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Longstanding theories in the field of neurophysiology have held that walking in rats is an unconscious, rhythmic locomotion that does not require cortical involvement. However, recent studies have suggested that the extent of cortical involvement during walking actually varies depending on the environmental conditions. To determine the impact of environmental conditions on cortical engagement in freely walking rats, we recorded limb kinematics and signals from implanted electroencephalography arrays in rats performing a series of natural behaviors. We found that rat gaits were significantly different across various locomotion terrains (e.g. walking on an upslope vs. downslope). Further, rat forelimbs and hindlimbs showed similar patterns of motion. The results also suggested that rat cortical engagement during walking varied across environmental conditions. Specifically, α band power significantly increased during 30° downslope walking in the posterior parietal, left secondary motor, and left somatosensory clusters. Additionally, during 30° upslope walking, the β band power was greater in the left primary motor and left and right secondary motor sources. Further, rats walking on up- or downslopes of varying steepness were found to have different cortical activities. Compared with 10° downslope walking, α band power was greater during 30° downslope locomotion in the left primary motor and somatosensory sources. These findings support the hypothesis that cortical contribution during walking in rats is influenced by environmental conditions, underlining the importance of goal-directed behaviors for motor function rehabilitation and neuro-prosthetic control in brain-machine interfaces.

Motor disorders in patients with diabetes are associated with diabetic peripheral neuropathy, which can lead to symptoms such as lower extremity weakness. However, it is unclear whether central motor system disorders can disrupt motor function in patients with diabetes. In a streptozotocin-induced rat model of type 1 diabetes, we used intracortical microstimulation to evaluate motor representations in the motor cortex, recorded antidromic motor cortex responses to spinal cord stimulation to evaluate the function of corticospinal tract (CST) axons, and used retrograde labeling to evaluate morphological alterations of CST neurons. The diabetic rats exhibited size reductions in the hindlimb area at 4 weeks and in trunk and forelimb areas after 13 weeks, with the hindlimb and trunk area reductions being the most severe. Other areas were unaffected. Additionally, we observed reduced antidromic responses in CST neurons with axons projecting to lumbar spinal segments (CST-L) but not in those with axons projecting to cervical segments (CST-C). This was consistent with the observation that retrograde-labeled CST-L neurons were decreased in number following tracer injection into the spinal cord in diabetic animals but that CST-C neurons were preserved. These results show that diabetes disrupts the CST system components controlling hindlimb and trunk movement. This disruption may contribute to lower extremity weakness in patients.

Low doses of histamine or H1R agonist 2-pyridylethylamine (2-PEA) into the knee-joint were found to decrease formalin-induced articular nociception in rats. In this study, we evaluated the participation of spinal NPY in the antinociceptive effect produced by 2-PEA. Injection of formalin (1.5%) into one of the knee-joints causes the limping of the respective limb due to nociception, which was registered each 5 min over 60 min. Neuropeptide Y1 receptor (Y1R) content in the spinal cord was evaluated by western-blotting. Intrathecal (i.t.) injection of Y1R agonist Leu31, Pro34-NPY (0.7–7 µmol) decreased nociception, while injection of the antagonist BIBO 3304 (4 μmol), increased nociception. Antinociception produced by 2-PEA was reversed by a sub-effective i.t. dose of the Y1R antagonist. Similarly, this antinociceptive effect was prevented by i.t. pretreatment with the neurotoxin NPY-saporin (750 ng), which also reduced immunoblotting for Y1R in spinal cord homogenates. These data support the idea that antinociception induced by H1R agonists in the knee-joint of rats may be mediated by the spinal release of NPY, and this peptide seems to be acting via Y1R.

Spinal cord injury (SCI) is a medical condition that currently lacks effective treatment. Galantamine is a reversible, competitive acetylcholinesterase inhibitor, used to treat patients with Alzheimeŕs disease. It has been demonstrated that galantamine increases cerebral neurogenesis and has a neuroprotective effect by binding to nicotinic receptors and has an anti-inflammatory effect due to its allosteric binding to the α7nAChR. In the present study, the effects of galantamine on functional recovery and histological outcome in a rat contusion model of SCI were analyzed. Male Wistar rats were submitted to SCI using a NYU/MASCIS impactor. The animals from the galantamine group were treated with 5 mg/kg galantamine intraperitoneally for 5 days. The Basso, Beattie and Bresnahan scale (BBB) was used to evaluate locomotor activity. The expression of beta3-tubulin, NFM, GFAP, O4, CD68 and CD3 was analyzed by flow cytometry. Rats that received galantamine had significantly higher BBB scores in comparison with the control lesion group. Galantamine treatment increased the percentage of NFM positive cells at 6 weeks post-injury and reduced the size of the lesion. The results indicate that galantamine increased tissue survival and accelerated hind limb motor function recovery. This is the first study that has shown the possibility of therapeutic use of galantamine in a model of acute spinal cord injury.

Spinal cord injury (SCI) often results in some form of paralysis. Recently, SCI therapy has been focused on preventing secondary injury to reduce both neuroinflammation and lesion size so that functional outcome after an SCI may be improved. Previous studies have shown that adenosine receptors (AR) are a major regulator of inflammation after an SCI. The current study was performed to examine the effect of caffeine, a pan-AR blocker, on spontaneous functional recovery after an SCI. Animals were assigned into 3 groups randomly, including sham, PBS and caffeine groups. The rat SCI was generated by an NYU impactor with a 10 g rod dropped from a 25 mm height at thoracic 9 spinal cord level. Caffeine and PBS were injected daily during the experiment period. Hind limb motor function was evaluated by the Basso, Beattie, Bresnahan (BBB) locomotor rating scale at 1 week and 4 weeks after the SCI. Spinal cord segments were collected after final behavior evaluation for morphological analysis. The tissue sparing was evaluated by luxol fast blue staining. Immunofluorescence stain was employed to assess astrocyte activation and neurofilament positioning, while microglia activation was examined by immunohistochemistry stain.The results showed that spontaneous functional recovery was blocked after the animals were subjected caffeine daily. Moreover, caffeine administration increased the demyelination area, promoted astrocyte and microglia activation and decreased the quantity of neurofilaments. These findings suggest that the neurotoxicity effect of caffeine may be associated with the inhibition of neural repair and the promotion of neuroinflammation.

Paraplegia following complex aortic intervention relies on crude evaluation of lower extremity strength such as whether the patient can lift their legs or flex the ankle. Little attention has been given to the possible long-term neurologic sequelae following these procedures in patients appearing functionally normal. We hypothesize that mice subjected to minimal ischemic time will have functional and histological changes despite the gross appearance of normal function. Male mice underwent 3 min of aortic occlusion (n=14) or sham surgery (n=4) via a median sternotomy. Neurologic function was graded by Basso Motor Score (BMS) preoperatively and at 24 h intervals after reperfusion. Mice appearing functionally normal and sham mice were placed on a walking beam and recorded on high-definition, for single-frame motion analysis. After 96hrs, spinal cords were removed for histological analysis. Following 3 min of ischemia, functional outcomes were split evenly with either mice displaying almost normal function n=7 or near complete paraplegia n=7. Additionally, single-frame motion analysis revealed significant changes in gait. Histologically, there was a significant stepwise reduction of neuronal viability, with even the normal function ischemic group demonstrating significant loss of neurons. Despite the appearance of normal function, temporary ischemia induced marked cyto-architectural changes and neuronal degeneration. Furthermore high-definition gait analysis revealed significant changes in gait and activity following thoracic aortic occlusion. These data suggest that all patients undergoing procedures, even with short ischemic times, may have spinal cord injury that is not evident clinically.

Friedreich ataxia is the most common autosomal recessive disorder of the cerebellum, causing degeneration of spinal sensory neurons and spinocerebellar tracts. The disease is caused by severely reduced levels of frataxin, a mitochondrial protein involved in iron metabolism. An experimental model has been generated by crossing mice homozygous for a conditional allele of the Fxn gene with mice heterozygous for a deleted exon 4 of Fxn carrying a tissue-specific Cre transgene under control of the muscle creatine kinase promoter. Relative to wild-type, Fxn null mutants were impaired on tests of motor coordination comprising horizontal bar, vertical pole, and the rotorod as well as displaying gait anomalies and the hindlimb clasping response. The Fxn KO/Mck model reproduces some key features of patients with Friedreich ataxia and provides an opportunity of ameliorating their symptoms with experimental therapies.

This paper probes into the participation of different concentrations of oxytocin in the pain regulation of lateral habenular nucleus on normal adult rats. The detailed experiment steps are as follows: microinject 1.25, 2.5, and 5nmol of oxytocin to three adult rats׳ lateral habenular nucleus, and measure the hind paw withdrawal latency (HWL) after 5, 10, 15, 20, 30, 45 and 60min with hot plate and pressure plate test as the indicator of algesia. At the same time, another group of adult rats were injected with normal saline as a control to observe the participation of oxytocin in the pain regulation of lateral habenular nucleus. Through the oxytocin microinjection, the HWL of rats was significantly prolonged, compared with the control group. According to the experimental results, the HWL of thermo stimulation on the left paw of the rats between OT group and the control group (injected with normal saline) showed no difference (p>0.05) after a microinjection of 1.25nmol in LHb, while the HWL of thermo stimulation on the left paw of the rats showed significant difference (p

The present study was designed to further compare the stepping-like movements generated via epidural (ES) and/or intraspinal (IS) stimulation. We examined the ability to generate stepping-like movements in response to ES and/or IS of spinal lumbar segments L1–L7 in decerebrate cats. ES (5–10 Hz) of the dorsal surface of the spinal cord at L3–L7 induced hindlimb stepping-like movements on a moving treadmill belt, but with no rhythmic activity in the forelimbs. IS (60 Hz) of the dorsolateral funiculus at L1–L3 (depth of 0.5–1.0 mm from the dorsal surface of the spinal cord) induced quadrupedal stepping-like movements. Forelimb movements appeared first, followed by stepping-like movements in the hindlimbs. ES and IS simultaneously enhanced the rhythmic performance of the hindlimbs more robustly than ES or IS alone. The differences in the stimulation parameters, site of stimulation, and motor outputs observed during ES vs. IS suggest that different neural mechanisms were activated to induce stepping-like movements. The effects of ES may be mediated more via dorsal structures in the lumbosacral region of the spinal cord, whereas the effects of IS may be mediated via more ventral propriospinal networks and/or brainstem locomotor areas. Furthermore, the more effective facilitation of the motor output during simultaneous ES and IS may reflect some convergence of pathways on the same interneuronal populations involved in the regulation of locomotion.

Moyamoya disease (MMD) is a chronic occlusive cerebrovascular disease with unknown etiology that is characterized by the development of abnormal vascular networks at the base of the brain. Recent genome-wide studies identified RNF213 as an important MMD susceptibility gene. However, the exact mechanism by which the RNF213 abnormality leads to MMD remains unknown. Thus, we sought to clarify the role of RNF213 in angiogenesis under ischemic conditions using conventional RNF213 knockout mice. We assessed the infarction volume, cerebral edema, and vascular density in the ischemic brain after transient middle cerebral artery occlusion (tMCAO). To further evaluate systemic angiogenesis following chronic ischemia, we investigated blood flow recovery using laser speckle flowmetry, the severity of ambulatory impairments, and vascular density in the hind-limb after permanent femoral artery ligation. Results were compared between homozygous RNF213 knockout mice (RNF213 -/-) and wild-type littermates (Wt). No significant differences were observed in infarction volume or the formation of edema following tMCAO, or in vascular density 28 days after tMCAO between RNF213 -/- and Wt. Blood flow recovery was significantly improved in RNF213 -/- from 3 to 28 days after femoral artery ligation, and angiogenesis as shown by vascular density in the hind-limb was significantly enhanced in RNF213 -/- at 28 days. The amelioration of ambulatory impairments was also evident in RNF213 -/-. Angiogenesis was enhanced in mice lacking RNF213 after chronic hind-limb ischemia, which suggested the potential role of the RNF213 abnormality in the development of pathological vascular networks in chronic ischemia.

In Parkinson disease (PD), a complex neurodegenerative disorder that affects nearly 10 million people worldwide, motor skills are significantly impaired. However, onset and progression of motor deficits and the neural correlates of these deficits are poorly understood. We used a genetic mouse model of PD (Pink1−/−), with phenotypic similarities to human PD, to investigate the manifestation of early-onset sensorimotor deficits. We hypothesized this mouse model would show early vocalization and gross motor dysfunction that would be progressive in nature. Pink1−/− mice, compared to wild type (WT) controls, were evaluated at 2, 3, 4, 5, and 6 months of age. To quantify deficit progression, ultrasonic vocalizations and spontaneous locomotor activity (cylinder test and pole test) were analyzed. Although somewhat variable, in general, Pink1−/− mice produced significantly more simple calls with reduced intensity as well as a larger percentage of cycle calls compared to WT counterparts. However, there were no significant differences in duration, bandwidth, or peak frequency for any of the ultrasonic call types between genotypes. Pink1−/− mice showed a significant impairment in limb motor skills with fewer hindlimb steps, forelimb steps, and rears and lands in the cylinder test compared to WT. Additionally, Pink1−/− mice took significantly longer to turn and traverse during the pole test. Immunohistochemical staining showed no significant difference in the number of tyrosine hydroxylase (TH) positive cells in the substantia nigra or density of TH staining in the striatum between genotypes. These data suggest the Pink1−/− mouse model may be instrumental in defining early motor biomarkers of PD in the absence of nigrostriatal dopamine loss.

The current study was performed to evaluate the mechanisms and therapeutic effects of overexpressing neuroglobin (Ngb) on spinal cord injury (SCI). Adeno-associated virus (AAV) was injected in the T12 section 7 days before SCI. Animals were randomly divided into four groups: a sham group, a vehicle group, an AAV-EGFP group and an AAV-Ngb group. Recovery of hind limb locomotor function was determined during the 3-week post operation period by the Basso, Beattie and Bresnahan locomotor rating scale. At 24 h after SCI and at the end of the study, the segments of spinal cord, centered with the lesion site were harvested for histopathological analysis. Immunofluorescence was performed using antibodies to recognize neuN in the lesion sections. At 24 h after SCI, the spinal cord tissue samples were removed to analyze tissue concentrations of superoxide dismutase (SOD) and malondialdehyde (MDA). Apoptotic cells were assessed using a terminal deoxynucleotidyl transferase, dUTP nick end labeling (TUNEL) kit. The expression of bcl-2, bax, cytochrome c, and cleaved caspase-3, were determined by Western blot assay and immunostaining analysis. The results showed that animals overexpressing Ngb had significantly greater recovery of locomotor function, less neuronal loss and fewer apoptotic cells. In addition, overexpressing Ngb significantly increased bcl-2 expression and SOD level, decreased bax expression, attenuated the release of cytochrome c from mitochondria to the cytosol fraction, and reduced the activity of caspase-3 and MDA level after SCI. These findings suggest, that overexpressing Ngb can significantly improve the recovery of locomotor function. This neuroprotective effect may be associated with the inhibition of neural apoptosis via the mitochondrial pathway.

Propriospinal pathways, consisting of axons from interneurons that project to other spinal segments, have been implicated as likely candidates to mediate interlimb coordination in developing and adult mammals during quadrupedal locomotion. The superficial thoracic ventrolateral funiculus (VLF) contains both ascending and descending axons, and when stimulated can induce alternating rhythmic locomotor-like activity in the lumbar ventral roots of the isolated neonatal rat spinal cord. The goal of this work was to characterize the synaptic inputs onto cervical neurons from ascending axons in the VLF. Sprague-Dawley rats (P4-P7) were deeply anesthesized with halothane and their spinal cords were isolated, removed, and maintained in vitro. Intracellular recordings were made from 68 cervical (C5-C7) neurons having 71 latency classifications in response to thoracic VLF stimulation. Antidromic (n=35), monosynaptic (n=2), di-or tri-synaptic (n=18) and long-latency polysynaptic (n=16) responses were recorded. Recordings from reduced preparations (mid-sagittal section at C5-C7) suggest that much of the delay in the long-latency polysynaptic responses require a bilaterally intact cervical spinal cord. Fifty-three percent (17/32) of the VLF responsive cervical interneurons tested also exhibited long latency excitatory responses to lumbar dorsal root stimulation suggesting that many of the cervical VLF responsive interneurons receive indirect input from lumbar primary afferents. We hypothesize that the VLF contains a population of ascending axons originating from lumbar propriospinal interneurons that can influence cervical inter- and motoneurons. These ascending VLF axons may participate in interlimb coordination by providing moment-by-moment feedback to the cervical enlargement of lumbar central pattern generator and/or hindlimb proprioceptive activity.

Glial cells play a crucial role in the pathomechanism of perinatal hypoxic-ischemic brain injury (HI) and are involved in the maintenance of a chronic state of inflammation that causes delayed neuronal damage. Activation of astrocytes is one factor prolonging brain damage and contributing to the formation of a glial scar that limits neuronal plasticity. In this context, the major astrocytic gap junction protein Connexin 43 (Cx43) has been ascribed various functions including regulation of astrocytic migration and proliferation. Here, we investigate glial responses like microglia/macrophages and astrocytic activation in a rat model of neonatal HI and characterize changes of these parameters upon transplantation of human umbilical cord blood cells (hUCB). As an alleviation of motor function in lesioned rats has previously been described in transplanted animals, we analyze the putative correlation between motor function and glial activation over time. The lesion-induced impairment of motor function, assessed by forelimb use bias, muscle strength and distal spasticity, was alleviated upon transplantation of hUCB short and long term. HI induced an acute inflammatory reaction with activation of microglia/macrophages and reactive astrogliosis associated with perilesional upregulation of Cx43 that slowly declined during the chronic post-ischemic phase. hUCB transplantation accelerated the regression of inflammatory events, narrowed the perilesional astrocytic wall and led to a downregulation of the investigated astrocytic proteins. Thus, in the immature brain, hUCB may indirectly reduce secondary cell death upon hypoxia-ischemia and facilitate post-ischemic plasticity through the attenuation of reactive gliosis. This article is part of a Special Issue entitled Electrical Synapses.

Considerable evidence supports the notion that on- and off-cells of the rostral ventromedial medulla (RVM) facilitate and depress, respectively, spinal nociceptive transmission. This notion stems from a covariation of on- or off-cell activities and spinal nocifensive reflexes. Such covariation could theoretically be due to their independently responding to a common source, or to an RVM-derived modulation of ventral horn neurons. Here, we tested whether on- and off-cells indeed modulate spinal nociceptive neurons. In deeply anesthetized rats, unitary recordings were simultaneously made from an RVM on-like or off-like cell and a spinal nociceptive neuron that shared a receptive field (RF) at a hind paw. Action potential firing in RVM/spinal neuron pairs was highly correlated, positively for on-like cells and negatively for off-like cells, both during ongoing activity and during application of calibrated noxious pressure to the RF. Microinjection of morphine into RVM induced a correlated decrease in on-like cell/spinal neuron ongoing activity and response to noxious stimulation. RVM morphine induced changes in off-like cell activity that were not correlated with spinal neuronal activity. These results suggest that on-cells exert a positive modulation upon spinal nociceptive neurons, upstream to ventral horn circuits and plausibly at the origin of nociceptive information that eventually reaches the cerebral cortex. On-cells may in this manner contribute to inflammation- and neuropathy-induced increases in withdrawal reflexes. Most significantly, on-cell modulation of nociceptive neurons may be a key factor in clinical pain conditions such as hyperalgesia and allodynia.

We examined gene expression in the lumbar spinal cord and the specific response of motoneurons, intermediate gray and proprioceptive sensory neurons after spinal cord injury and exercise of hindlimbs to identify potential molecular processes involved in activity dependent plasticity. Adult female rats received a low thoracic transection and passive cycling exercise for 1 or 4weeks. Gene expression analysis focused on the neurotrophic factors: brain-derived neurotrophic factor (BDNF), glial cell line-derived neurotrophic factor (GDNF), neurotrophin-3 (NT-3), neurotrophin-4 (NT-4), and their receptors because of their potential roles in neural plasticity. We also examined expression of genes involved in the cellular response to injury: heat shock proteins (HSP) -27 and -70, glial fibrillary acidic protein (GFAP) and caspases -3, -7, and -9. In lumbar cord samples, injury increased the expression of mRNA for TrkB, all three caspases and the HSPs. Acute and prolonged exercise increased expression of mRNA for the neurotrophic factors BDNF and GDNF, but not their receptors. It also increased HSP expression and decreased caspase-7 expression, with changes in protein levels complimentary to these changes in mRNA expression. Motoneurons and intermediate gray displayed little change in mRNA expression following injury, but acute and prolonged exercise increased levels of mRNA for BDNF, GDNF and NT-4. In large DRG neurons, mRNA for neurotrophic factors and their receptors were largely unaffected by either injury or exercise. However, caspase mRNA expression was increased by injury and decreased by exercise. Our results demonstrate that exercise affects expression of genes involved in plasticity and apoptosis in a cell specific manner and that these change with increased post-injury intervals and/or prolonged periods of exercise.

Amyotrophic lateral sclerosis GALS) is a neurodegenerative disorder characterized by the loss of upper and lower motoneurons. Clinically, it is manifested by weakness, muscle atrophy and progressive paralysis and ends up with patients' death 2-5 years after diagnosis. Although these symptoms lead in many cases to gait deficits in patients, an exhaustive locomotor profile of animal models mimicking the disease has not been assessed yet. In this work we evaluated the locomotor performance of the SOD1(G93A) mouse model of ALS using computerized treadmill gait analysis. SOD1(G93A) mice presented early (8 weeks of age) gait abnormalities, evidenced by an increase in the time of the propulsion phase of hindlimb stance. The alterations progressed during the disease until a complete disturbance of normal gait. This finding is meaningful to the field because the identification of a significant difference in a functional endpoint as early as 8 weeks might be a step forward resolving the debate about treatment of mice prior to the symptomatic phase in efficacy studies. These results also point out that digitizing analysis of treadmill locomotion may be useful to evaluate whether new therapeutic approaches are improving functional outcome of the animals. (C) 2011 Elsevier B.V. All rights reserved.

Attenuation of ischemic injury can be achieved by priming the brain with a sub-lethal ischemic insult, a phenomenon known as ischemic preconditioning (IP). We sought to determine if subjecting a distant organ, such as the lower limb, to a similar priming ischemic insult would result in protection of the brain from a subsequent severe ischemic injury, as induced by middle cerebral artery occlusion (MCAo) and if this protection is mediated via a neurogenic pathway. Adult Wistar male rats were subjected to either remote preconditioning (RPC) or sham surgery and then subsequently underwent 2 h MCAo 24, 48 or 72 h after the RPC/sham RPC stimulus. Of the animals undergoing RPC, only those that sustained MCAo 24 h later showed significantly smaller cerebral infarct volumes (150.34±30.91 mm(3)) and better clinical neurological outcomes (1.15±0.69) as compared to the sham RPC group (infarct volume 250.25±26.98mm(3); neurological score 1.80±0.87) (P

Research has shown that physical exercise may reduce degeneration in certain brain regions experiencing ataxia. Our laboratory utilized mutant spastic Han–Wistar rats (sHW) that display developmental abnormalities, including spastic paresis, fore limb tremors, hind limb rigidity, and a reduced life span (60–65 days of age). Concomitant neurodegeneration has been observed in the cerebellum (Purkinje cells). The purpose of this study was to investigate if moderate, aerobic exercise could reduce Purkinje cell neurodegeneration and improve the motor ability and survival of the mutant sHW rat. Mutant male littermates at the ages of 20 (n = 11 pairs) and 30 (n = 13 pairs) days old were divided into running groups and non-running groups. Mutant rats were run on a motorized treadmill at the rate of 15 m/min with a 10% slope. The “running” group ran for 30 min per day, 5 days a week; the “non-runners” remained nearby in the training facility. These conditions were held constant until the mutant runners could no longer run due to disease progression. Moderate exercise increased the lifespan of running mutant rats in both the 20-day start group (14% increase) and 30-day start group (13% increase). The rats exhibited improved motor function as open-field tests showed higher activity scores for runners after 50 days. Histological examination of the cerebellum revealed a 62% increase in Purkinje cell survival of the runners. These results suggest that aerobic exercise ameliorates, at least partially, cerebellar dysfunction in the sHW rat, an excellent model of ataxia.

The dorsal root reflex (DRR) and the axonal reflex (AR) are antidromic activities in primary afferents and are involved in neurogenic inflammation. DRRs and/or ARs lead to release of neuropeptides calcitonin gene-related peptide (CGRP) and substance P (SP). CGRP causes blood vessels to dilate leading to an increase in blood perfusion, whereas SP causes plasma extravasation, leading to edema. Both DRR and AR can be evoked by noxious stimuli. The goal of this study was to determine the role of DRR and AR in neurogenic inflammation by examining the blood perfusion (BP) change in hindpaws in response to formalin injection (an acute inflammatory agent). Laser Doppler images were collected simultaneously in both hindpaws in anesthetized rats to determine the level of BP. Local lidocaine was applied to the left sciatic nerve to block both orthodromic signals and antidromic DRRs without affecting ARs. All rats then received a subcutaneous formalin injection to the left hindpaw. Our results showed that (1) the mean BP of the left paw increased significantly following formalin injection, with or without lidocaine; (2) application of lidocaine in the left sciatic nerve alone significantly increased BP ipsilaterally; (3) formalin injection following lidocaine application significantly increased BP more than the group without lidocaine; and (4) there was delayed significant BP increase in the right (contralateral) hindpaw following formalin injection with or without lidocaine. It is concluded that ARs play a more important role than DRRs in formalin-induced neurogenic inflammation.

The study was aimed to investigate the protective effects of rosuvastatin on spinal cord ischemia in rats and to determine the effects of this agent on the expressions of nitric oxide synthase (NOS). Spinal cord ischemia was induced in male Sprague–Dawley rats by occluding the descending thoracic aorta. Experimental groups (n = 30 per group) were as follows: sham operation, control (receiving only normal saline), rosuvastatin (5 mg/kg/day for 10 days before occlusion), and rosuvastatin-mevalonate (5 mg/kg/day rosuvastatin and 5 mg/kg/day mevalonate for 10 days before occlusion). Neurological function was assessed at 6, 12, 24, 48, and 72 h after reperfusion. After 72 h reperfusion, spinal cords were harvested for 2,3,5,-triphenyltetrazolium chloride (TTC) staining, TUNEL staining, and nitric oxide (NO) assay. Immunohistochemistry, reverse transcription polymerase chain reaction and western blot were performed to determine the expressions of inducible, endothelial, and neuronal NOS (iNOS, eNOS, and nNOS) in rats with spinal cord ischemia. Spinal cord ischemia thus induced was marked by neurological dysfunction, spinal infarction, and neural cell apoptosis in animals. The results show that rosuvastatin significantly reduced the motor disturbance and the volume of infarctions and attenuated apoptotic neural cells death in the treated rats. Treatment with rosuvastatin remarkably decreased the NO level in spinal cord tissue. In addition, rosuvastatin inhibited iNOS mRNA and protein expression and increased eNOS mRNA and protein expression. However, rosuvastatin had no influence on nNOS mRNA and protein expression. Administrations of mevalonate completely reversed the changes caused by rosuvastatin. These results indicate that rosuvastatin can protect rat spinal cord against ischemia injury by modulation of NOS expressions.

Peripheral afferent denervation induces reorganization of somatotopic maps in the primary somatosensory cortex (S1). In the present study, we investigated somatotopic map plasticity after tail cut. Neonatal mice at postnatal days (P) 2-3 and adult mice at eight weeks of age were anesthetized with ether, and approximately two thirds of the tail was cut from the tip. Both groups of mice were anesthetized with urethane (1.7g/kg, i.p.) at 10weeks of age, and transcranial flavoprotein fluorescence imaging was performed in the S1. Neural activities in the S1 were elicited by vibratory stimulation applied to the contralateral hindpaw or the tail in control mice. The cortical areas activated by hindpaw, tail base, and tail tip stimuli were placed in this order according to the medial and posterior direction. In mice with tail cut, the tail base area moved to the more medial and posterior area corresponding to the tail tip in control mice. The shift of the tail base area was observed in both neonatal and adult tail cut mice, indicating the absence of a critical period before eight weeks. Medial and posterior shift of the tail base area with regard to the bregma was confirmed in tail cut mice. These data suggest that transcranial flavoprotein fluorescence imaging is a useful technique for investigating somatosensory map plasticity in mice.

Background: It is unclear whether spinal cord stimulation (SCS) at higher frequencies induces further increases in vasodilation and enhances clinical efficacy. Objectives: This study investigated effects of SCS at both a normal frequency (as used clinically) and two higher frequencies on peripheral vasodilation. Methods: A unipolar ball electrode was placed on the left dorsal column at the lumbar 2–3 spinal cord segments (L2–L3) in sodium pentobarbital anesthetized, paralyzed, and artificially ventilated rats. Cutaneous blood flow recordings from both ipsilateral (left) and contralateral (right) hind foot pads were measured with laser Doppler flow perfusion monitors. SCS at frequencies of 50, 200, or 500 Hz was applied at 30%, 60%, and 90% of motor threshold (MT) using standard square waves. Resiniferatoxin (RTX: an ultrapotent analog of capsaicin) and a calcitonin gene-related peptide (CGRP) receptor blocker (CGRP 8–37 ) was also used to elucidate mechanisms of SCS vasodilation at these higher frequencies. Results: SCS applied with the three frequencies produced similar MT ( n = 22). SCS at 500 Hz significantly increased cutaneous blood flow and decreased vascular resistance compared to changes induced by frequencies of 50 and 200 Hz ( P n = 8). RTX (2 μg/kg, i.v.) as well as CGRP 8–37 (2.37 mg/kg, i.v.) significantly reduced SCS-induced vasodilation at 500 Hz ( P n = 6) as compared to responses prior to administrations of these drugs. Conclusion: SCS at 500 Hz significantly increased SCS-induced vasodilation without influencing MT. Furthermore, effects of SCS at 500 Hz are mediated via activation of TRPV1-containing fibers and a release of CGRP.

Remote ischemic postconditioning (RIP) refers to an ischemia conducted in a distant organ that protects against a prior ischemia in another organ. We tested whether RIP protects against focal ischemia in the rat brain. Stroke was generated by a permanent occlusion of the left distal middle cerebral artery combined with a 30-min occlusion of the bilateral common carotid arteries (CCA) in male rats. After CCA release, RIP was generated by three cycles of 15-min occlusion/15-min release of the left-hind femoral artery. The results showed that rapid RIP performed immediately after CCA release reduced infarction by 67% measured at 2 days after stroke. In addition, delayed RIP initiated as late as 3 h, but not 6 h, still robustly reduced infarction by 43% 2 days after stroke. RIP's protective effect was abolished by injecting the protein synthesis inhibitor, cycloheximide, and the afferent nerve blocker, capsaicin, suggesting that RIP blocks ischemic injury by modulating protein synthesis and nerve activity. Nevertheless, rapid RIP did not reduce infarction size 2 months after stroke while it ameliorated the outcome of the behavioral test. In conclusion, RIP attenuates brain injury after focal ischemia.

Hindlimb unloading (HU) is known to induce changes in the neuromuscular system. However, no data describing the effects of HU on morphological characteristics of peripheral nerve have been reported so far. Therefore, we used soleus and radial nerves obtained from control and rats submitted to 14 days of HU to study the consequences of a decrease (soleus) or an increase (radial) in neural activity on its morphology. The mean number of fibers was not changed after HU. The soleus nerve axon diameter was weakly affected after HU, whereas the myelin thickness was reduced. For the radial nerve, both axon and fiber diameter were increased, and the myelin thickness and internodal distance were higher in HU rats. These results suggest that regulation of myelin maintenance undergoes plastic mechanisms. Neural activity and/or neural pattern might be essential in the maintenance of myelin sheath in adults. 1252

The purpose of the present study was to establish a rat bipedal walking model to examine the effects of bipedal walking on the central nervous system by training rats to perform bipedal walking over a period of 3 months. The characteristics of bipedal walking were investigated using kinematic and electromyographic methods in established bipedal walking models. Stable bipedal walking was achieved in rats by training them to stand with an upright posture and to walk with the hindlimbs using bipedal-walking training equipment to obtain a water reward. A stable head position in the rat bipedal walking model was attained primarily by closing the swing-phase period with a large angular change in the hip, knee, and ankle joints. The EMG burst pattern of the knee extensor (m. rectus femoris) and the erector muscle of the spine (m. longissimus) during bipedal walking was similar to that during quadrupedal walking in rats. We established two bipedal walking models using normal and forelimb-amputated rats. Comparative studies of these two bipedal walking models are expected to provide the information about the influence of forelimb movements on neuronal control of bipedal walking.

A possible treatment approach for chronic spinal cord injuries has been tested. We report that minced, autologous, pre-injured peripheral nerve administered as a single injection into an injury-induced cyst, resulting from a contusion injury of the thoracic spinal cord, stimulates recovery of hindlimb locomotor function in rats, as measured by the Basso, Beattie, Bresnahan Locomotor Rating Scale. This response was further enhanced by the addition of exogenous neurotrophic factors. Histological analysis showed axons of the corticospinal tract exhibited significant regeneration past the injury site, when quantified both by number and length. Results indicate that the use of a pre-injured peripheral nerve graft stimulates chronically injured descending nerves to overcome a local inhibitory environment. The resulting sprouting and growth past the injury site is associated with a significant improvement in locomotor function.

In the previous study we reported that morphine-induced conditioned place preference (CPP) could be suppressed by peripheral electric stimulation (PES), an effect related to the increased gene expression of opioid peptides in the central nervous system. Considering that opioids were known to elevate dopamine (DA) activity in the mesolimbic brain, the present study was designed to further analyze the possible involvement of the mesolimbic dopaminergic system (MLDS) in the suppressive effect of PES on the rewarding effects of morphine in SD male rats. We found that morphine-induced CPP can be successfully suppressed by PES, an effect accompanied by a reversal of the increased tissue contents of DA and its metabolites in the nucleus accumbens (NAc) of morphine-induced CPP rats. Our results suggest that MLDS seems to play important roles in the mechanisms underlying PES's suppression of the rewarding effect of drug-associated environmental cues in the rat.

Spinal cord stimulation (SCS) is used to relieve ischemic pain and improve peripheral blood flow in selected patients with peripheral arterial diseases. Our previous studies show that antidromic activation of transient receptor potential vanilloid-1 (TRPV1) containing sensory fibers importantly contributes to SCS-induced vasodilation.To determine whether peripheral terminals of TRPV1 containing sensory fibers produces vasodilation that depends upon the release of calcitonin gene-related peptide (CGRP) and nitric oxide (NO) during SCS.A unipolar ball electrode was placed on the left dorsal column at lumbar spinal cord segments 2-3 in sodium pentobarbital anesthetized, paralyzed and ventilated rats. Cutaneous blood flow from left and right hindpaws was recorded with laser Doppler flow perfusion monitors. SCS was applied through a ball electrode at 30%, 60%, 90% and 300% of motor threshold. Resiniferatoxin (RTX; 2 microg/ml, 100 microl), an ultra potent analog of capsaicin, was injected locally into the left hindpaw to functionally inactivate TRPV-1 containing sensory terminals. In another set of experiments, CGRP(8-37), an antagonist of the CGRP-1 receptor, was injected at 0.06, 0.12 or 0.6 mg/100 microl into the left hindpaw to block CGRP responses; N-omega-nitro-l-arginine methyl ester (L-NAME), a nonselective nitric-oxide synthase (NOS) inhibitor, was injected at 0.02 or 0.2 mg/100 microl into the left hindpaw to block nitric oxide synthesis; (4S)-N-(4-Amino-5[aminoethyl]aminopentyl)-N'-nitroguanidine, TFA, a neuronal NOS inhibitor, was injected at 0.02 or 0.1 mg/100 microl into the left hindpaw to block neuronal nitric oxide synthesis.SCS at all intensities produced vasodilation in the left hindpaw, but not in the right. RTX administration attenuated SCS-induced vasodilation at all intensities in the left hindpaw (P0.05, n=7) compared with responses before RTX. CGRP(8-37) administration attenuated SCS-induced vasodilation in the left hindpaw in a dose dependent manner (linear regression, P0.05) compared with responses before CGRP(8-37). In addition, L-NAME at a high dose, but not (4S)-N-(4-Amino-5[aminoethyl]aminopentyl)-N'-nitroguanidine, TFA, decreased SCS-induced vasodilation (P0.05, n=5).While TRPV1, CGRP and NO are known to be localized in the same nerve terminals, our data indicate that SCS-induced vasodilation depends on CGRP release, but not NO release. NO, released from endothelial cells, may be associated with vascular smooth muscle relaxation and peripheral blood flow increase in response to SCS.

The present experiments were done in the spinal female cat. In a prior work in the decerebrate female cat, stimulation of the sensory pudendal nerve (SPN) induced a depolarizing wave (LD) in hind limb motoneurons that outlasted the stimulus by up to 6 s. LD triggered self-sustained motoneuron firing (bistability). An intrinsic potential underlies bistable firing, which, in the cat, depends on two main factors; first, the integrity of pathways descending from the brain stem to the spinal cord and, second, the membrane potential of the motoneuron just before the stimulus; at high resting potential, excitatory short-lasting inputs induce transient but no sustained firing. Thus, no bistability occurs in the spinal cat or in hyperpolarized motoneurons of the decerebrate cat. LD might be an intrinsic potential that could also be absent in the spinal cat, or an extrinsic (synaptic) potential induced by spinal interneurons. In the latter case, the interneurons generating LD should show post-discharge as prolonged as LD. LD was produced in spinal cats and its amplitude did not change or increase slightly during hyperpolarizing pulses, which suggests that LD might be a synaptic response. Interneurons showing post-discharge to train of stimulation to SPN were located 100-200 microm above the pools of hind limb motoneurons. Some post-discharges were as prolonged as LD. We conclude that LD might be a synaptic response to local interneurons.

Transmission in spinal reflex pathways from the toes to the ankle flexor tibialis anterior (TA) and to the knee flexor semitendinosus (ST), and from the heel to the ankle extensor medial gastrocnemius (MG), has been studied during antigen-induced inflammation due to subcutaneous injection of ovalbumin at the heel or at the toes in pre-immunized, pentobarbitone-anesthetized rabbits. The ovalbumin challenge induced a mild swelling at the injection site that developed over 6 h. Inflammation at the toes facilitated both flexor reflexes evoked from the toes and inhibited MG extensor responses to stimulation at the heel, with effects usually developing within 75 min of the initial injection and persisting to the end of the 6 h post-injection recording period. When inflammation was induced at the heel, the heel-MG and toes-ST reflexes were enhanced, whereas the toes-TA reflex was inhibited. The findings support the view that long-lasting changes in reflexes evoked by noxious stimuli have evolved to enhance reflex protection of an injured site.

The aim of the present study was to investigate the effects of hindlimb unloading (HU) on the expression of neurotrophin-3 (NT-3) and brain-derived neurotrophic factor (BDNF), together with the expression of their high-affinity receptors tropomyosin receptor kinase C (TrkC) and tropomyosin receptor kinase B (TrkB), in lumbar (L4-6) segment of the spinal cord and in the soleus muscle. The mRNA and protein levels of the genes of interest were compared using quantitative PCR and western blot assays. Immunohistochemistry for NT-3 and BDNF was used to detect the levels of protein in the motoneurons in the lateral motor column. In this study, NT-3 and BDNF mRNA and protein expression were significantly increased in the spinal cord and soleus muscle after HU. NT-3 immunoreactivity, but not BDNF immunoreactivity, was significantly increased in the large motoneurons located in lateral motor column after 14 days of HU. The level of TrkC protein in the spinal cord and soleus muscle were significantly elevated after both 7 days and 14 days of HU. However, TrkC mRNA, TrkB mRNA and TrkB protein levels did not change significantly. Elevated BDNF, NT-3 and TrkC levels in the neuromuscular system indicate that neurotrophins are involved in HU-induced neuromuscular plasticity. NT-3 is a candidate to mediate the synaptic efficacy between alpha motoneurons and group Ia afferents.

Electromyographic (EMG) burst patterns of m. longissimus and the kinematics of the vertebral column were assessed in cats during treadmill walking for six downslope grades (5 degrees-30 degrees). The EMG bursts during downslope walking were weak between 5 degrees and 20 degrees. At steeper grades (20 degrees), EMG bursts were large. Bursts at T10 facilitated inward movements, and those at L1 decreased forward movements, while those at L5 decreased backward movements during downslope walking at steeper grades.

To gain insight into the neural mechanisms controlling vertebral column movement and its role in walking, we performed kinematic and electromyographic (EMG) studies on cats during level and upslope treadmill walking. Kinematic data of the limbs and vertebral column were obtained with a high-speed camera synchronized with EMG recordings from levels T10, L1, and L5 of m. longissimus dorsi (Long). During a single-step cycle at all upslope angles, vertebral movement in the lateral (left-right), cranial-caudal (forward-backward), and dorsal-ventral (upward-downward) directions was observed. Lateral movements were produced by forelimb take-off and hindlimb landing, and forward and upward movements were produced by hindlimb extension. During the single-step cycle, each of the three epaxial muscles, m. multifidus, m. iliocostalis, and Long, showed two bilateral EMG bursts. The onset of the EMG bursts coincided with the left-right movements, suggesting that epaxial muscle activity depresses lateral movement. The termination of the EMG bursts correlated with the forward and downward phase of the step cycle, suggesting that contraction of the epaxial muscles produces forward and downward movements. EMG bursts of the epaxial muscles increase the stiffness and produce inwardly movements to decrease the lateral movements of the vertebral column and the termination of EMG bursts control the movements into cranial and ventral direction of the vertebral column. The results suggest that the rhythmic EMG bursts in the epaxial muscles are produced by pattern generators, and the timing of EMG bursts among the different levels of the epaxial muscles are altered by walking condition input via peripheral afferents and descending pathways.

Amyotrophic lateral sclerosis (ALS) is a fatal disease caused by progressive degeneration of motor neurons in the spinal cord and motor cortex. Although the etiology of ALS remains unknown, a mutation of the gene encoding Cu,Zn-superoxide dismutase (SOD1) has been reported in 20% of familial cases of ALS (FALS). Transgenic mice that overexpress a mutated human SOD1 exhibit a phenotype and pathology similar to those observed in patients with FALS. Mitochondrial abnormality has been reported in patients with ALS and in animal models of FALS. We recently reported that l -carnitine, an essential cofactor for the β-oxidation of long-chain fatty acids, effectively inhibits various types of mitochondrial injury and apoptosis both in vitro and in vivo. The present study demonstrates that oral administration of l -carnitine prior to disease onset significantly delayed the onset of signs of disease (log-rank P = 0.0008), delayed deterioration of motor activity, and extended life span (log-rank P = 0.0001) in transgenic mice carrying a human SOD1 gene with a G93A mutation (Tg). More importantly, subcutaneous injection of l -carnitine increased the life span of Tg mice (46% increase in male, 60% increase in female) even when given after the appearance of signs of disease.

We tested the hypothesis that dorsal horn laminae III–IV cell receptive fields (RFs) are initially established in three steps: cutaneous axons penetrate the dorsal horn near their rostrocaudal (RC) levels of entry into the spinal cord. Their terminal branches distribute mediolaterally (ML) according to their relative distoproximal RF locations on the leg, and form nonselective synapses with nearby dorsal horn cell dendrites, establishing the initial dorsal horn cell RFs. Rootlet axon RFs in adult cats were used to approximate the RC entry levels of hindlimb skin input. Cord dorsum recordings of monosynaptic field potentials evoked by electrical skin stimulation provided the RC distributions of synaptic input. These were in close agreement. Simulated projections of all 22,000 hindlimb axons were similar to projections predicted from EPSP distributions, and with the observed projections of dorsal roots, cutaneous nerves, and individual axons. The simulated terminals were connected nonselectively to nearby dendrites of 135,000 simulated lamina III–IV cells whose dendritic surface area distributions were based on intracellularly stained cells. There was an overall similarity among pre- and postsynaptic embryonic and adult somatotopies, with a progressive transformation of RF angular location as a function of RC, ML dorsal horn location from an initial embryonic presynaptic concentric pattern to an adult postsynaptic radial one. The initial embryonic dorsal horn cell RF assembly hypothesis was supported by the simulations, as was the additional hypothesis that further refinement of connections would be necessary to establish sufficient selectivity to account for observed adult RFs and somatotopy.

The cholinergic agonist pilocarpine injected intraperitoneally (ip) increases mean arterial pressure (MAP) and superior mesenteric (SM) vascular resistance and reduces submandibular/sublingual gland (SSG) vascular resistance. In the present study, we investigated the effects of electrolytic lesions of the anteroventral third ventricle (AV3V) region on the changes in MAP, SM, and SSG vascular resistances induced by ip pilocarpine. Male Holtzman rats anesthetized with urethane (1.0 g/kg) and chloralose (60 mg/kg) were submitted to sham or electrolytic AV3V lesions and had pulsed Doppler flow probes implanted around the arteries. Contrary to sham rats, in 1-h and 2-day AV3V-lesioned rats, pilocarpine (4 micromol/kg) ip decreased MAP (-41 +/- 4 and -26 +/- 4 mm Hg, respectively, vs. sham: 19 +/- 4 mm Hg) and SM (-48 +/- 11 and -45 +/- 10%, respectively, vs. sham: 41 +/- 10%) and hindlimb vascular resistances (-65 +/- 32 and -113 +/- 29%, respectively, vs. sham: 19 +/- 29%). In 7-day AV3V-lesioned rats, pilocarpine produced no changes on MAP and SM and hindlimb vascular resistances. Similar to sham rats, pilocarpine reduced SSG vascular resistance 1 h after AV3V lesions (-46 +/- 6%, vs. sham: -40 +/- 6%), but it produced no effect 2 days after AV3V lesions and increased SSG vascular resistance (37 +/- 6%) in 7-day AV3V-lesioned rats. The responses to ip pilocarpine were similar in 15-day sham and AV3V-lesioned rats. The cholinergic antagonist atropine methyl bromide (10 nmol) iv slightly increased the pressor response to ip pilocarpine in sham rats and abolished for 40 min the fall in MAP induced by ip pilocarpine in 1-h AV3V-lesioned rats. The results suggest that central mechanisms dependent on the AV3V region are involved in the pressor responses to ip pilocarpine. Although it was impaired 2 and 7 days after AV3V lesions, pilocarpine-induced salivary gland vasodilation was not altered 1 h after AV3V lesions which suggests that this vasodilation is not directly dependent on the AV3V region.

The effect of experimental muscle pain on the amplitude and velocity sensitivity of muscle spindle primary afferent neurons in the trigeminal mesencephalic nucleus (Vmes) was examined. Extracellular recordings were made from 45 neurons designated as spindle primary- or secondary-like on the basis of their response to ramp-and-hold jaw movements. Velocity sensitivity was assessed in spindle primary-like afferents by calculating the mean dynamic index of each unit in response to three different velocities of jaw opening before and after intramuscular injection with hypertonic saline (HS, 5%, 100 microl). The amplitude sensitivity of all jaw muscle spindle afferents was assessed by calculating the mean firing rate of each unit in response to three different amplitudes of jaw openings during both the open and hold phases of the movement and with best-fit lines obtained, using linear regression analysis, before and after HS injection. The variance of the two regression lines obtained for each unit before and after the injection was compared using the coincidence test, and changes in intercept and slope were determined. Seventy-five percent of the primary-like units and 80% of the secondary-like units presented with changes in static behavior after HS injection. Thirty-six percent of the primary-like units showed changes in dynamic behavior. Injection of isotonic saline (control) did not alter the responses of the spindle afferent to jaw opening. Thus, our results demonstrate that the predominant effect of noxious stimulation was a shift in the amplitude sensitivity of both spindle primary-like and secondary-like afferents and, to a lesser extent, the velocity sensitivity of the spindle primary-like unit. In accordance with earlier studies in the cat hindlimb and neck muscles, these results suggest that the activation of masseter muscle nociceptor alters spindle afferent responses to stretch acting primarily through static gamma motor neurons.

Recent studies have reported that l-arginine-derived nitric oxide (NO) in the gracile nucleus modifies the hypotensive responses to electroacupuncture (EA) stimulation of Zusanli (ST 36). The purpose of this study was to examine the influence of EA stimulation of ST 36 on neuronal NO synthase (nNOS) expression in the brainstem nuclei in rats. EA stimulation of ST 36 and a non-acupoint was performed using 3 Hz of stimulation for 10 s every 2 min for a period of 120 min in rats anesthetized with ketamine. Rats in the sham-treated group received surgery and EA needles were placed into the acupoints without performing the stimulation. After 2-h stimulation and sham treatment, animals were perfused with 4% paraformaldehyde. Sections of rat medulla were examined by immunolabeling with a polyclonal antibody directed against nNOS. The brainstem nuclei were also visualized by NADPH-diaphorase histochemistry, a marker of nNOS activity. nNOS expression and NADPH-diaphorase reactivity were quantified by using a microscope with reticule grid to count the number of positive cells over a nucleus. Unilateral EA stimulation of ST 36 in rats caused increases in nNOS immunostained cells in the rostral region of the ipsilateral gracile nucleus, but was not altered in the contralateral gracile nucleus compared with sham-treated rats (P < 0.05, n = 6–7). NADPH-diaphorase-positive cells were also increased in the ipsilateral gracile nucleus of rats with EA stimulation. nNOS immunostaining and NADPH-diaphorase-positive neurons were significantly increased in both ipsilateral and contralateral sides of the medial nucleus tractus solitarius (mNTS) in rats receiving EA ST 36 compared with sham-treated animals (P < 0.05). nNOS immunostaining and NADPH-diaphorase reactivity was neither altered in the gracile nucleus and mNTS of non-acupoint stimulated rats nor other brainstem nuclei in rats with EA ST 36. These results show that nNOS immunoreactivity and NADPH-diaphorase reactivity are consistently increased in the gracile nucleus and the mNTS by EA ST 36. We conclude that EA ST 36 induces nNOS expression in the gracile nucleus and mNTS, and enhanced nNOS-NO in the nuclei may modify central cardiovascular regulation, which contribute to hypotensive effects of acupuncture.

We previously described a rat model where the injection of formalin in the tail induced a facilitation of the hindpaw withdrawal reflexes (hyperalgesia). In the present work, after injecting formalin in the tail, we measured the levels of pro-nociceptive mediators tumor necrosis factor-α (TNF) and substance P (SP) in the rat paws. A significant increase of SP levels was evident in the hindpaw, whereas no changes in SP were observed in the forepaw. Both in the hindpaw and in the forepaw the TNF levels were higher than normal at each stage of measurement. Our results indicate that a prolonged neuronal activation induced by formalin injection is associated with a change in nociceptive and inflammatory mediators in distal sites of the body. The fact that SP levels are changed in the hindpaw but not in the forepaw might point to the activation of a mechanism of retrograde signaling from central synapses to paw afferent nerves.

In limbed vertebrates, the capacity to generate rhythmic motor patterns for locomotion and scratching is distributed over spinal cord segments of the limb enlargement (e.g., lumbosacral segments), but within this region, rostral segments are more rhythmogenic than caudal segments. The underlying reasons for this rostrocaudal asymmetry are not clear. One possibility is that rostral and caudal segments receive distinct sets of propriospinal projections. To test this hypothesis, I injected horseradish peroxidase (HRP) into the ventral horn unilaterally in a rostral or caudal segment of the turtle hindlimb enlargement. I quantitatively assessed the distributions of retrogradely labeled neurons in six hindlimb enlargement and pre-enlargement segments. The cross-sectional distribution did not depend on which segment was injected. Ipsilateral labeling occurred predominantly in the deep dorsal horn, the lateral part of the intermediate zone, and the dorsal two-thirds of the ventral horn, while contralateral labeling occurred mainly in the medial part of the ventral horn and the lateral part of the intermediate zone. This cross-sectional distribution is similar to what has been seen in mammals. The rostrocaudal distribution of labeled cells, however, depended on which segment was injected. Rostral injections gave rise to rostrally skewed distributions, dominated by descending propriospinal neurons. Caudal injections gave rise to caudally skewed distributions, dominated by ascending propriospinal neurons. Thus, rostral segments of the hindlimb enlargement received more propriospinal inputs from immediately rostral than immediately caudal segments, while the reverse was true for inputs to caudal segments. This anatomical asymmetry may contribute to known functional asymmetries within the enlargement.

The TrkB-expressing sensory neurons seem to be involved in touch and other discriminative sensibilities. Thus, several slowly and rapidly adapting cutaneous mechanoreceptors, as well as muscle spindles, are reduced or absent in the territory of the trigeminal nerve in functionally TrkB-deficient mice. Whether this also occurs in the cutaneous or muscular territories of dorsal root ganglia has not been analyzed. Here we used immunohistochemistry and transmission-electron microscopy to analyze the impact of a mutation in the gene coding for TrkB on Meissner and Pacinian corpuscles, and muscle spindles. The animals were studied at the post-natal days 15 and 25, because at this time all the mechanoreceptors examined are fully developed. Typical Meissner's corpuscles, displaying S-100 protein immunoreactivity, were found in the digital pads of wild-type and TrkB+/- mice whereas they were absent in the TrkB-/- animals. Regarding Pacinian corpuscles, the mutation in the trkB gene does not alter either the immunohistochemical or the ultrastructural characteristics. Finally, in muscle spindles the arrangement of the intrafusal muscle fibers and nerve fibers was unchanged in the mutated animals. Nevertheless, about 10% of muscle spindles showed increased number of the intrafusal cells (between 6 and 12) and were supplied by more than one large myelinic nerve fiber. The present results strongly suggest that TrkB-expressing sensory neurons in dorsal root ganglia, like those of the trigeminal ganglion, are responsible for the development and maintenance of several rapidly adapting cutaneous mechanoreceptors, i.e. Meissner's corpuscles.

Although hemodynamic-based functional brain imaging techniques are powerful tools to explore the brain functions noninvasively, hemodynamic-based signal is strongly affected by spatial configuration of microvessels. Understanding the quantitative relations between microvascular structure and functional activity is therefore significant to make a valid signal interpretation for the imaging techniques. In the present study, we evaluated depth profiles of microvascular distributions in rat somatosensory subfields (barrel field, forelimb region, trunk region and hindlimb region) and characterized depth variations in microvascular structures, such as locations, lengths and directions of microvessels, throughout the cortical layers (I–VI). To obtain the accurate microvascular structure, we made a customized casting method by using confocal laser scanning microscope. We observed that microvascular distribution successively varied throughout the cortical layers (I–VI) and that the maximum number density of microvessels was consistently found in middle layers (III–V). In addition, superficial layers had relatively long microvessels, almost perpendicular to the cortical surface, whereas middle layers had short microvessels propagating in all directions. These regional differences in microvascular structures were closely related to the somatosensory subfields, e.g., barrel field was the greatest number density of microvessels among the investigated subfields. Based on these observations, we compared microvascular profiles with previously reported distribution patterns of tissue partial pressure of oxygen (pO2). The results showed that tissue pO2 was correlated with microvascular distribution in some but not all of the subfields. This finding shows that detailed microvascular profiles are helpful to investigate causal relationships between microvascular structure and functional activities in cerebral cortex.