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Adipose derived stem cells (ADSCs) are multipotent and can transdifferentiate into neural stem cells. We investigated the transdifferentiation of ADSCs to neural phenotype (NP) cells using selegiline and two-dimensional electrophoresis (2-DE). The perinephric and inguinal fat of rats was collected and used to isolate ADSCs that were characterized by immunophenotyping using flow cytometry. The ADSCs were differentiated into osteogenic and lipogenic cells. The NP cells were generated using 10 -9 mM selegiline and characterized by immunocytochemical staining of nestin and neurofilament 68 (NF-68), and by qRT-PCR of nestin, neurod1 and NF68. Total protein of ADSCs and NP cells was extracted and their proteome patterns were examined using 2-DE. ADSCs carried CD73, CD44 and CD90 cell markers, but not CD34. ADSCs were differentiated into osteocyte and adipocyte lineages. The differentiated NP cells expressed nestin, neuro d1 and NF-68. The proteome pattern of ADSCs was compared with that of NP cells and eight spots showed more than a two fold increase in protein expression. The molecular weights and isoelectric points of these highly expressed proteins were estimated using Melanie software. We compared these results with those of the mouse proteomic database using the protein isoelectric point database, and the functions of the eight proteins in differentiation of NP cells were predicted using the UniProt database. The probable identities of the proteins that showed higher expression in NP cells included cholinesterase, GFRa2, protein kinase C (PKC-eta) and RING finger protein 121. The sequences of the proteins identified from mouse database were aligned by comparing them with similar proteins in rat database using the Basic Local Alignment Search Tool (BLAST). The E values of all aligned proteins were zero, which indicates consistency of the matched protein. These proteins participate in differentiation of the neuron and their overexpression causes ADSCs transdifferentiation into NP cells.

Valproic acid (VPA) is an inhibitor of histone deacetylases (HDACs) that can regulate differentiation and proliferation of stem cells by epigenetic mechanisms. We investigated VPA induced histone H3 and H4 acetylation in adipose derived stem cells (ADSCs) transdifferentiated into neuron-like cells (NLCs). Rat ADSCs were transdifferentiated into neural stem cells (NSCs) that had been generated from neurospheres. The NSCs were differentiated into NLCs by induction with different concentrations of VPA at 24, 48 and 72 h. The NLCs were evaluated using anti-H3 and -H4 antibodies, and ADSCs, NSCs and NLCs were evaluated using immunofluorescence. The ADSCs were immunoreactive to CD90 and CD49d, but not to CD45 and CD31. Both the neurospheres and NSCs were immunostained with nestin and neurofilament 68. The neurospheres expressed Musashi1, Sox2 and Neu N genes as determined by RT-PCR. Our dose-response study indicated that the optimal concentration of VPA was 1 mM at 72 h. Histone acetylation levels of H3 and H4 immunostaining intensities in NLCs were significantly greater than for ADSCs and NSCs. VPA alters H4 and H3 acetylation immunoreactivities of ADSCs transdifferentiated into NLCs.

Poly-l-lysine-coated superparamagnetic iron oxide nanoparticles (SPIONs-PLL) were prepared and used as a novel-carrier for the transfer of brain-derived neurotrophic factor (BDNF) into neural stem cells (NSCs) under the beneficial influence of an external magnetic field. Pro-BDNF, a gene from human brain cDNA libraries, was obtained by polymerase chain reaction and constructed in a mammalian expression vector (PSecTag2/HygroB). The nanoparticles (NPs) were examined using Fourier transform infrared spectroscopy, zeta potential, and Transmission electron microscopy. From the results, the levels of BDNF among the transfected and untransfected cells were 30.326 ± 5.9 and 5.85 ± 3.11 pg/mL, respectively, as detected by an ELISA method. Moreover, the enhanced green fluorescent protein vector was used to evaluate the gene expression efficiency for SPIONs-PLL as a non-viral carrier in NSCs. This was performed under the influence of a magnetic field and the transfection reagents (such as Lipofectamine 2000), which served as a positive control. The histological analysis revealed that the concentration of intracellular NPs was significantly higher than intercellular NPs. These results suggest that SPIONs-PLL can serve as a novel alternative for the transfection of BDNF-NSCs and could be used in gene therapy.

Objectives: Growing body of evidence suggests that neurite outgrowth is a key determinant in the re-networking of damaged neuronal circuits as well as synaptogenesis. The essential molecule in this interesting process is microtubule-associated protein 2 (MAP2) studies demonstrated that inhibition of MAP2 by antisense Oligonucleotide hinders neurite outgrowth., Methods: In the current study, we evaluated the effects of valproic acid (VPA), a histone deacetylase inhibitor on the expression of MAP2 in the rat spinal cord injury model by real time RT-PCR and immunoreactivity assays., Results: We revealed that a significant increase in the MAP2 overexpression occurred in VPA-treated group compared to the sham group by quantitative real-time reverse transcription polymerase chain reaction (RT-PCR) and immunoreactivity assays for MAP2 gene expression., Discussion: The present findings indicated that systemic administration of VPA stimulated MAP2 gene expression and also supports the involvement of the MAP2-mediated de novo re-arborization and neurite outgrowth of neurons, which might contribute to successful neuronal re-wirings. Thus, VPA has promising applications in the treatment of many neurological problems, such as neurotrauma, Alzheimer and Parkinson's disease, and others.

Adult neural stem/precursor cells (NSPCs) residing in the subventricular zone of the lateral ventricles and the subgranular zone of the dentate gyrus of the hippocampus are involved in the memory formations and psychological problems. It is believed that basal levels of glucocorticoids are essential for neuronal development, plasticity, and survival, while stress-mediated levels of glucocorticoids produce neuronal loss. Degeneration of NSPCs by the apoptotic and necrotic stimuli have great devastating outcomes on the brain and contributes to the pathophysiology of neurological as well as psychological disorders. Using MTT assay, acridine orange, and TUNEL assay, we have demonstrated that cortisol at high and excessive (more than 5 μM) levels had anti-proliferative effects on the NSPCs derived from subventricular and subgranular zones in a dose- and time-dependent manner through apoptosis as well as necrosis. These outcomes can highlight the role of stress-mediated decline of adult neurogenesis in the aging brain and interconnect stress-mediated cortisol secretion with brain aging diseases.

Objective: Temporal lobe epilepsy is one of the most common types of epilepsy. Cytotherapy was tried for several neurological disorders. In this investigation, bone marrow stromal cells (BMSCs) were used in order to evaluate the recovery of epileptic rats induced by pilocarpine., Methods: The rats were divided into four groups: a negative control, treated groups, and epileptic rats treated with the vehicle only. The animals in the chronic phase were monitored for three weeks using Racine scale. BMSCs were labeled with BrdU and injected intravenously. After 6 weeks, the rats were killed and processed in paraffin as well as immunohistochemical techniques., Results: The results of the behavioral test showed that the number of seizures significantly decreased in treatment groups. Histopathology of the tissues from the untreated rats showed cell death and neurophagia. The numerical density of neurons per area was significantly higher in the treated groups than in the untreated ones. BMSCs localized in the hippocampus of the treated animals., Discussion: The results showed that there is structural and functional improvement in the epileptic rats treated with BMSCs.

Objective: Bone marrow stromal cells (BMSCs) were documented as a feasible candidate for cell therapy. Several protocols with one or more steps, using different chemicals, were used for inducing BMSCs differentiation into neuronal phenotype. Many of these chemicals were reported to be of mutagenic, teratogenic or carcinogenic properties. The purpose of this work was to evaluate the neuronal inductivity of selegiline to BMSCs., Methods: Selegiline was used to induce BMSCs following pre-induction with beta-mercaptoethanol and retinoic acid. The neuronal phenotype was evaluated using antineurofilament 200, neurofilament 68, anti-Map-2 and synapsin I antibodies. In addition, reverse transcription polymerase chain reaction was used for the expression of brain-derived neurotrophic factor (BDNF), nerve growth factor (NGF), neurotrophin 3 (NT-3), neuroligin 1 and post-synaptic density protein 95 (PSD-95)., Results: The result of the work showed that the induced BMSCs were immunoreactive for neurofilaments 200 and 68. Moreover, anti-Map-2 and synapsin I antibodies showed a structure morphologically consistent with synapses. Reverse transcription polymerase chain reaction showed that the induced BMSCs could express BDNF, NGF, NT-3, neuroligin 1 and PSD-95. The time course used was 1, 3, 6, 12, 24, 36 and 48 hours, and the percentages of neurofilament 200 immunoreactive cells were analysed using the logistic growth curve., Discussion: The results showed that selegiline was efficient and a potent inducer for BMSCs into neuronal phenotype, which can be used in the cell therapy in neurodegeneration and traumatic nervous tissue injuries.

Objectives: Bone marrow stromal cells (BMSCs) have been reported to improve movement deficit in adult rats with spinal cord injury (SCI). The purpose of this study is to determine the distribution of BMSCs in the spinal cord lesion of the contusion model of SCI., Methods: Laminectomy was carried out at L1 vertebra level and SCI was carried out using the weight drop method. BMSCs were isolated from adult rats, labeled with bromodeoxyuridine (BrdU) and administered intravenously to the rats 1 week after SCI, which were killed after 4 weeks. The non-treated animals were used as negative control, which showed cavitations of the spinal cord after 5 weeks of SCI. Rats in another group were killed immediately and used to study the hemorrhagic lesions. The volume densities (Vv) of the hemorrhage and cavitation were the highest at the site of direct trauma., Results: The numerical densities of the transmigrated cells per area (Nat) were as follows: 0.3 +/- 0.2, 3.9 +/- 0.4, 5.4 +/- 0.4, 8.4 +/- 0.5, 5.5 +/- 0.3, 3.6 +/- 0.3 and 0.4 +/- 0.2 at the end and the middle of the thoracic vertebra 13 (T13), the region between T13 and the first lumbar vertebra, the middle of L1, the region between L1 and L2, and the middle and the end of L2 vertebra, respectively. The distribution of Nat at the above regions was a Gaussian model. The volume densities of hemorrhage in the spinal cord taken from the above regions showed that hemorrhage with the highest volume density occurred at the impact site and the volume density declined as the samples taken were more distant from the impact site., Discussion: The migration of BMSCs in the injured region depends on the amount of the hemorrhage and damage to blood vessels of the spinal cord.

Objectives: To study the early effect of axotomy on spinal motoneurons and dendritic trees in the newborn rat., Methods: The left sciatic nerve of 50 neonatal Sprague--Dawley rats aged 5 days old was transected and the right area kept as a control. The operated animals were killed 2, 4, 8, 12 and 24 hours after axotomy. The L4--L6 segments of the spinal cord were sampled, and stained either with Golgi-Cox or Cresyl fast violet methods. The number of motoneurons, the largest soma diameter and the different parameters of the dendritic trees such as: number, length and thickness of the primary, secondary and tertiary branches in the axotomized sides were estimated and compared statistically with that of the intact sides., Results: The results indicated that in the axotomized sides, the number of motoneurons and the largest soma diameter were decreased, but these were significant only in 12- and 24-hour groups. The number of branches of the dendritic trees including primary, secondary and tertiary branches was not significantly decreased in the groups. The thickness of the dendrites showed a reduction; however, this was significant only for the tertiary branches in the 24-hour groups. The length of the primary, secondary and tertiary branches of the dendrites--especially the latter--were also decreased significantly in most of the groups., Conclusion: Axotomy at the early stages in newborn rats resulted in noticeable morphometrical changes in motoneurons and their dendrites.