Are paedomorphs actual larvae?

Amphibians display very diverse life cycles and development can be direct, where it occurs in ovo and a juvenile hatches directly, or biphasic, where an aquatic larva hatches and later undergoes metamorphosis followed by sexual maturation. In both cases, metamorphosis, corresponds to the post embryonic transition (PETr). A third strategy, only found in Urodeles, is more complex as larvae reach sexual maturity before metamorphosis, which can become accessory. The resulting paedomorphs retain their larval characters and keep their aquatic habitat. Does it mean that paedomorphs do not undergo PETr? Recent work using high throughput technologies coupled to system biology and developmental endocrinology revisited this question and provided novel datasets indicating that a paedomorph's “larval” tissue undergoes a proper developmental transition. Together with historical data, we propose that this transition is a marker of the PETr, which would be distinct from metamorphosis. This implies that (a) complex life cycles would result from the uncoupling of PETr and metamorphosis, and (b) biphasic life cycles would be a special cases where they occur simultaneously.     

Skull development in the Iberian newt,Pleurodeles waltl (Salamandridae: Caudata: Amphibia): timing,sequence, variations,and thyroid hormone mediation of bone appearance

The cranial ossification sequence in Pleurodeles waltl is widely used in phylogenetic analyses of amphibian origin and evolution. However, the patterns published to date are far from completely resolved and contain certain discrepancies. Based on a large sample of P. waltl specimens ranging from early post-hatching larvae to post-metamorphic newts, we determined the most common cranial ossification sequence and revealed its intraspecific variations. Since thyroid hormones (THs) are involved in the mediation of skull development in salamanders, we studied the role of THs in the cranial development of P. waltl. The normal sequence and timing of bone appearance were compared with those in larvae reared under conditions of high (in 1 and 2 ng mL^-1 triiodothyronine) and low [in 0.02% thiourea (TU), which inhibits thyroid gland activity] TH levels. Metamorphosis was greatly accelerated in the TH-treated larvae and was arrested in the TU-treated larvae, which retained the larval pattern of the palate and rudimentary external gills even after 2 years of the experiment. Early-appearing bones (the coronoid, vomer, palatine, dentary, squamosal, premaxilla, parasphenoid, pterygoid, prearticular, vomer, frontal, parietal, exoccipital, in this order) arise at the same stages and ages, and follow the same ossification sequence under different TH levels. The timing of the appearance of bones normally arising in the late larval and metamorphic periods (the quadratojugal, orbitosphenoid, prootic, maxilla, nasal, os thyroideum, prefrontal, quadrate, in this order) changes depending on the TH level. The maxilla and nasal display the most pronounced reaction to changes in the TH level: they appear precociously in TH-treated animals, while their appearance is postponed and they remain rudimentary in TU-treated animals. Because of different responses to THs, the order in which late-arising bones appear changes depending on the TH level. Although bones appearing early in larval ontogeny (e.g. the premaxilla, vomer, squamosal, palatine) display no TH-induced reaction when they start to develop, their further differentiation shows dependence on THs, and these bones become TH-inducible closer to metamorphosis. These findings indicate that TH involvement in the mediation of cranial development changes from minimal (if at all) in its early stages to maximal during metamorphosis. It is likely that the appearance of bones early in development is mediated by factors other than THs. Their further development is accompanied by changes in the mechanisms mediating their morphological differentiation. That is, likely non-hormonal mediation becomes replaced or/and complemented by hormonal mediation. The constituent parts of the same bone may exhibit differences in their reactions to changes in TH levels. Although in normal development, the overall cranial ossification sequence is constant, there was variation in the order in which late-appearing bones was recorded. These observations suggest that this variation results from individual variability in the internal TH level. Comparison with other salamanders suggests that (a) the pattern of TH mediation described in P. waltl is common for cranial development of metamorphosing urodeles and (b) the same bone may differ in its TH dependence in different salamanders, e.g. there are interspecific variations in the degree of TH dependence of individual cranial bones.     

Position discrimination in the salamander,Ambystoma tigrinum

Twenty larval tiger salamanders (Ambystoma tigrinum) were collected from a playa lake in West Texas and maintained in the laboratory for 7 days prior to the beginning of position training. Ten subjects in the larval state were trained on a position problem in a T-maze for 5 trials per day to a criterion of 80% correct responses, using a self-correction procedure. Following metamorphosis to the adult stage, these subjects were tested for retention. The remaining 10 subjects were allowed to metamorphose prior to training on the position problem. The results indicate reliably faster learning in the adult (terrestrial) subjects, and reliable retention of the response following metamorphosis. These results, taken in conjunction with informal observation of the behavior of the subjects in the experimental environment, suggest that the performance of the adult subjects was a result of their better perceptual apparatus.     

How thyroid hormones and their inhibitors affect cartilage growth and shape in the frog Xenopus laevis

Understanding how skeleton changes shape in ontogeny is fundamental to understanding how its shape diversifies in phylogeny. Amphibians pose a special case because their jaw and throat skeleton consists of cartilages that are dramatically reshaped midway through life to support new feeding and breathing styles. Although amphibian metamorphosis is commonly studied by immersing larvae in thyroid hormones (TH), how individual cartilages respond to TH is poorly understood. This study documents the effects of larval stage and TH type (T4 vs. T3), dose and deprivation on the size, shape and morphogenesis of the lower jaw and ceratohyal cartilages in the frog Xenopus laevis. It uses thyroid inhibitors to isolate the effects of each hormone at specific concentrations. It also deconstructs the TH responses into the effects on individual dimensions, and uses measures of percent change to eliminate the effects of body size and growth rate variation. As stage increases, T4 and T3 responses become increasingly similar to each other and to natural remodeling; the differences at low and intermediate stages result largely from abnormal responses to T3. Most notably, the beak‐like lower jaw commonly observed at the lowest stage in other studies results largely from arrested growth of cartilage. TH responses are superimposed upon the growth typical for each stage so that cartilages can attain postmetamorphic shapes through dimensional changes that exceed those of natural metamorphosis. Using thyroid inhibitors alters the outcome of TH‐induced remodeling, and T4 has almost the same capacity to induce metamorphic shape changes as T3. The results have implications for understanding how the starting shapes of larval elements affect morphogenesis, how chondrocytes behave to change cartilage shape, and how intracellular processing of TH might contribute to interspecific differences in shape change. Also, the data on animal mortality and which stages and doses most closely replicate natural remodeling have practical value for researchers who treat Xenopus tadpoles with TH.     

Localization of nitric oxide synthase-like immunoreactivity in the developing nervous system of the snail Ilyanassa obsoleta

Production of nitric oxide (NO), an evolutionarily conserved, intercellular signaling molecule, appears to be required for the maintenance of the larval state in the gastropod mollusc Ilyanassa obsoleta. Pharmacological inactivation of endogenous nitric oxide synthase (NOS), the enzyme that generates NO, can trigger metamorphosis in physiologically competent larvae of this species. Neuropils in the brains of these competent larvae display histochemical reactivity for NADPH diaphorase (NADPHd), an indication of neuronal NOS activity. The intensity of NADPHd staining is greatest in the neuropil of the apical ganglion (AG), a region of the brain that contains the apical sensory organ and that innervates the bilobed ciliated velum, the larval swimming and feeding organ. Once metamorphosis is initiated, the intensity of NADPHd staining in the AG and presumably, concomitant NO production, decline. The AG is finally lost by the end of larval metamorphosis, some 4 days after induction. To determine if the neurons of the AG are a source of larval NO, we conducted immunocytochemical studies on larval Ilyanassa with commercially available antibodies to mammalian neuronal NOS. We localized NOS-like immunoreactivity (NOS-IR) to 3 populations of cells in competent larvae: somata of the AG and putative sensory neurons in the edge of the mantle and foot. Immunocytochemistry on pre-competent larvae demonstrated that numbers of NOS-IR cells in the AG increase throughout the planktonic larval stage.     

Cranial Muscle Development in the Model Organism Ambystoma mexicanum: Implications for Tetrapod and Vertebrate Comparative and Evolutionary Morphology and Notes on Ontogeny and Phylogeny

There is still confusion about the homology of several cranial muscles in salamanders with those of other vertebrates. This is true, in part, because of the fact that many muscles present in early ontogeny of amphibians disappear during development and specifically during metamorphosis. Resolving this confusion is important for the understanding of the comparative and evolutionary morphology of vertebrates and tetrapods because amphibians are the phylogenetically most plesiomorphic tetrapods, concerning for example their myology, and include two often used model organisms, Xenopus laevis (anuran) and Ambystoma mexicanum (urodele). Here we provide the first detailed report of the cranial muscle development in axolotl from early ontogenetic stages to the adult stage. We describe different and complementary types of general muscle morphogenetic gradients in the head: from anterior to posterior, from lateral to medial, and from origin to insertion. Furthermore, even during the development of neotenic salamanders such as axolotls, various larval muscles become indistinct, contradicting the commonly accepted view that during ontogeny the tendency is mostly toward the differentiation of muscles. We provide an updated comparison between these muscles and the muscles of other vertebrates, a discussion of the homologies and evolution, and show that the order in which the muscles appear during axolotl ontogeny is in general similar to their appearance in phylogeny (e.g. differentiation of adductor mandibulae muscles from one anlage to four muscles), with only a few remarkable exceptions, as for example the dilatator laryngis that appears evolutionary later but in the development before the intermandibularis. Anat Rec, 296:1031–1048, 2013. © 2013 Wiley Periodicals, Inc.     

Organization of Foraging Behavior in Larvae of Cosmopolitan,Widespread, and Endemic <Emphasis Type="Italic">Drosophila</Emphasis> Species

To explore the evolution of higher order behavioral traits we investigated the organization of foraging patterns in first instar larvae of natural populations of Drosophila. We examined Drosophila simulans (cosmopolitan); D. mauritania (widespread); D. pavani, and D. gaucha (endemic). Larvae of these four species share the same behavior components that comprise foraging (locomotion, feeding, bending, and turning). D. simulans and D. mauritania larvae show uncoupled foraging patterns organized into partially independent behavioral elements. Larvae of D. pavani and D. gaucha exhibit coupled foraging behaviors based on a dependency between behavioral components. Hybrid larvae obtained from crosses of natural populations of D. simulans and D. mauritania show an organization of foraging patterns similar to that of the parental lines. In contrast, hybridization disrupts the organization of foraging patterns in D. pavani and D. gaucha intra- and inter-specific hybrid larvae. This suggests genetic co-adaptation for linkage between the behavioral components that comprise foraging. The organization of larval foraging patterns of the endemic species D. pavani seems readily affected by hybridization. The absence of linkage between behavioral components, as in the case of larval foraging patterns of D. simulans and D. mauritania could lead to an increase in the variability of organization of this higher order behavior. The possibility that larvae may use a variable and flexible behavioral integration of foraging patterns could contribute to their development and feeding in a diversity of substrates and climates. Edited by Yong-Kyu Kim     

Wide-scale application of Bti/Bs biolarvicide in different aquatic habitat types in urban and peri-urban Malindi,Kenya

Larval control is a major component in mosquito control programs. This study evaluated the wide-scale application of Bti/Bs biolarvicide (Bacillus thuringiensis var. israelensis [Bti] and Bacillus sphaericus [Bs]) in different aquatic habitats in urban and peri-urban Malindi, Kenya. This study was done from June 2006 to December 2007. The urban and peri-urban area of Malindi town was mapped and categorized in grid cells of 1 km². A total of 16 1-km² cells were selected based on presence Community Based Organization dealing with malaria control within the cells. Each of the 16 1-km² cells was thoroughly searched for the presence of potential larval habitats. All habitats, whether positive or negative for larvae, were treated and rechecked 24 h (1 day), 6 days, and 10 days later for the efficacy of Bti/Bs. Weekly larval sampling was done to determine the mosquito larval dynamics in the aquatic habitats during the study period. Morphological identification of the mosquito larvae showed that Anopheles gambiae s.l. Giles was the most predominant species of the Anopheles and while in the culicines, Cx. quinquefasciatus Say was the predominant species. Anopheles larvae were all eliminated in habitats within a day post-application. For culicine larvae, 38.1% (n = 8) of the habitat types responded within day 1 post-treatment and all the larvae were killed, they turned negative during the days of follow-up. Another 38.1% (n = 8) of the habitat types had culicine larvae but turned negative by day 6, while three habitats (14.3%) had larvae by 6th day but turned negative by 10th day. However during this Bti/Bs application studies, two habitat types, house drainage and cesspits (9.5%), remained positive during the follow-up although the mosquito larvae were significantly reduced. Both early and late instars of Anopheles larvae immediately responded to Bti/Bs application and reached 100% mortality. The early and late instars of culicine responded to the Bti/Bs application but not as fast as the Anopheles larval instars. The early instars Culex, responded with 90.8% mortality at day 1 post-treatment, and the mortality was 99.9% at day 10. Similarly, the late instars Culex followed the same trend and exhibited same mortalities. The weekly sampling in the aquatic habitats showed that there was a 36.3% mosquito larval reduction in the aquatic habitats over the 18-months study period. In conclusion, Bti/Bs biolarvicide are useful in reducing the mosquito larval densities in a wide range of habitats which have a direct impact of adult mosquito populations.     

Study of the relationship between thyroid hormones and lipid metabolism during KClO4-induced metamorphosis of landlocked lamprey, Petromyzon marinus.

This study examines the role of thyroid hormones (TH) (thyroxine and triiodothyronine) in regulating lipid metabolism of landlocked larval sea lampreys, Petromyzon marinus. Larvae were treated with either thyroxine (0.5 or 1 mg l(-1) water) or triiodothyronine (0.25 or 1 mg l(-1) water) in the presence or absence of the goitrogen, potassium perchlorate (KClO4) (0.05% w/v), for 4, 8, and 16 weeks. Treatment with KClO4 alone, which induced metamorphosis after 8 weeks and lowered plasma TH levels, reduced hepatic and renal total lipid content after 8 weeks of treatment. KClO4-induced lipid depletion after the 8-week treatment was supported by an increased rate of hepatic lipolysis, as indicated by increased triacylglycerol lipase activity. Furthermore, reduced lipogenesis in the liver was indicated by decreased hepatic acetyl-CoA carboxylase and diacylglycerol acyltransferase (DGAT) activities, and by decreased renal DGAT activity following 8 weeks of KClO4 treatment. Treatment of larvae for 4 weeks with TH alone resulted in either no change or a slight increase of lipid in the liver and kidney. TH treatments in combination with KClO4 failed to induce metamorphosis and, after up to 8 weeks, several TH treatments blocked changes in lipid content and enzyme activity associated with KClO4-induced metamorphosis. These experimental results suggest that TH deficiency during metamorphosis may promote lipid catabolism, while the presence of TH tends to protect/promote lipid reserves, perhaps favoring the larval condition. The actions of TH and KClO4 on metamorphosis-associated lipid metabolism in sea lampreys may be direct, permissive, and/or indirect via other factors.     

Nonreproductive role of gonadotropin‐releasing hormone in the control of ascidian metamorphosis

Background: Gonadotropin‐releasing hormones (GnRHs) are neuropeptides that play central roles in the reproduction of vertebrates. In the ascidian Ciona intestinalis, GnRHs and their receptors are expressed in the nervous systems at the larval stage, when animals are not yet capable of reproduction, suggesting that the hormones have non‐reproductive roles. Results: We showed that GnRHs in Ciona are involved in the animal′s metamorphosis by regulating tail absorption and adult organ growth. Absorption of the larval tail and growth of the adult organs are two major events in the metamorphosis of ascidians. When larvae were treated with GnRHs, they completed tail absorption more frequently than control larvae. cAMP was suggested to be a second messenger for the induction of tail absorption by GnRHs. tGnRH‐3 and tGnRH‐5 (the “t” indicates “tunicate”) inhibited the growth of adult organs by arresting cell cycle progression in parallel with the promotion of tail absorption. Conclusions: This study provides new insights into the molecular mechanisms of ascidian metamorphosis conducted by non‐reproductive GnRHs. Developmental Dynamics 243:1524–1535, 2014. © 2014 Wiley Periodicals, Inc.     

Thyroid and pituitary gland development from hatching through metamorphosis of a teleost flatfish, the Atlantic halibut

Fish larval development, not least the spectacular process of flatfish metamorphosis, appears to be under complex endocrine control, many aspects of which are still not fully elucidated. In order to obtain data on the functional development of two major endocrine glands, the pituitary and the thyroid, during flatfish metamorphosis, histology, immunohistochemistry and in situ hybridization techniques were applied on larvae of the Atlantic halibut (Hippoglossus hippoglossus), a large, marine flatfish species, from hatching through metamorphosis. The material was obtained from a commercial hatchery. Larval age is defined as day-degrees (D°=accumulated daily temperature from hatching). Sporadic thyroid follicles are first detected in larvae at 142 D° (27 days post-hatch), prior to the completion of yolk sack absorption. Both the number and activity of the follicles increase markedly after yolk sack absorption and continue to do so during subsequent development. The larval triiodothyronine (T₃) and thyroxine (T₄) content increases, subsequent to yolk absorption, and coincides with the proliferation of thyroid follicles. A second increase of both T₃ and T₄ occurs around the start of metamorphosis and the T₃ content further increases at the metamorphic climax. Overall, the T₃ content is lower than T₄. The pituitary gland can first be distinguished as a separate organ at the yolk sack stage. During subsequent development, the gland becomes more elongated and differentiates into neurohypophysis (NH), pars distalis (PD) and pars intermedia (PI). The first sporadic endocrine pituitary cells are observed at the yolk sack stage, somatotrophs (growth hormone producing cells) and somatolactotrophs (somatolactin producing cells) are first observed at 121 D° (23 days post-hatch), and lactotrophs (prolactin producing cells) at 134 D° (25 days post-hatch). Scarce thyrotrophs are evident after detection of the first thyroid follicles (142 D°), but coincident with a phase in which follicle number and activity increase (260 D°). The somatotrophs are clustered in the medium ventral region of the PD, lactotrophs in the anterior part of the PD and somatolactotrophs are scattered in the mid and posterior region of the pituitary. At around 600 D°, coinciding with the start of metamorphosis, somatolactotrophs are restricted to the interdigitating tissue of the NH. During larval development, the pituitary endocrine cells become more numerous. The present data on thyroid development support the notion that thyroid hormones may play a significant role in Atlantic halibut metamorphosis. The time of appearance and the subsequent proliferation of pituitary somatotrophs, lactotrophs, somatolactotrophs and thyrotrophs indicate at which stages of larval development and metamorphosis these endocrine cells may start to play active regulatory roles.     

Envirnmental conditions prerequisite for complete limb regeneration in the postmetamorphic adult land-phase salamander,Ambystoma

Historically, postmetamorphic adult land-phase salamanders have been shown to exhibit minimal to nonexistent limb regeneration. Hence, it has been generally accepted that these forms have lost the intrinsic capacity to regenerate a limb. Due to the experimental protocols used, an alternate explanation is also possible: that this intrinsic capacity cannot be expressed when the salamanders are maintained under adverse laboratory environmental conditions. Therefore, this study addresses two questions: (1) What are the optimal environmental conditions for long-term survival of adult land-phase salamanders; and (2) will complete limb regeneration occur in these salamanders if they are maintained under survival conditions. A mixed population of adult Ambystoma were tested under varying conditions of habitat, temperature, humidity, photoperiod, and food source. Complete limb regeneration was possible in 100% of four species of adult postmetamorphic land-phase Ambystoma salamanders given the proper environmental laboratory conditiions of a peat moss and potting soil habitat with a controlled temprature of 25°C ± 5°C, 70% or greater humidity, a 12/12 light/dark photoperiod, a diet including nightcrawlers released into their respective terraria, and an extended observation time of up to 370 days postamputation (dpa). Regeneration was completed during the following range periods for the adult salamanders: A. annulatum, 324 to 370 dpa; A. maculatum, 255 to 300 dpa; A. texanum, 215 to 250 dpa; and A. tigranum, 155 to 180 dpa.     

Expression of Class II Major Histocompatibility Complex Antigens on Adult T Cells in Xenopus is Metamorphosis- Dependent

Class II major histocompatibility complex (MHC) antigens are expressed predominantly on B lymphocytes and macrophages of tadpoles of the South African clawed frog, Xenopus laevis, as is the pattern in lymphocyte populations of most mammals. However, unlike most mammals, young postmetamorphic frogs show expression of class II MHC antigens on a high proportion of thymocytes and most peripheral T and B lymphocytes. Using the J-strain of Xenopus and the anticlass II monoclonal antibody, 14A2, we have studied, by indirect immunofluorescence, whether inhibition of metamorphosis would alter the pattern of expression of class II antigens during ontogeny. In control animals, class II antigens were virtually absent from thymic lymphocytes and peripheral T cells of normal untreated larvae, but could be found in increasing numbers in both populations after metamorphosis (10-12 weeks of age). In contrast, larvae, whose metamorphosis was inhibited by treatment with sodium perchlorate, had relatively few class II⁺ thymic lymphocytes throughout the 6-month period of study, and the proportion of class II⁺ splenic lymphocytes was approximately equal to that of IgM⁺ B lymphocytes. Thus, perchlorate-treated animals retained the larval pattern of class II epression, suggesting that emergence of class II⁺ T cells is dependent on metamorphosis.     

Gross morphological analysis of limb regeneration in postmetamorphic adult Ambystoma

Due to the great disparity between regeneration times for the larval salamander (40 days), axolotl (30+ days), newt (44 days), and adult salamander (155 to 370 days), a staging system was devised so correlative comparisons could be made between regenerative model systems. The sequence was based on two criteria: (1) the stages should be similar to previously reported sequences for the newt, axolotl, and larval salamander, and (2) the stages must be readily recognizable by examination of the external morphology in the living state. Postmetamorphic adult land-phase Ambystoma were amputated through the forearm, placed within survival conditions, and observed until regeneration was completed. Of the initial 160 salamanders, 157 (98%) progressed through 11 clearly defined stages of regeneration. A side-by-side comparison of the staging sequence for land and aquatic phase urodeles is given along with a summary of key external morphological characteristics for the adult salamander. It was noted that as the length of time for regeneration decreased, the relative ratio of the nerves innervating the limb (spinal nerves 4, 5, and 6) increased for the four species of Ambystoma examined: A. annulatum, 324 to 370 days postamputation (dpa) with a 1:1:1 neural tissue ratio; A. maculatum, 255 to 300 dpa with a 2:2:1 ratio; A. texanum, 215 to 250 dpa with a 2:2:2 ratio; and A. tigranum, 155 to 180 dpa with a 2:3:3 ratio.     

Die spektrale empfindlichkeit vonTriturus alpestris (Amphibia,Urodela) während des Wasser-und landlebens

Summary Spectral sensitivity of dark adapted eyes from aquatic and terrestrial animals ofTriturus alpestris was determined using the ERG. Spectral sensitivity changes with transition in a different habitat during metamorphosis and in adult animals in a seasonal course. In aquatic animals (larvae and adults in spring) data agree with the absorption curve for porphyropsin P 523₂. In juvenile and adult terrestrial animals in autumn data fit the absorption curve for rhodopsin P 502₁.

Mit Unterstützung des österreichischen Fonds zur Förderung der wissenschaftlichen Forschung.     

Evaluation of time toxicity, residual effect, and growth-inhibiting property of Carapa guianensis and Copaifera sp. in Aedes aegypti

Oils of Carapa guianensis and Copaifera spp. are well-known in the Amazonian region as natural insect repellents, and studies have reported their efficiency as larvicide against some mosquito species. However, toxicity persistence and effect on mosquito development have not yet been evaluated. Thus, the objective of this study was to evaluate the initial time of larvicidal activity, residual effect, and the effect of very low concentrations of these oils on Aedes aegypti. Different concentrations of the oils were used to evaluate the initial time of larval mortality and residual effect, as well as, the development of larvae, pupae, and adults. Results demonstrated that the lethal effect started mainly between the first 2 and 3 h of larvae exposure to oils, when using concentrations which ranged from 500 mg/L of C. guianensis and 90 mg/L of Copaifera sp. The toxic effect remained with total efficiency (100% mortality) until the sixth day for Copaifera sp. and 12th day for C. guianensis. When using sublethal dosages (ranging from 140 mg/L of C. guianensis to 26 mg/L of Copaifera sp.) mortality was observed after the larval molt. Also, imperfection of pupae and adult development and unsuccessful emergence of adults were observed. A product of botanical origin that could break the development of immature stage of mosquitoes and inhibit the emergence of adults should be essential in vector control. Thus, our results provide new information for a better understanding in using C. guianensis and Copaifera sp. oils with a potential to be used as a natural insecticide.     

Platelet-derived growth factor signaling as a cue of the epithelial-mesenchymal interaction required for anuran skin metamorphosis.

The anuran remodels the larval skin into the adult counterpart during metamorphosis. The construction of the precursor of adult epidermis (preadult epidermis) in Xenopus laevis larvae was coordinated with the development of the secondary connective tissue (s-ct) underneath the basement membrane, suggesting that the epithelial-mesenchymal interaction plays a critical role in the metamorphic conversion of the larval skin. mRNAs of platelet-derived growth factor A (PDGF-A) and PDGF receptor (PDGFR) -alpha were markedly up-regulated in the skin during spontaneous and thyroid hormone (TH) -induced metamorphosis. In situ hybridization experiments identified preadult epidermal basal cells and fibroblasts in developing subepidermal connective tissues at the late prometamorphic stage as PDGF-A and PDGFR-alpha mRNA-expressing cells, respectively. We developed an in vitro model of larval skin that was remodeled to the adult skin under the influence of TH. The presence of either of AG1296, a specific inhibitor of PDGFR tyrosine kinase autophosphorylation, or an excess of recombinant proteins of the soluble extracellular domain of PDGFR-alpha inhibited the following TH-induced processes, the proliferation of adult basal cells, the terminal differentiation of adult basal cells, and the activation of subepidermal fibroblasts. However, the inhibitors did not inhibit the TH-induced proliferation of preadult basal cells. We concluded that PDGF/PDGFR signaling is one of the prime cues in the epithelial-mesenchymal interaction required for the metamorphic skin remodeling.