Nerve fiber analysis and age-related changes of the human mandibular nerve

We morphometrically analyzed nerve fibers of the human mandibular nerve with a discriminative staining method that makes it possible to separate nerve fibers into myelinated and unmyelinated fibers. We counted numbers and transverse areas of myelinated axons under the microscope using an on-line image-analyzer. This study revealed the morphometric changes which affect the human mandibular nerve during the aging process.     

Unmyelinated nerve fibre analysis of the human lesser splanchnic nerve

The aim of the present study is to analyse unmyelinated nerve fibres of the human lesser splanchnic nerve in relation to the ageing process. With the help of an image-analyser, we examined 30 human lesser splanchnic nerves. The analysis was conducted with the use of a new staining method that makes it possible to discriminate various structures of the nervous tissue. Our report provides for the first time information on the ageing process of the human lesser splanchnic nerve fibres. The results indicate that a decrease in transverse area and perimeter of unmyelinated axons is one of the important changes occurring in the human lesser splanchnic nerve during the ageing process.     

Nerve fiber composition of the intracranial portion of the oculomotor, trochlear, and abducens nerves in the sheep

In the present investigation, the fiber content and the diameter spectra of the intracranial portion of the three oculomotor nerves (oculomotor, trochlear, and abducens nerves) were analysed in sheep by light and electron microscopy. It was determined that up to 14.98% of fibers in the oculomotor nerve, 17.01% in the trochlear nerve, and 11.87% in the abducens nerve were unmyelinated. The myelinated fibers showed a bimodal distribution in their size spectrum in all three nerves, with a majority of large myelinated axons, but a considerable proportion of small myelinated fibers, as well. The sensory function of the unmyelinated fibers present in the three oculomotor nerves is discussed also on the basis of our previous morphofunctional investigations.     

Unmyelinated nerve fibers of the human mandibular nerve

The aim of this research is to find and to evaluate morphometorically the unmyelinated nerve fibers in the human mandibular nerve using a light microscope. Our report demonstrates for the first time the presence of the unmyelinated nerve fibers of the human mandibular nerve stained by a special method. Our results also indicate that there is a morphometric change with aging in the unmyelinated axons of the nerve.     

Unmyelinated nerve fibers

The paper presents a critical review of various current concepts of the structure and kinetics of unmyelinated nerve fiber. A classification of nerve fibers, different from the earlier ones, is proposed, that demonstrates not only the morphological fiber types, but also the kinetics of their reversible transition stages from non-glial to myelinated fiber. Evidence is presented to show the erroneousness of conceptions, still appearing in many publications, that consider the unmyelinated nerve fiber as "the Remak's cable type fiber". According to the current data, "Remak's fiber" is a glial-neurite complex, i.e. a bundle of unmyelinated nerve fibers covered with a single glial cell. Using the electron microscope, it was demonstrated that comparable glial-neurite complexes of myelinated nerve fibers, formed in CNS in a similar way by a single oligodendrocyte, cannot be named a single fiber. Cutting the nerves makes visible that the single fibers forming "the Remak's fiber" stem from different cells, therefore they cannot be a single "fiber". It has been shown for the first time experimentally, that in extreme situations, as a result of contraction of gliocyte processes, unmyelinated fibers may "leave" the glial-neurite complexes and become the nonglial fibers. Some data are presented that may serve as criteria for differentiation of unmyelinated fiber with a stratified sheath from developing myelinated fiber.     

Morphological evidence for functional capsaicin receptor expression and calcitonin gene-related peptide exocytosis in isolated peripheral nerve axons of the mouse

Rat sciatic nerve axons express capsaicin, proton and heat sensitivity and respond to stimulation with a Ca2+-dependent and graded calcitonin gene-related peptide (CGRP) release. In this study we demonstrate that similar functions, including capsaicin-induced CGRP release, are to be found in the desheathed sciatic nerve of the mouse. We have morphologically investigated the mechanisms of this axonal release in regions away from the active zones of synapses. Capsaicin receptor 1 (TRPV1) and CGRP immunostaining was performed using electron microscopic visualization. TRPV1 was identified in the axoplasm and inside vesicles--presumably on axonal transport--as well as in considerable quantity in the axonal plasma membrane of unmyelinated nerve fibers. Most of the unmyelinated axons were immunopositive for CGRP and in unstimulated nerves CGRP-containing vesicles almost entirely filled the axoplasm. After capsaicin stimulation (10(-6) M for 5 min), the fibers appeared depleted of CGRP with only few vesicles remaining as well as some residual staining of the axoplasm. In addition a large number of vesicles were fused with the axonal membrane, forming classical exocytotic figures--the omega structures--lined with CGRP immunoreactive product. These results present morphological evidence for the distribution of TRPV1 along unmyelinated axons in peripheral nerve and also provide the first demonstration of vesicular neuropeptide exocytosis along unmyelinated axons in peripheral nerve.     

Structural and functional adaptation of the pulpal nerve fibers in the rat incisor

Special fine structure of the pulpal nerve fibers adapted to the continuously growing incisor was studied on 20 rats (25-d-old-males) electron microscopically and was compared with that of the molars of limited growth. The incisal pulp was innervated by 2 kinds of the nerve fibers derived from the cervical sympathetic and the trigeminal ganglia: polyaxonal unmyelinated and monoaxonal myelinated. In the apical pulp, however, the pulpal nerve fibers were unmyelinated, showing 3 different profiles in the fine structure: 1. polyaxonal, 2. polyaxonal with dial plate-like profile and 3. monoaxonal. The monoaxonal unmyelinated fiber reflected the transverse profile of the unmyelinated part of the myelinated axon derived from the trigeminal ganglion. It can be concluded that the pulpal nerve fibers derived from the trigeminal ganglion maintain the immatured nature in the apical proliferating pulp to adapt to the continuous growth of the incisor.     

Myelinated nerve fiber analysis of the human greater splanchnic nerve.

The purpose of the present study is to analyze the human greater splanchnic nerve in relation to aging. We adopted a new staining method which makes it possible to discriminate various structures of the nervous tissue. We examined 25 human greater splanchnic nerves from cadavers for anatomy dissection. We measured the number, area and perimeter of axons. The results reveal that: (1) there is no correlation between age and the number of axons; (2) the mean area and perimeter of axons increase with age, but not the total area and perimeter. We compared these results with those for the lesser splanchnic nerve. These morphological changes in the greater splanchnic nerve may indicate a kind of compensation through axon hypertrophy for hypofunctions in abdominal organ control.     

Quantitative study of the apical nerve fibers of adult and juvenile rat molars

The rat molar has become an important model for studies of interactions between nerves and the pulp-dentin complex, yet there is only limited quantitative information on the number and size distribution of axons entering the roots of this tooth. This study was undertaken to provide such a detailed characterization of the apical innervation of the rat molar. An additional objective was to compare the apical nerve composition of young, recently erupted rat molars with that of mature teeth in order to determine whether there is ongoing maturation of the innervation after the teeth have attained functional occlusion. A complete census was made of the nerve fibers entering the roots of both mature and recently erupted juvenile mandibular first molars in Sprague-Dawley rats. Each of the four roots of the first molars was processed for electron microscopy of thin sections near the apex. The majority of intradental nerve fibers entered the molar via the two larger (mesial and distal) roots. Within the apical root pulp, most, but not all, axons occurred within well-defined fascicles associated with blood vessels. Molars from adult animals (age 4 months) had a mean total of 232 (S.D. = 49, N = 7 teeth) myelinated fibers and 806 (S.D. = 143) unmyelinated axons entering the four roots. Fibers exceeding the Aδ size range (circumference ≥ 19 μm) accounted for only 4% of the myelinated axons at the apex. Molars from juvenile animals (age 4 weeks) had fewer myelinated fibers (mean 176, S.D. 18, N = 8), but more unmyelinated axons (mean 1,174, S.D. 160) than adults. The mean ratio of unmyelinated axons to myelinated axons was 6.6:1 for juveniles compared to 3.5:1 for adults. Juvenile teeth contained no myelinated fibers that exceeded 19 μm in circumference. These results indicate that the innervation of the rat molar resembles that of teeth of non-rodent mammals in that (1) innervation density is high, (2) there is a high ratio of unmyelinated axons, and (3) most of the myelinated fibers are of thin caliber. Furthermore, it appears that after the molar erupts, maturation of the nerve fiber composition continues with processes that include both a marked decrease in the number of unmyelinated axons and an increase in the number and size heterogeneity of myelinated fibers. © 1994 Wiley-Liss, Inc.     

自噬抑制剂3-甲基腺嘌呤可提高小鼠同种异体坐骨神经移植效率

文题释义： 细胞自噬：指细胞在受到创伤、饥饿、缺氧、感染等应激状态下的一种自我保护机制,这一过程可以无选择性地发生于所有真核细胞中,真核细胞通过自噬作用清除老化的细胞器和蛋白,以此来维持细胞生长发育的平衡。自噬过程主要的诱发因素是饥饿,即细胞营养物质的缺乏,此外也可通过一些感染、损伤、特定的蛋白如热休克蛋白、细胞因子等选择性地引发。 华勒氏变性：是指周围神经损伤后,残留的轴突及髓鞘结构迅速发生退化、崩解、吸收的过程。这一复杂过程有多种细胞因子及炎症细胞参与,是周围神经损伤后最重要的病理变化过程之一,影响损伤后续的修复再生。 背景：近年最新研究表明,华勒氏变性的发生与许旺细胞的自噬活动密切相关,对许旺细胞自噬活动进行调控,可以显著影响华勒氏变性的发生发展,从而改变后续的轴突再生及髓鞘化过程。 目的：在同种异体神经移植中对移植片段的细胞自噬过程进行抑制,观察是否影响移植后的修复效率。 方法：获取8只雌性C57BL/6J小鼠(购自北京维通利华)坐骨神经片段16条,分2组,分别于含自噬抑制剂3-甲基腺嘌呤的培养基及普通培养基中处理72 h。取16只雌性C57BL/6J小鼠,建立左侧坐骨神经缺损模型,实验组(n=8)植入含自噬抑制剂3-甲基腺嘌呤培养基处理过的坐骨神经片段,对照组(n=8)植入普通培养基处理的坐骨神经片段,术后2,4,6,8周,记录坐骨神经指数;术后8周取再生坐骨神经段,分别进行苏木精-伊红染色、免疫荧光染色、甲苯胺蓝染色、透射电镜观察等。动物实验通过北京协和医学院动物伦理委员会批准。 结果与结论：①实验组术后8周的坐骨神经指数高于对照组(P < 0.05),其余时间点两组间比较差异无显著性意义(P > 0.05);②苏木精-伊红染色显示,实验组神经组织完整,对照组神经组织可见大面积空洞;③免疫荧光染色显示,实验组可见较完整的神经束结构,对照组未见完整的神经束结构;④甲苯胺蓝染色显示,实验组可见有髓神经纤维和部分再生无髓神经纤维,对照组仅见少量有髓神经纤维与新生无髓轴突;⑤透射电镜显示,实验组髓鞘厚度及有髓纤维直径均大于对照组(P < 0.05);⑥结果表明应用3-甲基腺嘌呤处理移植前神经片段,可抑制许旺细胞自噬,有助于保留移植物髓鞘结构完整性,促进轴突再生及功能的恢复。 ORCID: 0000-0002-6259-2668(徐筑秋) 中国组织工程研究杂志出版内容重点：组织构建;骨细胞;软骨细胞;细胞培养;成纤维细胞;血管内皮细胞;骨质疏松;组织工程     

大鼠眼上直肌神经的生后发育

取生后两天、两周和两月的雄性Ｗｉｓｔａｒ大鼠双眼上直肌，进行硝酸银染色和乙酰胆碱酯酶染色，用光镜观察大鼠眼上直肌神经在生后的发育情况。在大鼠生后两天时，动眼神经的分支从眼上直肌的中后三分之一交界处由眶面进入该肌，它向该肌的球面区发出细小的神经纤维束，这些神经束抵达同一条肌纤维上。眶面层神经纤维细小，单条并且平行于肌纤维方向走行。此时，眼上直肌对乙酰胆碱酯酶染色着色浅，反应区未形成某一特定轮廓，说明此时运动神经的发育是不成熟的。在生后两周时，动眼神经的分支开始向眼上直肌的眶面层发出神经纤维；球面区中的神经纤维有明显的粗细两种。这时眼上直肌中的运动神经轴索对乙酰胆碱酯酶染色反应，出现葡萄状运动神经末梢及斑点状运动终板的雏形。生后两月时，眼上直肌中出现了肌梭及典型的葡萄状运动神经末梢和斑点状运动终板。生后两周是大鼠睁眼初期，由此可见，大鼠从闭眼至睁眼的时期，是运动神经发育的关键时期，此时，运动神经形成一些特定神经末梢，而睁眼后运动神经的发育主要在此基础上运动神经末梢日趋完善和成熟。     

Anodally focused polarization of peripheral nerve allows discrimination of myelinated and unmyelinated fiber input to brainstem nuclei

We investigated the ability of a novel direct current (DC) polarization technique to block selectively the conduction in peripheral myelinated nerve fibers and allowing propagation in only unmyelinated fibers. In anesthetized adult rats, distal branches of the sciatic nerve (caudal cutaneous sural and tibial nerves) were exposed for electrical stimulation of A- and C-fibers. Two specially fabricated trough electrodes of different size and surface area were placed onto the sciatic nerve. Through these proximal electrodes a controlled ramped DC was timed to coincide with the arrival of A- and C-fiber action potentials, evoked electrically at the distal nerves or naturally from the foot or ankle, with the intent of blocking propagation in A-fibers while allowing C-fiber throughput. Neuronal recordings were made both peripherally (proximal sciatic nerve fascicles or L5 dorsal roots) and centrally (single cells in the nucleus gracilis or nucleus reticularis gigantocellularis). The DC polarization was shown to block conduction in myelinated A-fibers effectively, while allowing conduction in the unmyelinated C-fibers, without activation of fibers via the DC polarization itself. This was dependent upon the following factors: electrode polarity, onset rate of polarization, peak amplitude of polarization, distance between polarizing electrodes, size difference between polarizing electrodes, and gross nerve size. These experiments demonstrate that anodally focused DC polarization, applied utilizing two trough electrodes of different sizes, is capable of effectively, reversibly, and reproducibly blocking conduction in myelinated A-fibers evoked either electrically or naturally, while still allowing conduction to occur in the unmyelinated C-fiber population. In the context of experimental usage, we have demonstrated blocking of low-threshold A-fiber, but not C-fiber, mediated inputs to the caudal brainstem. This technique should find wide application in studies involving the processing of information conveyed centrally by the unmyelinated C-fiber afferent population, including discriminating afferent responses to peripheral stimuli, the role of C-fiber input in reflex activity, and the plasticity following injury or other manipulations. Received: 14 November 1997 / Accepted: 3 March 1998     

Unmyelinated axons in a muscle nerve

Summary In continuation of earlier studies on the innervation of the sternomastoid muscle of the rat, a detailed morphometric analysis was performed on the unmyelinated axons of the nerve, in normal rats and after extensive cervical sympathectomy.In 4 normal rats an average of 314 myelinated and 319 unmyelinated axons were present. 42 days after surgery, the 3 animals showed Horner's syndrome and a highly significant 40% loss of unmyelinated axons. We therefore suggest that 40% of the C-fibers in this nerve are postganglionic sympathetic efferents and that the remaining 60% are type IV fibers, i.e., unmyelinated afferents. Our counts also indicate that part of the Remak bundles of the Schwann cells contain only sympathetic axons, whereas others contain mixed groups of sympathetic and afferent axons. Myelinated nerve fibers were not lost due to sympathectomy.Unexpectedly, the 3 animals analyzed 7–13 days after surgery showed Horner's syndrome but only a 16% loss of unmyelinated axons, which was not even statistically significant. Morphological signs of degeneration and sprouting did not provide any clue, but a possible explanation would be that a transitory sprouting of the remaining afterent C-fibers or Schwann cells occurred.     

Vagal nerve maturation in the fetal lamb: An ultrastructural and morphometric study

The maturation of the left vagal nerve was studied in the fetal lamb by transmission electron microscopy and by computer-assisted morphometry of sections of the entire nerve at seven gestational ages between 79 and 145 days (term is 147 days) and in the adult ewe. The number of unmyelinated axons per Schwann cell progressively decreased from 25 to 55 at 79 days to 1 to 5 at near-term. Unmyelinated axons of various sizes were enclosed within a single Schwann cell at all ages, but the mean axonal diameter increased in inverse relation to the number of unmyelinated axons. A few Schwann cells enclosed two myelinated axons, but in most instances myelination did not begin until a 1:1 ratio was achieved; some single axons with a Schwann cell remained unmyelinated in the adult. Myelinated fibers were rare at 79 days but myelination progressed rapidly thereafter until the adult ratio of myelinated: unmyelinated fibers was reached at about 100 days; myelinated axons were not uniformly distributed. The myelin sheaths and axons of small fibers progressively increased in diameter in late gestation, but new large fibers were not added. Early myelinating fibers and immature unmyelinated axons contained more microtubules than neurofilaments; neurofilaments predominated in mature axons with or without myelin. Cross-linkages between neurofilaments were already evident by 79 days. Maturation of the vagal nerve thus occurs first by an increase in number of myelinated fibers and then by an increase in the size of each fiber in this fixed population. The bimodal distribution in the size histogram of myelinated fibers is not achieved until 134 days gestation and correlates well with physiological maturation of respiratory patterns. © 1993 Wiley-Liss, Inc.     

Parasympathetic postganglionic cells in the glossopharyngeal nerve trunk and their relationship to unmyelinated nerve fibers in the fungiform papillae of the frog

The glossopharyngeal nerve of the frog is made up of afferent nerve fibers and efferent, parasympathetic and sympathetic nerve fibers. The precise origin and course of the parasympathetic efferent nerve fibers in the fungiform papillae of the frog's tongue were investigated. We found the ganglionic cells in the lingual branch of the frog glossopharyngeal nerve. The surface of the ganglionic cell bodies was partly covered by synaptic endings that impinged upon it. Synaptic endings contained clear synaptic vesicles and large dense-cored vesicles. After cutting of the glossopharyngeal nerve proximal to the jugular ganglion, synaptic endings were found to show definite signs of degeneration. These findings led us to the conclusion that the ganglionic cells in the lingual branch of the glossopharyngeal nerve are the parasympathetic postganglionic cells. After cutting of the glossopharyngeal nerve distal to the jugular ganglion, some unmyelinated nerve fibers in the fungiform papillae and postganglionic cells in the lingual branch remained intact. These results strongly suggest that the origin of some of the unmyelinated nerve fibers is the parasympathetic postganglionic cell in the lingual branch.     

Comparison between electrophysiologic and morphologic changes in lead induced peripheral neuropathy in rats

Compound nerve action potential (CNAP) of the mixed peripheral nerve is composed of A alpha beta, A delta, and C potentials. All components of CNAPs in the sciatic nerve were recorded by stimulating the tibial nerve of both control and lead-poisoned rats. Marked decrease of nerve conduction velocity and prolonged duration were found in A alpha beta and A delta fibers especially in large myelinated A alpha beta fibers. The amplitude decreased in A alpha beta potential, but the area did not change. In C potential produced by activation of unmyelinated fibers, nerve conduction velocity slightly decreased, but the amplitude and area did not significantly change. Pathologic correlates revealed prominent segmental demyelination with significant decrease of large myelinated fiber densities. Minimal axonal degeneration of unmyelinated fibers was present. We can conclude that electrophysiologic changes in the lead-poisoned rats correlate with pathologic changes in them.     

Neuropeptide Y- and substance P-like immunoreactive nerve fibers in the rat dura mater encephali

Summary Density and pattern of nerve fibers with neuropeptide Y-like immunoreactivity (NPY-LI) and substance P-like immunoreactivity (SP-LI) in the rat dura mater encephali were investigated by light and electron microscopy using whole-mount preparations. NPY-LI fibers are observed throughout the encephalic dura mater. A remarkable net of NPY-LI nerve fibers is located in the walls of the sagittal and transverse sinuses. Beyond that NPY-LI network, distinct NPY-LI nerve fibers or plexus occur in the rostral falx, parietal dura mater of the olfactory bulb, supratentorial dura mater, parietal dura mater of the cerebellum, tentorium cerebelli and the ventral dura mater. Electron microscopic studies reveal that NPY-LI is exclusively located in unmyelinated axons of small and large nerve fiber bundles, with or without a perineural sheath. Immunopositive C-fibers are predominantly associated with the vascular bed. SP-LI nerve fibers have a moderate and more uniform distribution in the encephalic dura mater. A distinct plexus of SP-LI fibers follows the branches of the middle meningeal artery and the adjacent dura mater. SP-LI fibers are most prominent in the parietal dura mater of the cerebellum. Fine beaded SP-LI fibers, arising from larger SP-LI fiber bundles, are observed in close association to the capillary bed. SP-LI axons are all unmyelinated. They are found in larger nerve fiber bundles with a perineural sheath or in Schwann cells lacking any perineural sheath. The function of NPY-LI and SP-LI nerve fibers in the rat dura mater is discussed in relation to their topography, density and termination.     

Role of transthyretin in posttraumatic regeneration of the sciatic nerve in mouse

Although thyroid hormones are known to have a significant influence on the development of nervous system, the absence of changes in the brain of mice deficient in transthyretin--a protein providing thyroid hormone transport across the blood-brain and blood-nerve barrier--remains unexplained. Therefore, the aim of this study was to determine the effect of transthyretin on the formation of myelinated and unmyelinated nerve fibers in sciatic nerve of mice. The myelinated and unmyelinated nerve fibers were counted in sciatic nerve of 3-months-old normal and transthyretin-knockout (transthyretin(-/-)) mice 15 and 30 days after nerve crushing. No differences were detected in the number of myelinated and unmyelinated nerve fibers in intact control (wild-type) animals group vs. transthyretin(-/-) mice. By days 15 and 30 after nerve crushing the number of myelinated nerve fibers was diminished by 54.7 and 71.8%, respectively, in transthyretin(-/-) mice, as compared to that in control animals. The number of unmyelinated nerve fibers at day 15 after the injury was not different in transthyretin(-/-) and control mice, however, by day 30 the number of these fibers in control group was found to increase significantly, exceeding that one in transthyretin(-/-) mice by 27.9%. These results indicate the important contribution of transthyretin, as a thyroxin carrier protein, to the process of posttraumatic regeneration of sciatic nerve. The absence of changes in nerve fiber numbers in transthyretin-knockout mice in postembryonic period suggests the presence of transthyretin-independent mechanism of thyroxin transport into the peripheral nervous system.     

Ultrastructural identification of a sympathetic component in the hypoglossal nerve of hamsters using experimental degeneration and horseradish peroxidase methods

We employed experimental degeneration, tract-tracing with wheatgerm agglutinin conjugated with horseradish peroxidase (WGA-HRP) and electron microscopy to explore the postganglionic sympathetic fibers in the hypoglossal nerve of hamsters. Quantitative results of normal untreated animals at the electron microscopic level showed the existence of unmyelinated fibers, which made up about 20% of the total fibers in the nerve, being more numerous on the left side. The nerve fibers were preferentially distributed at the periphery of the nerve. Following superior cervical ganglionectomy, most of the unmyelinated fibers underwent degenerative changes. Tract-tracing studies showed that some of the unmyelinated fibers were labeled by WGA-HRP injected into the superior cervical ganglion (SCG). It is suggested that the unmyelinated fibers represent the postganglionic sympathetic fibers originated from the SCG.     

Afferent aortic nerve fibers with their pathways in cardiac sympathetic nerves.

The effect of mechanical and chemical stimulation on activity of afferent aortic nerve fibers with pathways in the cardiac sympathetic nerves has been examined. Action potentials were derived from the second or third thoracic communicating ramus of the left side of anesthetized dogs. Thirty myelinated and 19 unmyelinated fibers responded to tapping the ascending aorta, aortic arch, and descending aorta. Both groups of fibers also responded to a rise as well as to a fall in aortic pressure. Spontaneous discharge of myelinated fibers was related to aortic pressure pulse whereas that of unmyelinated fibers was related to respiration. Asphyxia caused excitation of unmyelinated fibers but not of myelinated fibers. Both groups of fibers responded to topical application of lactic acid. Mechanical and chemical stimulation of the aorta after vagotomy caused a rise in systemic blood pressure and extension of the limbs. The results indicate the existence of afferent aortic fibers in the cardiac sympathetic nerves that cause circulatory and somatic responses.