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Myomesin and M protein: differential expression in embryonic fibers during pectoral muscle development.

By applying immunocytochemistry using monoclonal antibodies, we found that the myofibrillar M band of both presumptive type-I and -II fibers in the pectoralis major muscle of chickens contains two high-molecular-weight proteins, i.e., myomesin (Mr, 185,000) and M protein (Mr, 165,000), early in embryonic development (7 days in ovo), even though adult type-I fibers lack M protein. The developmental expression of M protein is unusual in that, from 10 to 14 days in ovo, it is gradually suppressed not only in presumptive type-I fibers but also in presumptive type-II fibers formed from primary-generation myotubes. This latter suppression is transient, as M protein is expressed in all adult type-II fibers derived from both the primary- and second-generation myotubes. Myomesin, on the other hand, is continuously expressed in all myotubes throughout development. This finding shows that myomesin and M protein expression is regulated independently in different myotube populations, and that the suppression of M protein in primary-generation myotubes accounts for the delayed accumulation of M protein during development, as previously revealed by biochemical analysis. Presumptive type-I fibers, which form in the deep portion of the muscle, become concentrated in a narrow band known as the red strip.

Aging↗

Changing glial organization relates to changing fiber order in the developing optic nerve of ferrets.

The structures of the developing eye-stalk and the relationships of early retinofugal fibers as they pass through the stalk, chiasm, and tract have been studied by light and electron microscopical methods in fetal ferrets aged 23-27 days. The early eye-stalk can be divided into two parts: a narrow extracranial part has a narrow lumen and is lined by few cells, whereas a thicker intracranial part has a wider lumen and is lined by several rows of cells. At the earliest stages no axon bundles are recognizable in the stalk, but fibers of the supraoptic commissure are already beginning to cross the midline in the diencephalon. Subsequently, as retinofugal axons invade the stalk, the glia of the extracranial part of the stalk have an interfascicular distribution and axon bundles are separately encircled by glial cytoplasm. In the intracranial part, as in the chiasm and tract, the glial cells occupy a periventricular position and send slender radial cytoplasmic processes to the subpial surface; these pass between groups of axons that here lie immediately deep to the subpial glia. Whereas axonal growth cones have no evident preferred distribution in the extracranial stalk, they tend to accumulate near the pial surface intracranially. The boundary between the two types of organization shifts as development proceeds so that the interfascicular glial structure of the early extracranial stalk first encroaches upon the intracranial parts and later appears in the chiasm. The characteristic adult arrangement of fibers in an age-related order in the optic chiasm and tract, but not in the optic nerve, can be understood if axonal growth cones are guided toward the pial surface by radial glia but not by interfascicular glia. From the distribution of the growth cones, this is what appears to happen.

Animals↗

Glial fiber pattern in the developing chicken cerebellum: vimentin and glial fibrillary acidic protein (GFAP) immunostaining.

The possible relation between glial fibers and the formation of longitudinal granule cell migration patterns that occur in the cerebellar anlage of the chicken was investigated by immunocytochemistry of vimentin (monoclonal antibody) and glial fibrillary acidic protein (polyclonal antibody against GFAP, PGF) on fixed and unfixed brain tissues. In addition, neuronal development was studied with a monoclonal antibody for neurofilament. Vimentin was present in radial and tangential fibers in the cerebellar anlage during granule cell migration in almost all parts of the anlage. However, no specific topographic relation of vimentin and GFAP to the migration pattern of granule cells was observed. In adults, Bergmann fibers and astroglia were stained with vimentin antiserum and not with GFAP antiserum. Conclusions are that radial fibers do not determine the formation of longitudinal cytoarchitectonic patterns in the chick cerebellum and that vimentin is the main cytoskeletal component of Bergmann fibers and astroglial cells in embryonic and adult chicken cerebellum.

Animals↗

Regeneration of spinal electrocyte fibers in Sternarchus albifrons: development of axon-Schwann cell relationships and nodes of Ranvier.

The electrocyte fibers in the gymnotid Sternarchus albifrons are highly differentiated myelinated axons which exhibit several types of nodes of Ranvier and characteristically short internode lengths. In the present study, regeneration of the electrocyte fibers following removal of the tail was examined by electron microscopy. By 36 days following extirpation, the regenerating electrocyte axons exhibit Type I nodes of Ranvier, with a normal morphology, and Type II nodes of Ranvier with a large nonmyelinated gap and a polypoid elaboration of the axon surface. Moreover, in the regenerating axons the internode length: diameter ratios are quite small. Thus, relatively normal axons-Schwann cell relations and a relatively normal differentiation of the axon surface are achieved during regeneration of the Sternarchus electrocyte fibers.

Animals↗

[Study on intratumor administration of lentinan--primary changes in cancerous tissues].

Lentinan was administered for gastric cancer in order to determine what type of changes would occur as a consequence. In an experimental study, the intraperitoneal administration of lentinan on cancerous tissues caused a marked development of reticular fibers, along with an enhanced interstitial response. Findings which support the relationship between a marked development of reticular fibers and an anti-tumor effect, were also obtained. Furthermore, lentinan was administered for human gastric cancer to evaluate whether or not an increase in interstitial response would be observed. As a result, reticular fibers developed in tumor sites with a resultant fragmentation of cancer cell nests. Many T lymphocytes infiltrated cancer sites where the lentinan was administered. These findings suggest that the intratumor administration of lentinan enhanced an interstitial response and also activated an anti-tumor immune response in the human gastric cancer tissues.

Animals↗

Organization of permanent and transient neuropeptide Y-immunoreactive neuron groups and fiber systems in the developing hamster diencephalon.

The development of neuropeptide Y-immunoreactive (NPY-IR) cell and fiber systems in the hamster diencephalon was studied. Eight perinatal groups of NPY-IR neurons develop into 12 distinct sets in nuclei of the adult diencephalon and mesencephalon. NPY-IR neurons of the thalamic precommissural nucleus, nucleus of the optic tract, and olivary pretectal nucleus are derived from the superior group. Those in the adult magnocellular nucleus of the posterior commissure and deep mesencephalic nucleus are from the dorsal group. An arcuate group contributes neurons to the arcuate nucleus and median eminence and a mammillary group transiently exists in the mammillary region. A medial group gives rise to two sets of neurons, one that migrates to the intergeniculate leaflet and another that develops in the medial nucleus reuniens. A very large ventral group provides NPY-IR neurons to the adult medial zona incerta and caudal reticular thalamus. Groups of NPY-IR neurons also appear in the bed nucleus of the stria terminalis and centromedian thalamic nucleus. Superior group neurons may undergo apoptosis. In several groups, neurons become fewer during development, and NPY-IR may disappear. NPY-IR neurons of several groups initially migrate away from the neuroepithelial zone with later emergence of a distinct, persistent set of NPY-IR neurons in the same neuroepithelial region. The data show that neuropeptide content can be used to identify particular sets of neurons early in development, thereby allowing migration patterns to be followed and principles of brain development to be elucidated.

Animals↗

Abnormal development of serotonin nerve fibers in the visual cortex in rats with methylazoxymethanol-induced microcephaly.

The postnatal development of serotonin (5HT)-immunoreactive axons was studied in the visual cortex of the cerebrum in both normal and microcephalic rats during early postnatal and young adult stages. Severe microcephaly in rat offspring was induced by prenatal exposure to methylazoxymethanol acetate (MAM), an anti-mitotic agent, on day 15 of gestation. From postnatal day 1 (PND 1) to PND 5, fine and short 5HT fibers were irregularly dispersed throughout the occipital cortex in both the control and MAM-treated rats (MAM-rats). A conspicuous aggregation of dot-like 5HT terminals was found in controls, but not in MAM-rats, in a shallow layer of the dorsomedial region of the occipital cortical plate. On PND 7, such an aggregation of 5HT terminals was found in both groups. The density of the aggregation increased up to PND 9, but then decreased gradually, finally becoming unrecognizable at around PND 15 in both groups. MAM-rats, however, always showed hyperaggregation of 5HT terminals when compared with controls on the same PND. The density of 5HT fibers gradually increased, and finally made up a network-like formation at PND 28 in both groups, its pattern was essentially identical to the abnormal distribution of 5HT fibers during the later stage. As a result, the network-like formation of 5HT fibers in the MAM-rats at PND 28 was markedly twisted and somewhat hyperdense. In Nissl-stained preparations from PND 9 to 15, the 5HT terminal aggregation in the control rats was precisely confined to the newly forming layer IV of the visual cortex. In the MAM-rats, on the other hand, the aggregation of 5HT terminals was not associated with a specific cortical layer because of a disarranged cytoarchitecture of the microcephaly.

Animals↗

Early struggles with single-fiber electromyography.

The development of single-fiber electromyography is described. This method, introduced in 1963, was met with doubts, and there were strong arguments that subunits, 10-30 synchronized muscle fibers, were in fact being recorded. The dispute ended in 1971, and single-fiber electromyography is now generally accepted.

Electromyography↗

Cylinder gratings in conical incidence with applications to modes of air-cored photonic crystal fibers.

We develop a formulation for cylinder gratings in conical incidence, using a multipole method. The theory, and its numerical implementation, is applied to two-dimensional photonic crystals consisting of a stack of one-dimensional gratings, each characterized by its plane wave scattering matrix. These matrices are used in combination with Bloch's theorem to determine the band structure of the photonic crystal from the solution of an eigenvalue problem. We show that the theory is well adapted to the difficult task of locating the complete band gaps needed to support air-guided modes in microstructured optical fibers, that is, optical fibers in which the confinement of light in a central air hole is achieved by photonic band-gap effects in a periodic cladding comprising a lattice of air holes in a glass matrix.

Journal Article↗

Short, thin asbestos fibers contribute to the development of human malignant mesothelioma: pathological evidence.

Based on animal studies, long and thin asbestos fibers (> or =8 microm in length and < or = 0.25 microm in width) have been postulated to be strongly carcinogenic inducing pleural malignant mesothelioma, while shorter, thicker fibers have been postulated to pose a lesser risk (Stanton hypothesis). The objective of this study is to test the validity of the Stanton hypothesis through direct pathologic analysis of human mesothelioma tissue. Digested bulk tissue samples, or ashed 25 microm thick sections, or both, were prepared from lung and mesothelial tissues taken from 168 cases of human malignant mesothelioma. In these tissues, 10,575 asbestos fibers (4820 in the lung and 5755 in mesothelial tissues (1259 in fibrotic serosa and 4496 in mesotheliomatous tissue)) were identified by high-resolution analytical electron microscopy. Dimensions of these asbestos fibers were measured in printed electron micrographs. Results were as follows: (1) long, thin asbestos fibers consistent with the Stanton hypothesis comprised only 2.3% of total fibers (247 / 10,575) in these tissues; (2) the majority (89.4%) of the fibers in the tissues examined were shorter than or equal to 5 microm in length (9454 of 10,575), and generally (92.7%) smaller than or equal to 0.25 microm in width (9808 of 10,575). (3) Among asbestos types detected in the lung and mesothelial tissues, chrysotile was the most common asbestos type to be categorized as short, thin asbestos fibers. (4) Compared with digestion technique of the bulk tissue, ashing technique of the tissue section was more effective to detect short, thin fibers. We conclude that contrary to the Stanton hypothesis, short, thin, asbestos fibers appear to contribute to the causation of human malignant mesothelioma. Such fibers were the predominant fiber type detected in lung and mesothelial tissues from human mesothelioma patients. These findings suggest that it is not prudent to take the position that short asbestos fibers convey little risk of disease.

Animals↗

Transitional expression of OX-2 and GAP-43 glycoproteins in developing rat cochlear nerve fibers.

The OX-2 and GAP-43 glycoproteins are two proteins involved in neuronal cell-to-cell interaction and/or growing of dendrites and axons. Therefore, for the auditory receptor the expression of these proteins could provide information on the afferent and efferent nerve fiber organization. The expression and distribution of OX-2 and GAP-43 were analyzed during the auditory receptor development and maturation (from embryonic day E13 to postnatal day P22). Both glycoproteins were early recognized in the cochleae of E13 rats. Then, they slowly but progressively disappeared, being absent when the animals reached the P22 postnatal day. At E13, a weak OX-2 expression was restricted to the perikaryon of the spiral ganglion neurons, while in the same period a strong GAP-43 immunostaining was found in both the neuronal perikaryon and the neurites. During the rat embryonic period (E13 to birth) the expression of both glycoproteins appeared progressively restricted to the neurites. During the rat postnatal period (P0 to P22), OX-2 and GAP-43 exhibited a dissimilar distribution pattern. The OX-2 glycoprotein appeared in the afferent, efferent and fibers of the auditory nerve, while the GAP-43 glycoprotein only appeared in the efferent nerve fibers. Present data suggest that OX-2 and GAP-43 could act as two complementary glycoproteins during the development, organization, and maturation of the cochlear nerve fibers. While both glycoproteins could participate in axonal growing and orientation, OX-2 could also be involved in a similar process for auditory dendrites.

Animals↗

Comparison between the predictions of diffusion-reaction models and localized Ca2+ transients in amphibian skeletal muscle fibers.

We developed a three-dimensional cylindrical diffusion-reaction model of a single amphibian myofibril in which Ca(2+) release occurred only at the Z-line. The model incorporated diffusion of Ca(2+), Mg(2+), and all relevant buffer species, as well as the kinetic binding reactions between the buffers and appropriate ions. Model data was blurred according to a Gaussian approximation of the point spread function of the microscope and directly compared with experimental data obtained using the confocal spot methodology. The flux parameters were adjusted until the simulated Z-line transient matched the experimental one. This model could not simultaneously predict key parameters of the experimental M- and Z-line transients, even when model parameters were adjusted to unreasonably extreme values. Even though the model was accurate in predicting the Z-line transient under conditions of high [EGTA], it predicted a significantly narrower Ca(2+) domain than observed experimentally. We modified the model to incorporate a broader band of release centered at the Z-line. This extended release model was superior both in simultaneously predicting critical features of the Z- and M-line transients as well as the domain profile under conditions of high [EGTA]. We conclude that a model of release occurring exclusively at the Z-line cannot explain our experimental data and suggest that Ca(2+) may be released from a broader region of the sarcoplasmic reticulum than just the T-tubule-sarcoplasmic reticulum junction.

Amphibians↗

Temporal regulation of VEID-7-amino-4-trifluoromethylcoumarin cleavage activity and caspase-6 correlates with organelle loss during lens development.

Lens fiber cell differentiation involves extensive reconstruction of the cell's architecture, including the degradation and elimination of all membrane-bound organelles via a process that has been likened to apoptosis. Using caspase reporter assays under conditions in which nonspecific cleavage of the reporter peptides by the proteasome has been inhibited, we investigated whether any specific caspase activities are temporally correlated with this process of organelle loss. Extracts from neonatal mouse lenses contained strong VEID-7-amino-4-trifluoromethylcoumarin (AFC) and minor IETD-AFC and LEVD-AFC cleavage activities, but no DEVD-AFC cleavage activity. Further testing suggested that the VEID-AFC and IETD-AFC cleavage activities were likely due to the same enzyme. In lens extracts from rat embryos, VEID-AFC cleavage activity increased during the period when organelles are eliminated, between embryonic days 15.5 and 18.5, whereas procaspase-6 protein levels decreased, suggesting that this enzyme is responsible for VEID-AFC cleavage. By contrast, in extracts from alpha AE7 transgenic mouse lenses in which apoptosis was induced, strong DEVD-AFC cleavage activity and activated caspase-3 protein were detected. Thus, within the same tissue, different caspase activities can predominate depending on the context, normal differentiation versus apoptosis. These results highlight the difference between normal fiber cell differentiation and apoptosis and the capacity of the lens to differentially regulate these two processes.

Animals↗

Development of the giant fiber neuron of Drosophila melanogaster.

The giant fiber system (GFS) of Drosophila melanogaster provides a convenient system in which to study neural development. It mediates escape behaviour through a small number of neurons, including the giant fibers (GFs), to innervate the tergotrochantral jump muscle (TTM) and the dorsal longitudinal flight muscles. The GFS has been intensively studied physiologically in both wild-type and mutant flies, and is often used as a system to study the effects of neural mutations on the physiology of the adult nervous system. Recently, much information has been gleaned as to how and when synaptogenesis, with its major target neurons, is achieved. However, little is known of the earlier development of this neuron. Here we have used an enhancer-trap, marking parts of the GFS, in conjunction with BrdU labelling, to attempt to follow the birth, axonogenesis, and the early morphological meeting of the GFs with their target neurons. From these anatomical observations we propose that the GF cell is not born during the larval or pupal stages and, therefore. appears to be a persistent embryonic cell. The axons of the GFs develop during the third instar. During the early pupal stages the GFs contact other identified neurons of the GFS. In addition, we see some aberrant development of the network, with some flies carrying only one GF, and yet others with extended axons. We present a model for the initial joining of the GFs and tergotrochanteral motorneurons (TTMns).

Animals↗

The development of the serotonergic fiber network of the lateral ventricles of the rat brain: a light and electron microscopic immunocytochemical analysis.

The development of the serotonergic innervation of the lateral ventricles of the rat brain during the first five postnatal weeks was studied with immunocytochemical techniques at the light and electron microscopic levels. In the lateral ventricles of newborn rats serotonergic fibers are only rarely seen. During the first postnatal week the number of serotonergic fibers increases but they are straight and thick, bearing only a few varicosities. By the end of the second postnatal week, however, they become finer, exhibit a large number of varicosities, and form a dense supraependymal network. During the following weeks this network becomes slightly denser but the morphology of fibers as well as their distribution pattern remain unchanged. Examination of sagittal vibratome sections revealed that a group of serotonergic fibers leaves the medial forebrain bundle and turning dorsocaudally between the corpus callosum and the caudate/putamen enter the lateral ventricle from its rostral pole. They then spread to form the supraependymal network of the lateral ventricles and probably of the rest of the ventricular system. Ultrastructural analysis showed that serotonin varicosities are located close to the ventricular surface of the ependymal lining but never make synapses with the ependymal cells. Examination of a large number of labeled fibers and varicosities showed that they are never located between the ependymal cells or in the subependymal layer. This finding was confirmed by examining series of semithin sections. On the basis of these and previous findings we suggest that serotonergic fibers arising in the midbrain raphe nuclei enter the lateral ventricle from its rostral pole, form a dense network within the ventricles, and release their content into the cerebrospinal fluid. This system, as judged with morphological criteria, matures by the end of the second postnatal week.

Aging↗

forked proteins are components of fiber bundles present in developing bristles of Drosophila melanogaster.

The forked (f) gene of Drosophila melanogaster encodes six different transcripts 6.4, 5.6, 5.4, 2.5, 1.9, and 1.1 kb long. These transcripts arise by the use of alternative promoters. A polyclonal antibody raised against a domain common to all of the forked-encoded products has been used to identify forked proteins on two-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis gels and in Drosophila pupal tissues. The antibody stains fiber bundles present in bristle cells for about 15 hr during normal pupal development. Electron microscopy shows that these fibers are present from 40 to 53 hr in bristles of wild-type flies but are absent in the null f36a mutant. The forked protein(s) thus appear to be an essential part of the bristle fibers. The phenotype of the f36a mutation can be rescued by a 13-kb fragment of the forked locus containing the coding regions for the 2.5, 1.9, and 1.1-kb transcripts, suggesting that the proteins encoded by the three large forked RNAs are dispensable during bristle development. Increasing the copy number of a P[w+,f+] construct containing the 13-kb fragment induces a hypermorphic bristle phenotype whose severity correlates with the number of copies of P[w+,f+] present. These results indicate that alterations in the ratios among the forked proteins, or between forked products and other components of the fiber, result in abnormal assembly of the fibrillar cytoplasmic structures necessary for bristle morphogenesis.

Animals↗

Development of an in situ fiber optic Raman system to monitor hydrothermal vents.

The development of a field portable fiber optic Raman system modified from commercially available components that can operate remotely on battery power and withstand the corrosive environment of the hydrothermal vents is discussed. The Raman system is designed for continuous monitoring in the deep-sea environment. A 785 nm diode laser was used in conjunction with a sapphire ball fiber optic Raman probe, single board computer, and a CCD detector. Using the system at ambient conditions the detection limits of SO(4)(2-), CO(3)(2-) and NO(3)(-) were determined to be approximately 0.11, 0.36 and 0.12 g l(-1) respectively. Mimicking the cold conditions of the sea floor by placing the equipment in a refrigerator yielded slightly worse detection limits of approximately 0.16 g l(-1) for SO(4)(-2) and 0.20 g l(-1) for NO(3)(-). Addition of minerals commonly found in vent fluid plumes also decreased the detection limits to approximately 0.33 and 0.34 g l(-1) respectively for SO(4)(-2) and NO(3)(-).

Animals↗

Neurotrophin 4/5 is required for the normal development of the slow muscle fiber phenotype in the rat soleus.

During normal postnatal development, rat soleus (SOL) muscle fibers undergo a dramatic fast-to-slow myosin heavy chain (MyHC) isoform transformation. We exploited this phenomenon to evaluate the role of neurotrophin 4/5 (NT-4/5) in the regulation of muscle fiber phenotype. Intramuscular injections of recombinant NT-4/5 into the SOL muscle of rat neonates significantly accelerated the normal fast-to-slow MyHC isoform transformation. Sequestration of endogenous NT-4/5 with TrkB-IgG prevented this transformation from occurring. Administration of the other TrkB ligand, brain-derived neurotrophic factor (BDNF), did not affect the normal course of the MyHC isoform transformation in this muscle, indicating that the observed effect is NT-4/5 specific. Botulinum toxin blockade of synaptic transmission significantly disrupted the normal fast-to-slow MyHC isoform switch. Because administration of NT-4/5 to paralyzed muscles failed to restore the normal course of this MyHC transformation, we believe that the effect of NT-4/5 is not directly on the muscle fibers but that it probably activates or forms a type of retrograde signal to motoneurons. The developmental upregulation of NT-4/5 mRNA in rat SOL muscle fibers occurred earlier than the upregulation of MyHC I/b mRNA associated with muscle fiber transformation. This timing is consistent with the idea that NT-4/5 is involved in early events that lead to the upregulation of the slow MyHC isoform in this muscle.

Animals↗