Xenopus (15 species), Hymenochirus (four species), and the poorly known genus Pseudhymenochirus (one species) occur in Africa; Pipa (seven species) is found in South America and lower Central America. Despite extensive work on the biology of Xenopus from various disciplines, the evolutionary relationships of Xenopus to other pipids have not been resolved. Phylogenetic analyis of morphological features of pipid frogs indicates that, contrary to earlier opinions, Hymenochirus and Pipa are closest relatives (sister-groups); these genera are placed in the subfamily Pipinae. Also, the currently recognized species of Xenopus do not form a natural group; the species tropicalis and epitropicalis are more closely related to Hymenochirus + Pipa than to the remaining species of Xenopus . The two discordant species are transferred to the genus Silurana , which is relegated to the new subfamily Siluraninae; it is the sister-group of the Pipinae. The remaining species of Xenopus constitute a monophyletic group that is placed in the subfamily Xenopodinae as the sister-group of the other genera of pipids.     

Loss of FilaminC (FLNc) results in severe defects in myogenesis and myotube structure

FilaminC (FLNc) is the muscle-specific member of a family of actin binding proteins. Although it interacts with many proteins involved in muscular dystrophies, its unique role in muscle is poorly understood. To address this, two models were developed. First, FLNc expression was stably reduced in C2C12 myoblasts by RNA interference. While these cells start differentiation normally, they display defects in differentiation and fusion ability and ultimately form multinucleated "myoballs" rather than maintain elongated morphology. Second, a mouse model carrying a deletion of last 8 exons of Flnc was developed. FLNc-deficient mice die shortly after birth, due to respiratory failure, and have severely reduced birth weights, with fewer muscle fibers and primary myotubes, indicating defects in primary myogenesis. They exhibit variation in fiber size, fibers with centrally located nuclei, and some rounded fibers resembling the in vitro phenotype. The similarity of the phenotype of FLNc-deficient mice to the filamin-interacting TRIO null mice was further confirmed by comparing FLNc-deficient C2C12 cells to TRIO-deficient cells. These data provide the first evidence that FLNc has a crucial role in muscle development and maintenance of muscle structural integrity and suggest the presence of a TRIO-FLNc-dependent pathway in maintaining proper myotube structure.     

Glomerular development and growth of the renal blood vascular system in Xenopus laevis (Amphibia: Anura: Pipidae) during metamorphic climax.

Microcorrosion casts of the renal vascular system of tadpoles of the Clawed Frog, Xenopus laevis, were observed by scanning electron microscopy. Glomerular differentiation was studied qualitatively and quantitatively during developmental stages 56-66 (metamorphic climax). The general structure of the renal vascular system corresponds to the pattern commonly found in anurans; however, the arterial supply has conspicuous connecting vessels that supply groups of glomeruli. In the dorsal part of the kidney, qualitative differentiation of glomerular structures precedes quantitative growth. The ventral part of the kidney has larger, well-developed renal corpuscles of nearly adult appearance. Four developmental stages of glomerulogenesis are distinguished morphologically and their glomerular and vascular growth is analyzed.     

Speciation of Protopolystoma Bychowsky, 1957 (Monogenea: Polystomatidae) in hosts of the genus shape Xenopus (Anura: Pipidae)

The taxonomy, geographical distribution and hostrange of the polystomatid genus ProtopolystomaBychowsky, 1957 are reviewed. P. xenopodis(Price, 1943) and five new species are recognised,which occur in clawed toads ( Xenopus spp.)throughout subsaharan Africa. Of the two clawed toadsubgenera, Xenopus and Silurana, only theformer is infected. Protopolystoma spp. aredifferentiated by morphological variation of the gut,large hamulus and penis armature. P.xenopodis is found in Xenopus laevis subspeciesin South Africa, Transkei, Zimbabwe, DemocraticRepublic of Congo (D.R.C.), Rwanda, Uganda, Kenya andCameroon ( X. l. poweri and X. l.sudanensis are new host records). It also occurs inintroduced populations of X. l. laevis in theUnited States (southern California) and United Kingdom(South Wales). In subsaharan Africa the speciesdisplays significant, but continuous, geographicalvariation of penis spine size between southernpopulations in X. l. laevis and those in morenortherly host subspecies. Data on the natural hostrange of this parasite were complemented by anexperimental study of host-specificity in the southernform. This can produce patent infections in X.l. victorianus and X. gilli, but not X.wittei nor X. (Silurana) tropicalis. P.simplicis n. sp. is endemic to central and eastAfrican areas, infecting X. laevis subspecies ineastern D.R.C., Rwanda, Uganda and western Kenya, X. wittei-like hosts in eastern D.R.C., westernUganda, Rwanda and Burundi, X. vestitus inwestern Uganda and Xenopus sp. at Nairobi,Kenya. P. ramulosus n. sp. occurs in X.fraseri-like toads in eastern D.R.C. (Gabon andCameroon are also possible literature records), and P. fissilis n. sp. is found in X. fraseri-and X. wittei-like species in Cameroon andeastern D.R.C., and in southern Rwanda, respectively. Two Protopolystoma taxa are found in X.muelleri populations now suspected to representdistinct species: P. occidentalis n. sp. occursin X. muelleri (western form) in Ghana, Togo,Nigeria and Cameroon, while P. orientalis n. sp.is found in X. muelleri (eastern form) in SouthAfrica, Zimbabwe and Tanzania. The allopatricallydistributed species P. ramulosus, P.simplicis, P. occidentalis and P.orientalis form a relatively homogenous grouping withsome interspecific morphological overlap. These taxaare distinguished from P. xenopodis by penisspine morphology and from P. fissilis by hamulusroot form and aspects of gut morphology. Unidentified Protopolystoma sp. have been recorded in X. clivii in Ethiopia, X. fraseri aff. inCameroon and Xenopus sp. in Kenya and Tanzania. At some localities, single host species were infectedby two representatives of Protopolystoma. P. fissilis was recorded in eastern D.R.C. with P. ramulosus, with Protopolystoma sp. inCameroon in X. fraseri-like hosts and with P. simplicis in X. wittei-like hosts in Rwanda. P. xenopodis co-occurred with P. simplicisin X. laevis subspecies through central and eastAfrica.     

The Bearing of Filter Feeding on the Water Pumping Mechanism of Xenopus Tadpoles (Anura: Pipidae)

Abstract New findings on the functional anatomy of filter feeding have permitted some clarification of water flow through tadopoles. The dorsal velar apparatus projects downward from the roof of the buccopharynx and consists of three mucosal pads and three vanes on both sides of the midline. At least four functions may be ascribed to the dorsal velar apparatus. 1. Each pad and its complementary vane fit so flush into each of the three bilateral filter cavities that exhalent water must flow in thin sheets over the filter plates before its exit via the pharyngeal clefts. 2. Like hydrofoils, the vanes deflect water and prevent strong currents from disorganizing the mucous food cords. 3. When tadpoles cease water pumping, the plugging effect of the dorsal velar apparatus in the filter cavities helps to occlude the pharyngeal clefts. 4. Secretions of dorsal velar glands, especially on the surface of the velar pads, may assist the mucus of the ventral velum to entrap suspended food. Other findings concern the M3b muscle, which helps to elevate the buccopharyngeal floor during expiration; the lack of cilia, but presence of low glandular elevations on the ventral velum; the reduction of the exits from the pharyngeal clefts 2 and 3 into a single valvular exit controlled by the B2b and B2c muscles; and the probable absence of a mechanical feeding function for the oral tentacles.     

Characterization of insulin and atypically processed proglucagon-derived peptides from the surinam toad Pipa pipa (Anura:Pipidae)

Electrospray mass spectrometry was used to identify insulin, glucagon and two peptides related to glucagon-like peptide-1 (GLP-1) in an extract of the pancreas of the Surinam toad, Pipa pipa, a species belonging to the same family as the African clawed frog, Xenopus laevis. Purification and characterization of the peptides established the primary structure of Pipa insulin as A-chain: GIVEQCCHSS(10)CTLLQLETYC(20) N and B-Chain: FSNQR LCGSH(10) LVEALHLVCG(20) DRGFFYYPKA(30). This amino acid sequence contains several substitutions (B5 His --> Arg, B16 Tyr --> His, A12 Ser --> Thr, A14 Tyr--> Leu, A18Asn --> Thr) of residues that have otherwise been quite strongly conserved during vertebrate evolution. Pipa glucagon comprises 37 amino acid residues (HSQGTFTSDY(10) SKYLDSRRAQ(20) DFVQWLMNTK(30)QSGGLSS) and the 29 amino-acid-residue peptide was not identified in the extract. In Xenopus and mammalian preproglucagons, the glucagon-29 sequence is followed by Lys-Arg which functions as a recognition site for a prohormone convertase. We propose that a point mutation in the gene encoding Pipa preproglucagon has transformed the Lys(30)-Arg(31) processing site into Lys-Gln with the result that the site in no longer recognized by the processing enzyme. In contrast, Pipa GLP-32 and GLP-37 are of the same molecular size as the corresponding peptides from Xenopus.     

Patterns of spatial and temporal cranial muscle development in the African clawed frog, Xenopus laevis (Anura: Pipidae)

The African Clawed Toad, Xenopus laevis, has been a major vertebrate model organism for developmental studies for half a century. Because most studies have focused on the early stages of development, this has had the effect that many aspects of organogenesis and later development remain relatively poorly known in this species. In particular, little is known about cranial muscle development even at the level of morphology and histological differentiation of muscle anlagen and muscle fibers. In this study, we document the morphogenesis and histological differentiation of cranial muscles in X. laevis. We provide a detailed account of the timing of development for each of the cranial muscles, and also describe a new muscle, the m. transversus anterior. The cranial musculature of X. laevis larvae generally develops in a rostrocaudal sequence. The first muscles to differentiate are the extrinsic eye muscles. Muscles of the mandibular and hyoid arches develop almost simultaneously, and are followed by the muscles of the branchial arches and the larynx, and by the mm. geniohyoideus and rectus cervicis. Despite the fact that differentiation starts at different stages in the different muscles, most are fully developed at Stage 14. These baseline data on the timing of muscle differentiation in the X. laevis can serve as a foundation for comparative studies of heterochronic changes in cranial muscle development in frogs and other lissamphibians.     

The free swimming Pipa larvae,with a review of pipid larvae and pipid phylogeny (Anura: Pipidae)

This paper describes the morphology of the free swimming Pipa larvae, compares them with Xenopus, Hymenochirus, and to some extent, Rhinophrynus larvae, and presents a morphological diagnosis of pipid larvae. Pipa and Xenopus have very similar chondrocrania. Hymenochirus is superficially different but has the same diagnostic features. The differences appear related to its small size and predatory habitus. Other aspects of anatomy, especially the filter apparatus are very different in each genus. The filter apparatus of Pipa is somewhat reduced and seems modified for the retention of relatively large (20+microns) particles. Similar adaptations may have been annectant to predations in Hymenochirus, which lacks a filter apparatus. However, varying states of seven character complexes, which cut across the varying ecology, show that there are two basic pipid lineages, each currently confined to Africa or South America, respectively. Recent finds of fossil South American Xenopus indicate that these two lineages separated before the continents did. This does not warrant the recognition of two subfamilies because Xenopus and Hymenochirus are too different. Pseudhymenochirus is not an intermediate between them; it is a primitive Hymenochirus. Eight character states separate pipid and rhynophrynid larvae.     

Anomalous rates of evolution of pancreatic polypeptide and peptide tyrosine-tyrosine (PYY) in a tetraploid frog, Xenopus laevis (Anura:Pipidae)

The South African clawed frog Xenopus laevis is believed to have arisen as a result of a tetraploidization event occurring approximately 30 million years ago. Two molecular forms of pancreatic polypeptide (PP) have been isolated from an extract of the pancreas of this species and two molecular forms of peptide tyrosine-tyrosine (PYY) from the intestine. Despite the fact that the amino acid sequence of PP has, in general, been very poorly conserved during the evolution of tetrapods (only Pro(5), Pro(8), Gly(9), Ala(12), Tyr(27), Arg(33) and Arg(35) are invariant among species studied so far), the two Xenopus PPs differ by only a single amino acid substition (Asp(22)-->Glu). In contrast the two molecular forms of PYY differ by six amino acid substitutions (Glu(15)-->Gln, Thr(18)-->Ala, Leu(21)-->Met, Ile(22)-->Thr, Ile(28)-->Val, Val(31)-->Ile). The data imply that strong evolutionary pressure is acting to conserve the functional domain in both genes encoding PP and so suggest that PP may have an important physiological role in amphibians (although the nature of this role has yet to be determined). The more rapid mutation of the functional domain in the genes encoding PYY, a peptide whose amino acid sequence has been quite well conserved in tetrapods and whose physiological significance is well established, suggests that one of the PYY genes may be evolving towards a new function or towards becoming a pseudogene.     

The biology and systematics of a new species of Xenopus (Anura: Pipidae) from the highlands of Central Africa

A new species of clawed toad, Xenopus wittei sp. nov. is described from highland areas overlapping the borders of Rwanda, Uganda and Zaire. A comprehensive morphological analysis of the species has been carried out. X. wittei sp. nov. is very closely related to X. vestitus and both have a tetraploid chromosome complement (2 n = 72), the only examples within the genus. Nevertheless, the two species may be distinguished by easily observed features, including head morphology and dorsal body colours and patterns, and are also identified by their karyotypes and mating calls.
Data on distribution, ecology and parasite infection are derived from both museum collections and fieldwork in Central Africa. X. wittei sp. nov. is sympatric in parts of its range with X. vestitus, X. laevis victorianus and X. I. bunyoniensis , with up to three of the taxa sharing the same habitats. The implications of potential species interactions are discussed. Experimental hybridization has confirmed the reproductive isolation of the two tetraploid species. Parasitological studies show that they may be distinguished from one another by host specific species of the nematode Chitwoodchabaudia and from other Xenopus species by the absence of the tapeworm genus Cephalochlamys . Large scale population movements of the Xenopus species have apparently occurred in the Central African highlands during the period of available records. Sympatry observed amongst the present day representatives of Xenopus may have been an important factor in the evolution of these toads since X. wittei sp. nov. and X. vestitus are interpreted as allopolyploids, the products of interspecies hybridization.     

Evidence from peptidomic analysis of skin secretions that allopatric populations of Xenopus gilli (Anura:Pipidae) constitute distinct lineages

The International Union for Conservation of Nature (IUCN) Endangered Cape Platanna Xenopus gilli inhabits disjunct ranges at the tip of Cape Peninsula and near the town of Kleinmond on opposite sides of False Bay in the extreme southwest of Africa. Peptidomic analysis of host-defense peptides in norepinephrine-stimulated skin secretions from frogs from the Cape Peninsula range resulted in the identification of two magainins, two peptide glycine–leucine–amide (PGLa) peptides, two xenopsin-precursor fragment (XPF) peptides, nine caerulein-precursor fragment (CPF) peptides, and a peptide related to peptide glycine–glutamine (PGQ) previously found in an extract of Xenopus laevis stomach. The primary structures of the peptides indicate a close phylogenetic relationship between X. gilli and X. laevis but only magainin-1, PGLa and one CPF peptide are identical in both species. Consistent with previous data, the CPF peptides show the greatest antimicrobial potency but are hemolytic. There are appreciable differences in the expression of host-defense peptide genes in frogs from the population of animals sampled near Kleinmond as peptides corresponding to magainin-G2, XPF-G1, XPF-G2, and four CPF peptides, present in secretions from the Cape Peninsula frogs, were not identified in the skin secretions from Kleinmond frogs. Conversely, PGLa-G3, XPF-G3, and three CPF peptides were identified in the Kleinmond frogs but not in the Cape Peninsula animals. The data support the conclusion from morphometric analyses and comparisons of the nucleotide sequences of mitochondrial genes that the disjunct populations of X. gilli have undergone appreciable genetic, morphological, and phenotypic divergence.