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Distal neuropathy in experimental diabetes mellitus.

Although nerve conduction slowing is a well-accepted abnormality in rats with acute experimental diabetes, reports of neuropathological changes in diabetic rat nerves have been inconsistent. To examine this further, we studied electrophysiological and morphological features of posterior tibial nerves and their distal branches from four-week streptozotocin-induced diabetic rats and matched controls. Diabetic rat posterior tibial motor conduction was slowed (mean +/- 1 SD, 34.8 +/- 3.1 m/sec; controls, 41.2 +/- 2.5 m/sec), and evoked muscle response amplitudes were only half of control values. Using quantitative techniques, we documented a diminution in number of the largest myelinated fibers in otherwise normal mid posterior tibial nerves, with an increase in the smaller sizes, indicating either a degree of axonal atrophy or impaired fiber growth during development. The principal pathological finding was active breakdown of myelinated fibers in the most distal motor twigs of hindfoot muscles supplied by posterior tibial branches, with preservation of fibers in more proximal segments of these nerves. This anatomical lesion in diabetic nerves could account for both oberved conduction slowing and lowered muscle response amplitudes. A consistent feature in both diabetic and control mid lateral plantar nerves was a zone of demyelination that apparently occurs at this natural site of nerve entrapment in rats. Taken together, the pathological abnormalities of peripheral nerve in acute experimental diabetes are best explained as resulting from a distal axonopathy.

Animals↗

Estimate of gas transfer rates of an intravascular membrane oxygenator.

An asymmetric hollow fiber membrane was prepared from a newly synthesized fluorinated aromatic polyimide (6FDA-6FAP) by using a dry/wet phase inversion process. The membrane was used in a membrane oxygenator over a long period of time. In this study, the potential of the membrane for intravascular membrane oxygenation (IVOX) was studied in respect to oxygen transfer. The gas permeance of the membrane and three commercially available hollow fiber membranes for membrane oxygenators was measured in a gas-gas system and a gas-liquid system and discussed relative to the membrane structures. The oxygen transfer rates of the IVOX devices using these four membranes were estimated by a mathematical kinetic model, with the oxygen permeance measured in the gas-liquid system. The results showed that the device using the 6FDA-6FAP hollow fiber membrane has the highest oxygen transfer rate and is believed to be applicable to IVOX. The methods to determine oxygen transfer rate of a hollow fiber membrane and the mathematical kinetic model, are useful for developing a hollow fiber membrane and a device for oxygenation.

Humans↗

The regenerative potential of oxytalan fibers. An experimental study in the monkey.

Although a number of studies have described the oxytalan fibers as being a natural component of the periodontal ligament, little information exists about the regenerative potential of these connective tissue fibers. The aim of the present study was to examine whether oxytalan fibers have the capacity to reform after regenerative periodontal therapy. Intrabony defects were produced surgically at the mesial aspects of teeth 37, 35, 45, 47 and at the distal aspects of teeth 11, 21, 31, 41 in one monkey (Macaca fascicularis). After 3 months, the defects were exposed using a full-thickness flap procedure. The root surfaces were debrided and subsequently PDGF-growth factors were placed in the defects. 4 of the 8 sites were covered with a bioresorbable membrane before closure of the wound. Post-surgically, antibiotics were given systemically for 1 week, and tooth cleaning was carried out 1x a week during the entire experimental period. After 5 months, the animal was sacrificed and the oral tissues were fixed by perfusion with 10% buffered formalin. Specimens containing the defects and surrounding tissues were dissected free and histological sections were cut in the mesio-distal direction, parallel to the long axes of the teeth. The sections were stained with hematoxylin and eosin or with the oxone-aldehyde-fuchsin-Halmi staining method and subsequently examined in the light and in the electron microscope. The results revealed that new oxytalan fibers oriented mainly in an apico-occlusal direction had developed in the regenerated periodontal ligament. Many of the newly-formed fibers were inserted into the new cementum, thus suggesting a strong relationship between this tissue and the oxytalan fibers. It is concluded that the regenerated periodontal ligament connective tissue formed after surgery contains oxytalan fibers similar to those present in the original tissue. These results demonstrate that oxytalan fibers develop de novo in the newly-formed periodontal ligament.

Alveolar Bone Loss↗

Microanatomical and biochemical origins of normal and precancerous cervical autofluorescence using laser-scanning fluorescence confocal microscopy.

Clinical studies have shown that in vivo fluorescence spectroscopy can improve the diagnosis of cervical precancer. Recent work suggests that epithelial fluorescence increases, whereas stromal fluorescence decreases, with precancer. However, the microanatomic and biochemical sources of fluorescence in living cervical tissue have not yet been established. This study aims to characterize the origins of living normal and precancerous cervical fluorescence at microscopic levels using laser-scanning fluorescence confocal microscopy. Ten pairs of colposcopically normal and abnormal biopsies were obtained; transverse, 200 microm thick, short-term tissue cultures were prepared and imaged when viable with UV (351-364 nm) and 488 nm excitation before and after addition of the vital dye, Mitotracker Orange. In normal epithelium basal epithelial cells showed cytoplasmic fluorescence; parabasal, intermediate and superficial cells showed fluorescence only at the periphery of the cell. In low-grade precancers cytoplasmic fluorescence was visible in the bottom one-third of the epithelium; in high-grade precancers cytoplasmic fluorescence was visible throughout the lower two-thirds of the epithelium. Cytoplasmic fluorescence was colocalized with the MitoTracker probe and is attributed to mitochondrial reduced form of nicotinamide adenine dinucleotide at UV excitation and mitochondrial flavin adenine dinucleotide at 488 nm excitation. Stromal fluorescence originated from matrix fibers; with the development of precancer the density and fluorescence intensity of matrix fibers decrease. Autofluorescence properties of precancerous cervix reflect an increased number of metabolically active mitochondria in epithelial cells and a reduced stromal fluorescence, which can be an indicator for altered communication between precancerous epithelium and stroma. These changes can explain differences in in vivo fluorescence spectra of normal and precancerous cervical tissue.

Cervix Uteri↗

Is myoelectric activity transmittable from one muscle to another: an experimental study.

BACKGROUND: In contrast to other striated muscles in the body, pelvic floor muscles possess resting electric activity. We have demonstrated that this activity was due to smooth muscle fibers detected in the levator ani muscle (LAM) which presumably develop in adaptation to variations in the intraabdominal pressure and visceral weight action. AIMS: The current communication tries to answer the question whether this resting myoelectric activity is transmittable to other muscles which lack such activity. METHODS: The LAM and obturator internus muscle (OIM) were exposed in 14 mongrel dogs through a paraanal incision. The resting EMG activity of both muscles was recorded. The border of the LAM closest to its origin at the white line, was released and sutured to the obturator fascia. The EMG activity of both the LAM and OIM was recorded at rest and on LAM stimulation. The EMG recording was repeated after suturing of the LAM to the OIM whose fascia had been removed. The test was repeated after LAM anesthetization. RESULTS: Resting and stimulated EMG activity was recorded in the LAM, but not in the OIM, before and after suturing the LAM to the obturator fascia. On removing the obturator fascia, both muscles recorded similar EMG activity at rest and on LAM stimulation (p > 0.05, p > 0.05, respectively). Thirty minutes from LAM anesthetization both muscles showed no EMG activity at rest or on LAM stimulation; after 3 hours, the 2 muscles recorded an activity similar to that before anesthetization (p > 0.05). CONCLUSION: The myoelectric activity can be transmitted to defasciated muscles. This finding may be useful in electrostimulation therapy and in developing type 1 fibers in striated muscles.

Animals↗

Morphologic and electrophysiologic identification of multiply innervated fibers in rat extraocular muscles.

Mammalian extraocular muscles comprise singly and multiply innervated fibers (SIFs, MIFs) that are distributed in both the global and orbital layers of the muscles. Previously it has been concluded solely on the basis of electrophysiologic analysis that in rat, MIFs of the global layer show graded responses or slow peak potentials instead of the action potentials exhibited by SIFs. To confirm directly this conclusion, a protocol that allows the combined electrophysiologic-morphologic characterization of muscle fibers has been developed. It is based on electrophysiological analysis of the muscle fibers followed by intracellular labeling with Lucifer yellow and staining of nerve endings with a modified Koelle stain for cholinesterase activity. Labeled fibers have been recognized and followed in sequential 20-micron cross-sections of the whole muscle.

Action Potentials↗

Tyrosine hydroxylase and neuropeptide-Y immunoreactivity in pineal glands developing in situ and in pineal grafts.

Postnatal development of the innervation of the pineal gland in situ as well as the reinnervation of pineal grafts by tyrosine hydroxylase (TH)- and neuropeptide Y (NPY)-immunoreactive nerve fibers were examined using the avidin-biotin-peroxidase immunohistochemical technique. TH-immunoreactive nerve fibers appeared in the pineal gland on the second postnatal day (P2) in both hamsters and gerbils. NPY-immunoreactive nerve fibers first appeared in the pineal gland of gerbils on P2 and in the hamsters on P3. By the seventh postnatal day (P7), the pineal glands of both hamsters and gerbils were richly innervated by TH- and NPY-fibers that appeared as smooth fibers or fibers with sporadic varicosities. By the age of 4 weeks, the innervation of the pineal glands of hamsters and gerbils by TH- and NPY-fibers was fully developed. Abundant TH- and NPY-fibers formed a dense meshwork in the parenchyma of the superficial and deep pineals. The great majority of the fibers bore a large number of varicosities. More NPY-fibers were found in the pineal glands of gerbils than hamsters. NPY fibers were distributed evenly throughout the pineal glands of the gerbil, but they were more often located in the central region of the superficial pineal of the hamster. For the pineal grafts, superficial pineals from neonatal and 4-week-old hamsters were transplanted to different sites in the third cerebral ventricle (infundibular recess, posterior third ventricle) or beneath the renal capsule. The pineal grafts from 4-week-old donors appeared to undergo severe degeneration and eventually disappeared. The pineal grafts from neonatal hamsters, however, successfully survived and became well integrated into their new locations. Abundant TH- and NPY-fibers in the host brain were found surrounding the pineal grafts placed in the third cerebral ventricle, but were only rarely seen entering the parenchyma of the grafts. A few TH-fibers were demonstrated in the renal grafts 4 weeks after transplantation. These studies describe the postnatal development of the innervation of the pineal glands in situ by TH- and NPY-nerve fibers, and demonstrate a lack of reinnervation of cerebroventricular pineal grafts by TH and NPY fibers from adjacent host brain.

Animals↗

Transient patterns of serotonergic innervation in the rat visual cortex: normal development and effects of neonatal enucleation.

The transient aggregation of serotonin (5-HT)-containing fibers in the early development of rat visual cortex was examined immunohistochemically. The aggregation of 5-HT immunoreactive (IR) fibers consisted of three stages which were classified according to the course of time and degree of space occupied. The primary aggregation appeared in the subplate and moved upward along the development of the cortex. The aggregation proceeded to the secondary stage in presumptive layer IV. The fibers extended in a column-like structure following the secondary aggregation and formed the tertiary aggregation. The upper edge of the tertiary aggregation formed a lattice-like pattern in layer I and its structure was recognized to be similar to the structure of a 'blob' which characterizes the primary visual cortex in monkey. This transient aggregation of 5-HT-IR fibers began in the subplate of the anterior visual cortex on postnatal day 2 (PND 2) and progressed towards the posterior. On PND 11, the secondary and tertiary aggregations were completed in the entire region. No further aggregation of 5-HT-IR fibers was observed on PND 15. The anterior-to-posterior axis in the aggregation process corresponds to the direction of differentiation in the layer structure of cortex. In order to investigate the relationship between the transient aggregation of 5-HT-IR fibers and the development of the visual pathway, the secondary and tertiary aggregation on PND 11 were observed after postnatal monocular or binocular enucleation. Enucleation of eye balls did not affect either the area occupied by the 5-HT-IR fibers in the secondary aggregation or the number of column structures in the tertiary aggregation. However, the contralateral and ipsilateral cortices of monocularly enucleated cases were irregularly shaped in the secondary aggregation. The distribution of 5-HT-IR fiber terminals in the binocular area (Oc1B) increased in density on the contralateral side in the monocular enucleation, while that of both sides in the binocular enucleation was of non-homogeneous density and were shaped irregularly. The above results suggest that the transient aggregation of 5-HT-IR fibers observed in the early stage of development of visual cortex is regulated primarily by the intrinsic factors, and that extrinsic factors, such as visual pathway input, affect the aggregation within the boundary of such intrinsic factors. That is, the visual pathway input and the input balance from both eyes affect the distribution density of 5-HT-IR fibers and the shape of the visual cortex, respectively.

Animals↗

[Development of the mitochondrial apparatus and blood supply of skeletal muscle fibers during ontogenesis of domestic fowl].

The diameter, length, and numerical density of capillaries, diameter of muscle fibers, size and numerical density of their profiles, and relative volume of mitochondria in them were determined in the chicken red oxidative gastrocnemius and white glycolytic pectoral muscle during development from day 10 of embryogenesis to six month of postnatal life. The bulk blood flow was measured in these muscles by hydrogen clearance during postembryonic development. During embryogenesis, the fibers of gastrocnemius muscle develop and grow at a higher rate, while during postembryonic development, those of the pectoral muscle develop faster. The density of mitochondrial profiles increases during embryogenesis and decreases after hatching, while their mean size increases, especially in the oxidative fibers, but it somewhat decreases in 6-month old chicks. Redistribution of mitochondria by the fiber section during development takes place in both muscles: they are localized predominantly in the center in 18-day embryos and in the periphery, especially in the gastrocnemius fibers, in 6-month old fowl. At hatching, the lengths of capillaries are similar in both muscles, but as chicks grow, the proportion of longer (more than 600 microm) capillaries in the pectoral muscle sharply increases, while their density and bulk blood flow decrease. Ratios were determined between structural parameters of the capillary bed and mitochondria, on the one hand, and oxygen consumption (ml/min per 1 mm fiber and 100 g muscle mass), on the other.

Animals↗

Early areal differentiation of the human cerebral cortex: entorhinal area.

The early cytoarchitectonic specialization and area-specific differentiation of the prospective entorhinal cortex were studied in the postmortem human fetal brains (9.5-13.5 postovulatory weeks). At 10 weeks, using the Golgi method, we saw the appearance of area-specific large neurons (promoter neurons) with widely bifurcating apical dendrites situated at the outer margin of the cortical plate of the prospective entorhinal cortex. The analysis of the serial Nissl-stained sections revealed the concomitant appearance of a one-cell-thick layer (monolayer) at the interface between the cortical plate and the marginal zone and multilaminated spread of the deep part of the cortical plate. This is the earliest sign of area-specific cytoarchitectonic differentiation of all pallial regions characterized by the presence of the typical cortical plate. The first subareal differentiation within the entorhinal cortex begins at 13 postovulatory weeks with uneven development of fiber-rich lamina dissecans, which separates two cellular laminae principals (externa and interna), and with the appearance of characteristic cell islands of the prospective layer II. At rostral levels, cell islands begin to develop in the rostromedial entorhinal area at the subpial depths where large promoter neurons reside. At intermediate levels, both lamina dissecans and lamina principalis interna are well delineated. At caudal levels, lamina principalis interna is continuous with the upper subplate zone of the adjacent neocortex. Both area-specific neurons (promoters) and fiber-rich (afferent) strata develop synchronously during the earliest areal differentiation of the cerebral cortex. The precocious lamination of the cortical plate is the crucial event in the histogenesis of the entorhinal cortex.

Afferent Pathways↗

[Differentiation of neurons and synapses of mossy fibers in transplants of fascia dentata developing in the rat neocortex].

Embryonic fascia dentata tissue isolated from the hippocampus was transplanted heterotopically into the neocortex of adult rats. Ultrastructural characteristics of neurons and synapses in transplants were studied nine months later. It has been found that the main types of neurons present in fascia dentata undergo differentiation in the transplants, and a dense neuropile containing various types of synapses is produced. A characteristic feature of the transplanted neurons is the presence of additional microspines on somatic and dendrite surfaces; this appears to be due to a deficiency of external and internal afferents. Gigantic synaptic terminals of granule cell axons (mossy fibers) in transplants possess unique morphological characteristics, which allow their identification in a complex neuropile. Just as in situ, they form two types of contacts: chemical asymmetric contacts with dendrite spines and desmosome-like ones with dendrite surface characteristics. However, accumulations of large vesicles with electron-dense centers can often be observed near the active zones of the synapses, and desmosome-like connections are more prominent. The most important feature is that gigantic synapses in transplants use midsize and small dendrites as postsynaptic targets up to terminal branches, and they contact with spines of the usual shape and size, whereas in situ terminal synaptic contacts of mossy fibers are formed only with gigantic processes of initial segments of the large apical dendrites. Thus, in the absence of normal synaptic targets, mossy fibers can produce contacts having all features of functional synapses, but with atypical postsynaptic structures.

Animals↗

The extracellular matrix protein betaIG-H3 is expressed at myotendinous junctions and supports muscle cell adhesion.

Molecules of the extracellular matrix (ECM) play important roles in the development and maintenance of myotendinous junctions (MTJs), specialized regions of muscle to bone union. In this report we provide evidence that skeletal muscle cells synthesize the collagen- and fibronectin-binding ECM protein betaIG-H3 and that betaIG-H3 is localized to MTJs. In situ hybridization experiments revealed that during E16.5-E18.5 of murine development, betaIG-H3 RNA transcripts were expressed where developing skeletal muscle fibers contact primordial cartilage and bone. Immunohistochemical analysis verified that the betaIG-H3 protein itself localized distinctively at MTJs, and ultrastructural analysis suggested that betaIG-H3 associates with extracellular fibers and the surface of cells. In vitro, recombinant betaIG-H3 functioned as an adhesion substratum for skeletal muscle cells. Adhesion was significantly reduced by anti-integrin alpha7 and beta1 antibodies, suggesting that betaIG-H3 binds to skeletal muscle cells via alpha7beta1 integrin. Localization of betaIG-H3 to the termini of skeletal muscle fibers and the binding of betaIG-H3 to cells and to molecules of the ECM suggests that betaIG-H3 may play an organizational and structural role in developing MTJs, linking skeletal muscle to components of the ECM.

Animals↗

Autoradiographic evidence that septohippocampal fibers reinnervate fascia dentata denervated by entorhinal lesion during development.

The projection from the ventromedial septum to the fascia dentata was investigated autoradiographically in normal adult rats and in adult rats whose entorhinal cortex had been removed unilaterally at the age of 11 days. In the fascia dentata of normal rats and in the fascia dentata contralateral to the entorhinal lesion septohippocampal fibers and terminals were distributed just below and, to a lesser extent, just above the granular layer. The molecular layer above the supragranular zone was lightly and more or less uniformly innervated. Ipsilateral to the entorhinal lesion, however, the outer part of the dentate molecular layer received an anomalously dense septal projection (average 3-4 times the contralateral projection). The entorhinal lesion did not consistently affect the density of this projection in any other lamina. These results confirm that septohippocampal fibers increase their density of innervation when synaptic sites are made available by degeneration of lateral perforant path fibers during development. This represents a net increase in total septal innervation of the fascia dentata, not merely a change in the distribution of the projection among its target zones.

Afferent Pathways↗

Unraveling the neuron jungle: the 1879-1886 publications by Wilhelm His on the embryological development of the human brain.

The neuron doctrine, one of the central paradigms of the neurological sciences, states that neurons are individual units. Wilhelm His helped to lay the scientific foundation for the neuron doctrine. This Swiss researcher's experimental approach, which was based on the examination of embryological material, is of particular interest because few investigators at that time were using it. Only selected excerpts from His' seminal 1886 work have been translated into English. Therefore, details regarding his experimental materials and primary observations are unavailable to the American reader. Our objective was to translate additional sections of the seminal 1886 publication and, when relevant, portions of previously untranslated works dating back to 1879 and 1883 so that we could better understand this embryologist's experimental materials, his primary observations, and the basis for his conclusions. By 1886, His had compiled a collection of 12 human embryos of various measured lengths and estimated gestational ages ranging from 2.15 mm to 24 mm, and 2 weeks to 8.5 weeks. He had studied the embryological development of nerve fibers in successively older specimens. For example, he contrasted the young embryo N, whose "nerves. stop only halfway and the distal part [of the extremity] is completely nerve-free," with the older embryo Zw, in which the nerves had already reached "the root of the fingers but none [were] in the territory of the end phalanges." In the 1870s and 1880s, using a discrete collection of 12 human embryos of consecutively older ages, His observed the development of nerve fibers and, from this pattern of growth, eventually suggested that nerve cells were individual units and that the transmission of impulses was possible without direct continuity between neurons. Although this age-based embryological approach seems relatively straightforward by today's standards, few other researchers were using it at the time.

Brain↗

Zebrafish slow muscle cell migration induces a wave of fast muscle morphogenesis.

The specification and morphogenesis of slow and fast twitch muscle fibers are crucial for muscle development. In zebrafish, Hedgehog is required for slow muscle fiber specification. However, less is known about signals that promote development of fast muscle fibers, which constitute the majority of somitic cells. We show that when Hedgehog signaling is blocked, fast muscle cell elongation is disrupted. Using genetic mosaics, we show that Hedgehog signal perception is required by slow muscle cells but not by fast muscle cells for fast muscle cell elongation. Furthermore, we show that slow muscle cells are sufficient to pattern the medial to lateral wave of fast muscle fiber morphogenesis even when fast muscle cells cannot perceive the Hedgehog signal. Thus, the medial to lateral migration of slow muscle fibers through the somite creates a morphogenetic signal that patterns fast muscle fiber elongation in its wake.

Animals↗

Animal models of malignant mesothelioma.

Animal models of diffuse malignant mesothelioma have historically been used to assess carcinogenicity of various fiber types and to study the pathogenesis of this unusual neoplasm. Pleural and peritoneal mesotheliomas have been induced in rodents following exposure to erionite or asbestos fibers, radionuclides, particulate nickel compounds, and chemicals such as 3-methylcholanthrene. The role of SV40 virus as a cofactor with asbestos fibers in the development of diffuse malignant mesotheliomas in humans has been explored in animal models. SV40 virus alone induces mesotheliomas in hamsters. Generation of new transgenic mouse strains with targeted expression of SV40 large T and small t antigens in the mesothelium would be very useful for mechanistic studies. Human malignant mesotheliomas frequently show hypermethylation or deletions at the Cdkn2a/Arf and Cdkn2b gene loci and deletions or mutations at the NF2 gene locus. Heterozygous Nf2 (+/-) mice exposed to crocidolite asbestos fibers exhibited accelerated development of malignant mesotheliomas compared to wild-type littermates. Loss of the wild-type Nf2 allele, leading to biallelic inactivation, was observed in nine mesothelioma cell lines derived from Nf2 (+/-) mice. Similar to human malignant mesotheliomas, tumors from Nf2 (+/-) mice showed frequent homozygous deletions of the Cdkn2a/Arf locus and adjacent Cdkn2b tumor suppressor gene. As in the human disease, murine mesotheliomas also showed constitutive activation of Akt. This murine model of asbestos carcinogenesis recapitulates the molecular and histopathological features of the human disease and has significant implications for preclinical testing of novel preventive or therapeutic modalities.

Animals↗

Myocardial tissue engineering with autologous myoblast implantation.

OBJECTIVE: Implanting myoblasts derived from autologous skeletal muscle, that is, satellite cells, for myocardial replacement has many advantages when compared with implanting either fetal cardiac myocytes (ethical and donor availability issues) or established cell lines (oncogenicity). Furthermore, autologous myoblasts do not require immunosuppression. The feasibility of satellite cell differentiation into muscle fibers, after implantation into the myocardium, was confirmed by means of a unique cell-labeling technique. METHODS: Myoblasts (satellite cells) isolated from the skeletal muscle of adult rats are labeled with 4',6-diamidino-2-phenylindone, which binds to DNA and to the protein tubulin to form a fluorescent complex, and implanted into the left ventricular wall of isogenic rats. The specimens are harvested 1 to 4 weeks after myoblast implantation. Histologic sections are examined under a fluorescent microscope. RESULTS: The labeling efficiency of satellite cells with 4',6-diamidino-2-phenylindole is nearly 100%. In 4 specimens, the progressive differentiation of implanted myoblasts into fully developed striated muscle fibers can be observed. CONCLUSION: Our earlier studies of autologous myoblast implantation into the cryoinjured myocardium of dogs suggested that these cells could differentiate into cardiac myocytes. However, it had been difficult to firmly establish these findings with the use of cell markers, thereby proving that the neomyocardium had indeed been derived from the implanted myoblasts. In this study, using 4',6-diamidino-2-phenylindole as a satellite cell marker, we were able to demonstrate that the implanted satellite cells did in fact differentiate into fully developed, labeled muscle fibers. Because of the obvious advantages of using autologous donor myoblasts, the clinical application of this approach may provide a novel strategy for the future management of heart failure.

Animals↗

Normal and abnormal pathfinding of facial nerve fibers in the chick embryo.

Development of the facial nerve was studied in normal chicken embryos and after surgical disruption of ingrowing sensory facial nerve fibers at 38-72 h of incubation. Disruption of facial nerve fibers by otocyst removal often induced a rostral deviation of the facial nerve and ganglion to the level of the trigeminal ganglion. Cell bodies of the geniculate ganglion trailed their deviating neurites and occupied an abnormal rostral position adjacent to the trigeminal ganglion. Deviating facial nerve fibers were labeled with the carbocyanine fluorescent tracer DiI in fixed tissue. Labeled fibers penetrated the cranium adjacent to the trigeminal ganglion, but they did not follow the trigeminal nerve fibers into the brain stem. Rather, after entering the cranium, they projected caudally to their usual site of entrance and proceeded towards their normal targets. This rostral deviation of the facial nerve was observed only after surgery at 48-72 h of incubation, but not in cases with early otocyst removal (38-48 h). A rostral deviation of the facial nerve was seen in cases with partial otocyst removal when the vestibular nerve was absent. The facial nerve followed its normal course when the vestibular nerve persisted. We conclude that disruption of the developing facial pathway altered the routes of navigating axons, but did not prevent pathfinding and innervation of the normal targets. Pathfinding abilities may not be restricted to pioneering axons of the facial nerve; later-developing facial nerve fibers also appeared to have positional information. Our findings are consistent with the hypothesis that navigating axons may respond to multiple guidance cues during development. These cues appear to differ as a function of position of the navigating axon.

Acetylcholinesterase↗