Trochlear-oculomotor nerve interactions in Xenopus laevis tadpoles: a temporal study.

When the trochlear nerve (NIV), which innervates the superior oblique muscle (SOM), is crushed or cut at stages 48-49 in Xenopus tadpoles, fibers from the oculomotor nerve (NIII) sprout and invade the SOM. The maximal percentage of specimens having at least one oculomotor nerve fiber on the SOM on a given day increased from 9.1% following a single crushing of NIV to 84.2% following three successive severings of NIV and the average number of silver-impregnated NIII fibers per specimen increased from 0.23 +/- 0.16 (mean +/- S.E.M.) in the single-crush experiment to 7.35 +/- 1.33 in the triple-cut experiment. This increase directly reflects the delay in the return of NIV. As NIV returns to the SOM, a portion of the inappropriate innervation is lost; while another portion appears to be stable and is in evidence 90 days after a single sectioning of NIV. The more rapidly NIV returns to the SOM, the more complete is the displacement of the NIII fibers. This suggests that the association between NIII and the SOM changes with time so that easy displacement of the inappropriate innervation is likely only when the reinnervation by the appropriate nerve fibers is rapid.     

Subdivisions of the terminal nerve in Xenopus laevis

The majority of recent studies on the terminal nerve (nt) in various vertebrates either involved tracer injections into the nasal cavity or made use of the LHRH-/FMRFamide-like immunoreactivity (ir) of a portion of its fibers. The present investigation was designed to determine the extent of overlap between data rendered by the two methods in Xenopus. The findings reveal no overlap of nt projections visualized by the two experimental techniques. This result sheds doubt on the validity of current definitions of the nt system.     

Origin and segregation of cranial placodes in Xenopus laevis

Cranial placodes are local thickenings of the vertebrate head ectoderm that contribute to the paired sense organs (olfactory epithelium, lens, inner ear, lateral line), cranial ganglia and the adenohypophysis. Here we use tissue grafting and dye injections to generated fate maps of the dorsal cranial part of the non-neural ectoderm for Xenopus embryos between neural plate and early tailbud stages. We show that all placodes arise from a crescent-shaped area located around the anterior neural plate, the pre-placodal ectoderm. In agreement with proposed roles of Six1 and Pax genes in the specification of a panplacodal primordium and different placodal areas, respectively, we show that Six1 is expressed uniformly throughout most of the pre-placodal ectoderm, while Pax6, Pax3, Pax8 and Pax2 each are confined to specific subregions encompassing the precursors of different subsets of placodes. However, the precursors of the vagal epibranchial and posterior lateral line placodes, which arise from the posteriormost pre-placodal ectoderm, upregulate Six1 and Pax8/Pax2 only at tailbud stages. Whereas our fate map suggests that regions of origin for different placodes overlap extensively with each other and with other ectodermal fates at neural plate stages, analysis of co-labeled placodes reveals that the actual degree of overlap is much smaller. Time lapse imaging of the pre-placodal ectoderm at single cell resolution demonstrates that no directed, large-scale cell rearrangements occur, when the pre-placodal region segregates into distinct placodes at subsequent stages. Our results indicate that individuation of placodes from the pre-placodal ectoderm does not involve large-scale cell sorting in Xenopus.     

Androgen receptor mRNA expression in Xenopus laevis CNS: Sexual dimorphism and regulation in laryngeal motor nucleus

Using Northern analysis, in situ hybridization, and nuclease protection assays, the expression and regulation of androgen receptor messenger RNA (AR mRNA) was examined in the CNS of juvenile Xenopus laevis. Only one of the AR mRNA isoforms expressed in X. laevis is transcribed in the CNS as shown by Northern blot analysis. Nuclease protection assays demonstrate that the expression of AR mRNA is higher in the brain stem than in the telencephalon and diencephalon. Although expression of AR mRNA is widespread throughout the CNS, cells of cranial nerve nucleus IX-X (N. IX-X) and spinal cord display the highest in situ hybridization signals in their cytoplasm. Double labeling using horseradish peroxidase and digoxigenin labeled AR probes reveals that laryngeal and anterior spinal cord motor neurons express AR mRNA. More cells express AR mRNA in N. IX-X of males than of females. The number of AR expressing cells in N. IX-X decreases following gonadectomy in both sexes, and dihydrotestosterone (DHT) treatment for 1 month reverses this effect. Increased expression of AR mRNA in the brain of DHT treated animals is also apparent in nuclease protection assays. Sex differences in number of AR expressing cells and hormone regulation of AR mRNA expression in motor nuclei may influence neuromuscular systems devoted to sexually differentiated behaviors. © 1996 John Wiley & Sons, Inc.     

Regulation of neurons in the suprachiasmatic nucleus of Xenopus laevis

In the amphibian Xenopus laevis, suprachiasmatic melanotrope-inhibiting neurons (SMINs) play an important role in the regulation of the background adaptation process. In this study, we investigated the innervation of the SMINs at the light- and electron- microscopical level. Immunocytochemistry in combination with confocal laser scanning microscopy revealed co-existence of neuropeptide Y (NPY) and synaptobrevin in spots in the direct vicinity of the SMINs, suggesting the existence of NPY-containing synapses on these cells. At the ultrastructural level, the SMINs showed a high degree of plasticity, containing more electron-dense vesicles and a larger extent of RER in white- than in black-adapted animals. In black-adapted animals, symmetric synapses (Gray type II) were observed on the soma of the SMINs, suggesting an inhibitory input to these cells. The synaptic profiles contained electron-lucent and electron-dense vesicles, indicating the involvement of both a classical neurotransmitter and a neuropeptide (possibly NPY) in this input. In white-adapted animals, synapses were only found at some distance from the SMIN somata. Our findings indicate a striking plasticity of the innervation of the SMINs in relation to background adaptation and support the hypothesis that the SMINs are innervated by NPY-containing interneurons that inhibit SMIN activity in black-adapted animals.     

Development of the Xenopus laevis viiith cranial nerve: Increase in number and area of axons of the saccular and papillar branches

Development of three branches of the VIIIth cranial nerve was examined in the anuran, Xenopus laevis. Sectioned tissue from the saccular, amphibian papillar, and basilar papillar branches of stage 52 larvae, 1 day postmetamorphosis juveniles, and 2-year adult animals was analyzed under the light microscope with a digital image analysis system. Numbers and cross-sectional areas of myelinated axons were measured in five to six nerve sections at each developmental age for each of the three branches. In all three branches, results show a significant increase in axon numbers between larval stage 52 and juvenile ages and negligible increase in axon number between the juvenile and adult ages. There were differences in the average number of axons between the saccular (704.4 ± 39.5; n = 5), amphibian papillar (508.4 ± 35.0; n = 5), and basilar papillar (316.0 ± 7.0; n = 5) branches of adult animals. Myelinated axons increase at an estimated rate of 11.7, 15.1, and 6.2 axons per day for the saccular, amphibian papillar, and basilar papillar branches, respectively. Axonal cross-sectional areas increased throughout the developmental ages of this study, with the greatest increase taking place between juvenile and adult ages. In adult animals, 98% of axons in all three branches have diameters between 2–10 μm. Ratios of axons to hair cells in adult animals were estimated at 0.3, 1.1, and 5.3 for the sacculus, amphibian papilla, and basilar papilla, respectively. The higher axon to hair cell ratio correlates with the increasing acoustical frequency sensitivity of the end organ. J. Morphol. 234:263–276, 1997. © 1997 Wiley-Liss, Inc.     

Development of functional sex differences in the larynx of Xenopus laevis

Three laryngeal properties associated with the production of masculine song--laryngeal muscle tension, fiber twitch type, and fiber recruitment--are markedly sexually dimorphic in adult Xenopus laevis frogs. To elucidate the pattern of sexual differentiation, tension and fiber recruitment in male and female larynges and fiber twitch type in male larynges were examined throughout postmetamorphic development. Masculinization of male laryngeal properties begins early in postmetamorphic development and continues until adulthood. In contrast, tension and fiber recruitment in females do not change after the end of metamorphosis. Laryngeal muscle tension and fiber type are gradually and progressively masculinized; the temporal pattern of masculinization is very similar for these properties. Fiber recruitment, on the other hand, appears to masculinize in a stepwise manner. Masculinization of all three properties is highly correlated with larynx weight in males. We have used this relation to divide postmetamorphic development into seven stages associated with key events in sexual differentiation. This staging scheme provides an important experimental tool for studying the hormonal regulation of sexual differentiation, the subject of the accompanying paper.     

Sex and side differences in the minor non-metrical cranial variants

Thirty-three epigenetic traits were examined on 260 skulls from present-day Sardinian population. Side and sex differences were studied, together with possible differences between the methods commonly used. The Authors concluded that side and sex differences are practically irrelevant. They also recommend research-workers to use one method only in comparing the traits studied.     

Histological development of the cement gland in Xenopus laevis: a light microscopic study

The differentiation and degeneration of the cement gland in Xenopus laevis is described. The gland is first observed histologically at stage 19 (neural tube stage) as a packed group of apical ectoderm cells heavily laden with oocyte pigment granules, lying ventral to the cranial neural fold. By tailbud stage 35/36, the gland cells have increased in height and are approximately ten times taller than nonglandular apical ectoderm cells. The nuclei divide the gland cells into an apical region that is eosinophilic and contains oocyte pigment granules, and a basal region that contains clear droplets. The cells are decreasing in height by stage 40 (early tadpole) and begin to lose their pigment granules. Between stages 45 and 48, the pigment is extruded and the clear basal droplets diminish in number. From stage 48 to 49 the cells become vacuolated and the histotypic characteristics of the functional gland are lost. The gland is not vascularized, nor do phagocytic cells appear in its vicinity during any stage of its development. It remains bordered at its base by subjacent basal ectoderm during its entire life cycle of 10 to 12 days at room temperature.     

Multinucleation during myogenesis of the myotome of Xenopus laevis: a qualitative study

Ultrastructural studies of myogenesis in the myotome of Xenopus laevis reveal that the myotubes developed by stage 33/34 have peripheral myofibrils but are still uninucleate with a single large nucleus. By stage 45, the cytoplasm of the muscle cells is filled with myofibrils and there are many small peripheral nuclei, resulting in multinucleate muscle fibres. With the electron microscope, we have examined myotomes from stages 33/34 to 59 of development and some stages were also investigated by autoradiography. There was no evidence from autoradiographic studies for DNA synthesis in muscle cells, and the increase in the number of myonuclei was accompanied by a decrease in their size. Satellite cells were not seen at the myotube stage but were first seen after the cells had become multinucleate, with many small nuclei close together forming rows. Constrictions were frequently observed in the large single nuclei. It is concluded that division of the myonuclei by amitosis is mainly responsible for the multinucleation that occurs during development of the myotome muscle in Xenopus laevis.     

Sex differences in motor unit behaviour: A review

The lack of systematic investigations on sex-related differences in motor unit behaviour poses a challenge in understanding and optimizing health and performance in males and females. Limited investigations revealed that sex differences in motor unit behaviour might be present in human muscles. This review summarizes the current knowledge on sex differences in motor unit behaviour and potential factors that may contribute to these differences. We show significant under-representation of female participants in motor unit studies and a limited number of studies investigating sex differences in motor unit behaviour. We place the current insights within the context of methodological limitations and outline several recommendations and future directions to improve female representation in this research area. We conclude that there is an urgent need to gather more data in females and investigate sex differences in motor unit behaviour. The knowledge gained could be used to develop sex-specific approaches to improve neuromuscular performance and rehabilitation.     

Xenopus laevis oocyte as a model for the study of the cytoskeleton

At the beginning of diplotene, the oocyte of Xenopus laevis is a cell of about 10–20 microns destined to increase 10,000-fold its size when the oocyte becomes filled with yolk platelets and has accumulated a great number of pigment granules in a half of its periphery. Its internal architecture is gradually accomplished during growth because of several factors, especially because of cytoskeletal changes. In the fully-grown oocyte, the cytoskeleton appears to sustain the eccentrically located germinal vesicle through arms radiating from the cortex to the germinal vesicle, a unique organization not to be found in other Amphibians. In this report, we summarized and analysed steps of cytoskeletal proteins and related mRNAs organization and function throughout diplotene stage, highlighting our studies in this animal model. The cytoskeletal proteins appear to exploit their activity with respect to ribosomal 60S subunit maturation and during translation. Most importantly, the polarity of the oocyte is achieved through a sophisticated and highly organized localization of mRNAs and cytoskeletal proteins in one side of the cell. This asymmetry will start the construction of the oocyte polarity that is instrumental for determining the characteristic of this cell, which will become an embryo. Moreover, in the same time membrane composition, conditioned by the underlying cytoskeletal organization, will acquire the prerequisites for sperm binding and fusion.     

The reaction of the preoptic nucleus of Xenopus laevis tadpoles to osmotic stimulation

Summary The development of the preoptic nucleus of Xenopus laevis tadpoles during metamorphosis was studied and the effect of osmotic stimulation on this process investigated. The development of this region was not affected by treatment for one or more days in hypertonic media. It was found that at the end of metamorphosis the neurosecretory cells in the preoptic nucleus are localized in three regions: the rostro-dorsal, the caudo-dorsal and the ventral region. After osmotic stimulation only the neurosecretory cells of the caudo-dorsal region appeared to have reacted, as indicated by their loss of neurosecretory (PIC positive) material. It is concluded that the cells of this region may be involved in the synthesis of the posterior lobe hormones.The author thanks Prof. Dr. J. C. van de Kamer and Dr. F. C. G. van de Veerdonk for their interest and many helpful discussions, Dr. L. Boomgaart and Dr. A. P. van Overbeeke for correcting the English text and Miss C. M. G. van Bemmel for technical assistance.     

Laryngeal muscle and motor neuron plasticity in Xenopus laevis: Testicular masculinization of a developing neuromuscular system

In Xenopus laevis, the sexual differentiation of the neuromuscular system responsible for courtship song is controlled by testicular androgen secretion. To explore the sensitivity of this system to androgenic stimulation, male and female frogs were gonadectectomized and given testis transplants at seven different developmental stages between the end of metamorphosis and adulthood, grown to sexual maturity, and the laryngeal muscle fibers and motor axons were counted. Muscle fiber and axon numbers in males were not affected by the testicular transplant at any stage. In females, testicular transplants at all developmental stages increased muscle fiber numbers in adulthood. Values attained were, however, significantly less than those of adult intact or testis-transplanted males. Testis transplantation increased laryngeal axon numbers in females to levels equivalent to those of intact males; this effect was obtained at every stage of postmetamorphic development including adulthood. To further explore androgen regulation in adults, males and females were gonadectomized and implanted with silicone tubes containing testosterone propionate for 1.5–3 years and laryngeal muscle fibers and axon numbers compared to those of gonadectomized or sham-operated adult controls. Neither treatment with exogenous androgen nor gonadectomy had any effect on laryngeal muscle fiber or axon number in either males or females; values did not differ from those of sham-operated controls. We conclude that testicular secretions can induce laryngeal muscle fiber and axon addition in females throughout postmetamorphic life. This degree of plasticity, exhibited after the period when adult values are normally attained, stands in contrast to the effects of administration of synthetic androgen and suggests that the degree of plasticity in adult females may be underestimated if exogenous hormones rather than testicular transplants are provided. © 1993 John Wiley & Sons, Inc.