Thoracic Spinal Cord Segments Research Articles

Response of ependymal progenitors to spinal cord injury or enhanced physical activity in adult rat.

Ependymal cells (EC) in the spinal cord central canal (CC) are believed to be responsible for the postnatal neurogenesis following pathological or stimulatory conditions. In this study, we have analyzed the proliferation of the CC ependymal progenitors in adult rats processed to compression SCI or enhanced physical activity. To label dividing cells, a single daily injection of Bromo-deoxyuridine (BrdU) was administered over a 14-day-survival period. Systematic quantification of BrdU-positive ependymal progenitors was performed by using stereological principles of systematic, random sampling, and optical Dissector software. The number of proliferating BrdU-labeled EC increased gradually with the time of survival after both paradigms, spinal cord injury, or increased physical activity. In the spinal cord injury group, we have found 4.9-fold (4 days), 7.1-fold (7 days), 4.9-fold (10 days), and 5.6-fold (14 days) increase of proliferating EC in the rostro-caudal regions, 4 mm away from the epicenter. In the second group subjected to enhanced physical activity by running wheel, we have observed 2.1-2.6 fold increase of dividing EC in the thoracic spinal cord segments at 4 and 7 days, but no significant progression at 10-14 days. Nestin was rapidly induced in the ependymal cells of the CC by 2-4 days and expression decreased by 7-14 days post-injury. Double immunohistochemistry showed that dividing cells adjacent to CC expressed astrocytic (GFAP, S100beta) or nestin markers at 14 days. These data demonstrate that SCI or enhanced physical activity in adult rats induces an endogenous ependymal cell response leading to increased proliferation and differentiation primarily into macroglia or cells with nestin phenotype.

Three-element phased-array coil for imaging of rat spinal cord at 7T

To overcome some of the limitations of an implantable coil, including its invasive nature and limited spatial coverage, a three-element phased-array coil is described for high-resolution magnetic resonance imaging (MRI) of rat spinal cord. This coil allows imaging both thoracic and cervical segments of rat spinal cord. In the current design, coupling between the nearest neighbors was minimized by overlapping the coil elements. A simple capacitive network was used for decoupling the next neighbor elements. The dimensions of individual coils in the array were determined based on the signal-to-noise ratio (SNR) measurements performed on a phantom with three different surface coils. SNR measurements on a phantom demonstrated higher SNR for the phased array coil relative to two different volume coils. In vivo images acquired on rat spinal cord with our coil demonstrated excellent gray and white matter contrast. To evaluate the performance of the phased array coil under parallel imaging, g-factor maps were obtained for acceleration factors of 2 and 3. These simulations indicate that parallel imaging with an acceleration factor of 2 would be possible without significant image reconstruction-related noise amplifications.

Erythropoietin during porcine aortic balloon occlusion-induced ischemia/reperfusion injury

Aortic occlusion causes ischemia/reperfusion injury, kidney and spinal cord being the most vulnerable organs. Erythropoietin improved ischemia/reperfusion injury in rodents, which, however, better tolerate ischemia/reperfusion than larger species. Therefore, we investigated whether erythropoietin attenuates porcine aortic occlusion ischemia/reperfusion injury. Before occluding the aorta for 45 mins by inflating intravascular balloons, we randomly infused either erythropoietin (n = 8; 300 IU/kg each over 30 mins before and during the first 4 hrs of reperfusion) or vehicle (n = 6). During aortic occlusion, mean arterial pressure was maintained at 80% to 120% of baseline by esmolol, nitroglycerine, and adenosine 5'-triphosphate. During reperfusion, noradrenaline was titrated to keep mean arterial pressure >80% of baseline. Kidney perfusion and function were assessed by fractional Na-excretion, p-aminohippuric acid and creatinine clearance, spinal cord function by lower extremity reflexes and motor evoked potentials. Blood isoprostane levels as well as blood and tissue catalase and superoxide dismutase activities allowed evaluation of oxidative stress. After 8 hrs of reperfusion, kidney and spinal cord specimens were taken for histology (hematoxylin-eosin, Nissl staining) and immunohistochemistry (TUNEL assay for apoptosis). Parameters of oxidative stress and antioxidative activity were comparable. Erythropoietin reduced the noradrenaline requirements to achieve the hemodynamic targets and may improve kidney function despite similar organ blood flow, histology, and TUNEL staining. Neuronal damage and apoptosis was attenuated in the thoracic spinal cord segments without improvement of its function. During porcine aortic occlusion-induced ischemia/reperfusion erythropoietin improved kidney function and spinal cord integrity. The lacking effect on spinal cord function was most likely the result of the pronounced neuronal damage associated with the longlasting ischemia.

Immunohistochemical detection of connexin36 in sympathetic preganglionic and somatic motoneurons in the adult rat

Gap junctional communication in the adult CNS plays an important role in the synchronization of neuronal activities. In vitro studies have shown evidence of electrotonic coupling through gap junctions between sympathetic preganglionic motoneurons and between somatic motoneurons in the neonatal and adult rat spinal cord. Electrotonic transmission of membrane oscillations might be an important mechanism for recruitment of neurons and result in the generation of rhythmic sympathetic and somato-motor activity at the population level. Gap junctions in the adult spinal cord are constituted principally by connexin36 (Cx36). However, the distribution of Cx36 in specific neuronal populations of the spinal cord is unknown. Here, we identify Cx36-like immunoreactivity in sympathetic preganglionic and somatic motoneurons in thoracic spinal cord segments of the adult rat. For this purpose, double immunostaining against Cx36 and choline acetyltransferase (ChAT) was performed on transverse sections (20 μm) taken from spinal segments T6–T8. Cx36 punctate immunostaining was detected in the majority of ChAT-immunoreactive (-ir) neurons from lamina VII [intermediolateral cell column (IML) and intercalated cell group (IC)], lamina X [central autonomic nucleus (CA)] and in ventral horn neurons from laminae VIII and IX. Cx36 puncta were distributed in the neuronal somata and along dendritic processes. The presence of Cx36 in ChAT-ir neurons is consistent with electrical coupling between sympathetic preganglionic motoneurons and between somatic motoneurons through gap junctions in the adult spinal cord.

Target muscles for retrograde gene delivery to specific spinal cord segments

Targeted retrograde gene delivery into the injured spinal cord is less invasive for the damaged tissue. One of the advantages of this approach is the possible selection of target organs according to the level of spinal cord injury. We evaluated nine candidate target organs for retrograde delivery of an adenovirus vector carrying beta-galactosidase (AdV-LacZ) gene to cervical, thoracic and lumbar spinal cord segments. One week after vector injection into each muscle, we assessed the LacZ gene expression in the spinal cord by X-gal staining. The most appropriate target organs with high transduction efficacy were the sternomastoid and clavotrapezius muscles for cervical spinal cord, tibialis anterior and the gastrocnemius muscles for the lumbar spinal cord. Retrograde gene delivery to the thoracic spinal cord was inefficient probably due to the small number of anterior horn neurons in the region. Gene expression was mainly identified over the anatomical area of innervation and not into other body organs. Our results suggested that retrograde delivery of adenovirus genome to the cervical and lumbar spinal cord segments seems feasible by injection of an adenoviral vector into the appropriate target organ. Adenovirus vector is an efficient retrograde tracer since it can deliver the carried gene to a wide area of the spinal cord and not to other body organs.

Low-frequency electroacupuncture suppresses zymosan-induced peripheral inflammation via activation of sympathetic post-ganglionic neurons

Electroacupuncture (EA) is used to treat a variety of inflammatory diseases; however, the neurophysiological mechanisms underlying EA's anti-inflammatory effect remain unclear. Accumulating evidence suggests that the sympathetic nervous system regulates immunologic and inflammatory responses and thus we hypothesized that this system could be involved in EA's anti-inflammatory effect (EA-AI). The goal of the present study was to evaluate whether the sympathetic nervous system plays a critical role in EA-AI using a mouse air pouch inflammation model. We found that bilateral low-frequency (1 Hz) EA applied to the Zusanli acupoint significantly suppressed the number of zymosan-induced leukocytes migrating into the air pouch. Furthermore, double-labeling immunohistochemical experiments showed that EA stimulation increased Fos expression in choline acetyltransferase (ChAT)-positive sympathetic pre-ganglionic neurons in the intermediolateral region of thoracic spinal cord segments. Chemical sympathetic denervation by intraperitoneal injection of 6-hydroxydopamine (which spares sympathetic adrenal medullary innervation) significantly inhibited EA-AI. In contrast, adrenalectomy did not alter EA-AI. Finally, systemic propranolol administration significantly inhibited EA's anti-inflammatory effect, suggesting that β-adrenoceptors are involved. Collectively, these results suggest that EA produces an anti-inflammatory effect in this mouse air pouch model by activating the sympathetic nervous system leading to the release of catecholamines from post-ganglionic nerve terminals, which act on β-adrenoceptors on immune cells to inhibit their migration.

Distribution of glycinergic neuronal somata in the rat spinal cord

Glycine transporter 2 (GlyT2) mRNA is exclusively expressed in glycinergic neurons, and is presently considered a reliable marker for glycinergic neuronal somata. In this study, we have performed non-radioactive in situ hybridization to localize GlyT2 mRNA in fixed free-floating sections of cervical (C2 and C6), thoracic (T5), lumbar (L2 and L5) and sacral (S1) segments of the rat spinal cord. The results showed that in all segments the majority of the GlyT2 mRNA labeled (glycinergic) neuronal somata was present in the deep dorsal horn and the intermediate zone (laminae III–VIII), with around 50% (range 43.7–70.9%) in laminae VII&VIII. In contrast, the superficial dorsal horn, the motoneuronal cell groups and the area around the central canal contained only few glycinergic neuronal somata. The density (number of glycinergic neuronal somata per mm 2) was also low in these areas, while the highest densities were found in laminae V to VIII. The lateral spinal nucleus and the lateral cervical nucleus also contained a limited number of glycinergic neurons. Our findings showed that the distribution pattern of the glycinergic neuronal somata is similar in all the examined segments. The few differences that were found in the relative laminar distribution between some of the segments, are most likely due to technical reasons. We therefore conclude that the observed distribution pattern of glycinergic neuronal somata is present throughout the spinal cord. Our findings further showed that the non-radioactive in situ hybridization technique for identifying GlyT2 mRNA in fixed free-floating sections is a highly efficient tool for identifying glycinergic neurons in the spinal cord.

Marek's Disease Virus–Induced Transient Paralysis Is Associated with Cytokine Gene Expression in the Nervous System

Marek's disease (MD)-associated transient paralysis (TP) was experimentally induced in chickens by intraperitoneal inoculation of RB1B strain of Marek's disease virus (MDV). Between 7 and 11 days post-infection (d.p.i.), neck and limb paralysis was observed in 18% of infected chickens, which was associated with various degrees of edema, vacuolation, perivascular cuffing of mononuclear cells, and glial cell infiltration mainly in the cerebrum, cerebellum, and brain stem. The chickens that were infected but did not progress to develop TP until 12 d.p.i. also had similar lesions suggestive of encephalitis in the cerebrum, cerebellum, and brain stem. Chickens infected with MDV had more interleukin (IL)-6, IL-12, and interferon (IFN)-gamma in their brain tissues compared to uninfected chickens. Moreover, IL-18 was significantly increased in brain tissues of birds showing clinical signs of TP compared to uninfected birds. Importantly, the expression of IL-6, IL-18, and IFN- gamma in brain tissues of MDV-infected chickens with signs of TP was significantly increased compared to that in asymptomatic MDV-infected birds. MDV genome load in the brain of chickens showing clinical signs of TP was higher than that in asymptomatic MDV-infected chickens but was not statistically significant. The lesions in the cervical, thoracic, and lumbar spinal cord segments in MDVinfected chickens were characterized mainly by perivascular cuffing of mononuclear cells irrespective of the group. The expression of mRNA for IL-18 and IFN-gamma genes was not significantly different in spinal cord tissues of chickens with TP compared to clinically normal, MDV-infected and noninfected chickens. These results suggest possible underlying immunologic mechanisms for MDV-induced TP.

CENTRAL AUTONOMIC INNERVATION OF THE KIDNEY. WHAT CAN WE LEARN FROM A TRANSNEURONAL TRACING STUDY IN AN ANIMAL MODEL?

Renal sympathetic innervation is involved in the maintenance of fluid homeostasis, modulation of renal secretion from juxtaglomerular cells, sodium resorption from renal tubular cells and renal hemodynamics. The understanding of central innervation and neuronal connections is important for studying the consequences of renal disease and surgical interventions compromising renal nerves. A total of 38 individual adult male Sprague-Dawley rats were used for retrograde transneuronal mapping of the spinal cord and brain stem after pseudorabies virus (PRV) injection into the left kidney in 30 and control experiments in 8. After a survival time of 72, 96 or 120 hours the animals were sacrificed. Exploration of the abdominal and pelvic visceral organs was done, and the brain and spinal cord were harvested via dorsal laminectomy. After cutting on a freezing microtome the tissue was immunostained for PRV. After kidney injection inspection of the abdominal and pelvic cavity revealed an enlarged bladder with hemic urine. The urine was sterile and the bladder wall showed signs of neurogenic inflammation. Other organs were not affected. PRV positive cells were primarily found within the ipsilateral nucleus intermediolateralis of thoracic spinal cord segments T6 to T13. At the supraspinal level PRV positive cells were found within certain regions, namely the nuclei raphes, rostral ventromedial and ventrolateral medulla, A5 noradrenergic cell region, locus coeruleus and nucleus paraventricularis of the hypothalamus. This investigation demonstrates the anatomical basis for broad central sympathetic innervation of the kidney. The neurogenic inflammation within the spinal cord inherent to the PRV tracing method causes an inflammatory reaction within the bladder. This can be due to increased sympathetic nerve activity, followed by peripheral, neurogenically mediated inflammation.

Nerve-pathways of acupoint Fengch’ih in rat by anterograde transport of HRP

To study the nervous-pathways of Fengch'ih acupuncture by means of anterograde transport of aqueous solution of horseradish peroxidase (HRP). Fifty Wistar rats were randomly divided into 1, 2, 3, 4, and 5 d groups, and every group had 10 animals. HRP (30% aqueous solution) was injected into a Fengch'ih. Serial, transverse or capital, 40 microm sections of the cervical spinal ganglia, cervical and thoracic spinal cord segment and brain were cut on a cryotome. Sections were incubated for HRP histochemistry according to the tetramethylbenzidine (TMB). Part of the sections were counterstained with neutral red. After 1 d of survival times, many labeled cell bodies were found in 1-4 cervical spinal ganglia, anterior horn of 1-4 cervical spinal cord, ventromedial division of facial nucleus, accessory facial nucleus ipsilaterally. With increasing survival times, the intensity of labeled cells were slightly decreased. Fengch'ih may bring into full play its effect by correlation of posterior ear branch of facial nerve and anterior branch of 2-3 cervical nerve with 1-4 cervical the anterior horn of the spinal cord, ventromedial division of facial nucleus, accessory facial nucleus.

Patterns of relationship between activity of sympathetic nerves in rabbits and rats

We used frequency domain analysis (power spectra, ordinary and partial coherence and phase spectra) of simultaneously recorded activity of postganglionic sympathetic nerves to investigate the construction of their central generators in rabbits and rats anesthetized with urethane. As found earlier in the cat, power spectra of sympathetic nerve discharge (SND) consisted of a wide-band component (1 to 10 Hz in rabbits and 1 to 15–20 Hz in rats) and superimposed cardiac and respiratory related peaks. The coherence between pairs of SNDs in the cardiac, vertebral, and renal nerves was significant over a wide range of frequencies, from 0 to 6–10 Hz in rabbits, and except for a sharp peak at the heart rate, was not explained by baroreceptor feedback. In rats, the coherence between distant nerves was relatively low (<0.2) except at the cardiac and respiratory frequencies. Analysis of partial coherences for the three nerves in rabbits revealed two main patterns; one characterized by dominance of the cardiac SND generator, and the other by strong coupling of the vertebral and cardiac SNDs, as compared with renal SND. Phase spectra of distant nerves contained a well-defined transportation lag corresponding to a delay of ∼70 ms between upper and lower thoracic spinal cord segments. At frequencies close to heart rate however, the phase was constant in most experiments indicating that different mechanisms are involved in transmitting wide band and oscillatory components of resting SND. The similarities between sympathetic oscillators in cats, studied previously in great detail, and rabbits preferred in recent behavioral studies allow the translation of knowledge between these two species.

Postsynaptic dorsal column neurons express NK1 receptors following colon inflammation

Recent clinical and experimental studies have suggested that the dorsal column pathway and specifically postsynaptic dorsal column neurons play an important role in the transmission of visceral pain. In our study we have mapped the distribution of postsynaptic dorsal column neurons in thoracic, lumbar and sacral spinal cord segments. The presence of immunoreactivity for neurokinin 1 receptors on these postsynaptic dorsal column neurons was examined under control conditions and after colon inflammation. The largest number of postsynaptic dorsal column neurons was found in the lumbar enlargement. They were mostly located in laminae III–IV, but depending on the spinal segment, about 7–15% of them were in the deep medial dorsal horn and in the central canal area. Under control conditions none of the 1438 postsynaptic dorsal column neurons examined expressed neurokinin 1 receptors. However, after induction of colon inflammation about 1.4% of the 2015 postsynaptic dorsal column neurons observed in the experimental group showed immunoreactivity for neurokinin 1 receptors. These neurons were preferentially found in the lower thoracic and lumbosacral spinal segments where they represented about 3–4% of the total population of postsynaptic dorsal column neurons examined. The de novo expression of neurokinin1 receptors on postsynaptic dorsal column neurons after colon inflammation suggests that substance P released from visceral primary afferents under inflammatory conditions could help produce central sensitization of these neurons.

Suckling-induced activation of neural c-fos expression at lower thoracic rat spinal cord segments

Suckling stimulation is essential for neuroendocrine and sympathetic reflex activation during lactation. In the present study, the induction of c-fos gene expression was used to identify neuronal populations in the spinal cord activated by acute 5 min suckling or by electrical stimulation of the central stump of the first abdominal mammary nerve in lactating rats previously separated from their litters for 6 or 18 h. In addition, to investigate whether spinal sympathetic preganglionic neurons are activated by suckling, dual immunostaining (Fos and choline acetyltransferase) was performed. Fos was expressed at low levels in continuously suckled and 6 h nonsuckled mothers, but no expression was found after 18 h of nonsuckling. On the other hand, in 6 h nonsuckled rats, significant increments in Fos expression occurred in several regions after acute suckling and after electrical stimulation. Also, the pattern of Fos expression in each spinal laminae was different for the two stimuli, i.e. more intense effects of suckling in deep laminae V–X and more intense effects in laminae I–IV with electrical stimulation. Double-labeling after suckling was found only in sympathetic preganglionic neurons from the intermedio-medial cell column, whereas after electrical stimulation, double label was observed only in neurons from the intermedio-lateral cell column. On the other hand, no effect upon Fos protein expression was observed after suckling and only a minor effect after electrical stimulation of mammary nerve in 18 h nonsuckled rats. These results are consistent with previous findings on the sympathetic reflex regulation of the mammary gland, as well as on the importance of the nonsuckling interval for optimal functioning of lactation.

Changes in synaptic inputs to sympathetic preganglionic neurons after spinal cord injury

Spinal cord injury (SCI) leads to plastic changes in organization that impact significantly on central nervous control of arterial pressure. SCI causes hypotension and autonomic dysreflexia, an episodic hypertension induced by spinal reflexes. Sympathetic preganglionic neurons (SPNs) respond to SCI by retracting and then regrowing their dendrites within 2 weeks of injury. We examined changes in synaptic input to SPNs during this time by comparing the density and amino acid content of synaptic input to choline acetyltransferase (ChAT)-immunoreactive SPNs in the eighth thoracic spinal cord segment (T8) in unoperated rats and in rats at 3 days or at 14 days after spinal cord transection at T4. Postembedding immunogold labeling demonstrated immunoreactivity for glutamate or gamma-aminobutyric acid (GABA) within presynaptic profiles. We counted the number of presynaptic inputs to measured lengths of SPN somatic and dendritic membrane and identified the amino acid in each input. We also assessed gross changes in the morphology of SPNs using retrograde labeling with cholera toxin B and light microscopy to determine the structural changes that were present at the time of evaluation of synaptic density and amino acid content. At 3 days after SCI, we found that retrogradely labeled SPNs had shrunken somata and greatly shortened dendrites. Synaptic density (inputs per 10-microm membrane) decreased on ChAT-immunoreactive somata by 34% but increased on dendrites by 66%. Almost half of the inputs to SPNs lacked amino acids. By 14 days, the density of synaptic inputs to dendrites and somata decreased by 50% and 70%, respectively, concurrent with dendrite regrowth. The proportion of glutamatergic inputs to SPNs in spinal cord-transected rats ( approximately 40%) was less than that in unoperated rats, whereas the GABAergic proportion (60-68%) increased. In summary, SPNs participate in vasomotor control after SCI despite profound denervation. An altered balance of excitatory and inhibitory inputs may explain injury-induced hypotension.

Intrathecal anaesthesia alters intracellular Ca2+/Mg2+ homeostasis in the spinal cord neurones of experimental rats.

The effect of anaesthesia induced by intrathecal injection of 6.3% MgSO4 or 4% lidocaine on intracellular electrolyte homeostasis in spinal cord neurones of a rat model was investigated. Intracellular Ca2+, Mg2+, Na+ and K+ concentrations were determined at different times after intrathecal administration of NaCl (saline, a control group), MgSO4 or lidocaine. In both thoracic and lumbar spinal cord segments, Ca2+ concentrations rose significantly 30 min and 2 h after 6.3% MgSO4 injection, and after 24 h were still significantly increased compared with the values obtained from the control group which were subjected to sham 'anaesthesia' by saline injection (172, 121 and 108 ng mg-1 protein vs. control 23 ng mg-1 protein, respectively, in the thoracic segment and 222, 229 and 176 ng mg-1 protein vs. control 43 ng mg-1 protein, respectively, in the lumbar segment). Lidocaine injection also produced a significant increase in intracellular Ca2+ in the thoracic and lumbar spinal cord segments (69, 64 and 53 ng mg-1 protein vs. control 33.4 ng mg-1 protein and 26, 94 and 46 ng mg-1 protein vs. 23 ng mg-1 protein respectively). Only a modest rise in intracellular Mg2+ was observed after intrathecal MgSO4 or lidocaine injection (27 ng mg-1 protein vs. 23 ng mg-1 protein). Na+ and K+ concentrations decreased 24 h after MgSO4 and 1 h after lidocaine injection. Intrathecal anaesthesia by MgSO4 or lidocaine alters intracellular electrolyte homeostasis in spinal cord neurones of experimental rats. A possible common mechanism of action via Ca2+ ion channels is discussed.

Changes in NMDA receptor subunit expression in response to contusive spinal cord injury.

Differential assembly of N-methyl-D-aspartate (NMDA) receptor subunits determines their functional characteristics. Using in situ hybridization, we found a selective increase of the subunits NR1 and NR2A mRNA at 24 h in ventral motor neurons (VMN) caudal to a standardized spinal cord contusion injury (SCI). Other neuronal cell populations and VMN rostral to the injury site appeared unaffected. Significant up-regulation of NR2A mRNA also was seen 1 month after SCI in thoracic and lumbar VMN. The selective effects on VMN caudal to the injury site suggest that the loss of descending innervation leads to increased NMDA receptor subunit expression in these cells after SCI, which may alter their responses to glutamate. In contrast, protein levels determined by western blot analysis show decreased levels of NR2A 1 month after SCI in whole thoracic segments of spinal cord that included the injury sites. No effects of injury were seen on subunit levels in cervical or lumbar segments. Taken together with our previous study showing alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionate receptor subunit down-regulation after injury, our data suggest that glutamate receptor composition is significantly altered after SCI. These changes need to be taken into account to properly understand the function of, and potential pharmacotherapy for, the chronically injured spinal cord.

Ultrastructural localization of the binding fragment of tetanus toxin in putative gamma-aminobutyric acidergic terminals in the intermediolateral cell column: a potential basis for sympathetic dysfunction in generalized tetanus.

Tetanus toxin (TeTx) causes sympathetic hyperactivity, a major cause of mortality in generalized tetanus, apparently by obstructing the inhibition of sympathetic preganglionic neurons (SPNs). Neuroanatomic tracing and immunohistochemistry were used to investigate whether axon terminals in the intermediolateral cell column (IML) that synapse on SPNs and use the inhibitory neurotransmitter gamma-aminobutyric acid (GABA) may be infected transsynaptically with TeTx. The binding fragment of TeTx (TTC; an atoxic surrogate of TeTx) and the cholera toxin B subunit (CTB; a retrograde tracer) were injected into the rat superior cervical ganglion and, over 16-48 hours, were transported to the ipsilateral IML in the caudal half of the last cervical and first three thoracic spinal cord segments. With light microscopy, diffuse CTB immunolabeling extended throughout SPN perikarya and dendrites. Punctate TTC and GABA immunolabeling were accumulated densely in the neuropil between and surrounding SPN processes. With electron microscopy, 54% of the axon terminals in the IML (n = 1,337 terminals) were TTC immunolabeled (TTC(+)), and 25% contained putative neurotransmitter levels of GABA immunolabeling (GABA(+)). On average, GABA(+) terminals had a 76% chance of also being TTC(+) and a 62% greater chance of being TTC(+) than GABA(-) terminals (P < 0.000001). Axon terminals were just as likely to be TTC(+) and/or GABA(+) regardless of whether the dendrites they synapsed on were large (>1 microM) or small in cross-sectional area or were labeled retrogradely. Sympathetic hyperactivity in tetanus may involve 1) retrograde and transsynaptic transport of TeTx by SPNs and 2) at least in part, an infection of GABAergic terminals in the IML.

BOLD MRI of the human cervical spinal cord at 3 tesla

The feasibility of functional MRI of the spinal cord was investigated by carrying out blood oxygen-level dependent (BOLD) imaging of the human cervical spinal cord at a field of 3 T. BOLD imaging of the cervical spinal cord showed an average intensity increase of 7.0% during repeated exercise with the dominant hand with a return to baseline during rest periods. The areas of activation were predominantly on the same side of the spinal cord as the hand performing the exercise, between the levels of the sixth cervical and first thoracic spinal cord segments. The direct correspondence between these areas and those involved with the transmission of motor impulses to the hand, and reception of sensory information from the hand, demonstrates that spinal functional magnetic resonance imaging is feasible. Magn Reson Med 42:571–576, 1999. © 1999 Wiley-Liss, Inc.

BOLD MRI of the human cervical spinal cord at 3 tesla.

The feasibility of functional MRI of the spinal cord was investigated by carrying out blood oxygen-level dependent (BOLD) imaging of the human cervical spinal cord at a field of 3 T. BOLD imaging of the cervical spinal cord showed an average intensity increase of 7.0% during repeated exercise with the dominant hand with a return to baseline during rest periods. The areas of activation were predominantly on the same side of the spinal cord as the hand performing the exercise, between the levels of the sixth cervical and first thoracic spinal cord segments. The direct correspondence between these areas and those involved with the transmission of motor impulses to the hand, and reception of sensory information from the hand, demonstrates that spinal functional magnetic resonance imaging is feasible. Magn Reson Med 42:571-576, 1999.

Segmental Specificity of Chick Sympathetic Preganglionic Projections Is Influenced by Preganglionic Neurons from Neighboring Spinal Cord Segments

Sympathetic preganglionic neurons of the chick are located between the brachial and lumbosacral enlargements of the spinal cord. Their axons exit the spinal cord via their adjacent ventral roots and project rostrally or caudally along the sympathetic trunk to innervate sympathetic ganglia. The projections of sympathetic preganglionic neurons are segmentally specific. Neurons from the 16th cervical (C16) and the first thoracic (T1) spinal cord segments project predominantly in the rostral direction, whereas those from the fifth thoracic (T5) to the first lumbar (L1) spinal segments project predominantly in the caudal direction. Neurons from intervening spinal cord segments (T2-T4) project in rostral and caudal directions. In the present study, neural tube manipulations show that the direction of preganglionic projections is altered by both the elimination and addition of preganglionic neurons projecting into the sympathetic trunk from neighboring segments. The present study also compares the projections of preganglionic neurons from transplants of multiple neural tube segments with those from transplants of single neural tube segments reported in a previous study (Yip, 1987). In the previous study when single thoracic neural tube segments were transplanted to the cervical level, preganglionic neurons did not maintain their original projection patterns. The present study found that, when contiguous neighboring segments were transplanted to the cervical level, preganglionic neurons maintained projection patterns characteristic of their original segmental levels. These results indicate that the direction of preganglionic projections can be influenced by neurons from neighboring segments, suggesting that the formation of segmentally specific preganglionic projections during embryogenesis may involve the interactions of preganglionic neurons with those from neighboring spinal cord segments.