Ventral Horn Neurons Research Articles

Induction of repulsive guidance molecule in neurons following sciatic nerve injury

Repulsive guidance molecule (RGM) is a protein implicated in both axonal guidance and neural tube closure. We examined the expression of RGMa in the spinal cord after the sciatic nerve crush by immunohistochemistry. Although there was no RGMa immunoreactivity under naïve conditions in the dorsal horn, a weak signal for RGMa was found at 24 h after the nerve crush, and this signal was progressively increased in the NeuN-positive neurons in the ipsilateral dorsal horn from superficial to deep layers at 10 days after surgery. In the neurons of the ipsilateral ventral horn, RGMa was also induced at 10 days after surgery, whereas no RGMa signal could be observed in naïve conditions or at 24 h after surgery. Thus, RGMa expression is upregulated both in the ipsilateral dorsal and ventral horns in response to the sciatic nerve injury. We next examined the effects of complete Freund's adjuvant (CFA)-induced inflammation on RGMa expression in the spinal cord. However, no RGMa expression was observed at 24 h and 10 days after the CFA injection in the dorsal horn, suggesting that RGMa is not involved in inflammation-induced gyperalgesia. Our present study demonstrates that induction of RGMa is associated with the peripheral nerve injury.

Activity of Renshaw cells during locomotor-like rhythmic activity in the isolated spinal cord of neonatal mice.

In the present study, we examine the activity patterns of and synaptic inputs to Renshaw cells (RCs) during fictive locomotion in the newborn mouse using visually guided recordings from GABAergic cells expressing glutamic acid decarboxylase 67-green fluorescent protein (GFP). Among the GFP-positive neurons in the lumbar ventral horn, RCs were uniquely identified as receiving ventral root-evoked short-latency EPSPs that were markedly reduced in amplitude by nicotinic receptor blockers mecamylamine or tubocurarine. During locomotor-like rhythmic activity evoked by bath application of 5-HT and NMDA, 50% of the recorded RCs fired in-phase with the ipsilateral L2 flexor-related rhythm, whereas the rest fired in the extensor phase. Each population of RCs fired throughout the corresponding locomotor phase. All RCs received both excitatory and inhibitory synaptic inputs during the locomotor-like rhythmic activity. Blocking nicotinic receptors with mecamylamine markedly reduced the rhythmic excitatory drive, indicating that these rhythmic inputs originate mainly from motor neurons (MNs). Inhibitory synaptic inputs persisted in the presence of the nicotinic blocker. Part of this inhibitory drive and remaining excitatory drive could be from commissural interneurons because the present study also shows that RCs receive direct crossed inhibitory and excitatory synaptic inputs. However, rhythmic synaptic inputs in RCs were also observed in hemicord preparations in the presence of mecamylamine. These results show that, during locomotor activity, RC firing properties are modulated not only by MNs but also by the ipsilateral and contralateral locomotor networks.

Transient expression of A‐type K+ channel α subunits Kv4.2 and Kv4.3 in rat spinal neurons during development

A-type K(+) currents (I(A)s) have been detected from the ventral horn neurons in rat spinal cord during embryonic day (E) 14 to postnatal day (P) 8 but not in adulthood. It is not known which types of neurons and which A-type K(+) channel alpha subunits express the I(A)s and what the possible function might be. Here, we examined the expression of two A-type K(+) channel alpha subunits, Kv4.2 and Kv4.3, in rat spinal cord at various developmental stages by immunohistochemistry. We found a transient expression of Kv4.2 in somatic motoneurons during E13.5-P8 with a peak around E17.5, which coincides temporally with the natural selection of motoneurons. Transient expression of Kv4.2 and Kv4.3 was also observed in the intermediate gray (IG) interneurons. During E19.5-P14, some IG interneurons express Kv4.2, some express Kv4.3 and a subset co-express Kv4.2 and Kv4.3. Peak expression of Kv4.2 and Kv4.3 in the IG interneurons was detected around P1, which coincides temporally with the developmental selection of IG interneurons. In contrast to the I(A)-expressing subunits Kv4.2 and Kv4.3, a delayed-rectifier K(+) channel alpha subunit Kv1.6 is persistently expressed in somatic motoneurons and IG interneurons. Together, these data support the hypothesis that expression of I(A)s may protect I(A)-expressing somatic motoneurons, and possibly also IG interneurons, from naturally occurring cell death during developmental selection.

Re‐growth of catecholaminergic fibers and protection of cholinergic spinal cord neurons in spinal repaired rats

The extent of re-growth of catecholaminergic fibers, the survival of cholinergic neurons and the degree of autonomic dysreflexia were assessed in complete spinal cord-transected adult rats that received a repair treatment of peripheral nerve grafts and acidic fibroblast growth factor (aFGF). The rats were randomly divided into three groups: (1) sham control group (laminectomy only); (2) spinal cord transection at T8 (transected group); and (3) spinal cord transection at T8, followed by aFGF treatment and peripheral nerve graft (repaired group). The spinal cords and brains of all rats were collected at 6 months post-surgery. Immunohistochemistry for tyrosine hydroxylase (TH) and dopamine-beta-hydroxylase (DBH), and fluoro-gold (FG) retrograde tracing were used to evaluate axon growth across the damage site, and immunocytochemistry for choline acetyl transferase (ChAT) was used to evaluate cholinergic neuronal cell survival following the injury and treatment. When comparing with the transected group, the repaired group showed: (1) lower elevation of mean arterial pressure during colorectal distension; (2) retrogradely labeled neurons in the hypothalamus, zona incerta, subcoeruleus nuclei and rostral ventrolateral medulla following application of FG below the repair site; (3) the presence of TH- and DBH-labeled axons below the lesion site; (4) higher numbers of ChAT-positive neurons in ventral horn and intermediolateral column near the lesion site. We conclude that peripheral nerve graft and aFGF treatments facilitate the re-growth of catecholaminergic fibers, also protect sympathetic preganglionic neurons and spinal motor neurons, and reduce autonomic dysfunction in a T-8 spinal cord-transected rat model.

Protective effects of extract of Ginkgo biloba (EGb 761) on nerve cells after spinal cord injury in rats

An experimental animal model was used to assess spinal cord injury following lateral hemitransection at thoracic spinal cord level. To determine whether extract of Ginkgo biloba (EGb) could have a neuroprotective effect in spinal cord injury (SCI) in rats. Department of Biological Sciences and Biotechnology, Tsinghua University, China. A total of 72 adult rats were divided randomly into three groups: the EGb group, normal saline (NS) group, and sham operation group (sham group). After thoracic spinal cord hemitransection was performed at the level of the 9th thoracic vertebra (T9), rats in the EGb group were given 100 mg/kg EGb 761 daily, while rats in the NS group received NS. The rats in the sham group only underwent laminectomy without spinal cord hemitransection. At various time points after surgery, thoracic spinal cords were sampled and sliced for histochemistry, immunohistochemistry of inducible nitric oxide synthase (iNOS), and terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) of apoptotic cells. Myelin staining showed that the area of cavities was small and the demyelinated zones were limited at and around the injury site of the spinal cord in the EGb group, while the area of cavities was large and the demyelinated zones were serious in the NS group. Nissl staining showed that the ratio of bilateral ventral horn neurons (transection side/uninjured side) in the EGb group was higher than that in the NS group (P<0.05). The apoptotic index and the percentage of iNOS-positive cells were lower in the EGb group than in the NS group. Furthermore, the percentage of iNOS-positive cells positively correlated with the apoptotic index (r( 2)=0.729, P<0.01) after SCI. This study demonstrated that EGb 761 could inhibit iNOS expression and have neuroprotective effect by preventing nerve cells from apoptosis after SCI in rats.

Parkinson's Disease α-Synuclein Transgenic Mice Develop Neuronal Mitochondrial Degeneration and Cell Death

Alpha-synuclein (alpha-Syn) is enriched in nerve terminals. Two mutations in the alpha-Syn gene (Ala53--> Thr and Ala30--> Pro) occur in autosomal dominant familial Parkinson's disease. Mice overexpressing the human A53T mutant alpha-Syn develop a severe movement disorder, paralysis, and synucleinopathy, but the mechanisms are not understood. We examined whether transgenic mice expressing human wild-type or familial Parkinson's disease-linked A53T or A30P mutant alpha-syn develop neuronal degeneration and cell death. Mutant mice were examined at early- to mid-stage disease and at near end-stage disease. Age-matched nontransgenic littermates were controls. In A53T mice, neurons in brainstem and spinal cord exhibited large axonal swellings, somal chromatolytic changes, and nuclear condensation. Spheroid eosinophilic Lewy body-like inclusions were present in the cytoplasm of cortical neurons and spinal motor neurons. These inclusions contained human alpha-syn and nitrated synuclein. Motor neurons were depleted (approximately 75%) in A53T mice but were affected less in A30P mice. Axonal degeneration was present in many regions. Electron microscopy confirmed the cell and axonal degeneration and revealed cytoplasmic inclusions in dendrites and axons. Some inclusions were degenerating mitochondria and were positive for humanalpha-syn. Mitochondrial complex IV and V proteins were at control levels, but complex IV activity was reduced significantly in spinal cord. Subsets of neurons in neocortex, brainstem, and spinal cord ventral horn were positive for terminal deoxynucleotidyl transferase-mediated biotinylated UTP nick end labeling, cleaved caspase-3, and p53. Mitochondria in neurons had terminal deoxynucleotidyl transferase-mediated biotinylated UTP nick end labeling-positive matrices and p53 at the outer membrane. Thus, A53T mutant mice develop intraneuronal inclusions, mitochondrial DNA damage and degeneration, and apoptotic-like death of neocortical, brainstem, and motor neurons.

Immunohistochemical localisation of the voltage gated potassium ion channel subunit Kv3.3 in the rat medulla oblongata and thoracic spinal cord

Voltage gated K + channels (Kv) are a diverse group of channels important in determining neuronal excitability. The Kv superfamily is divided into 12 subfamilies (Kv1–12) and members of the Kv3 subfamily are highly abundant in the CNS, with each Kv3 gene (Kv3.1–Kv3.4) exhibiting a unique expression pattern. Since the localisation of Kv subunits is important in defining the roles they play in neuronal function, we have used immunohistochemistry to determine the distribution of the Kv3.3 subunit in the medulla oblongata and spinal cord of rats. Kv3.3 subunit immunoreactivity (Kv3.3-IR) was widespread but present only in specific cell populations where it could be detected in somata, dendrites and synaptic terminals. Labelled neurones were observed in the spinal cord in laminae IV and V, in the region of the central canal and in the ventral horn. In the medulla oblongata, labelled cell bodies were numerous in the spinal trigeminal, cuneate and gracilis nuclei whilst rarer in the lateral reticular nucleus, hypoglossal nucleus and raphe nucleus. Regions containing autonomic efferent neurones were predominantly devoid of labelling with only occasional labelled neurones being observed. Dual immunohistochemistry revealed that some Kv3.3-IR neurones in the ventral medullary reticular nucleus, spinal trigeminal nucleus, dorsal horn, ventral horn and central canal region were also immunoreactive for the Kv3.1b subunit. The presence of Kv3.3 subunits in terminals was confirmed by co-localisation of Kv3.3-IR with the synaptic vesicle protein SV2, the vesicular glutamate transporter VGluT2 and the glycine transporter GlyT2. Co-localisation of Kv3.3-IR was not observed with VGluT1, tyrosine hydroxylase, serotonin or choline acetyl transferase. Electron microscopy confirmed the presence of Kv3.3-IR in terminals and somatic membranes in ventral horn neurones, but not motoneurones. This study provides evidence supporting a role for Kv3.3 subunits in regulating neuronal excitability and in the modulation of excitatory and inhibitory synaptic transmission in the medulla oblongata and spinal cord.

Adenosine-induced neuroprotective actions in ventral horn neurons of the rat spinal cord.

Adenosine-induced neuroprotective actions in ventral horn neurons of the rat spinal cord.

Immunohistochemical localization of calbindin‐D28k and calretinin in the spinal cord of Xenopus laevis

Immunohistochemical techniques were used to investigate the distribution and morphology of neurons containing the calcium-binding proteins calbindin-D28k (CB) and calretinin (CR) in the spinal cord of Xenopus laevis and determine the extent to which this organization is comparable to that of mammals. Most CB- and CR-containing neurons were located in the superficial dorsal gray field, but with distinct topography. The lateral, ventrolateral, and ventromedial fields also possessed abundant neurons labeled for either CB or CR. Double immunohistofluorescence demonstrated that a subpopulation of dorsal root ganglion cells and neurons in the dorsal and ventrolateral fields contained CB and CR. By means of a similar technique, a cell population in the dorsal field was doubly labeled only for CB and nitric oxide synthase (NOS), whereas in the ventrolateral field colocalization of NOS with CB and CR was found. Choline acetyltransferase immunohistochemistry revealed that a subpopulation of ventral horn neurons, including motoneurons, colocalized CB and CR. The involvement of CB- and CR-containing neurons in ascending spinal projections was demonstrated combining the retrograde transport of dextran amines and immunohistochemistry. Cells colocalizing the tracer and CB or CR were quite numerous, primarily in the dorsal and ventrolateral fields. Similar experiments demonstrated supraspinal projections from CB- and CR-containing cells in the brainstem and diencephalon. The distribution, projections, and colocalization with neurotransmitters of the neuronal systems containing CB and CR in Xenopus suggest that CB and CR are important neuromodulator substances with functions conserved in the spinal cord from amphibians through mammals.

Expression of cyclooxygenase in rat spinal cord with acute contusion

目的 观察正常和急性打击损伤大鼠脊髓组织中环氧化酶( COX)的表达.方法 48只 Sprague- Dawley大鼠,随机平均分为正常组,手术对照组,脊髓损伤后 2、 4、 8、 16、 24、 48 h组.采用改良 Allen' s打击脊髓损伤动物模型.应用免疫组织化学染色的方法观察两种 COX蛋白的表达变化.结果 COX- 1蛋白在正常胸椎脊髓组织中有基础性表达,主要分布在 脊髓灰质后角和白质后索的神经纤维网中; COX- 2蛋白在正常胸椎脊髓组织中也存在基础性表达,主要分布于脊髓灰质神经元细胞;急性打击脊髓损伤后 COX- 1蛋白的表达没有明显变化;急性打击脊髓损伤后 4 h, COX- 2蛋白的表达开始增高,损伤后 24 h染色程度达到高峰, 并维持至损伤后 48 h,表达增高主要出现在脊髓灰质的神经元细胞中.结论与环氧化酶在外周组织中的表达特点不同,正常胸段脊髓组织中同时存在 COX- 1和 COX- 2蛋白的基础性表达 .急性机械打击脊髓损伤后,参与继发损伤的主要环氧化酶亚型是 COX- 2。

Modulation of calcium currents in mouse ventral horn neurons by extracellular pH

Neuronal activity has been shown to modulate the pH of the extracellular environment. Since neuronal circuits in the ventral horn of the spinal cord are highly active during patterned movements, and voltage-gated calcium channels play an important role in the production of spinal motoneuron output, the effects of changes in extracellular pH (pH(e)) on calcium currents in ventral horn neurons of the mouse spinal cord were examined. It is demonstrated that these channels are sensitive to modulation by pH(e). The amplitude of the current mediated by these channels increased as the pH(e) was elevated. The elevated pH(e) also led to a hyperpolarizing shift in the voltage dependence of both activation and inactivation. The opposite effects were seen for a decrease in pH(e). It was also noted that a decrease in pH(e) was associated with a faster inactivation of the current. It is concluded that voltage-gated calcium currents in ventral horn neurons are modulated by changes in pH(e), and that this modulation may play a physiologically important role in determining motoneuronal excitability during behaviors such as locomotion.

A semi-quantitative analysis of Fos expression by mustard oil

In the present study, a semi-quantitative analysis of Fos expression by mustard oil was performed. For this purpose, mustard oil was applied to the skin of the right hind foot of Wistar rats at various concentrations: 5%, 10%, 30%, 50%, 80% and 100% in liquid paraffin. The distribution and number of activated Fos-positive cells in the stimulated side (ipsilateral) and contralateral side of the spinal cord were investigated following the application. The ED50 of the response was also determined. The number of Fos-labelled cells gradually increased in a dose dependent manner in both sides of superficial layers (laminae I–II) of the spinal cord with increasing concentration of mustard oil. The increase between the doses was found significant in the ipsilateral superficial layers. The increase was significant in the contralateral superficial layers at concentrations above 50%. Very few Fos-labelled cells were observed around the central canal region in all concentrations. Higher doses of the mustard oil did not increase the number of activated cells in the deeper layers. However, the expression in the deeper layers (laminae III–X) does not show a consistent trend. Also, none of the concentrations used produced labelling in neurons of the deep ventral horn neurons or in motor neurons. Forty percent (40%) of mustard oil gave an approximately 1/2 maximum response i.e. an approximate ED 50. This may be important for studies using intrathecal application of antagonist following the mustard oil activation of skin nerve fibres.

Presynaptic Angiotensin II AT1 Receptors Enhance Inhibitory and Excitatory Synaptic Neurotransmission to Motoneurons and Other Ventral Horn Neurons in Neonatal Rat Spinal Cord

In neonatal spinal cord, we previously reported that exogenous angiotensin II (ANG II) acts at postsynaptic AT1 receptors to depolarize neonatal rat spinal ventral horn neurons in vitro. This study evaluated an associated increase in synaptic activity. Patch clamp recordings revealed that 38/81 thoracolumbar (T7-L5) motoneurons responded to bath applied ANG II (0.3-1 microM; 30 s) with a prolonged (5-10 min) and reversible increase in spontaneous postsynaptic activity, selectively blockable with Losartan (n = 5) but not PD123319 (n = 5). ANG-II-induced events included both spontaneous inhibitory (IPSCs; n = 6) and excitatory postsynaptic currents (EPSCs; n = 5). While most ANG induced events were tetrodotoxin-sensitive, ANG induced a significant tetrodotoxin-resistant increase in frequency but not amplitude of miniature IPSCs (n = 7/13 cells) and EPSCs (n = 2/7 cells). In 35/77 unidentified neurons, ANG II also induced a tetrodotoxin-sensitive and prolonged increase in their spontaneous synaptic activity that featured both IPSCs (n = 5) and EPSCs (n = 4) when tested in the presence of selective amino acid receptor antagonists. When tested in the presence of tetrodotoxin, ANG II was noted to induce a significant increase in the frequency but not the amplitude of mIPSCs (n = 9) and mEPSCs (n = 8). ANG also increased spontaneous motor activity from isolated mouse lumbar ventral rootlets. Collectively, these observations support the existence of a wide pre- and postsynaptic distribution of ANG II AT1 receptors in neonatal ventral spinal cord that are capable of influencing both inhibitory and excitatory neurotransmission.

Are paragriseal cells in the avian lumbosacral spinal cord displaced ventral spinocerebellar tract neurons?

In lumbosacral segments the spinal cord of birds contains numerous paragriseal neurons lying in the white matter of lateral and ventral funiculus. These paragriseal cells project to the cerebellum. Neurons of the dorsal horn (mainly Clarke's column) make up a dorsal spinocerebellar tract and neurons of the ventral horn (mainly spinal border cells) are at the origin of a ventral spinocerebellar tract. It was the aim of this investigation to look for the distribution of spinocerebellar ventral horn neurons and paragriseal cells in the thoracolumbosacral spinal cord of pigeons and to compare this distribution with that of the cervical enlargement. Neuroanatomical tracers were injected into the anterior cerebellum of pigeons and labeled spinal neurons were counted throughout the length of the spinal cord. In the cervical enlargement the number of spinocerebellar ventral horn neurons increases more rostral than that of dorsal horn neurons but the number of both groups of neurons decreases simultaneously at the caudal end of the enlargement. In the ventral horn of thoracolumbosacral segments the number of spinocerebellar ventral horn neurons and paragriseal cells increases again more rostrally than that of dorsal horn cells. However, the number of ventral horn cells decreases whereas that of paragriseal cells and of dorsal horn cells is maintained. This shows that the number of ventral horn cells decreases in favor of paragriseal cells, which supports the suggestion that paragriseal cells are displaced ventral horn spinocerebellar neurons. It is discussed whether the paragriseal neurons migrate toward their input.

Motor Neuron Disease in Mice Expressing the Wild Type-Like D90A Mutant Superoxide Dismutase-1

Mutant human CuZn-superoxide dismutases (hSOD1s) cause amyotrophic lateral sclerosis (ALS). The most common mutation is the wild type-like D90A and to explore its properties, transgenic mice were generated and compared with mice expressing wild-type hSOD1. D90A hSOD1 was both in vivo in mice and in vitro under denaturing conditions nearly as stable as the wild-type human enzyme. It appeared less toxic than other tested mutants, but mice homozygous for the transgene insertion developed a fatal motor neuron disease. In these mice, the disease progression was slow and there were bladder disturbances similar to what is found in human ALS cases homozygous for the D90A mutation. The homozygous D90A mice accumulated detergent-resistant hSOD1 aggregates in spinal cords, and abundant hSOD1 inclusions and vacuoles were seen in the ventral horns. Mice expressing wild-type hSOD1 at a comparable rate showed similar pathologic changes but less and later. Hemizygous D90A mice showed even milder alterations. At 600 days, the wild-type hSOD1 transgenic mice had lost more ventral horn neurons than hemizygous D90A mice (38% vs 31% p < 0.01). Thus, wild-type hSOD1 shows a significant neurotoxicity in the spinal cord, that is less than equal but more than half as large as that of D90A mutant enzyme.

Comparison of the membrane dysfunction induced by in vitro ischemia in ventral and dorsal horn neurons of the rat spinal cord.

Comparison of the membrane dysfunction induced by in vitro ischemia in ventral and dorsal horn neurons of the rat spinal cord.

Systematic variation in effects of serotonin and norepinephrine on repetitive firing properties of ventral horn neurons

Spinal interneurons are essential integrators of descending and peripheral input that receive profuse monoaminergic influence from brainstem nuclei. In this study, the effects of the monoamines serotonin and norepinephrine on the intrinsic properties of ventral horn interneurons were investigated in a slice preparation of the lumbar cord of 7–19 day old rats. Three cell groups with distinct firing patterns in response to steps of injected current were observed and classified as repetitive-firing, initial-burst or single-spiking. Input conductance tended to be largest in single-spiking cells whereas repetitive-firing cells showed the greatest tendency for spontaneous firing and had the fastest rate of rise for the action potential. Rhythmic firing behaviors were defined by the frequency-current relation evoked by linearly increasing current ramps. The monoaminergic modulation of firing patterns and frequency-current relations was primarily studied in repetitive-firing cells. The frequency-current threshold current was decreased in cells with high pre-drug values and increased in cells with low pre-drug values. Therefore, monoamine administration decreased the input-output heterogeneity of the repetitive-firing cells by compressing the range of frequency-current threshold currents. This action of monoamines may have a key role in the suppression of sensory-evoked reflexes and the production of coordinated movement.

Neuronal plasticity after spinal cord injury: identification of a gene cluster driving neurite outgrowth

Functional recovery after spinal cord injury (SCI) may result in part from axon outgrowth and related plasticity through coordinated changes at the molecular level. We employed microarray analysis to identify a subset of genes the expression patterns of which were temporally coregulated and correlated to functional recovery after SCI. Steady-state mRNA levels of this synchronously regulated gene cluster were depressed in both ventral and dorsal horn neurons within 24 h after injury, followed by strong re-induction during the following 2 wk, which paralleled functional recovery. The identified cluster includes neuritin, attractin, microtubule-associated protein 1a, and myelin oligodendrocyte protein genes. Transcriptional and protein regulation of this novel gene cluster was also evaluated in spinal cord tissue and in single neurons and was shown to play a role in axonal plasticity. Finally, in vitro transfection experiments in primary dorsal root ganglion cells showed that cluster members act synergistically to drive neurite outgrowth.

ErbB transmembrane tyrosine kinase receptors are expressed by sensory and motor neurons projecting into sciatic nerve.

Adult spinal cord motor and dorsal root ganglion (DRG) sensory neurons express multiple neuregulin-1 (NRG-1) isoforms that act as axon-associated factors promoting neuromuscular junction formation and Schwann cell proliferation and differentiation. NRG-1 isoforms are also expressed by muscle and Schwann cells, suggesting that motor and sensory neurons are themselves acted on by NRG-1 isoforms produced by their peripheral targets. To test this hypothesis, we examined the expression of the NRG-1 receptor subunits erbB2, erbB3, and erbB4 in rat lumbar DRG and spinal cord. All three erbB receptors are expressed in these tissues. Sciatic nerve transection, an injury that induces Schwann cell expression of NRG-1, alters erbB expression in DRG and cord. Virtually all DRG neurons are erbB2- and erbB3-immunoreactive, with erbB4 also detectable in many neurons. In spinal cord white matter, erbB2 and erbB4 antibodies produce dense punctate staining, whereas the erbB3 antibody primarily labels glial cell bodies. Spinal cord dorsal and ventral horn neurons, including alpha-motor neurons, exhibit erbB2, erbB3, and erbB4 immunoreactivity. Spinal cord ventral horn also contains a population of small erbB3+/S100beta+/GFAP- cells (GFAP-negative astrocytes or oligodendrocytes). We conclude that sensory and motor neurons projecting into sciatic nerve express multiple erbB receptors and are potentially NRG-1 responsive.

Mitochondrial accumulation in the distal part of the initial segment of chicken spinal motoneurons

The axonal initial segment is the initiation site of action potentials and is characterized morphologically by a dense undercoating and fascicles of microtubules connected by cross-bridges. In order to analyze subcellular structures in the initial segment, we made serial transverse sections of initial segments of identified chicken motoneurons by retrograde transport of horseradish peroxidase (HRP) injected into the muscle. The mean (±SD) length of the initial segment was 28.1±2.3 μm ( n=6). Mitochondria accumulated in the distal part of the initial segment, which was 1.4–6.9 μm in length (5–23% of the total length of the initial segment). In the transverse section of the distal part, mitochondrial density was 15.8±6.2% ( n=5), while in the middle and proximal parts it was 6.1±1.6% and 5.6±1.4%, respectively. Mitochondrial accumulation was observed in common in phasic and tonic motoneurons in the chicken, and also observed in the distal part of the initial segment of the large ventral horn neurons of the chicken without HRP injection. These findings suggest that accumulated mitochondria play an important role in maintaining the physiological function of the distal part of the motoneuron initial segment.