Number Of Anterior Horn Cells Research Articles

Cryopreserved human umbilical cord patch for in-utero spina bifida repair.

To identify a patch system to repair surgically created spina bifida in a sheep model for its efficacy in healing the skin defect, protecting the underlying spinal cord and reducing the Chiari II malformation. Spina bifida was created surgically in 16 fetuses from eight timed-pregnant sheep at gestational age of 75 days. Two fetuses did not survive the procedure. Repeat hysterotomy was performed at 95 days' gestation to cover the defect with either biocellulose film with underwater adhesive (BCF-adhesive) (n = 7) or human umbilical cord with suture (HUC-suture) (n = 7). Three fetuses without formation of the defect served as reference controls. The skin healing was examined by direct visualization after a planned Cesarean section at term, followed by histological analysis using hematoxylin and eosin and Masson's trichrome stains. Mid-sagittal sections of the fetal cranium and upper cervical spine were analyzed by a pediatric neuroradiologist who was blinded to the type of patch received. Three fetuses that received the BCF-adhesive and six fetuses that received the HUC-suture survived to term for final analysis. As a result of dislodgment of the BCF-adhesive, all spina bifida defects repaired using BCF-adhesive were not healed and showed exposed spinal cord with leakage of cerebrospinal fluid. In contrast, all spinal defects repaired by HUC-suture were healed with complete regrowth of epidermal, dermal and subdermal tissue components, with no exposed spinal cord. The maximal skin wound width was 21 ± 3.6 mm in the BCF-adhesive group but 3 ± 0.8 mm in the HUC-suture group (P < 0.001). The spinal cord area (P = 0.001) and the number of anterior horn cells (P = 0.03) was preserved to a greater degree in the HUC-suture group than in the BCF-adhesive group, whilst psammoma bodies, signifying neuronal degeneration, were only observed in the BCF-adhesive group. Anatomic changes, indicative of Chiari II malformation, were seen in all three fetuses of the BCF-adhesive group but in none of the HUC-suture group (P < 0.01). Cryopreserved umbilical cord graft is a promising regenerative patch for intrauterine repair of spina bifida.

Improved neurological outcome by intramuscular injection of human amniotic fluid derived stem cells in a muscle denervation model.

PurposeThe skeletal muscle develops various degrees of atrophy and metabolic dysfunction following nerve injury. Neurotrophic factors are essential for muscle regeneration. Human amniotic fluid derived stem cells (AFS) have the potential to secrete various neurotrophic factors necessary for nerve regeneration. In the present study, we assess the outcome of neurological function by intramuscular injection of AFS in a muscle denervation and nerve anastomosis model.Materials and MethodsSeventy two Sprague-Dawley rats weighing 200–250 gm were enrolled in this study. Muscle denervation model was conducted by transverse resection of a sciatic nerve with the proximal end sutured into the gluteal muscle. The nerve anastomosis model was performed by transverse resection of the sciatic nerve followed by four stitches reconnection. These animals were allocated to three groups: control, electrical muscle stimulation, and AFS groups.ResultsNT-3 (Neurotrophin 3), BDNF (Brain derived neurotrophic factor), CNTF (Ciliary neurotrophic factor), and GDNF (Glia cell line derived neurotrophic factor) were highly expressed in AFS cells and supernatant of culture medium. Intra-muscular injection of AFS exerted significant expression of several neurotrophic factors over the distal end of nerve and denervated muscle. AFS caused high expression of Bcl-2 in denervated muscle with a reciprocal decrease of Bad and Bax. AFS preserved the muscle morphology with high expression of desmin and acetylcholine receptors. Up to two months, AFS produced significant improvement in electrophysiological study and neurological functions such as SFI (sciatic nerve function index) and Catwalk gait analysis. There was also significant preservation of the number of anterior horn cells and increased nerve myelination as well as muscle morphology.ConclusionIntramuscular injection of AFS can protect muscle apoptosis and likely does so through the secretion of various neurotrophic factors. This protection furthermore improves the nerve regeneration in a long term nerve anastomosis model.

Numerous FUS-positive inclusions in an elderly woman with motor neuron disease

We report an autopsy case of a 75-year-old Japanese woman with motor neuron disease (MND) showing numerous neuronal and glial inclusions immunostained with anti-fused in sarcoma (FUS) antibody. At 73 years, she received a diagnosis of MND and died of respiratory insufficiency 2 years later. No mutation was found in all exons of the FUS gene. Neuropathological examination revealed a reduced number of anterior horn cells and degeneration of the pyramidal tracts. Neither Bunina bodies nor inclusions positive for ubiquitin/phosphorylated TAR DNA binding protein of 43 kD (pTDP-43), such as skein-like or round inclusions, were observed. However, basophilic inclusions (BIs) were frequently observed in the remaining neurons of the anterior horns, facial nuclei, hypoglossal nuclei, vestibular nuclei, dentate nuclei and inferior olivary nuclei. In an immunohistochemical analysis, the BIs showed strong immunoreactivity with anti-FUS and anti-ubiquitin-binding protein p62 (p62) antibodies. The nuclear staining of FUS was preserved in some neurons with FUS-positive inclusions, and a few FUS-positive glial inclusions were found. FUS-positive inclusions were more common than p62-positive inclusions in some anatomical regions, and in some neurons, p62 immunoreactivity was observed in only parts of the BIs. These results suggest that BI formation and TDP-43 aggregation have different pathogenic mechanisms, and FUS may play an important role in the pathogenesis of MND with BIs. This patient has the oldest reported age of onset for MND with BIs, and clinical features observed in this patient were indistinguishable from those of classic sporadic MND. Therefore, we consider that the age of onset and clinical features of FUS-related disorders may be variable.

Local administration of glial cell line-derived neurotrophic factor improves behavioral and histological deficit of neonatal Erb's palsy in rats.

We sought to evaluate from a behavioral and histological viewpoint the effect of local administration of glial cell line-derived neurotrophic factor (GDNF) on neonatal preganglionic Erb's palsy in rats. The Erb's palsy model was produced by transecting the anterior and posterior roots of the left C5-C7 nerves of 7-day-old rats. The rats were divided into GDNF-treated (n = 10) and vehicle-treated groups (n = 11). After we transected the roots, contact in the proximal and distal stumps of the transected nerves was maintained, and the transected point and the entire intraspinal portion of the transected roots were enclosed by Gelfoam soaked with 10 micro g GDNF or saline. The behavioral evaluation consisted of a foot-fault test and a forepaw muscle strength test, all of which were performed from the third to the seventh weeks after the operation. Seven weeks after the operation, all rats were killed, the number of anterior horn cells was counted at C5-C7, and the differences on each side were compared. In the vehicle-treated group, the foot-fault test indicated an abnormality in forelimb function on the root transection side. In the GDNF-treated group, however, significant improvement in forelimb function was observed on the basis of the foot-fault test results obtained in the third to sixth weeks after the operation. In the histological evaluation, the number of anterior horn cells from the side in which the operation took place in the vehicle-treated group was significantly less than that taken from the contralateral side at each segment. In the GDNF-treated group, however, there was no difference in any of the segments, regardless of the side from which they were taken. Local administration of GDNF in a neonatal preganglionic Erb's palsy model resulted in significant improvement in deficits on the basis of behavioral and histological evaluations.

Development of a novel experimental rat model for neonatal pre-ganglionic upper brachial plexus injury

A neonatal upper brachial plexus injury, referred to as Erb's palsy, is a serious obstetric problem. Some surgical methods are used to treat this injury, but they are inadequate. To seek new treatments for Erb's palsy, we used a model for cervical preganglionic root transection in neonate rats and evaluated the behavioral and histological compatibility of this model with Erb's palsy. Two groups were used in this study. In the group, receiving the Erb operation, the left anterior and posterior roots of spinal vertebra C5–C7 were transected at the preganglionic level, and the results were compared with those of a group that received a sham operation. In the group, receiving the Erb operation, walking difficulties and behavioral abnormalities were observed. These observations were noted on the side where the transection took place, and the problems were attributed to proximal muscle weakness in the forelimb. Additionally, the forepaw grip was not impaired. Furthermore, in this group, the number of anterior horn cells in the cervical cord on the transected side was significantly lower than that on the contralateral side ( P<0.001). The results of this study indicate that the model fulfills the criteria for the clinical symptoms of Erb's palsy and that it may also serve as a new method for enabling treatment of the condition.

Increased expression of growth-associated protein 43 on the surface of the anterior horn cells in amyotrophic lateral sclerosis.

This study examined axonal terminal alterations in the anterior horn of amyotrophic lateral sclerosis (ALS) patients. An antibody against growth-associated protein 43 (GAP43), a phosphoprotein which is expressed in elongating terminals of neurites, was employed for immunohistochemical staining. Lumbar spinal cords taken at autopsy from five ALS patients and from six control adults were examined. In control patients, there were numerous GAP43-positive granules diffusely dispersed throughout the anterior horn neuropil, and individual large anterior horn cells (AHCs) showed numerous tiny immunoreactive granules and small dots on the surface. A small number of AHCs showed dense accumulation of GAP43 immunoreactivity on the surface of the cell body and proximal processes. In all ALS patients, similar accumulation of GAP43 immunoreactivity was seen on the surface of a large number of remaining AHCs. Statistical analysis revealed a significant increase in number of AHCs with such accumulation in ALS patients. These results suggest that during the ALS disease process there may be plastic alterations or a compensatory mechanism of the axonal terminals located on the surface of some AHCs for ongoing anterior horn presynaptic terminal degeneration.

Microscopic morphometry of the spinal cord with cervical spondylotic myelopathy

Quantitative studies with morphometric assessments were carried out on the slpinal cord of four cases (aged 71–88 years) with cervical spondylotic myelopathy (CSM), and compared with four normal spinal cords from age‐matched cadaver specimens (aged 76–84 years). The histological preparation of the slpinal cord was made after embedding in celloidin and staining with luxol fast blue‐periodic acid‐Schiff‐hematoxylin and Klüver‐Barrera methods. the number, average transverse area and perimeter of the anterior horn cells (AHC) in the Rexed's lamina IX at C3, C5, C6 and L1 levels were counted and measured; the number, average transverse area and perimeter of axon in the posterior funiculus at C3 level and in the lateral corticospinal tracts at L1 level were also counted and measured under high magnification with oil immersion (1340 times), using a combination of an electronic optical planimeter, a microscope with a drawing tube, and a personal computer. The results can be summarized as: (i) The numbers of AHC in CSM at C5, C6 and L1 levels were significantly decreased compared with normal controls, and the average transverse area of AHC at C6 and L1 levels showed a decrease in varying degrees (p&lt;0.05 or P&lt;0.01, respectively). However, the number and average area of AHC in CSM at C3 level did not show any difference. (ii) The number and average transverse area of axons in the posterior funiculus with CSM at C3 level showed a significant reduction compared with normal controls (P&lt;0.01 and P&lt;0.05, respectively). (iii) There was an obvious decrease in the number of axons in the lateral corticospinal tracts of CSM at L1 level (P&lt;0.01), but the average area of axons showed no statistical reduction. It can be deduced from these results that there were ascending degeneration of the posterior funciculus and descending degeneration of the lateral corticospinal tracts above and below the segments of comparession caused by CSM. The loss in number and reduction in transverse area of AHC mainly occurred in those segments involved with compression. However, the result of AHC at L1 level revealed that CSM may have a distal influence on AHC below thesegments of compression.

Age-related reductions in number and size of anterior horn cells at C6 level of the human spinal cord.

We examined numbers and sizes of anterior horn cells (AHCs) of the human spinal cord at C6 level with aging process. Spinal cords were obtained from twenty-four male cadavers, age ranged from 41 to 97 years without any accompanying pathological changes of the spinal cord. For quantitative investigation of the AHCs, spinal cord segments were embedded in celloidin after secondary fixation and dehydration; sections (20 microns thick) were stained with luxol fast blue-periodic acid-Schiff-hematoxylin and Klüver-Barrera methods; the neuronal number, the cell body area and the perimeter of AHCs in the gray matter (equivalent to the Rexed's lamina IX) were counted and measured using a combination of a microscope with a drawing tube, a digitizer and a computer at 400 times magnification. A marked reduction in number of AHCs was found in the aged subjects. Similarly, there were significantly age-related decreases in the cross-sectional area and perimeter of AHCs at C6 segment of the spinal cord. These findings indicate that the reduction in number and size of AHCs in the human spinal cord is associated with degenerative neuronal changes and loss of motoneurons during aging process.

Adult spinal muscular athrophy: A report of four cases

Four cases of spinal muscular atrophy (SMA) are reported, 3 with detailed autopsy findings. These are compared with 2 cases of typical amyotrophic lateral sclerosis (ALS) with neuropathological data. The 3 autopsy cases of SMA only showed decreased numbers of anterior horn cells in the spinal cord, with no change in the cortical pyramidal cells, including the Betz cells, and no degeneration of the pyramidal tracts, while the ALS cases showed loss both of lower and upper motor neurons and degeneration of the pyramidal tracts. In our opinion, the infantile, juvenile, adult, and late-life forms of SMA are really a single disease entity that occurs at varying ages and is separate from ALS. The term lower motor neuron disease would be preferable because the lesions are not limited to the spinal cord, but also occur in the brain stem.

Morphological study on the hereditary neurogenic amyotrophic dogs: accumulation of lipid compound-like structures in the lower motor neuron.

A morphological study was performed on hereditary neurogenic amyotrophic dogs, the clinical features of which especially resembled spinal progressive muscular atrophy (SPMA), a human motor neuron disease. The skeletal muscles showed obvious neurogenic atrophy with endomysial fibrosis. The peripheral nerves revealed axonal degeneration mainly limited to the motor nerve. In the spinal cord, the number of anterior horn cells seemed normal but, interestingly enough, numerous accumulated granules were detected in these anterior horn cells. Histochemically, these granules were interpreted as a lipid compound. Under the electron microscope, the granules were disclosed as multi-lamellar structures, arranged concentrically or in parallel, resembling membranous cytoplasmic bodies (MCBs) or zebra bodies. This finding strongly suggests that hereditary abnormality of lipid metabolism may underlie SPMA in these dogs. However, unlike other metabolic disorders where accumulations of granules are diffusely distributed, in the dogs we examined accumulations were found only in the anterior horn cells of the spinal cord and in the hypoglossal and spinal accessory nuclei. We are unable to explain this occurrence at the present time. Further investigations should be made on dogs because they serve as an important animal model of human motor neuron disease.

Electromyographic and histopathologic correlations in arthrogryposis.

PREVIOUS interpretations of the pathologic processes underlying arthrogryposis multiplex congenita (AMC) have varied markedly. Recently, Smith et al1have concluded from electromyographic (EMG) observations in 14 patients and Drachman and Banker2,3have reported from necropsy studies that the underlying process was neurogenic. Adams et al4reported both neurogenic and myopathic cases confirmed by necropsy. There were decreased numbers of anterior horn cells in the spinal cord of cases considered to be neurogenic. Skeletal muscles showed small myofibers. In myopathic cases there were no spinal cord changes, but diffuse myopathic changes were present in muscle. Banker et al5reported the autopsy findings in a case of congenital muscular dystrophy, and Pearson and Fowler6described a necropsy of hereditary nonprogressive muscular dystrophy as causes of arthrogryposis. Bargeton et al7found changes indicating a collagen disease in their autopsied patient, and in ther opinon the collagen

Lower motor neuron deficit in arthrogryposis. An EMG study.

The pathogenesis of arthrogryposis multiplex congenita has continued to be obscure since the disorder was first described by Otto in 1841. Although there is fairly uniform agreement today that the multiplex congenital contractures found in this disease are related to abnormality of muscle and to reduced intrauterine movement, the nature and cause of the muscle abnormality remain in doubt. 1-3 After reviewing the literature, Drachman and Banker 4 concluded that all cases labeled arthrogryposis which have had microscopic examination of muscle showed changes attributable either to denervation or to a primary nonprogressive myopathy. They point out that in 12 of the 13 known cases in which the spinal cords of arthrogrypotic patients have also been studied, the one consistent finding has been significant reduction in the number of anterior horn cells, particularly at cervical and lumbosacral levels. In 1 of the 12 spinal cords neuronophagia was observed, and this was