Density Of Motoneurons Research Articles

Intraventricular Administration of Exosomes from Patients with Amyotrophic Lateral Sclerosis Provokes Motor Neuron Disease in Mice.

Amyotrophic lateral sclerosis (ALS) is a severe disease of the central nervous system (CNS) characterized by motor neuron damage leading to death from respiratory failure. The neurodegenerative process in ALS is characterized by an accumulation of aberrant proteins (TDP-43, SOD1, etc.) in CNS cells. The trans-synaptic transmission of these proteins via exosomes may be one of the mechanisms through which the pathology progresses. The aim of this work was to study the effect of an intraventricular injection of exosomes obtained from the cerebrospinal fluid (CSF) of ALS patients on the motor activity and CNS pathomorphology of mice. The exosomes were obtained from two ALS patients and a healthy donor. Exosome suspensions at high and low concentrations were injected into the lateral brain ventricles of male BALB/c mice (n = 45). Motor activity and physiological parameters were evaluated twice a month; morphological examination of the spinal cord was performed 14 months after the start of the experiment. Nine months after administration of exosomes from the ALS patients, the animals started exhibiting a pathological motor phenotype; i.e., altered locomotion with paresis of hind limbs, coordination impairment, and increasing episodes of immobility. The motor symptoms accelerated after administration of a higher concentration of exosomes. The experimental group showed a significant decrease in motor neuron density in the ventral horns of the spinal cord, a significant increase in the number of microglial cells, and microglia activation. The TDP43 protein in the control animals was localized in the nuclei of motor neurons. TDP43 mislocation with its accumulation in the cytoplasm was observed in the experimental group. Thus, the triggering effect of the exosomal proteins derived from the CSF of ALS patients in the development of a motor neuron pathology in the experimental animals was established. This confirms the pathogenetic role of exosomes in neurodegenerative progression and makes it possible to identify a new target for ALS therapy.

Motor neuron involvement threatens survival in spinocerebellar ataxia type 1.

Motor neuron involvement threatens survival in spinocerebellar ataxia type 1.

Local host response of commercially available dural patches for fetal repair of spina bifida aperta in rabbit model.

Fetal surgery for spina bifida aperta (SBA) by open hysterotomy typically repairs anatomical native tissue in layers. Increasingly, fetoscopic repair is performed using a dural patch followed by skin closure. We studied the host response to selected commercially available patches currently being used in a fetal rabbit model for spina bifida repair. SBA was surgically induced at 23-24days of gestation (term=31days). Fetal rabbits were assigned to unrepaired (SBA group), or immediate repair with Duragen™ or Durepair™. Non-operated littermates served as normal controls. At term, spinal cords underwent immunohistochemical staining including Nissl and glial fibrillary acidic protein. We hypothesized that spinal cord coverage with a dural patch and skin closure would preserve motor neuron density within the non-inferiority limit of 201.65cells/mm2 and reduce inflammation compared to unrepaired SBA fetuses. Motor neuron density assessed by Nissl staining was conserved both by Duragen (n=6, 89.5; 95% CI -158.3 to -20.6) and Durepair (n=6, 37.0; 95% CI -132.6 to -58.5), whereas density of GFAP-positive cells to quantify inflammation was lower than in unrepaired SBA-fetuses (SBA 2366.0±669.7cells/mm2 vs. Duragen 1274.0±157.2cells/mm2 ; p=0.0002, Durepair 1069.0±270.7cells/mm2 ; p<0.0001). Covering the rabbit spinal cord with either Duragen or Durepair followed by skin closure preserves motor neuron density and reduces the inflammatory response.

Additives migrating from 3D-printed plastic induce developmental toxicity and neuro-behavioural alterations in early life zebrafish (Danio rerio)

The environmental impact of exposure to 3D-printed plastics as well as potential migration of toxic chemicals from 3D-printed plastics remains largely unexplored. In this work we applied leachates from plastics fabricated using a stereolithography (SLA) process to early developmental stages of zebrafish (Danio rerio) to investigate developmental toxicity and neurotoxicity. Migration of unpolymerized photoinitiator, 1-hydroxycyclohexyl phenyl ketone (1−HCHPK) from a plastic solid phase to aqueous media at up to 200 mg/L in the first 24 h was detected using gas chromatography–mass spectrometry. Both plastic extracts (LC50 22.25% v/v) and 1−HCHPK (LC50 60 mg/L) induced mortality and teratogenicity within 48 h of exposure. Developmental toxicity correlated with in situ generation of reactive oxygen species (ROS), an increase in lipid peroxidation and protein carbonylation markers and enhanced activity of superoxide dismutase (SOD) and glutathione-S-transferase (GST) in embryos exposed to concentrations as low as 20% v/v for plastic extracts and 16 mg/L for 1−HCHPK. ROS-induced cellular damage led to induction of caspase-dependent apoptosis which could be pharmacologically inhibited with both antioxidant ascorbic acid and a pan-caspase inhibitor. Neuro-behavioral analysis showed that exposure to plastic leachates reduced spontaneous embryonic movement in 24–36 hpf embryos. Plastic extracts in concentrations above 20% v/v induced rapid retardation of locomotion, changes in photomotor response and habituation to photic stimuli with progressive paralysis in 120 hpf larvae. Significantly decreased acetylcholinesterase (AChE) activity with lack of any CNS-specific apoptotic phenotypes as well as lack of changes in motor neuron density, axonal growth, muscle segment integrity or presence of myoseptal defects were detected upon exposure to plastic extracts during embryogenesis. Considering implications of the results for environmental risk assessment and the growing usage of 3D-printing technologies, we speculate that some 3D-printed plastic waste may represent a significant and yet very poorly uncharacterized environmental hazard that merits further investigation on a range of aquatic and terrestrial species.

Transplantation of clinical-grade human neural stem cells reduces neuroinflammation, prolongs survival and delays disease progression in the SOD1 rats

Stem cells are emerging as a therapeutic option for incurable diseases, such as Amyotrophic Lateral Sclerosis (ALS). However, critical issues are related to their origin as well as to the need to deepen our knowledge of the therapeutic actions exerted by these cells. Here, we investigate the therapeutic potential of clinical-grade human neural stem cells (hNSCs) that have been successfully used in a recently concluded phase I clinical trial for ALS patients (NCT01640067). The hNSCs were transplanted bilaterally into the anterior horns of the lumbar spinal cord (four grafts each, segments L3–L4) of superoxide dismutase 1 G93A transgenic rats (SOD1 rats) at the symptomatic stage. Controls included untreated SOD1 rats (CTRL) and those treated with HBSS (HBSS). Motor symptoms and histological hallmarks of the disease were evaluated at three progressive time points: 15 and 40 days after transplant (DAT), and end stage. Animals were treated by transient immunosuppression (for 15 days, starting at time of transplantation). Under these conditions, hNSCs integrated extensively within the cord, differentiated into neural phenotypes and migrated rostro-caudally, up to 3.77 ± 0.63 cm from the injection site. The transplanted cells delayed decreases in body weight and deterioration of motor performance in the SOD1 rats. At 40DAT, the anterior horns at L3–L4 revealed a higher density of motoneurons and fewer activated astroglial and microglial cells. Accordingly, the overall survival of transplanted rats was significantly enhanced with no rejection of hNSCs observed. We demonstrated that the beneficial effects observed after stem cell transplantation arises from multiple events that counteract several aspects of the disease, a crucial feature for multifactorial diseases, such as ALS. The combination of therapeutic approaches that target different pathogenic mechanisms of the disorder, including pharmacology, molecular therapy and cell transplantation, will increase the chances of a clinically successful therapy for ALS.

Electrical stimulation of human corticospinal axons at the level of the lumbar spinal segments.

Electrical stimulation over the mastoids or thoracic spinous processes has been used to assess subcortical contribution to corticospinal excitability, but responses are difficult to evoke in the resting lower limbs or are limited to only a few muscle groups. This might be mitigated by delivering the stimuli lower on the spinal column, where the descending tracts contain a greater relative density of motoneurons projecting to lower limb muscles. We investigated activation of the corticospinal axons innervating tibialis anterior (TA) and rectus femoris (RF) by applying a single electrical stimulus over the first lumbar spinous process (LS). LS was paired with transcranial magnetic stimulation (TMS) at interstimulus intervals (ISIs) of -16 (TMS before LS) to 14ms (LS before TMS). The relationship between muscle contraction strength (10%-100% maximal) and the amplitude of single-pulse TMS and LS responses was also investigated. Compared to the responses to TMS alone, responses to paired stimulation were significantly occluded in both muscles for ISIs ≥-8ms (p≤0.035), consistent with collision of descending volleys from TMS with antidromic volleys originating from LS. This suggests that TMS and LS activate some of the same corticospinal axons. Additionally, the amplitude of TMS and LS responses increased with increasing contraction strengths with no change in onset latency, suggesting responses to LS are evoked transsynaptically and have a monosynaptic component. Taken together, these experiments provide evidence that LS is an alternative method that could be used to discern segmental changes in the corticospinal tract when targeting lower limb muscles.

Chronic ethanol forced administration from adolescence to adulthood reduces cell density in the rat spinal cord.

Ethanol (EtOH) consumption is a risk factor for central nervous system damage, especially during adolescence. This study aimed to investigate the possible effects of chronic EtOH forced administration on gray and white matter of the spinal cord, from adolescence to adulthood. For this, male Wistar rats were administered EtOH by gavage (6.5 g/kg/day; 22.5% w/v) from the 35th to the 90th day of life, while control animals received only distilled water. After exposure, animals were euthanized and their spinal cords processed to obtain cervical and thoracic segments for histological analyses. Quantitative analyses of total cell density and motor neurons of white and gray matter from the ventral horns were evaluated. Forced EtOH administration model showed a decrease in the motoneuron density in the spinal cord in both segments evaluated. Analyses of total cell density showed that the cervical segment was more susceptible to damages promoted by EtOH, with a significant decrease in cell density. Our results showed that chronic EtOH exposure during adolescence could promote injuries to the spinal cord, with neurodegeneration of motoneurons and other cell types present in neural parenchyma.

The differences in the structure of the motor nucleus of the medial gastrocnemius muscle in male and female rats

There are many reports describing sexual dissimilarities in the CNS, particularly in the brain and cortical regions. However, knowledge regarding sexual dissimilarities in the spinal cord and in particular in the hindlimb muscle-motoneuron connectivity controlling locomotion is limited. In order to recognise sex differences in the architecture of the medial gastrocnemius (MG) motor nucleus in rats of the same age, retrograde-labelled motoneurons were identified following a bath of the proximal stump of the transected MG nerve in horseradish peroxidase. The rostrocaudal distribution of motoneurons along the spinal cord and on transverse sections as well as the size and density of motoneurons in the motor nucleus were determined from serial microscopic images. It was shown that the MG pool extended throughout the L4–L6 segments, with a length that was 32% greater in males. The position of the moto pool within the ventral horn of the spinal cord transversal sections was also different in both sexes: the pool was located more dorsally and laterally in males. Three size categories of motoneurons with different soma diameters were distinguished: α1 (27.5–40.0μm), α2 (>40.0μm) and γ (<27.5μm). The density of α (α1 and α2) motoneurons as well as γ motoneurons was higher in females, by about 13% and 23%, respectively. The number of α1 motoneurons was 8% higher in females, whereas there were 46% more α2 motoneurons in males. The most significant differences in the distribution concerned α2 motoneurons, which revealed a lower density in the rostral parts of the MG motor nucleus in females. It was therefore concluded that the length of the MG motor nucleus was shorter, whereas the mean density of α and γ motoneurons was higher in females.

MR Imaging-based Estimation of Upper Motor Neuron Density in Patients with Amyotrophic Lateral Sclerosis: A Feasibility Study.

Purpose To determine if magnetic resonance (MR) imaging metrics can estimate primary motor cortex (PMC) motor neuron (MN) density in patients with amyotrophic lateral sclerosis (ALS). Materials and Methods Between 2012 and 2014, in situ brain MR imaging was performed in 11 patients with ALS (age range, 35-81 years; seven women and four men) soon after death (mean, 5.5 hours after death; range, 3.2-9.6 hours). The brain was removed, right PMC (RPMC) was excised, and MN density was quantified. RPMC metrics (thickness, volume, and magnetization transfer ratio) were calculated from MR images. Regression modeling was used to estimate MN density by using RPMC and global MR imaging metrics (brain and tissue volumes); clinical variables were subsequently evaluated as additional estimators. Models were tested at in vivo MR imaging by using the same imaging protocol (six patients with ALS; age range, 54-66 years; three women and three men). Results RPMC mean MN density varied over a greater than threefold range across patients and was estimated by a linear function of normalized gray matter volume (adjusted R2 = 0.51; P = .008; <10% error in most patients). When considering only sporadic ALS, a linear function of normalized RPMC and white matter volumes estimated MN density (adjusted R2 = 0.98; P = .01; <10% error in all patients). In vivo data analyses detected decreases in MN density over time. Conclusion PMC mean MN density varies widely in end-stage ALS possibly because of disease heterogeneity. MN density can potentially be estimated by MR imaging metrics. © RSNA, 2018 Online supplemental material is available for this article.

The Neuroprotective Effects of Alcoholic Extract of Levisticum Officinale on Alpha Motoneurons’ Degeneration After Sciatic Nerve Compression in Male Rats

As peripheral neurons are injured, several inflammatory factors will be released from the fiber of the injured nervous leading to morphological and biochemical changes in the injured point. This changes ultimately results in the Wallerian degeneration of the cell bodies of these neurons in the spinal cord which are involved in retrograde reaction. Levisticum officinale is one of the herbs that has been traditionally used for treatment of many diseases. Many studies have confirmed the anti-inflammatory and anti-oxidant effects of this plant. It is expected that the extract of Levisticum officinale would have a significant role in the restoration of nervous system injuries. The purpose of this study was to investigate the effects of the alcoholic extract of Levisticum officinale on the neuron protection of the sciatic nerves of injured rats. In this study, the ethanolic extract was obtained from Levisticum officinale. Then the rats were randomly divided into 5 groups, each containing 6 rats, including two control and compression groups and three treatment groups treated with 50, 75 and 100 mg/kg doses of the extract. Sciatic nerve in compression and treatment groups was exposed to compression for 60 seconds, and the ethanolic extract was injected intraperitoneally in the first, second and third weeks. The spinal cord of the rats were sampled and the neurons of the samples were counted using a dissector method after tissue passaging and staining. Meanwhile, the density of motoneurons was measured. T-test analysis was performed by using SPSS software. The density of α motoneurons in compression group decreased significantly as compared to the control group (p<0.001). The density of α motoneurons in all treatment groups with 50,75 and 100 extract doses was significantly increased as compared to the compression group (p<0.01). The highest neuron protection was observed in the treatment group with 100 mg/kg dose as compared to the compression group. Conclusion: Ethanolic extract of Levisticum officinale has neuron protective and restorative effects on the injuries of the peripheral nerves such as compression.

Polysialic Acid Glycomimetic Promotes Functional Recovery and Plasticity After Spinal Cord Injury in Mice

Regeneration after injury of the central nervous system is poor due to the abundance of molecules inhibiting axonal growth. Here we pursued to promote regeneration after thoracic spinal cord injury in young adult C57BL/6J mice using peptides which functionally mimic polysialic acid (PSA) and human natural killer cell-1 (HNK-1) glycan, carbohydrate epitopes known to promote neurite outgrowth in vitro. Subdural infusions were performed with an osmotic pump, over 2 weeks. When applied immediately after injury, the PSA mimetic and the combination of PSA and HNK-1 mimetics, but not the HNK-1 mimetic alone, improved functional recovery as assessed by locomotor rating and video-based motion analysis over a 6-week observation period. Better outcome in PSA mimetic-treated mice was associated with higher, as compared with control mice, numbers of cholinergic and glutamatergic terminals and monaminergic axons in the lumbar spinal cord, and better axonal myelination proximal to the injury site. In contrast to immediate post-traumatic application, the PSA mimetic treatment was ineffective when initiated 3 weeks after spinal cord injury. Our data suggest that PSA mimetic peptides can be efficient therapeutic tools improving, by augmenting plasticity, functional recovery when applied during the acute phase of spinal cord injury.

Moderate exercise delays the motor performance decline in a transgenic model of ALS

The relationship between exercise and amyotrophic lateral sclerosis (ALS), a neurodegenerative disorder characterized by motor neuron loss, rapidly progressive weakness and early death has been controversial. We studied the effect of a high (HEX) and moderate-level exercise (MEX) on body weight, motor performance and motor neuron counts in the ventral horn of spinal cords in a transgenic mouse model of ALS (G93A-SOD1) that overexpresses a mutated form of the human SOD1 gene that is a cause of familial ALS. These transgenic mice show several similarities to the human disease, including rapid progressive motor weakness from 100 days of age and premature death at around 135 days of age. Mice were exposed to high or mid-level exercise of left sedentary (SED). At 70, 95 and 120 days of age, spinal cords were processed following euthanasia. Motor neurons larger than 15 μm in diameter were counted with a design-based stereological protocol using an optical fractionator probe in the ventral horn of different regions of the cord and compared to wild-type littermates. Moderate exercise delayed the onset of motor deficit by over a week. High exercise slightly but significantly hastened the onset of motor performance deficits. Motor neuron density in the lumbar cord was significantly higher in MEX group compared to SED at 95 days of age. These results show the beneficial effects of moderate exercise on the preservation of motor performance that correlates with higher motor neuron density in the ventral horn of the lumbar spinal cord in G93A mice.

Neuroprotetive Effect of Curcuma longa Alcoholic Extract on Peripheral Nerves Degeneration after Sciatic Nerve Compression in Rats

Summary There are extensive evidences those axonal processes of the nervous system (peripheral and/or central) may degenerate after nerve injuries. Wallerian degeneration and choromatolysis are the most conspicuous phenomena that occur in response to injuries. In this research the effect of root alcoholic extract of Salvia staminea on neuronal density of motoneurons in spinal cord anterior horn in rats was studied. Forty adult male wistar rats were used and divided to five groups (control, compression, three experimental groups). In compression and experimental groups right sciatic nerve were highly compressed for 60 s, assigned to experimental groups (Compression + alcoholic extract of salvia root injections (25 ,50,75 mg/kg -1 ,

O.619 How does nicotine affect bone regeneration?

Maternal cigarette use during pregnancy is associated with increased incidence of neural impairments in offspring, but nicotine's unique contribution to any neuropathology remains unclear, and nicotine's neurodevelopmental effects assessed in animal models vary with concentration. During ontogenesis, the chick oculomotor complex (OMN) is regulated by central nervous system (CNS) afferent-derived and target-derived trophic factors, allowing assessment of nicotine's potential interference in receptor-mediated CNS trophic phenomena, unconfounded by myriad other compounds in cigarette smoke. In the current study, 100 ng nicotine applied daily in ovo to yolk during embryonic days (E) 1–7 mimicked maternal plasma nicotine concentrations during fetal cranial nerve development. Nicotine-treated embryos exhibited a 15% decrease in whole body weight and 7% decrease in brain weight at E16. However, at E16, nicotine-treated embryos had 37% and 15% increases in the combined ventromedial+lateral (v) OMN motoneuron density and soma area, respectively, effects not observed in the optic tectum, in which nicotine cholinergic receptor expression is delayed until E8–12. Incorporation of tritiated thymidine into whole brain DNA demonstrated that the nicotine treatment did not cause increased rates of whole brain mitosis, suggesting that the dosage regimen did not elicit a cytotoxic, wound-healing, response of differentiating cells. As determined by DNA fragment-labeling assay during the normal period of cell death, vOMN apoptosis occurs maximally on E11 during a normal period of declining cell density, and a dose–response study demonstrated 78% E11 vOMN apoptotic suppression at approximately 0.30 μM cumulative yolk nicotine with an inhibition threshold between 0.10 and 0.20 μM. These results suggest that plasma nicotine concentrations resulting from tobacco use or nicotine replacement therapy (NRT) are sufficient to inhibit motoneuron apoptosis and enhance neuronal growth.

Establishing a standardized therapeutic testing protocol for spinal muscular atrophy

Several mice models have been created for spinal muscular atrophy (SMA); however, there is still no standard preclinical testing system for the disease. We previously generated type III-specific SMA model mice, which might be suitable for use as a preclinical therapeutic testing system for SMA. To establish such a system and test its applicability, we first created a testing protocol and then applied it as a means to investigate the use of valproic acid (VPA) as a possible treatment for SMA. These SMA mice revealed tail/ear/foot deformity, muscle atrophy, poorer motor performances, smaller compound muscle action potential and lower spinal motoneuron density at the age of 9 to 12 months in comparison with age-matched wild-type littermate mice. In addition, VPA attenuates motoneuron death, increases spinal SMN protein level and partially normalizes motor function in SMA mice. These results suggest that the testing protocol developed here is well suited for use as a standardized preclinical therapeutic testing system for SMA.

Sertoli cells improve survival of motor neurons in SOD1 transgenic mice, a model of amyotrophic lateral sclerosis

Cell replacement therapy has been widely suggested as a treatment for multiple diseases including motor neuron disease. A variety of donor cells have been tested for treatment including isolated preparations from bone marrow and embryonic spinal cord. Another cell source, Sertoli cells, have been successfully used in models of diabetes, Parkinson's disease and Huntington's disease. The ability of these cells to secrete cytoprotective proteins and their role as ‘nurse cells’ supporting the function of other cell types in the testes suggest their potential use as neuroprotective cells. The current study examines the ability of Sertoli cells injected into the parenchyma of the spinal cord to protect motor neurons in a mouse model for amyotrophic lateral sclerosis. Seventy transgenic mice expressing the mutant (G93A) human Cu–Zn superoxide dismutase (SOD1) received a unilateral spinal injection of Sertoli-enriched testicular cells into the L4–L5 ventral horn (1 × 10 5 cells total) prior to the onset of clinical symptoms. The animals were euthanized at the end stage of the disease. Histological and morphometric analyses of the transplant site were performed. A significant increase in the number of surviving ChAT positive motor neurons was found ipsilateral to the injection compared with contralateral and uninjected spinal cord. The ipsilateral increase in motor neuron density was dependent upon proximity to the injection site. Sections rostral or caudal to the injection site did not display a similar difference in motor neuron density. Implantation of a Sertoli-cell-enriched preparation has a significant neuroprotective benefit to vulnerable motor neurons in the SOD1 transgenic model. The therapeutic benefit may be the result of secreted neurotrophic factors present at a critical stage of motor neuron degeneration in this model.

Adenoviral clostridial light chain gene-based synaptic inhibition through neuronal synaptobrevin elimination

Clostridial neurotoxins have assumed increasing importance in clinical application. The toxin's light chain component (LC) inhibits synaptic transmission by digesting vesicle-docking proteins without directly altering neuronal health. To study the properties of LC gene expression in the nervous system, an adenoviral vector containing the LC of tetanus toxin (AdLC) was constructed. LC expressed in differentiated neuronal PC12 cells was shown to induce time- and concentration-dependent digestion of mouse brain synaptobrevin in vitro as compared to control transgene products. LC gene expression in the rat lumbar spinal cord disrupted hindlimb sensorimotor function in comparison to control vectors as measured by the Basso-Beattie-Bresnahan (BBB) scale (P<0.001) and rotarod assay (P<0.003). Evoked electromyography (EMG) showed increased stimulus threshold and decreased response current amplitude in LC gene-transferred rats. At the peak of functional impairment, neither neuronal TUNEL staining nor reduced motor neuron density could be detected. Spontaneous functional recovery was observed to parallel the cessation of LC gene expression. These results suggest that light chain gene delivery within the nervous system may provide a nondestructive means for focused neural inhibition to treat a variety of disorders related to excessive synaptic activity, and prove useful for the study of neural circuitry.

6-WEEK AD LIBITUM ADMINISTRATION OF NICOTINE SIGNIFICANTLY ALTERS THE GENE EXPRESSION PROFILE IN THE BONE MARROW AND SPLEEN OF C57BL6 MICE

Maternal cigarette use during pregnancy is associated with increased incidence of neural impairments in offspring, but nicotine's unique contribution to any neuropathology remains unclear, and nicotine's neurodevelopmental effects assessed in animal models vary with concentration. During ontogenesis, the chick oculomotor complex (OMN) is regulated by central nervous system (CNS) afferent-derived and target-derived trophic factors, allowing assessment of nicotine's potential interference in receptor-mediated CNS trophic phenomena, unconfounded by myriad other compounds in cigarette smoke. In the current study, 100 ng nicotine applied daily in ovo to yolk during embryonic days (E) 1–7 mimicked maternal plasma nicotine concentrations during fetal cranial nerve development. Nicotine-treated embryos exhibited a 15% decrease in whole body weight and 7% decrease in brain weight at E16. However, at E16, nicotine-treated embryos had 37% and 15% increases in the combined ventromedial+lateral (v) OMN motoneuron density and soma area, respectively, effects not observed in the optic tectum, in which nicotine cholinergic receptor expression is delayed until E8–12. Incorporation of tritiated thymidine into whole brain DNA demonstrated that the nicotine treatment did not cause increased rates of whole brain mitosis, suggesting that the dosage regimen did not elicit a cytotoxic, wound-healing, response of differentiating cells. As determined by DNA fragment-labeling assay during the normal period of cell death, vOMN apoptosis occurs maximally on E11 during a normal period of declining cell density, and a dose–response study demonstrated 78% E11 vOMN apoptotic suppression at approximately 0.30 μM cumulative yolk nicotine with an inhibition threshold between 0.10 and 0.20 μM. These results suggest that plasma nicotine concentrations resulting from tobacco use or nicotine replacement therapy (NRT) are sufficient to inhibit motoneuron apoptosis and enhance neuronal growth.

Exposure to low concentrations of nicotine during cranial nerve development inhibits apoptosis and causes cellular hypertrophy in the ventral oculomotor nuclei of the chick embryo

Maternal cigarette use during pregnancy is associated with increased incidence of neural impairments in offspring, but nicotine's unique contribution to any neuropathology remains unclear, and nicotine's neurodevelopmental effects assessed in animal models vary with concentration. During ontogenesis, the chick oculomotor complex (OMN) is regulated by central nervous system (CNS) afferent-derived and target-derived trophic factors, allowing assessment of nicotine's potential interference in receptor-mediated CNS trophic phenomena, unconfounded by myriad other compounds in cigarette smoke. In the current study, 100 ng nicotine applied daily in ovo to yolk during embryonic days (E) 1–7 mimicked maternal plasma nicotine concentrations during fetal cranial nerve development. Nicotine-treated embryos exhibited a 15% decrease in whole body weight and 7% decrease in brain weight at E16. However, at E16, nicotine-treated embryos had 37% and 15% increases in the combined ventromedial+lateral (v) OMN motoneuron density and soma area, respectively, effects not observed in the optic tectum, in which nicotine cholinergic receptor expression is delayed until E8–12. Incorporation of tritiated thymidine into whole brain DNA demonstrated that the nicotine treatment did not cause increased rates of whole brain mitosis, suggesting that the dosage regimen did not elicit a cytotoxic, wound-healing, response of differentiating cells. As determined by DNA fragment-labeling assay during the normal period of cell death, vOMN apoptosis occurs maximally on E11 during a normal period of declining cell density, and a dose–response study demonstrated 78% E11 vOMN apoptotic suppression at approximately 0.30 μM cumulative yolk nicotine with an inhibition threshold between 0.10 and 0.20 μM. These results suggest that plasma nicotine concentrations resulting from tobacco use or nicotine replacement therapy (NRT) are sufficient to inhibit motoneuron apoptosis and enhance neuronal growth.

Neuronal survival following remote adenovirus gene delivery.

Virus-mediated central nervous system gene delivery is a promising means of treating traumatized tissue or degenerative diseases. In the present study, the authors examined gene expression and neuronal survival in the spinal cord after sciatic nerve administration of an adenovirus vector expressing a LacZ reporter gene. The time course of adenovirus gene expression, DNA fragmentation, and neuronal density were quantified in rat lumbar spinal cord by staining for beta-galactosidase (beta-Gal), terminal deoxynucleotidyl transferase, and cresyl violet after microinjection of either saline or the reporter virus into rat sciatic nerve. The expression of beta-Gal following remote vector delivery peaked at 7 days and declined thereafter but was not accompanied by neuronal cell death, as measured by DNA fragmentation. No significant difference in spinal motor neuron density was detected between virus-treated and control rats at any time point examined. Although the spinal cords removed from rats treated with cyclosporine prior to adenovirus injection contained substantially more neurons staining for beta-Gal at 7 days (67% of total neurons), the decay in the number of stained neurons was not paralleled by a decline in motor neuron density. The authors conclude that remote gene expression is suppressed by a noncytolytic process.