颞叶癫(癎)大鼠海马轴突导向分子Sema3F及其受体Np2表达的变化

目的研究颞叶癫大鼠海马轴突导向分子Sema3F及其受体Np2表达的变化。方法给SD大鼠腹腔注射匹罗卡品、氯化锂制作颞叶癫模型。用免疫组化法和原位杂交技术对致后不同时间点大鼠海马CA1区、CA3区、齿状回的Sema3F mRNA、Np2 mRNA和蛋白表达进行检测,并与正常对照组比较。结果颞叶癫大鼠致后7d、15d,海马CA1区、CA3区Sema3F mRNA、Np2 mRNA和蛋白的表达明显低于正常对照组(P<0.05~0.01),致后30d、60d表达与正常对照组差异无统计学意义;而齿状回Sema3F mRNA、Np2 mRNA和蛋白的表达与正常对照组的差异无统计学意义。结论颞叶癫大鼠海马CA1区、CA3区Se-ma3F、Np2表达在致后早期明显下调,而在慢性期恢复正常。     

颞叶癫大鼠海马轴突导向分子Sema3F及其受体Np2表达的变化

目的 研究颞叶癫(癎)大鼠海马轴突导向分子Sema3F及其受体Np2表达的变化.方法 给SD大鼠腹腔注射匹罗卡品、氯化锂制作颞叶癫(癎)模型.用免疫组化法和原位杂交技术对致(癎)后不同时间点大鼠海马CA1区、CA3区、齿状回的Sema3F mRNA、Np2 mRNA和蛋白表达进行检测,并与正常对照组比较.结果 颞叶癫(癎)大鼠致(癎)后7 d、15 d,海马CA1区、CA3区Sema3F mRNA、Np2 mRNA和蛋白的表达明显低于正常对照组(P＜0.05～0.01), 致(癎)后30 d、60 d表达与正常对照组差异无统计学意义;而齿状回Sema3F mRNA、Np2 mRNA和蛋白的表达与正常对照组的差异无统计学意义.结论 颞叶癫(癎)大鼠海马CA1区、CA3区Sema3F、Np2表达在致(癎)后早期明显下调,而在慢性期恢复正常.     

氯化锂-匹罗卡品致痫大鼠海马EphA5及ephrinA3的表达及意义

目的探讨颞叶癫痫发作后海马EphA5及ephrinA3基因的表达变化和轴突出芽的关系。方法建立氯化锂-匹罗卡品颞叶癫痫大鼠模型,利用原位杂交方法检测致痫后12h、24h、7d、15d、30d、60d海马CA3区、CA1区EphA5及ephrinA3 mRNA的表达,快速Golgi染色观察CA1区的轴突出芽。结果致痫后,EphA5 mRNA在CA3区表达下调,ephrinA3 mRNA在CA1区表达下调,均在7d降至最低点,与对照组相比差异有显著意义(P<0.01),此后逐渐回升,但15d时仍低于对照组(P<0.05),在30d和60d与对照组相比差异无统计学意义(P>0.05)。快速Golgi染色显示,对照组大鼠CA1区轴突走行正常,匹罗卡品致大鼠SE后7dCA1区锥体细胞层出现显著增多的轴突染色。结论CA3区的EphA5和CA1区的ephrinA3的表达下调可能与CA1区的轴突出芽、突触重建有关。     

颞叶癫(癎)大鼠模型内嗅皮质和齿状回内Sema3A及其受体Np1的表达变化

目的 探讨颞叶癫(癎)大鼠脑内Sema3A及其受体Np1的表达变化在癫(癎)发病中的作用.方法 SD大鼠制成颞叶癫(癎)模型,Neo-Timm染色证实苔藓纤维出芽,免疫组化和原位杂交技术对致(癎)后不同时间点内嗅皮质和齿状回的Sema3A mRNA、Np1 mRNA和蛋白表达进行分析.结果 与对照组比较,颞叶癫(癎)大鼠海马齿状回内分子层存在苔藓纤维出芽(7 d:0.70±0.42,15 d:1.50±0.52,30 d:2.20±0.41,60 d:2.50±0.51,P＜0.05);在匹罗卡品致(癎)后7 d,实验组内嗅皮质区Sema3A mRNA的表达(0.3006±0.0675)明显低于对照组(0.4562±0.0457,P＜0.01);齿状回内Np1mRNA(0.2337±0.0358)及蛋白(0.2706±0.0389)的表达亦明显低于对照组(P＜0.01).结论 内嗅皮质区Sema3A mRNA、齿状回内Np1 mRNA和蛋白的表达下调,可能参与了苔藓纤维的出芽机制.     

红藻氨酸致颞叶癫(癎)大鼠脑内Semaphorin3A、Semaphorin4C基因表达的研究

目的研究轴索导向分子Semaphorin3A(Sema3A)、4C(Sema4C)对癫大鼠海马苔藓纤维重建的调控作用及对皮层神经元的保护作用。方法大鼠侧脑室内注射红藻氨酸制备颞叶癫模型,原位杂交法检测致痫间后1d,1、2、3、4周大鼠脑内Sema3A/Sema4C mRNA表达。结果致痫间后1周Sema3A、Sema4CmRNA分别在齿状回(DG),CA3区表达明显下降(P<0.01),持续至3、4周时恢复至正常(P>0.05);致痫间后1d Sema3A mRNA在皮层表达明显下降(P<0.01),持续至1、2周后恢复至正常(P>0.05)。结论红藻氨酸致痫间后DG及CA3区神经元分别下调Sema3A/Sema4C mRNA的表达,促进癫大鼠苔藓纤维重建;皮层神经元通过下调Sema3A mRNA的表达来维持自身存活。     

颞叶癫痫神经元型钙粘素mRNA表达研究

目的研究匹罗卡品诱导的颞叶癫痫神经元型钙粘素的表达变化.方法用氯化锂加匹罗卡品诱导SD大鼠颞叶癫痫模型,特异性探针原位杂交检测神经元型钙粘素mRNA表达.结果对照组动物神经元型钙粘素mRNA在皮层和海马各区均有表达,给药后4 h和12 h神经元型钙粘素mRNA在CA1、CA3和齿状回均表达下降,24 h开始恢复,注药后3 d神经元型钙粘素mRNA表达显著高于对照组,7 d表达最高.结论神经元型钙粘素mRNA在颞叶癫痫表达呈双相变化.     

颞叶癫痫大鼠海马TrkB mRNA及其蛋白表达的动态变化

目的 探讨颞叶癫痫发作大鼠海马TrkB mRNA及其蛋白表达的动态变化特征。方法 建立匹罗卡品(PILO)颞叶癫痢大鼠模型,应用原位杂交及免疫组织化学方法分别检测致(?)大鼠海马齿状回、CA3区及CA1区TrkB nRNA及其蛋白质表达的变化。结果 PILO致(?)后3～6 h,海马齿状回颗粒细胞层、CA1、CA3区锥体细胞层TrkB mRNA表达显著增高(P<0.01),稍后TrkB蛋白表达也随之增高。第7-30 d,TrkB mRNA及其蛋白在齿状回、CA3区呈现第二次表达增强。结论在癫(?)发作早期,TrkB表达增强,提示其可能参与急性癫痫状态的发生;后期表达增强则可能参与了海马的可塑性反应而与慢性自发性发作形成有关。     

颞叶癫痈大鼠海马内Rnd1 mRNA及蛋白表达变化

目的：动态观察小分子GTPase Rho家族的Rnd1 mRNA及其蛋白在氯化锂-毛果芸香碱（匹罗卡品）致痫大鼠模型海马中的表达变化，探讨其在颞叶癫痫发生发展中的作用。方法：在氯化锂-毛果芸香碱颞叶癫痫模型中应用逆转录聚合酶链反应（RT—PCR）检测癫痫持续发作（SE）后各时间点海马内Rnd1 mRNA的表达变化，并运用免疫组织化学染色法及Neo—Timm染色法分别检测齿状回门区、CA1区及CA3区中该蛋白在不同时间点的表达变化及苔藓纤维出芽（MFS）情况。结果：实验发现模型组于SE后8h内即出现Rnd1表达上调，SE后约1d达高峰，7d左右回复至对照组水平，此后其mRNA表达水平与对照组相似；而免疫组化染色发现Rnd1蛋白表达从SE后8h内即开始上调，约3d达高峰，至7d虽略有回落，但仍高于对照组水平，且这种情况可一直持续至慢性期。结论：急性期海马齿状回门区Rnd1表达上调可能通过促进MFS的发生参与了颞叶癫痫的发生。     

氯化锂-匹罗卡品致痫幼鼠海马结构损伤的形态学及苔藓纤维发芽研究

目的 观察幼鼠致痫后海马的组织病理学改变。方法 采用氯化锂－匹罗卡品腹腔注射制成幼鼠癫痫持续状态模型，应用常规病理及电镜观察海马结构的形态学改变，同时应用Timm组织化学染色方法进行苔藓纤维发芽的研究。结果 海马区神经元可见变性、坏死改变，以CA1区、CA3区为重。Timm染色见齿状回内分子层和CA3区下锥体层苔藓纤维发芽增加。结论 （1）氯化锂－匹罗卡品诱导的幼鼠癫痫持续状态可造成海马区神经元损伤；（2）幼鼠癫痫持续状态后海马CA1、CA3区神经元损伤较重；（3）幼鼠癫痫持续状态后可致苔藓纤维发增加。     

颞叶癫痫大鼠海马XIAP相关因子-1蛋白表达变化研究

目的动态观察颞叶癫痫大鼠海马神经元XIAP相关因子-1(XAF1)的表达变化,探讨其在癫痫发生发展中的作用。方法清洁级SD雄性成年大鼠36只,随机分为正常对照组、致痫(SE)后3h、6h、12h、24h、72h组。用氯化锂(LiCl)和匹罗卡品(PILO)制备癫痫动物模型,应用免疫组化染色技术检测致痫后各时间点XAF1蛋白表达情况。结果对照组海马CA1、CA3区神经元XAF1蛋白表达极少,SE组CA1、CA3区XAF1蛋白表达在3～24h表达逐渐增高,72h表达有所下降,但仍高于对照组,各时间点两组间差异均有统计学意义(P0.01)。结论SE大鼠海马神经元XAF1表达增高,它可能参与了SE后脑神经元凋亡的调控。     

Genetics of neurofibromatosis 1 and the NF1 gene

Neurofibromatosis 1 serves as a paradigm for understanding the principles of human genetics. The concepts of gene mutation, penetrance of the condition, variable clinical expressivity, mosaicism, age-dependent expression of clinical manifestations, and pleiotropy are evident in this autosomal dominant condition. The lack of genotype-phenotype correlation, except the whole-gene deletion phenotype, leads to speculation on modifiers of the haploinsufficient state of the NF1 gene product neurofibromin. The variant form of neurofibromatosis, neurofibromatosis Noonan's syndrome, suggests potential interaction of independent biochemical pathways. Identification of the NF1 gene led to the discovery of its role in ras signal transduction. Neurofibromin is a negative regulator of intracellular ras signaling. This observation now provides the framework for the development of rational medical therapies. In addition, knowledge of the molecular basis of the variable expression of clinical manifestations could provide better anticipatory guidance and more effective management of the medical complications that are associated with this condition.     

Genetic polymorphisms of glutathione S-transferases GSTM1, GSTT1, GSTP1 and GSTA1 as risk factors for schizophrenia

Oxidative damage is thought to play a role in the predisposition to schizophrenia. We determined if the polymorphisms of the GSTP1, GSTM1, GSTT1 and GSTA1 genes, which affect the activity of these enzymes against oxidative stress, have a role as susceptibility genes for schizophrenia, analyzing 138 schizophrenic patients and 133 healthy controls. We found that the combination of the absence of GSTM1 gene with the of the GSTM1 gene with the polymorphism GSTA1*B/*B, and the presence of the GSTT1 gene, represents a risk factor for schizophrenia, indicating that the combination of different GST polymorphisms has a role in the predisposition to schizophrenia, probably affecting the capacity of the cell to detoxify the oxidized metabolites of catecholamines.     

Cardiovascular effects of centrally applied endothelin-1 1-31 and its relationship to endothelin-1 1-21 in rats

Endothelin-1(1-31) (ET-1(1-31)) is a novel member of the endothelin family, which comprises 31 amino acids and derived from the selective hydrolysis of big ET-1 by chymase. Although ET-1(1-31) has been reported to be involved in biological effects via direct or indirect (converting to ET-1(1-21)) mechanisms, the cardiovascular effects of central ET-1(1-31) are not fully identified. The present study was designed to comparatively investigate the cardiovascular effects of intracerebroventricular (icv) application of ET-1(1-31) or ET-1(1-21) in anesthetized rats. Injection (icv) of ET-1(1-31) (500 pmol) produced a biphasic blood pressure response: an initial increase (from 118+/-8 to 138+/-14 mmHg, P<0.05) followed by a sustained decrease in BP (from 118+/-8 to 58+/-9 mmHg, P<0.05), which was very similar to BP response to icv injection of big ET-1 (500 pmol) or ET-1(1-21) (25 pmol)(.) The cardiovascular effects of icv injection of ET-1(1-31) or ET-1(1-21) were completely antagonized by ET(A) receptor antagonist BQ123 but not ET(B) receptor antagonist BQ788. Furthermore, pretreatment with ET converting enzyme inhibitor phosphoramidon (10 nmol) abolished the cardiovascular effects evoked by icv injection of ET-1(1-31) or big ET-1. In conclusion, the current data showed that central ET-1(1-31) produced the similar cardiovascular effects as those of central ET-1(1-21), and suggesting that the central cardiovascular effects of ET-1(1-31) resulted from it converting to ET-1(1-21) and then activating ET(A) receptors.     

High prevalence of ADHD symptoms in unmedicated youths with post-H1N1 narcolepsy type 1

ObjectivesTo characterize attention deficit-hyperactivity disorder (ADHD) symptoms in unmedicated post-H1N1 narcolepsy type 1 (NT1) youths, and explore associations between ADHD symptoms and the narcolepsy phenotype.MethodsA total of 50 consecutively enrolled post-H1N1 NT1 youths (7–20 years, 62% females, 98% HLA-DQB1106:02-positive, 98% CSF hypocretin-1 deficient, 88% vaccinated) were assessed after two weeks off medication for ADHD (ADHD diagnosis pre/post-narcolepsy, parent-rated ADHD symptoms) and narcolepsy-phenotyped (semi-structured interview, Stanford Sleep Questionnaire, Epworth Sleepiness Scale, polysomnography (PSG), Multiple Sleep Latency Test (MSLT)).ResultsIn sum, 26 (52%) and 15 (30%) of participants had ADHD symptoms above and below the clinical significant cut-off, respectively, while 9 (18%) had no ADHD symptoms. High values were found for ADHD total score (mean (SD), 17.9 (9.5)) and ADHD subscores (inattentive score, 11.0 (6.3); hyperactive/impulsivity score, 6.9 (4.7)). These were significantly higher than previously reported in a mainly medicated narcolepsy cohort (p < 0.0001). Age, gender and disease duration did not influence scores. Two participants (4%) had ADHD diagnosis prior to narcolepsy onset. ADHD symptoms were correlated with parent-rated, but not with patient rated ESS scores, objective sleepiness (mean sleep latency), sleep fragmentation (sleep stage shift index, awakening index), or CSF hypocretin-1 level.ConclusionComorbid ADHD symptoms were more prevalent in unmedicated post-H1N1 NT1 youths than previously reported in mainly medicated pediatric narcolepsy cohorts. The high prevalence was not due to pre-existing ADHD and generally not correlated with core narcolepsy sleep/wake phenotype characteristics, indicating that the ADHD symptoms were not a direct consequence of disturbed sleep or daytime sleepiness.     

The wmN1 enhancer region in intron 1 is required for expression of human PLP1

The myelin proteolipid protein gene (PLP1) encodes the most abundant protein present in myelin from the central nervous system (CNS). Its expression must be tightly controlled as evidenced by mutations that alter PLP1 dosage; both overexpression (elevated PLP1 copy number) and lack thereof (PLP1 deletion) result in X‐linked genetic disorders in man. However, not much is known about the mechanisms that govern expression of the human gene. To address this, transgenic mice were generated which utilize human PLP1 (hPLP1) sequences (proximal 6.2 kb of 5′‐flanking DNA to the first 38 bp of exon 2) to drive expression of a lacZ reporter cassette. LoxP sites were incorporated around a 1.5‐kb section of hPLP1 intron 1 since it contains sequence orthologous to the wmN1 region from mouse which, previously, was shown to augment expression of a minimally‐promoted transgene coincident with the active myelination period of CNS development. Eight transgenic lines were generated with the parental, 6.2hPLP(+)Z/FL, transgene. All lines expressed the transgene appropriately in brain as evidenced by staining with X‐gal in white matter regions and olfactory bulb. Removal of the “wmN1” region from 6.2hPLP(+)Z/FL with a ubiquitously expressed Cre‐driver caused a dramatic reduction in transgene activity. These results demonstrate for the first time that the wmN1 enhancer region: (1) is functional in hPLP1; (2) works in collaboration with its native promoter—not just a basal heterologous promoter; (3) is required for high levels of hPLP1 gene activity; (4) has a broader effect, both spatially and temporally, than originally projected with mPlp1.