Tadpole Tail Muscle Research Articles

Effects of lithium and selenium in the tail muscle of American bullfrog tadpoles (Lithobates catesbeianus) during premetamorphosis

The amphibian populations have faced a drastic decline over the past decades. This decline has been associated with the presence of contaminants in the environment, among other environmental stressors. The present study tested the responses following the exposure to lithium (2.5 mg L-1) and selenium (10μg L-1), both isolated and as a mixture, on the metabolic status of the tail muscle of premetamorphic American bullfrog (Lithobates catesbeianus) through the assessment of the total protein content, mobilization of glucose and triglycerides, and the activity of lactate dehydrogenase (LDH). The exposure followed a 21-day assay with two sampling periods (on the 7th and 21st day after the onset of exposure) to evaluate the effects over time. The group exposed to the mixture presented a statistically decreased LDH activity (P < 0.05) in both sampling periods. The presence of selenium elicited a statistically significant increase (P < 0.05) in the glucose mobilization after 7 days of exposure. After 21 days, the animals exposed to selenium presented levels of glucose mobilization comparable to the control group. The mobilization of glucose and triglycerides remained similar to the control group for the animals exposed to lithium and to the mixture in both periods of sampling (P > 0.05). The total protein content did not show any statistical difference in the treated groups throughout the experiment (P > 0.05). The presented results highlight the importance of the assessment of mixtures that can occur in the environment, since the combination of contaminants may elicit distinct toxicity compared with the effects triggered by the chemicals isolated.

In Vivo Transfection of Naked DNA into Xenopus Tadpole Tail Muscle.

In vivo gene transfer systems are important to study foreign gene expression and promoter regulation in an organism, with the benefit of exploring this in an integrated environment. Direct injection of plasmids encoding exogenous promoters and genes into muscle has numerous advantages: the protocol is easy, efficient, and shows time-persistent plasmid expression in transfected muscular cells. After injecting naked-DNA plasmids into tadpole tail muscle, transgene expression is strong, reproducible, and correlates with the amount of DNA injected. Moreover, expression is stable as long as the tadpoles remain, or are maintained, in premetamorphic stages. By directly expressing genes and regulated promoters in Xenopus tadpole muscle in vivo, one can exploit the powerful experimental advantages of gene transfer systems in an intact, physiologically normal animal.

Insulin-like growth factor 1 regulation of proliferation and differentiation of Xenopus laevis myogenic cells in vitro.

To understand the mechanism of muscle remodeling during Xenopus laevis metamorphosis, we examined the in vitro effect of insulin-like growth factor 1 (IGF-1) on growth and differentiation of three different-fate myogenic cell populations: tadpole tail, tadpole dorsal, and young adult leg muscle. IGF-1 promoted growth and differentiation of both tail and leg myogenic cells only under conditions where these cells could proliferate. Inhibition of cell proliferation by DNA synthesis inhibitor cytosine arabinoside completely canceled the IGF-1's cell differentiation promotion, suggesting the possibility that IGF-1's differentiation-promotion effect is an indirect effect via IGF-1's cell proliferation promotion. IGF-1 promoted differentiation dose dependently with maximum effect at 100-500ng/ml. RT-PCR analysis revealed the upregulation (11-fold) of ifg1 mRNA expression in developing limbs, suggesting that IGF-1 plays a role in promoting muscle differentiation during limb development. The combined effect of triiodo-L-thyronine (T3) and IGF-1 was also examined. In adult leg cells, IGF-1 promoted growth and differentiation irrespective of the presence of T3. In larval tail cells, cell count was 76% lower in the presence of T3, and IGF-1 did not promote proliferation and differentiation in T3-containing medium. In larval dorsal cells, cell count was also lower in the presence of T3, but IGF-1 enhanced proliferation and differentiation in T3-containing medium. This result is likely due to the presence among dorsal cells of both adult and larval types (1:1). Thus, IGF-1 affects only adult-type myogenic cells in the presence of T3 and helps accelerate dorsal muscle remodeling during metamorphosis.

Structural origin of the drastic modification of second harmonic generation intensity pattern occurring in tail muscles of climax stages xenopus tadpoles.

Second harmonic generation (SHG) microscopy is a powerful tool for studying submicron architecture of muscles tissues. Using this technique, we show that the canonical single frequency sarcomeric SHG intensity pattern (SHG-IP) of premetamorphic xenopus tadpole tail muscles is converted to double frequency (2f) sarcomeric SHG-IP in metamorphic climax stages due to massive physiological muscle proteolysis. This conversion was found to rise from 7% in premetamorphic muscles to about 97% in fragmented muscular apoptotic bodies. Moreover a 66% conversion was also found in non-fragmented metamorphic tail muscles. Also, a strong correlation between predominant 2f sarcomeric SHG-IPs and myofibrillar misalignment is established with electron microscopy. Experimental and theoretical results demonstrate the higher sensitivity and the supra resolution power of SHG microscopy over TPEF to reveal 3D myofibrillar misalignment. From this study, we suggest that 2f sarcomeric SHG-IP could be used as signature of triad defect and disruption of excitation-contraction coupling. As the mechanism of muscle proteolysis is similar to that found in mdx mouse muscles, we further suggest that xenopus tadpole tail resorption at climax stages could be used as an alternative or complementary model of Duchene muscular dystrophy.

Non-viral Expression of Mouse Oct4, Sox2, and Klf4 Transcription Factors Efficiently Reprograms Tadpole Muscle Fibers in Vivo

Adult mammalian cells can be reprogrammed into induced pluripotent stem cells (iPSCs) by a limited combination of transcription factors. To date, most current iPSC generation protocols rely on viral vector usage in vitro, using cells removed from their physiological context. Such protocols are hindered by low derivation efficiency and risks associated with genome modifications of reprogrammed cells. Here, we reprogrammed cells in an in vivo context using non-viral somatic transgenesis in Xenopus tadpole tail muscle, a setting that provides long term expression of non-integrated transgenes in vivo. Expression of mouse mOct4, mSox2, and mKlf4 (OSK) led rapidly and reliably to formation of proliferating cell clusters. These clusters displayed the principal hallmarks of pluripotency: alkaline phosphatase activity, up-regulation of key epigenetic and chromatin remodeling markers, and reexpression of endogenous pluripotent markers. Furthermore, these clusters were capable of differentiating into derivatives of the three germ layers in vitro and into neurons and muscle fibers in vivo. As in situ reprogramming occurs along with muscle tissue repair, the data provide a link between these two processes and suggest that they act synergistically. Notably, every OSK injection resulted in cluster formation. We conclude that reprogramming is achievable in an anamniote model and propose that in vivo approaches could provide rapid and efficient alternative for non-viral iPSC production. The work opens new perspectives in basic stem cell research and in the longer term prospect of regenerative medicine protocols development.

The cellular basis for phenotypic plasticity of body size in Western Spadefoot toad (Spea hammondi) tadpoles: patterns of cell growth and recruitment in response to food and temperature manipulations

Body size is highly variable within and among populations, both as a result of genetic variation and as a plastic response to environmental variation. From a proximate perspective, body size depends upon cell size, cell number, and extracellular matrix, but we know little about their independent contributions to size nor how these contributions vary with environmental influences. Here, I introduce the tail muscle of anuran tadpoles as a new system for studying this issue. Body size and tail size of tadpoles is sensitive to variation in food and temperature. I show first that tail muscle size is strongly correlated with overall body size, thus making it a good tissue to study size regulation. Second, the relative role of cell size and cell number, but not extracellular matrix, shows an interaction between food and temperature treatments and across ages. For example, in young tadpoles food effects on size are due exclusively to cell size at low temperatures but both cell size and number at high temperatures. This pattern partially reverses for older tadpoles. Despite the complexity of this interaction, the two populations compared show nearly identical patterns, suggesting that the plastic response is robust.

CDNA cloning and functional characterization of Xenopus laevis DNase γ

We report here the cDNA cloning and functional analysis of Xenopus DNase gamma (xDNase gamma). Two forms of cDNAs are isolated from adult spleen: one composing a 933 bp open reading frame for the enzymatically active xDNase gamma protein, and the other encoding an inactive short alternative form. Northern blot analysis revealed that the xDNase gamma mRNA is expressed in spleen, liver, testis, and ovary. xDNase gamma expression is scarcely detected in the tail muscle of tadpoles; however, it increases during metamorphosis and reaches a maximum during the late metamorphic climax. The ectopic expression of xDNase gamma results in the appearance of extensive DNA fragmentation in C2C12 myoblasts after the induction of apoptosis. In contrast, Xenopus DNase I fails to induce apoptotic DNA ladder formation under the same conditions. Our results suggest a possible involvement of xDNase gamma in apoptosis during amphibian metamorphosis.

Nuclear receptor corepressor recruitment by unliganded thyroid hormone receptor in gene repression during Xenopus laevis development.

Thyroid hormone receptors (TR) act as activators of transcription in the presence of the thyroid hormone (T(3)) and as repressors in its absence. While many in vitro approaches have been used to study the molecular mechanisms of TR action, their physiological relevance has not been addressed. Here we investigate how TR regulates gene expression during vertebrate postembryonic development by using T(3)-dependent amphibian metamorphosis as a model. Earlier studies suggest that TR acts as a repressor during premetamorphosis when T(3) is absent. We hypothesize that corepressor complexes containing the nuclear receptor corepressor (N-CoR) are key factors in this TR-dependent gene repression, which is important for premetamorphic tadpole growth. To test this hypothesis, we isolated Xenopus laevis N-CoR (xN-CoR) and showed that it was present in pre- and metamorphic tadpoles. Using a chromatin immunoprecipitation assay, we demonstrated that xN-CoR was recruited to the promoters of T(3) response genes during premetamorphosis and released upon T(3) treatment, accompanied by a local increase in histone acetylation. Furthermore, overexpression of a dominant-negative N-CoR in tadpole tail muscle led to increased transcription from a T(3)-dependent promoter. Our data indicate that N-CoR is recruited by unliganded TR to repress target gene expression during premetamorphic animal growth, an important process that prepares the tadpole for metamorphosis.

Larval-to-adult conversion of a myogenic system in the frog, Xenopus laevis, by larval-type myoblast-specific control of cell division, cell differentiation, and programmed cell death by triiodo-L-thyronine

Abstract For the clarification of larval-to-adult muscle conversion, the authors established primary culture methods for adult- and larval-type myoblasts in the frog, Xenopus laevis, and examined the hormonal response in each case. The cell types were enzymatically dissociated from adult frog leg and tadpole tail muscles, respectively. The cells became attached to culture plates, proliferated, and fused with each other to form multinucleated myotubes within one week. Five significant differences between the two cell types were noted. (1) Adult cells showed greater proliferation activity than larval cells, the former increasing 5.5-fold over 6 days while the latter increase only 2.5-fold. (2) Differentiation (fusion) of larval type myoblasts started earlier. Cell fusion began on day 2 or 3 in larval cells and on day 4 in adult cells. (3) The metamorphic hormone, triiodo-L-thyronine (T3) decreased larval cell numbers to 56 % of that of control-cultures on day 7 but had no effect on adult cell number. DNA synthetic activity (3H-thymidine incorporation) in larval cells decreased under T3 (10− 8M) to 45 % of the control level on day 7. (4) Differentiation of adult myoblasts into myotubes was promoted by T3, whereas that of larval cells diminished by half. (5) Myotube death was induced by T3 specifically in larval but not in adult cultures. In addition to the myotube death, double staining with TUNEL (in situ DNA nick end labeling) and anti-desmin antibody indicated that T3 induces myoblast (desmin+ cell) death specifically in larval but not in adult cells. It is thus evident that the conversion of a larval-type myogenic system during metamorphosis becomes possible through nearly totally specific control of cell division, cell differentiation, and programmed cell death at a precursor cell level by T3.

Larval-to-adult conversion of a myogenic system in the frog, Xenopus laevis, by larval-type myoblast-specific control of cell division, cell differentiation, and programmed cell death by triiodo-L-thyronine

For the clarification of larval-to-adult muscle conversion, the authors established primary culture methods for adult- and larval-type myoblasts in the frog, Xenopus laevis, and examined the hormonal response in each case. The cell types were enzymatically dissociated from adult frog leg and tadpole tail muscles, respectively. The cells became attached to culture plates, proliferated, and fused with each other to form multinucleated myotubes within one week. Five significant differences between the two cell types were noted. (1) Adult cells showed greater proliferation activity than larval cells, the former increasing 5.5-fold over 6 days while the latter increase only 2.5-fold. (2) Differentiation (fusion) of larval type myoblasts started earlier. Cell fusion began on day 2 or 3 in larval cells and on day 4 in adult cells. (3) The metamorphic hormone, triiodo-L-thyronine (T 3) decreased larval cell numbers to 56 % of that of control-cultures on day 7 but had no effect on adult cell number. DNA synthetic activity ( 3H-thymidine incorporation) in larval cells decreased under T 3 (10 − 8 M) to 45 % of the control level on day 7. (4) Differentiation of adult myoblasts into myotubes was promoted by T 3, whereas that of larval cells diminished by half. (5) Myotube death was induced by T 3 specifically in larval but not in adult cultures. In addition to the myotube death, double staining with TUNEL (in situ DNA nick end labeling) and anti-desmin antibody indicated that T 3 induces myoblast (desmin + cell) death specifically in larval but not in adult cells. It is thus evident that the conversion of a larval-type myogenic system during metamorphosis becomes possible through nearly totally specific control of cell division, cell differentiation, and programmed cell death at a precursor cell level by T 3.

Postembryonic expression of the myosin heavy chain genes in the limb, tail, and heart muscles of metamorphosing amphibian tadpoles

Thyroid hormone is presumed to play a role in initiating and/or orchestrating the postembryonic expression of the genes encoding isoforms of the myosin heavy chains (MHCs) that characterize the muscle fibres in an adult organism. The fact that the postembryonic development of a free-living amphibian tadpole takes place during its thyroid hormone-dependent metamorphosis has made the metamorphosing tadpole an ideal system for elucidating the molecular mechanism(s) by which this hormone affects these postembryonic changes. In this review, we summarize the results from recent studies focused on the postembryonic expression of the MHC genes in the skeletal muscles and hearts of metamorphosing anuran (Rana catesbeiana) tadpoles. The demonstration that mRNAs encoding at least five of the MHC isoforms present in the tadpole tail muscles are also present in the adult hind-limb muscles and that an mRNA encoding a cardiac-specific MHC isoform is present in the heart of both the tadpole and adult organism, rules out the possibility that thyroid hormone initiates the expression of these MHC genes. Instead, it seems more likely that this hormone acts by modulating the expression of one or more of the genes encoding these particular MHC isoforms. Whatever the case, the fact that sequence homology suggests that the five distinct skeletal muscle-specific MHCs are all “fast” isoforms raises the question of how these MHCs are distributed among the three different fibre types described for Rana. On the other hand, the possibility exists that the mRNAs for one or more of these fast MHC isoforms encode developmental isoforms that are present but not translated in the muscles of the tadpole and/or adult frog. Finally, an evaluation of the evolutionary relatedness of the R. catesbeiana MHCs to the MHCs in another species of Rana and to the MHCs in other vertebrates discloses, among other things, that the nucleotide sequence in the R. catesbeiana cardiac MHC isoform is more closely related to the chicken ventricular MHC isoform than it is to any of the other MHC isoforms examined. Microsc. Res. Tech. 50:458–472, 2000. © 2000 Wiley-Liss, Inc.

Postembryonic expression of the myosin heavy chain genes in the limb, tail, and heart muscles of metamorphosing amphibian tadpoles.

Thyroid hormone is presumed to play a role in initiating and/or orchestrating the postembryonic expression of the genes encoding isoforms of the myosin heavy chains (MHCs) that characterize the muscle fibres in an adult organism. The fact that the postembryonic development of a free-living amphibian tadpole takes place during its thyroid hormone-dependent metamorphosis has made the metamorphosing tadpole an ideal system for elucidating the molecular mechanism(s) by which this hormone affects these postembryonic changes. In this review, we summarize the results from recent studies focused on the postembryonic expression of the MHC genes in the skeletal muscles and hearts of metamorphosing anuran (Rana catesbeiana) tadpoles. The demonstration that mRNAs encoding at least five of the MHC isoforms present in the tadpole tail muscles are also present in the adult hind-limb muscles and that an mRNA encoding a cardiac-specific MHC isoform is present in the heart of both the tadpole and adult organism, rules out the possibility that thyroid hormone initiates the expression of these MHC genes. Instead, it seems more likely that this hormone acts by modulating the expression of one or more of the genes encoding these particular MHC isoforms. Whatever the case, the fact that sequence homology suggests that the five distinct skeletal muscle-specific MHCs are all "fast" isoforms raises the question of how these MHCs are distributed among the three different fibre types described for Rana. On the other hand, the possibility exists that the mRNAs for one or more of these fast MHC isoforms encode developmental isoforms that are present but not translated in the muscles of the tadpole and/or adult frog. Finally, an evaluation of the evolutionary relatedness of the R. catesbeiana MHCs to the MHCs in another species of Rana and to the MHCs in other vertebrates discloses, among other things, that the nucleotide sequence in the R. catesbeiana cardiac MHC isoform is more closely related to the chicken ventricular MHC isoform than it is to any of the other MHC isoforms examined.

Expression of the myosin heavy chain genes in the tail muscle of thyroid hormone-induced metamorphosingRana catesbeiana tadpoles

In tadpoles of the North American bullfrog, Rana catesbeiana, spontaneous and thyroid hormone (T3)-induced metamorphosis is characterized by regression of the tail, which is preceded by a decrease in total protein synthesis in tail tissues. We have demonstrated that thyroid hormone treatment of a tadpole does not affect the synthesis of all proteins equally in the tadpole tail muscle. For example, the synthesis of myosin heavy chains (MHCs) is depressed within 1 day and decreases to 45% of control values after 5 days of T3 treatment, whereas the decreased synthesis of soluble muscle proteins is transient and returns to above control levels by day 5. To determine whether the hormone-induced decrease in MHC synthesis is the result of changes in the transcription of translation of MHC mRNAs, we isolated cDNAs complementary to five different MHC mRNAs from a tail muscle cDNA library and used them to examine the levels of each MHC mRNA in the tail muscle of T3-treated tadpoles. mRNAs that recognize the cDNAs for these five different MHCs are all expressed in the tadpole tail and limb muscles, as well as in the adult leg muscles. MHC mRNAs unique to tadpole tail were not detected. Interestingly, the relative amounts of mRNA for four of the five MHCs increase in tail muscle after T3 treatment of the tadpole, suggesting that repression of MHC gene expression at the protein level does not result from a decrease in the amount of MHC mRNAs. Rather, these results support the contention that the decreased synthesis of MHCs in the tail muscle of T3-treated tadpoles is caused by this hormone, either directly or indirectly, depressing the translation of the MHC mRNAs in this tissue. These results, coupled with the observation that the synthesis of soluble muscle proteins is depressed only in a transient fashion, suggest that T3 may be initiating the expression of a gene(s) that encodes a protein(s) responsible for inhibiting the translation of the MHCs and, perhaps, other structural proteins in the tadpole tail muscle. Whatever the case, the translational regulation of MHC synthesis occurs well before any degradation of the tail tissue is evident and appears to be one of the earliest events in the hormone-induced cell death program of the tadpole tail muscle.

Apoptosis in Xenopus tadpole tail muscles involves Bax-dependent pathways.

Apoptosis is a fundamental mechanism implicated in normal development. One of the most spectacular developmental events involving apoptosis is tail regression during amphibian metamorphosis. We analyzed how thyroid hormone (3, 5, 3'-triiodothyronine, T3), the orchestrator of metamorphosis, affects expression and function of the proapoptotic gene Bax in the tail muscle of free-living Xenopus tadpoles. During natural metamorphosis Bax mRNA was expressed in tail muscles and was spatially correlated with apoptosis. Precocious treatment of tadpoles with T3 induced Bax expression and apoptosis. To verify that Bax expression was causally related to apoptosis we used a naked DNA gene transfer method to express Bax in the dorsal tail muscle. This induced apoptosis, and the process was exacerbated by T3 treatment. To determine whether T3 effects on Bax expression involved transcriptional regulation, we injected a Bax promoter sequence into dorsal and caudal tail muscles. In the dorsal muscle, T3 treatment did not affect transcription from the Bax promoter. However, in the caudal muscle, T3 treatment significantly increased Bax transcription. We conclude that T3-induced apoptosis in Xenopus tadpole tail muscles involves Bax-activating and Bax-synergis tic mechanisms. These programs are induced in spatially and temporally distinct manners.

Differential responses to ligands of overexpressed thyroid hormone and retinoid X receptors in a Xenopus cell line and in vivo

In an attempt to explain the contrasting patterns of expression of Xenopus thyroid hormone (xTR) and retinoid X (xRXR) receptor genes and to extend our understanding of the role of heterodimerization of these receptors during amphibian metamorphosis, we have investigated the response to their respective ligands of cells in which xTR and xRXR were overexpressed. Results obtained with two separate approaches are now described. In the first, 3,3′,5-triiodothyronine (T 3) was found to strongly upregulate xTR β mRNA in XTC-2 cells, but not of xTR α or xRXR α mRNAs, while xRXR γ transcripts could not be detected. 9- cis-retinoic acid (9- cis-RA) did not substantially influence the expression of any of these four receptor genes. When transcription from three different thyroid response elements (TREs) (a palindromic TREpal, an inverted repeat +6 [F2] and a direct repeat +4 [DR+4] as present in the promoter of xTR β gene) was measured in XTC-2 cells in which xTR β and xRXR α were overexpressed, only T 3 upregulated transcription while 9- cis-RA, alone or together with T 3, was ineffective. 9- cis-RA however enhanced transcription from an RXR responsive element (RXR-RE). The second approach involved overexpression of xTR β and xRXR α in premetamorphic Xenopus tadpole tail muscle followed by measuring the response of the tails to T 3 in organ culture. After validating the microinjection/culture procedure histochemically, we found that T 3 enhanced transcription from the xTR β DR+4 TRE in tails in which xTR β was overexpressed but the overexpression of xRXR α failed to modify this response. It is concluded that in both XTC-2 cells and tadpole tails, overexpressed xRXR fails to modify the enhanced transcriptional response of endogenous and overexpressed xTR β to T 3 and that exogenous 9- cis-RA is ineffective.

Muscle weakness during tricaine anesthesia

An isolated preparation of tadpole tail muscle was used to assess the peripheral effects of tricaine (3-aminobenzoic acid ethyl ester) at anesthetic concentrations and under physiological conditions. The drug effect on the electrically-evoked twitch was tested using short-pulse durations that elicited synaptically mediated effects or longer-duration pulses that stimulated the muscle directly. Tricaine reduced both types of response at anesthetic and even subanesthetic concentrations. At steady state concentrations that produced surgical anesthesia in vivo, tricaine reduced the directly evoked response by about half. It is concluded that tricaine anesthesia has a pronounced peripheral effect on neuromuscular function and that direct effect(s) on muscle are a major component.

Talin and vinculin in the oocytes, eggs, and early embryos of Xenopus laevis: A developmentally regulated change in distribution

We have investigated the expression and distribution of talin and vinculin in the oocytes, eggs, and embryos of Xenopus laevis. Antibodies to the previously characterized avian proteins stain several different Xenopus cell types identically by immunofluorescence: adhesion plaques of cultured kidney (A6) cells, the cell peripheries of oviduct cells, and the postsynaptic neuromuscular junctions of tadpole tail muscle fibers. These antibodies also identify cognate proteins of the appropriate sizes on immunoblots of A6 cell and oviduct lysates. Using these antibodies on ovarian tissue, we find talin to be highly localized at the cortices of oocytes and vinculin to be in the oocyte cytoplasm and absent from the oocyte cortex. In the cells of the ovarian layers that surround the oocytes, talin and vinculin can be detected as soluble and cytoskeletal components. Vinculin is first detectable as a cytoskeletal component in eggs, appearing some time during or between oocyte maturation and oviposition. During early embryo development, talin and vinculin are colocalized in the cortex of cleavage furrows and blastomeres. Thus, Xenopus oocytes and eggs display different distributions of talin and vinculin. The change from unlinked localization to colocalization appears to be developmentally regulated, occurring during the transition from oocyte to egg.

Identification of cells by backscattered electron imaging of silver stained bulk tissues in scanning electron microscopy.

Anuran tadpole tail muscle was stained en bloc by a modified light microscope silver stain for light microscopy and freeze-fractured in liquid nitrogen after partial dehydration with ethanol. The fractured specimens were observed in both secondary electron and backscattered electron modes in a scanning electron microscope. Since the cell nuclei specifically stained with silver provided high contrast against the unstained background due to atomic number contrast of backscattered electron image, various cells were easily identified by a comparison of secondary electron images and compositional images of backscattered electron signals.

Tail muscle hydrolases during metamorphosis of the anuran tadpole. I. Histochemical studies.

Naphthylamidase, beta-glucuronidase and trimetaphosphatase have been studied histochemically in the tail muscles of Rana catesbeiana, Rana japonica and Xenopus laevis tadpoles. During the early stages of growth, tadpole tail muscles showed very low activity of these hydrolases, whereas during the prometamorphic and metamorphic stages, they exhibited a marked increase in enzyme activity. This was localized in the subepidermal and subnotochordal mesenchymous tissue and in the myoseptum, i.e. between the muscle bundles.

Changes in the satellite cells of the anuran tadpole tail muscles during metamorphosis.

The incidence and the ultrastructure of the satellite cells associated with red and white muscle fibres of the anuran (Rana japonica) tadpole tail were examined in three stages (hindlimb-bud, prometamorphic, and climax) during metamorphosis. In these three stages the satellite cells are found more frequently in the red fibres than in the white fibres. As metamorphosis proceeds, the incidence of the satellite cells decreases in both red and white fibres. The satellite cells in the hindlimb-bud stage have abundant cytoplasm rich in organelles. Cisternae of rough endoplasmic reticulum are often dilated. The intercellular space between the satellite cell and the muscle fibre is narrow and relatively constant (15-25 nm). Some satellite cells appear to be in the process of fusion with the muscle fibre. Cytoplasmic volume and organellar content are gradually reduced during metamorphosis. In the prometamorphic and climax stages, the intercellular space is in general gradually extended. It often exceeds 0.5 microns, and usually contains the basal lamina and fibrous materials. Some satellite cells have cytoplasmic processes that project into the interstitial space through a gap in the basal lamina.