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OBJECTIVE: To investigate trends in the characteristics of postoperative C5 palsy following anterior decompression and fusion associated with advancements in this surgical procedure to treat cervical degenerative disorders. METHODS: We included 801 consecutive patients who underwent anterior decompression and fusion for cervical degenerative disorders from 2006 to 2019 and investigated the incidence, onset, and prognosis of C5 palsy. In addition, we compared the incidence of C5 palsy with that found in our previous investigation. RESULTS: The cases of 42 (5.2%) patients were complicated by C5 palsy. For patients with ossification of the longitudinal ligament (OPLL), 22 (12.4%) of 177 were complicated with C5 palsy, and the incidence was significantly higher than that in patients without OPLL (20 [3.2%] of 624, P < 0.01). The incidence of C5 palsy in patients without OPLL was significantly lower than that found in our previous investigation (P < 0.01). The incidence of C5 palsy in patients that required contiguous multilevel corpectomy was significantly higher in patients that required within a single corpectomy (P < 0.01). At 1-year follow-up, muscle strength in 3 (6.1%) of 49 limbs had not improved sufficiently. CONCLUSIONS: With advancements in surgical techniques which allowed necessary and sufficient spinal cord decompression and avoided unnecessary corpectomy, the incidence of C5 palsy in patients without OPLL was decreased significantly. By contrast, for patients with OPLL, the incidence of C5 palsy was similar to the incidence found previously, perhaps because a broad and contiguous multilevel corpectomy was usually needed to decompress the spinal cord sufficiently.
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Ossificação do Ligamento Longitudinal Posterior , Fusão Vertebral , Humanos , Vértebras Cervicais/cirurgia , Descompressão Cirúrgica/efeitos adversos , Descompressão Cirúrgica/métodos , Pescoço/cirurgia , Paralisia/epidemiologia , Paralisia/etiologia , Paralisia/cirurgia , Fusão Vertebral/efeitos adversos , Fusão Vertebral/métodos , Ossificação do Ligamento Longitudinal Posterior/cirurgia , Ossificação do Ligamento Longitudinal Posterior/complicações , Resultado do TratamentoRESUMO
Attenuation of the secondary injury of spinal cord injury (SCI) can suppress the spread of spinal cord tissue damage, possibly resulting in spinal cord sparing that can improve functional prognoses. Granulocyte colony-stimulating factor (G-CSF) is a haematological cytokine commonly used to treat neutropenia. Previous reports have shown that G-CSF promotes functional recovery in rodent models of SCI. Based on preclinical results, we conducted early phase clinical trials, showing safety/feasibility and suggestive efficacy. These lines of evidence demonstrate that G-CSF might have therapeutic benefits for acute SCI in humans. To confirm this efficacy and to obtain strong evidence for pharmaceutical approval of G-CSF therapy for SCI, we conducted a phase 3 clinical trial designed as a prospective, randomized, double-blinded and placebo-controlled comparative trial. The current trial included cervical SCI [severity of American Spinal Injury Association (ASIA) Impairment Scale (AIS) B or C] within 48 h after injury. Patients are randomly assigned to G-CSF and placebo groups. The G-CSF group was administered 400 µg/m2/day × 5 days of G-CSF in normal saline via intravenous infusion for five consecutive days. The placebo group was similarly administered a placebo. Allocation was concealed between blinded evaluators of efficacy/safety and those for laboratory data, as G-CSF markedly increases white blood cell counts that can reveal patient treatment. Efficacy and safety were evaluated by blinded observer. Our primary end point was changes in ASIA motor scores from baseline to 3 months after drug administration. Each group includes 44 patients (88 total patients). Our protocol was approved by the Pharmaceuticals and Medical Device Agency in Japan and this trial is funded by the Center for Clinical Trials, Japan Medical Association. There was no significant difference in the primary end point between the G-CSF and the placebo control groups. In contrast, one of the secondary end points showed that the ASIA motor score 6 months (P = 0.062) and 1 year (P = 0.073) after drug administration tend to be higher in the G-CSF group compared with the placebo control group. Moreover, in patients aged over 65 years old, motor recovery 6 months after drug administration showed a strong trend towards a better recovery in the G-CSF treated group (P = 0.056) compared with the control group. The present trial failed to show a significant effect of G-CSF in primary end point although the subanalyses of the present trial suggested potential G-CSF benefits for specific population.
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Fator Estimulador de Colônias de Granulócitos/uso terapêutico , Recuperação de Função Fisiológica/efeitos dos fármacos , Traumatismos da Medula Espinal/tratamento farmacológico , Adolescente , Adulto , Idoso , Método Duplo-Cego , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Adulto JovemRESUMO
PURPOSE: To elucidate the independent preoperative factors that have a significant impact on poor surgical outcome after laminoplasty for K-line (+) ossification of the posterior longitudinal ligament (OPLL). Analyses in K-line (+) patient population can exclude the influence by mal-alignment and thick OPLL, both of which are well known two major factors that have significant impact on clinical outcome. METHODS: The present study included 72 patients (50 male and 22 female) who underwent laminoplasty for K-line (+) cervical OPLL and were followed-up for at least 1 year. Recovery of Japanese Orthopedic Association score (JOA score) for cervical myelopathy was used as the measure of clinical outcome. For radiographic assessment, the type of OPLL, the maximum OPLL occupation ratio, the C2-C7 angle, and the segmental range of motion at the peak of OPLL (segmental ROM) were assessed. To elucidate the factors that are significantly associated with a poor clinical outcome after laminoplasty for K-line (+) OPLL, statistical analyses were conducted. RESULTS: The mean preoperative JOA score was 8.9 points and improved to 12.8 points after surgery. The recovery of JOA score was 47 ± 35%. Stepwise logistic regression following univariate analyses revealed that preoperative segmental ROM at the peak of OPLL is an independent factor associated with a poor outcome (p = 0.04, odds ratio = 1.15). CONCLUSIONS: Large preoperative segmental ROM at the peak of the OPLL is an independent factor that has significant impact on poor surgical outcome after laminoplasty for K-line (+) OPLL.
Assuntos
Vértebras Cervicais/cirurgia , Laminoplastia/métodos , Ossificação do Ligamento Longitudinal Posterior/patologia , Ossificação do Ligamento Longitudinal Posterior/cirurgia , Ossificação Heterotópica , Amplitude de Movimento Articular , Adulto , Idoso , Idoso de 80 Anos ou mais , Mau Alinhamento Ósseo , Vértebras Cervicais/diagnóstico por imagem , Vértebras Cervicais/patologia , Vértebras Cervicais/fisiopatologia , Feminino , Seguimentos , Humanos , Laminoplastia/efeitos adversos , Masculino , Pessoa de Meia-Idade , Ossificação do Ligamento Longitudinal Posterior/diagnóstico por imagem , Ossificação do Ligamento Longitudinal Posterior/fisiopatologia , Período Pré-Operatório , Fatores de Risco , Resultado do TratamentoRESUMO
Despite extensive research on primate cognitive function, understanding how anatomical connectivity at a neural circuit level relates to information transformation across different cortical areas remains primitive. New technology is needed to visualize inter-areal anatomical connectivity in living monkeys and to tie this directly to neurophysiological function. Here, we developed a novel method to investigate this structure-function relationship, by combining optical intrinsic signal imaging (OISI) with optogenetic stimulation in living monkeys (opto-OISI). The method involves expressing channelrhodophsin-2 in one area (source) followed by optical imaging of optogenetic activations in the other area (target). We successfully demonstrated the potential of the method with interhemispheric columnar projection patterns between V1/V2 border regions. Unlike the combination of optogenetics and functional magnetic resonance imaging (opto-fMRI), opto-OISI has the advantage of enabling us to detect responses of small clusters of neurons, even if the clusters are sparsely distributed. We suggest that opto-OISI can be a powerful approach to understanding cognitive function at the neural circuit level, directly linking inter-areal circuitry to fine-scale structure and function.
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Córtex Cerebral , Conectoma , Imageamento por Ressonância Magnética , Neurônios , Imagem Óptica , Optogenética , Animais , Córtex Cerebral/diagnóstico por imagem , Córtex Cerebral/metabolismo , Channelrhodopsins/biossíntese , Macaca mulatta , Masculino , Neurônios/citologia , Neurônios/metabolismoRESUMO
INTRODUCTION: Granulocyte colony-stimulating factor (G-CSF) is generally used for neutropaenia. Previous experimental studies revealed that G-CSF promoted neurological recovery after spinal cord injury (SCI). Next, we moved to early phase of clinical trials. In a phase I/IIa trial, no adverse events were observed. Next, we conducted a non-randomised, non-blinded, comparative trial, which suggested the efficacy of G-CSF for promoting neurological recovery. Based on those results, we are now performing a phase III trial. METHODS AND ANALYSIS: The objective of this study is to evaluate the efficacy of G-CSF for acute SCI. The study design is a prospective, multicentre, randomised, double-blinded, placebo-controlled comparative study. The current trial includes cervical SCI (severity of American Spinal Injury Association (ASIA) Impairment Scale B/C) within 48 hours after injury. Patients are randomly assigned to G-CSF and placebo groups. The G-CSF group is administered 400 µg/m2/day×5 days of G-CSF in normal saline via intravenous infusion for 5 consecutive days. The placebo group is similarly administered a placebo. Our primary endpoint is changes in ASIA motor scores from baseline to 3 months. Each group includes 44 patients (88 total patients). ETHICS AND DISSEMINATION: The study will be conducted according to the principles of the World Medical Association Declaration of Helsinki and in accordance with the Japanese Medical Research Involving Human Subjects Act and other guidelines, regulations and Acts. Results of the clinical study will be submitted to the head of the respective clinical study site as a report after conclusion of the clinical study by the sponsor-investigator. Even if the results are not favourable despite conducting the clinical study properly, the data will be published as a paper. TRIAL REGISTRATION NUMBER: UMIN000018752.
Assuntos
Fator Estimulador de Colônias de Granulócitos/administração & dosagem , Neuroproteção/efeitos dos fármacos , Traumatismos da Medula Espinal/tratamento farmacológico , Doença Aguda , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Ensaios Clínicos Fase III como Assunto , Método Duplo-Cego , Feminino , Humanos , Japão , Masculino , Pessoa de Meia-Idade , Estudos Multicêntricos como Assunto , Estudos Prospectivos , Ensaios Clínicos Controlados Aleatórios como Assunto , Recuperação de Função Fisiológica , Índice de Gravidade de Doença , Fatores de Tempo , Resultado do Tratamento , Adulto JovemRESUMO
Cerebellar malformations cause changes to the sleep-wake cycle, resulting in sleep disturbance. However, it is unclear how the cerebellum contributes to the sleep-wake cycle. To examine the neural connections between the cerebellum and the nuclei involved in the sleep-wake cycle, we investigated the axonal projections of Purkinje cells in the mouse posterior vermis by using an adeno-associated virus (AAV) vector (serotype rh10) as an anterograde tracer. When an AAV vector expressing humanized renilla green fluorescent protein was injected into the cerebellar lobule IX, hrGFP and synaptophysin double-positive axonal terminals were observed in the region of medial parabrachial nucleus (MPB). The MPB is involved in the phase transition from rapid eye movement (REM) sleep to Non-REM sleep and vice versa, and the cardiovascular and respiratory responses. The hrGFP-positive axons from lobule IX went through the ventral spinocerebellar tract and finally reached the MPB. By contrast, when the AAV vector was injected into cerebellar lobule VI, no hrGFP-positive axons were observed in the MPB. To examine neurons projecting to the MPB, we unilaterally injected Fast Blue and AAV vector (retrograde serotype, rAAV2-retro) as retrograde tracers into the MPB. The cerebellar Purkinje cells in lobules VIII-X on the ipsilateral side of the Fast Blue-injected MPB were retrogradely labeled by Fast Blue and AAV vector (retrograde serotype), but no retrograde-labeled Purkinje cells were observed in lobules VI-VII and the cerebellar hemispheres. These results indicated that Purkinje cells in lobules VIII-X directly project their axons to the ipsilateral MPB but not lobules VI-VII. The direct connection between lobules VIII-X and the MPB suggests that the cerebellum participates in the neural network controlling the sleep-wake cycle, and cardiovascular and respiratory responses, by modulating the physiological function of the MPB.
Assuntos
Vermis Cerebelar/citologia , Núcleos Parabraquiais/citologia , Células de Purkinje/citologia , Amidinas , Animais , Dependovirus/genética , Proteínas de Fluorescência Verde/genética , Proteínas de Fluorescência Verde/metabolismo , Masculino , Camundongos Endogâmicos ICR , Vias Neurais/citologia , Técnicas de Rastreamento Neuroanatômico , Marcadores do Trato NervosoRESUMO
STUDY DESIGN: A retrospective multicenter survey. OBJECTIVE: To investigate the perioperative complications of oblique lateral interbody fusion (OLIF) surgery. SUMMARY OF BACKGROUND DATA: OLIF has been widely performed to achieve minimally invasive, rigid lumbar lateral interbody fusion. The associated perioperative complications are not yet well described. METHODS: The participants were patients who underwent OLIF surgery under the diagnosis of degenerative lumbar diseases between April 2013 and May 2015 at 11 affiliated medical institutions. The collected data were classified into intraoperative and early-stage postoperative (≤1 mo) complications. The intraoperative complications were then subcategorized into organ damage (neural, vertebral, vascular, and others) and other complications, mainly related to instrumental failure. The collected data were also divided and analyzed based on whether the surgeon was certified to perform the surgery and the incidence of complications in the early (April 2013-March 2014) and late stages (April 2014-May 2015) of OLIF introduction. RESULTS: In the 155 included patients, 75 complications were reported (incidence rate, 48.3%). The most common complication was endplate fracture/subsidence (18.7%), followed by transient psoas weakness and thigh numbness (13.5%) and segmental artery injury (2.6%). Almost all these complications were transient, except for three patients who had permanent damage: one had ureteral injury and two had neurological injury. Postoperative complications included surgical site infection (1.9%) and reoperation (1.9%). Whether the primary operator was experienced did not affect the incidence of complications. Regarding the introductory stage, the incidence of complications was 50% in the early stage and 38% in the late stage. CONCLUSION: The overall incidence of perioperative complications of OLIF surgery reached 48.3%, of which only 1.9% resulted in permanent damage. Our analysis based on surgeon experience indicated that the OLIF procedure could be performed without increasing incidence of complications, under the guidance of experienced supervisors. LEVEL OF EVIDENCE: 3.
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Degeneração do Disco Intervertebral/cirurgia , Complicações Intraoperatórias/epidemiologia , Dor Lombar/cirurgia , Vértebras Lombares/cirurgia , Complicações Pós-Operatórias/epidemiologia , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Feminino , Humanos , Incidência , Masculino , Pessoa de Meia-Idade , Período Pós-Operatório , Reoperação , Estudos Retrospectivos , Fusão Vertebral/métodos , Resultado do Tratamento , Adulto JovemRESUMO
During cerebral development, many types of neurons are sequentially generated by self-renewing progenitor cells called apical progenitors (APs). Temporal changes in AP identity are thought to be responsible for neuronal diversity; however, the mechanisms underlying such changes remain largely unknown. Here we perform single-cell transcriptome analysis of individual progenitors at different developmental stages, and identify a subset of genes whose expression changes over time but is independent of differentiation status. Surprisingly, the pattern of changes in the expression of such temporal-axis genes in APs is unaffected by cell-cycle arrest. Consistent with this, transient cell-cycle arrest of APs in vivo does not prevent descendant neurons from acquiring their correct laminar fates. Analysis of cultured APs reveals that transitions in AP gene expression are driven by both cell-intrinsic and -extrinsic mechanisms. These results suggest that the timing mechanisms controlling AP temporal identity function independently of cell-cycle progression and Notch activation mode.
Assuntos
Linhagem da Célula/genética , Córtex Cerebral/metabolismo , Células-Tronco Neurais/metabolismo , Neurogênese/genética , Neurônios/metabolismo , Animais , Pontos de Checagem do Ciclo Celular/genética , Diferenciação Celular , Córtex Cerebral/citologia , Córtex Cerebral/crescimento & desenvolvimento , Embrião de Mamíferos , Perfilação da Expressão Gênica , Regulação da Expressão Gênica no Desenvolvimento , Proteínas de Membrana/genética , Proteínas de Membrana/metabolismo , Camundongos , Camundongos Endogâmicos ICR , Camundongos Transgênicos , Nucleosídeo NM23 Difosfato Quinases/genética , Nucleosídeo NM23 Difosfato Quinases/metabolismo , Células-Tronco Neurais/citologia , Neurônios/citologia , Receptor Notch1/genética , Receptor Notch1/metabolismo , Transdução de Sinais , Análise de Célula Única , Proteínas com Domínio T/genética , Proteínas com Domínio T/metabolismo , Fatores de TempoRESUMO
Solute carrier (SLC) transporters play important roles in absorption and disposition of drugs in cells; however, the expression pattern of human SLC transporters in the skin has not been determined. In the present study, the expression patterns of 28 human SLC transporters were determined in the human skin. Most of the SLC transporter family members were either highly or moderately expressed in the liver, while their expression was limited in the skin and small intestine. Treatment of human keratinocytes with a reactive metabolite of ibuprofen significantly reduced cell viability. Expression array analysis revealed that S100 calcium binding protein A7A (S100A7A) was induced nearly 50-fold in dermal cells treated with ibuprofen acyl-glucuronide. Determination of the expression of drug-metabolizing enzymes as well as drug transporters prior to the administration of drugs would make it possible to avoid the development of idiosyncratic skin diseases in individuals.
Assuntos
Proteínas de Transporte/metabolismo , Proteínas de Membrana Transportadoras/metabolismo , Dermatopatias/metabolismo , Pele/metabolismo , Transporte Biológico/fisiologia , Linhagem Celular , Sobrevivência Celular/fisiologia , Glucuronatos/metabolismo , Humanos , Ibuprofeno/análogos & derivados , Ibuprofeno/metabolismo , Intestino Delgado/metabolismo , Queratinócitos/metabolismo , Fígado/metabolismo , Proteína A7 Ligante de Cálcio S100 , Proteínas S100/metabolismoRESUMO
STUDY DESIGN: An open-labeled multicenter prospective nonrandomized controlled clinical trial. OBJECTIVE: To confirm the feasibility of using granulocyte colony-stimulating factor (G-CSF) for treatment of acute spinal cord injury (SCI). SUMMARY OF BACKGROUND DATA: We previously reported that G-CSF promotes functional recovery after compression-induced SCI in mice. On the basis of these findings, we conducted a multicenter prospective controlled clinical trial to assess the feasibility of G-CSF therapy for patients with acute SCI. METHODS: The trial ran from August 2009 to March 2011, and included 41 patients with SCI treated within 48 hours of onset. Informed consent was obtained from all patients. After providing consent, patients were divided into 2 groups. In the G-CSF group (17 patients), G-CSF (10 µg/kg/d) was intravenously administered for 5 consecutive days, and in the control group (24 patients), patients were similarly treated except for the G-CSF administration. We evaluated motor and sensory functions using the American Spinal Cord Injury Association score and American Spinal Cord Injury Association impairment scale at 1 week, 3 months, 6 months, and 1 year after onset. RESULTS: Only 2 patients did not experience American Spinal Cord Injury Association impairment scale improvement in the G-CSF group. In contrast, 15 patients in the control group did not experience American Spinal Cord Injury Association impairment scale improvement. In the analysis of increased American Spinal Cord Injury Association motor score, a significant increase in G-CSF group was detected from 1 week after the administration compared with the control group. After that, some spontaneous increase of motor score was detected in control group, but the significant increase in G-CSF group was maintained until 1 year of follow-up. CONCLUSION: Despite the limitation that patient selection was not randomized, the present results suggest the possibility that G-CSF administration has beneficial effects on neurological recovery in patients with acute SCI. LEVEL OF EVIDENCE: 3.
Assuntos
Fator Estimulador de Colônias de Granulócitos/administração & dosagem , Recuperação de Função Fisiológica/efeitos dos fármacos , Recuperação de Função Fisiológica/fisiologia , Traumatismos da Medula Espinal/diagnóstico , Traumatismos da Medula Espinal/tratamento farmacológico , Doença Aguda , Adulto , Idoso , Idoso de 80 Anos ou mais , Estudos de Viabilidade , Feminino , Seguimentos , Humanos , Masculino , Pessoa de Meia-Idade , Estudos Prospectivos , Traumatismos da Medula Espinal/fisiopatologia , Fatores de Tempo , Resultado do Tratamento , Adulto JovemRESUMO
We produced a miniaturized, multicode, multiband, and programmable light-emitting diode (LED) stimulator for wireless control of optogenetic experiments. The LED stimulator is capable of driving three independent LEDs upon reception of an infrared (IR) signal generated by a custom-made IR transmitter. Individual LED photopulse patterns are assigned to different codes of the IR signals (up to 256 codes). The photopulse patterns can be programmed in the on-board microcontroller by specifying the parameters of duration ([Formula: see text]), frequency ([Formula: see text]), and pulse width ([Formula: see text]). The IR signals were modulated at multiple carrier frequencies to establish multiband IR transmission. Using these devices, we could remotely control the moving direction of a Thy1-ChR2-YFP transgenic mouse by transcranially illuminating the corresponding hemisphere of the primary motor cortex. IR transmitter and LED stimulator will be particularly useful in experiments where free movement or patterned concurrent stimulation is desired, such as testing social communication of rodents.
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STUDY DESIGN: An open-labeled multicenter prospective controlled clinical trial. OBJECTIVE: To confirm the feasibility of granulocyte colony-stimulating factor (G-CSF) administration for patients with thoracic myelopathy. SUMMARY OF BACKGROUND DATA: Although G-CSF is best known as an important cytokine commonly used to treat neutropenia, it also has nonhematopoietic functions. Previous experimental studies have shown that G-CSF can enhance tissue regeneration of several organs, such as the heart and the brain. We previously reported that G-CSF promotes functional recovery after spinal cord injury in rodents. On the basis of those findings, we started a clinical trial of neuroprotective therapy, using G-CSF for patients with worsening symptoms of thoracic myelopathy. METHODS: Patients whose Japanese Orthopaedic Association (JOA) score for thoracic myelopathy had decreased 2 points or more during a recent 1-month period were eligible for entry. After giving informed consent, patients were assigned to G-CSF and control groups. The G-CSF group (n = 10) received G-CSF 10 µg/kg per day intravenously for 5 consecutive days. The control group (n = 14) received similar treatments as the G-CSF group except for G-CSF administration. The primary outcome was JOA recovery rate at 1 month after G-CSF administration or initial treatment. RESULTS: There was greater improvement in neurological functioning between baseline and 1-month follow-up in the G-CSF group (JOA recovery rate: 29.1 ± 20.5%) than in the control group (JOA recovery rate: 1.1 ± 4.2%) (P < 0.01). No serious adverse events occurred during or after the G-CSF administration. CONCLUSION: The results provide evidence that G-CSF administration caused neurological recovery in patients with worsening symptoms of thoracic compression myelopathy.
Assuntos
Fator Estimulador de Colônias de Granulócitos/uso terapêutico , Fármacos Neuroprotetores/uso terapêutico , Recuperação de Função Fisiológica/efeitos dos fármacos , Doenças da Medula Espinal/tratamento farmacológico , Administração Intravenosa , Adulto , Idoso , Esquema de Medicação , Estudos de Viabilidade , Feminino , Fator Estimulador de Colônias de Granulócitos/administração & dosagem , Fator Estimulador de Colônias de Granulócitos/efeitos adversos , Humanos , Contagem de Leucócitos , Masculino , Pessoa de Meia-Idade , Fármacos Neuroprotetores/administração & dosagem , Fármacos Neuroprotetores/efeitos adversos , Estudos Prospectivos , Doenças da Medula Espinal/patologia , Doenças da Medula Espinal/fisiopatologia , Vértebras Torácicas , Resultado do Tratamento , Adulto JovemRESUMO
The cerebellum controls smooth and skillful movements and it is also involved in higher cognitive and emotional functions. The cerebellum is derived from the dorsal part of the anterior hindbrain and contains two groups of cerebellar neurons: glutamatergic and gamma-aminobutyric acid (GABA)ergic neurons. Purkinje cells are GABAergic and granule cells are glutamatergic. Granule and Purkinje cells receive input from outside of the cerebellum from mossy and climbing fibers. Genetic analysis of mice and zebrafish has revealed genetic cascades that control the development of the cerebellum and cerebellar neural circuits. During early neurogenesis, rostrocaudal patterning by intrinsic and extrinsic factors, such as Otx2, Gbx2 and Fgf8, plays an important role in the positioning and formation of the cerebellar primordium. The cerebellar glutamatergic neurons are derived from progenitors in the cerebellar rhombic lip, which express the proneural gene Atoh1. The GABAergic neurons are derived from progenitors in the ventricular zone, which express the proneural gene Ptf1a. The mossy and climbing fiber neurons originate from progenitors in the hindbrain rhombic lip that express Atoh1 or Ptf1a. Purkinje cells exhibit mediolateral compartmentalization determined on the birthdate of Purkinje cells, and linked to the precise neural circuitry formation. Recent studies have shown that anatomy and development of the cerebellum is conserved between mammals and bony fish (teleost species). In this review, we describe the development of cerebellar neurons and neural circuitry, and discuss their evolution by comparing developmental processes of mammalian and teleost cerebellum.
Assuntos
Evolução Biológica , Cerebelo/crescimento & desenvolvimento , Regulação da Expressão Gênica no Desenvolvimento , Animais , Padronização Corporal , Núcleo Celular/fisiologia , Proliferação de Células , Cerebelo/anatomia & histologia , Cerebelo/citologia , Cerebelo/fisiologia , Desenvolvimento Embrionário , Peixes/anatomia & histologia , Peixes/fisiologia , Neurônios GABAérgicos/citologia , Neurônios GABAérgicos/fisiologia , Mamíferos/anatomia & histologia , Mamíferos/fisiologia , Vias Neurais , Tubo Neural/citologia , Tubo Neural/crescimento & desenvolvimento , Tubo Neural/fisiologia , Neurogênese , Células de Purkinje/citologia , Células de Purkinje/fisiologia , Especificidade da Espécie , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismoRESUMO
OBJECTIVE: Granulocyte colony-stimulating factor (G-CSF) is a cytokine that is clinically used to treat neutropenia. G-CSF also has non-hematopoietic functions and could potentially be used to treat neuronal injury. To confirm the safety and feasibility of G-CSF administration for acute spinal cord injury (SCI), we have initiated a phase I/IIa clinical trial of neuroprotective therapy using G-CSF. METHODS: The trial included a total of 16 SCI patients within 48 h of onset. In the first step, G-CSF (5 µg/kg/day) was intravenously administered for 5 consecutive days to 5 patients. In the second step, G-CSF (10 µg/kg/day) was similarly administered to 11 patients. We evaluated motor and sensory functions of patients using the American Spinal Cord Injury Association (ASIA) score and ASIA impairment scale (AIS) grade. RESULTS: In all 16 patients, neurological improvement was obtained after G-CSF administration. AIS grade increased by one step in 9 of 16 patients. A significant increase in ASIA motor scores was detected 1 day after injection (P < 0.01), and both light touch and pin prick scores improved 2 days after injection (P < 0.05) in the 10 µg group. No severe adverse effects were observed after G-CSF injection. CONCLUSION: These results indicate that intravenous administration of G-CSF (10 µg/kg/day) for 5 days is essentially safe, and suggest that some neurological recovery may occur in most patients. We suggest that G-CSF administration could be therapeutic for patients with acute SCI.
Assuntos
Fator Estimulador de Colônias de Granulócitos/uso terapêutico , Fármacos Neuroprotetores/uso terapêutico , Recuperação de Função Fisiológica/efeitos dos fármacos , Traumatismos da Medula Espinal/tratamento farmacológico , Adulto , Idoso , Feminino , Humanos , Masculino , Pessoa de Meia-IdadeRESUMO
OBJECTIVE: Based on the neuroprotective effects of granulocyte colony-stimulating factor (G-CSF) on experimental spinal cord injury, we initiated a clinical trial that evaluated the safety and efficacy of neuroprotective therapy using G-CSF for patients with worsening symptoms of compression myelopathy. METHODS: We obtained informed consent from 15 patients, in whom the Japanese Orthopaedic Association (JOA) score for cervical myelopathy decreased two points or more during a recent 1-month period. G-CSF (5 or 10 µg/kg/day) was intravenously administered for five consecutive days. We evaluated motor and sensory functions of the patients and the presence of adverse events related to G-CSF therapy. RESULTS: G-CSF administration suppressed the progression of myelopathy in all 15 patients. Neurological improvements in motor and sensory functions were obtained in all patients after the administration, although the degree of improvement differed among the patients. Nine patients in the 10-µg group (n=10) underwent surgical treatment at 1 month or later after G-CSF administration. In the 10-µg group, the mean JOA recovery rates 1 and 6 months after administration were 49.9±15.1 and 59.1±16.3%, respectively. On the day following the start of G-CSF therapy, the white blood cell count increased to more than 22,700 cells/mm3. It varied from 12,000 to 50,000 and returned to preadministration levels 3 days after completing G-CSF treatment. No serious adverse events occurred during or after treatment. CONCLUSION: The results indicate that G-CSF administration at 10 µg/kg/day is safe for patients with worsening symptoms of compression myelopathy and may be effective for their neurological improvement.
Assuntos
Fator Estimulador de Colônias de Granulócitos/administração & dosagem , Fármacos Neuroprotetores/administração & dosagem , Compressão da Medula Espinal/tratamento farmacológico , Compressão da Medula Espinal/fisiopatologia , Medula Espinal/efeitos dos fármacos , Espondilose/fisiopatologia , Adulto , Idoso , Progressão da Doença , Feminino , Humanos , Injeções Intravenosas/métodos , Masculino , Pessoa de Meia-Idade , Parestesia/tratamento farmacológico , Parestesia/etiologia , Parestesia/fisiopatologia , Quadriplegia/tratamento farmacológico , Quadriplegia/etiologia , Quadriplegia/fisiopatologia , Recuperação de Função Fisiológica/efeitos dos fármacos , Recuperação de Função Fisiológica/fisiologia , Medula Espinal/fisiologia , Compressão da Medula Espinal/etiologia , Espondilose/complicações , Resultado do TratamentoRESUMO
The guidance of axonal projections to ipsilateral and contralateral regions is essential for integration of bilateral sensory information and coordination of movement. In the development of olivocerebellar projections, newborn neurons of inferior olivary (IO) nuclei ventrally migrate from the hindbrain rhombic lip to the floor plate (FP). The cell bodies of IO neurons cannot cross the FP but their axons can, and thus IO neurons project their axons only to the contralateral cerebellar cortex. The molecular mechanisms determining the contralateral axonal projections of IO neurons, however, are obscure. The IO neurons and their axons express EphA4, whereas the FP expresses an EphA4 ligand, EphrinB3, from embryonic day 12.5. Therefore, we tested whether EphA4-deficient mice (EphA4(-/-) ) would show impairment in the development of olivocerebellar projections. We found that, in EphA4(-/-) embryos, some of the IO neurons projected their axons to the ipsilateral cerebellar cortex because the cell bodies of the IO neurons abnormally crossed the FP. Furthermore, even in adults, EphA4(-/-) cerebella were bilaterally innervated by unilateral IO subnuclei. These observations indicate that EphA4 is involved in the contralateral axonal projections of IO neurons by preventing their cell bodies from crossing the midline FP.
Assuntos
Movimento Celular/fisiologia , Lateralidade Funcional/fisiologia , Neurogênese/fisiologia , Neurônios/citologia , Núcleo Olivar/embriologia , Receptor EphA4/metabolismo , Animais , Axônios/metabolismo , Axônios/ultraestrutura , Padronização Corporal/fisiologia , Imuno-Histoquímica , Hibridização In Situ , Camundongos , Camundongos Endogâmicos ICR , Camundongos Knockout , Neurônios/metabolismo , Núcleo Olivar/crescimento & desenvolvimento , Receptor EphA4/deficiênciaRESUMO
Genetically encoded Ca(2+) indicators (GECIs) are promising tools for cell type-specific and chronic recording of neuronal activity. In the mammalian central nervous system, however, GECIs have been tested almost exclusively in cortical and hippocampal pyramidal cells, and the usefulness of recently developed GECIs has not been systematically examined in other cell types. Here we expressed the latest series of GECIs, yellow cameleon (YC) 2.60, YC3.60, YC-Nano15, and GCaMP3, in mouse cortical pyramidal cells as well as cerebellar Purkinje cells using in utero injection of recombinant adenoviral vectors. We characterized the performance of the GECIs by simultaneous two-photon imaging and whole-cell patch-clamp recording in acute brain slices at 33 ± 2°C. The fluorescent responses of GECIs to action potentials (APs) evoked by somatic current injection or to synaptic stimulation were examined using rapid dendritic imaging. In cortical pyramidal cells, YC2.60 showed the largest responses to single APs, but its decay kinetics were slower than YC3.60 and GCaMP3, while GCaMP3 showed the largest responses to 20 APs evoked at 20 Hz. In cerebellar Purkinje cells, only YC2.60 and YC-Nano15 could reliably report single complex spikes (CSs), and neither showed signal saturation over the entire stimulus range tested (1-10 CSs at 10 Hz). The expression and response of YC2.60 in Purkinje cells remained detectable and comparable for at least over 100 days. These results provide useful information for selecting an optimal GECI depending on the experimental requirements: in cortical pyramidal cells, YC2.60 is suitable for detecting sparse firing of APs, whereas GCaMP3 is suitable for detecting burst firing of APs; in cerebellar Purkinje cells, YC2.60 as well as YC-Nano15 is suitable for detecting CSs.
RESUMO
The adult cerebellum is organized into longitudinal compartments that are revealed by specific axonal projections (olivocerebellar and corticonuclear projections). These compartments in the adult cerebellum are closely correlated with the striped expression of zebrin II (aldolase C), a late-onset marker of Purkinje cells. Similarly, the embryonic cerebellum is organized into longitudinal compartments that are revealed by striped expression of other genes (early-onset markers). The cerebellar compartments are thought to be the basic and functional subdivisions of the cerebellum. However, the relationship between the embryonic (early-onset) and the adult (late-onset) compartments has remained unknown, because the pattern of the embryonic compartments is distinct from that of the adult compartments. To examine this issue, we labeled Purkinje cells (PCs) born at embryonic day (E) 10.5, E11.5, and E12.5 by using an adenoviral vector and traced their fated positions in the adult cerebellum. By comparing the striped distribution of each cohort of birth date-related PCs with the striped pattern of zebrin II immunoreactivity (zebrin II bands) in the entire adult cerebellum, we found that the striped distribution of PCs correlated strikingly with zebrin II bands. Generally, a single early-onset compartment was transformed directly into a single late-onset compartment. Therefore, our observation also indicated the close correlation between the compartments formed by birth date-related PCs and olivocerebellar projections. Furthermore, we found that the cerebellum was composed of three units showing lateral-to-medial developmental gradients, as revealed by the birth dates of PCs. The results suggest that PC birth dates play an important role in organizing cerebellar compartmentalization.
Assuntos
Cerebelo/anatomia & histologia , Cerebelo/embriologia , Células de Purkinje/fisiologia , Adenoviridae , Animais , Feminino , Vetores Genéticos/genética , Vetores Genéticos/metabolismo , Camundongos , Proteínas do Tecido Nervoso/metabolismo , Gravidez , Células de Purkinje/citologiaRESUMO
We describe a low-cost, small, remotely triggerable LED device for wireless control of transcranial optical stimulation of cortical neurons, for use in freely moving mice. The device is easily mountable on the head of a mouse with a high-polymer block. Using the Thy1-ChR2-YFP transgenic mice, we demonstrate that the device is capable of remotely triggering muscle twitches upon activation of the primary motor cortex in freely moving conditions.
Assuntos
Eletrônica Médica/instrumentação , Eletrônica Médica/métodos , Estimulação Luminosa/instrumentação , Estimulação Luminosa/métodos , Imagens com Corantes Sensíveis à Voltagem/instrumentação , Imagens com Corantes Sensíveis à Voltagem/métodos , Animais , Potencial Evocado Motor/fisiologia , Potencial Evocado Motor/efeitos da radiação , Masculino , Camundongos , Camundongos Transgênicos , Córtex Motor/citologia , Córtex Motor/fisiologia , Córtex Motor/efeitos da radiação , Células Piramidais/fisiologia , Células Piramidais/efeitos da radiaçãoRESUMO
Adenoviral (Ad) vectors are useful for in vivo gene transfer into the brain. If Ad vectors are injected into the ventricle of mouse embryonic brain, Ad vectors introduce a foreign gene into neural progenitor cells on the surface of ventricle. However, Ad vectors were unable to deliver a foreign gene to a targeted region of the embryonic brain because Ad vectors evenly infected the neural progenitor cells on the surface of ventricle. Therefore, the Ad infection to the neural progenitor cells was uncontrollable. To develop a directional gene-transfer with Ad vector, we generated Ad vector tagged with magnetic nanoparticles (Ad-mag) by linking a biotinylated adenovirus vector with a streptavidin-conjugated magnetic nanoparticle. Ad-mags were attracted by magnetic force in vitro and in vivo. When Ad-mags were injected into the ventricle of mouse embryo and a strong magnet was attached to the head of the embryo, Ad-mags were attracted to the restricted direction or region where the magnet was placed. As a result, Ad-mags efficiently introduced a foreign gene into the restricted region of the brain.