Temporal and compartmental restriction of neuron-specific enolase expression in the rat mesostriatal system.

The striatum and the mesencephalic dopamine neurons which innervate it, are each organized into developmentally and biochemically distinct compartments. Striatal patches, characterized in the neonate by high concentrations of opiate receptors and substance P, are innervated prenatally by fibers originating in one group of midbrain dopamine neurons, the ventral tier. By the third postnatal day, a dense dopamine projection from neurons in the dorsal tier of the mesostriatal group innervates non-patch areas of the striatum, i.e. the matrix, and is followed by the appearance there of neurotensin, somatostatin and calcium binding protein. We have recently observed that the period of establishment of connections between dorsal tier dopamine neurons and their target cells in the striatal matrix is accompanied by a surge in expression of the gene coding for tyrosine hydroxylase (TH). In order to determine the overall metabolic state of mesencephalic and striatal neurons during the period of up-regulation of TH gene expression, we have applied immunocytochemistry for neuron specific enolase (NSE), and cytochrome oxidase histochemistry, known markers for neuronal activity, as well as TH immunohistochemistry to the mesencephalon and striatum of postnatally developing rats. At birth, both NSE and cytochrome oxidase were expressed almost exclusively in the patches, appearing in the matrix only after the 2nd postnatal day. Patches of NSE remained visible thru the 14th day. In the mesencephalon, cytochrome oxidase and immunoreactive NSE cells in adjacent sections, were present only in the pars reticulata (i.e. ventral tier). By day 8, both techniques identified nigral cells in the dorsal as well as ventral tiers.(ABSTRACT TRUNCATED AT 250 WORDS)     

Sequences in the proximal 5′ flanking region of the rat neuron-specific enolase (NSE) gene are sufficient for cell type-specific reporter gene expression

We investigated the regulation of the rat neuron-specific enolase gene using a transient transfection approach. Recent transgenic mouse studies have shown that a 1.8-kb segment of the ratNSE gene 5′ flanking region, including the first (noncoding) exon but not the first intron, is able to drive expression of a reporter gene in parallel with endogenousNSE. These data suggest thatcis-acting elements responsible for the spatial and temporal pattern ofNSE gene expression are located within the proximal 1.8 kb of the 5′ flanking sequence. To further investigate this region, we joined the 1.8-kb regulatory cassette to thecat reporter gene and generated a number of constructs in which the flanking sequence was progressively deleted from the 5′ end. These constructs were tested by transient transfection into neuronal and nonneuronal cells, followed by an assay for CAT activity. We found that as little as 255 bp of 5′ flanking sequence was able to confer cell type-specificity on the reporter gene. Further truncation to 120 bp of 5′ sequence resulted in a sharp downregulation of reporter activity in PC12 cells but a significant rise in both Neuro-2A neuroblastoma cells and nonneuronal Ltk- cells, indicating thatcis-acting elements controlling the regulation ofNSE in Ltk-, Neuro-2A, and PC12 cells may lie within the 135 bp region covered by this deletion. This region contains an AP-2 site and an element similar in sequence and position to a motif identified in the proximal promoter region of the neuron-specific peripherin gene. Reduction to 95 bp of 5′ sequence resulted in a slight downregulation of CAT activity in all cell lines tested, and further truncation to 65 bp of 5′ sequence caused a universal reduction to background levels of CAT activity, concomitant with the disruption of the basalNSE promoter. Our results show that the 5′ flanking region of theNSE gene is capable of conferring cell type-specificity on a heterologous gene in transfected cells and that elements responsible for this are located within the proximal 255 bp.     

Activity of neuron-specific enolase in normal and lesioned rat brain

Rat brain contains 3 forms of enolase, a neuron-specific form (NSE), a hybrid form, and a non-neuronal form (NNE) which were separated by DEAE-cellulose column chromatography. The enolase activity corresponding to each form of the enzyme eluted from the columns was determined spectrophotometrically. Using this assay procedure, an activity ratio (%NNE/%NSE) was calculated for cerebellum, brainstem, sciatic nerve, adrenals and liver. The results indicated excellent agreement between this enzymatically determined ratio and published values of a similar ratio (NNE/NSE) determined by radioimmunoassay for enzyme protein. Following in vivo destruction of neurons by intracerebral injection of the selective neurotoxin, kainic acid, there was a significant decrease in the activity of NSE and hybrid enolase (neuronal forms) and no change in the activity of NNE (glial form). These data indicate that separation and measurement of enolase species is useful to determine levels of these species in normal tissue and to estimate neuronal damage biochemically in brain lesions.     

The development of laminar staining for neuron-specific enolase in the rat somatosensory cortex

The expression of the enzyme neuron-specific enolase (NSE) in the central nervous system (CNS) has been used as a developmental marker based on observations that it is expressed shortly after the arrival of afferent inputs. The immunostaining pattern of NSE was examined in the laminae of the somatosensory cortex of the rat and the relationship of this staining pattern with previous data on the timing of afferent and efferent arrival was determined. Male rat pups were sacrificed on postnatal days 1 (24 h after birth), 3, 5, 8, 10, 12, 15 and 20, and as an adult (over 90 days of age). Sections were stained with an anti-NSE antibody using the avidin-biotin immunocytochemical method. Sections from day 1 animals revealed stained cells in the subplate layer and cortical plate, presumably in cells destined to form layers VI and V. By day 8 there was staining in layers II, III, V and VI, the same layers that exhibited staining in the adult rat. This appears consistent with the arrival of afferents and efferents which is completed by approximately postnatal day 7. On day 10, there was a change in the staining pattern: cell staining in layer VI was decreased and then increased gradually up to adult levels by day 20. A stable pattern of NSE staining was not observed previous to day 20. These results suggest that changes in NSE expression following the initial arrival of afferents may relate to maturation of the neurons.     

神经元特异性烯醇化酶与精神障碍研究进展

回顾了近期国内外学者对精神障碍与神经元特异性烯醇化酶（neuron—specificenolase;NSE）变化的观察,提出NSE在精神障碍方面应当深入研究其变化机制及应用,并及时开展心境障碍和神经症方面的研究。     

Proximal regions of the olfactory marker protein gene promoter direct olfactory neuron-specific expression in transgenic mice

Olfactory marker protein (OMP) expression is highly restricted to mature olfactory neurons (ON). Less than 0.3 kb of upstream 5′ flanking sequence of the OMP gene directs lacZ expression preferentially to ON in several independently derived lines of transgenic mice. A larger transgene with 0.8 kb of upstream flanking sequence also gave lacZ expression in ON and in a few ectopic sites in the central nervous system (CNS). In addition to the main olfactory epithelium, endogenous OMP is also expressed in chemosensory neurons of the vomeronasal and septal organs, and lacZ expression was detected in neurons of these sites as well. This confirmed the presence of regulatory sequences in the proximal portion of the OMP gene. Endogenous OMP expression in ON was normal in all transgenic lines. Strikingly, in several transgenic lines lacZ expression was restricted to subsets of ON. In one such line, ON axons were intensely stained for lacZ and projected to a subset of olfactory bulb glomeruli. Although identifiable subsets of ON and their termination fields have been described previously, this is the first demonstration of this phenomenon in transgenic mice. These lines of transgenic mice thus provide in vivo models for characterization of genetic elements regulating developmental and functional organization of the olfactory neuroepithelium. © 1996 Wiley-Liss, Inc.     

戊四氮致痫大鼠脑脊液神经元特异性烯醇化酶的水平测定

目的通过测定戊四氮致痫大鼠脑脊液中的神经元特异性烯醇化酶(NSE)水平的变化,探讨痫性发作与脑损伤之间的关系。方法110只Wistar大鼠分为实验组和对照组。实验组给予戊四氮(PIZ)腹腔注射。根据痫性发作的强度分为轻度组和重度组。用酶联免疫吸附法(ELISA)测定不同痫性发作强度、不同时间点大鼠脑脊液中NSE水平的变化,并与对照组进行对比。结果重度组脑脊液NSE水平在1h、2h、4h、6h均显著高于轻度组和对照组(P<0.01);轻度组脑脊液NSE含量在1h高于对照组(P<0.05),在2h、4h、6h显著高于轻度组;脑脊液NSE水平在痫性发作后1h迅速升高。4h达到高峰,24h恢复到正常水平。结论痫性发作可对大鼠神经细胞造成明显的损害。损害的程度与痫性发作的程度有关。痫性发作后4h可能是进行早期干预、防止脑细胞损伤的时间窗。     

Serum neuron-specific enolase in the major subtypes of status epilepticus

OBJECTIVES: To determine the relative magnitudes of neuron-specific enolase (NSE) levels after complex partial status epilepticus (SE), absence SE, generalized convulsive SE, and subclinical generalized convulsive SE (frequently referred to as acute symptomatic myoclonic status epilepticus). BACKGROUND: NSE is a marker of acute brain injury and blood-brain barrier dysfunction, which is elevated in SE. METHODS: Serum NSE levels were drawn in 31 patients 1, 2, 3, and 7 days after SE. Patients were classified as acute symptomatic or remote symptomatic, and the duration and outcome of SE were determined and correlated with the peak NSE level. RESULTS: NSE was elevated significantly in all four subtypes of SE, but NSE levels were highest in complex partial and subclinical SE. The mean peak NSE level for the complex partial SE group was 23.88 ng/mL (n = 12), 21.5 ng/mL for absence SE (n = 1), 14.10 ng/mL for the generalized convulsive SE group (n = 12), and 37.83 ng/mL for the subclinical SE group (n = 6), all of which was significantly higher than normal control subjects (5.02 ng/mL). Outcome was significantly different between the three groups (p = 0.0007), and was significantly worse for subclinical SE (p = 0.0005, subclinical versus generalized convulsive SE). CONCLUSION: Serum NSE levels were highest in complex partial and subclinical generalized convulsive SE. The extremely high levels of NSE in subclinical SE reflect the severity of the acute neurologic insults and poor outcome common to subclinical SE. High NSE levels in complex partial SE reflects the long duration of SE in this subgroup, and potential for brain injury.     

CSF neuron-specific enolase as a quantitative marker of neuronal damage in a rat stroke model

A technique for chronic cisternal cerebrospinal fluid (CSF) sampling in conscious rats was used to obtain multiple 50 microliters samples before and up to 7 days after middle cerebral artery occlusion. Neuron-specific enolase (NSE) concentrations were measured by radioimmunoassay using a readily available kit. The volume of infarction was measured by integrating the area of damage on 9 evenly spaced histological sections of the forebrain. This correlated well (r = 0.97, P less than 0.001) with the concentration of CSF neuron-specific enolase integrated over the first 5 days post occlusion, in animals with pure cortical and mixed cortical and striatal lesions. The correlation was maintained in animals given the NMDA antagonist MK-801. There was also a good correlation between the CSF NSE concentration 3 days post-MCAO and the volume of infarction (r = 0.92, P less than 0.01). It is therefore possible that CSF neuron-specific enolase may be useful as a quantitative marker of ischaemic damage in humans and provide a useful adjunct in the assessment of neuroprotective drugs in stroke.     

Significance of serum neuron-specific enolase in transient global amnesia

Neuron-specific enolase (NSE) is a glycolytic enzyme, which is associated with neuronal cell dysfunction in the brain. This study evaluated the role of serum NSE levels of patients with transient global amnesia (TGA). In addition, the relationship between serum NSE levels and the clinical features of TGA was explored. Forty-eight patients with TGA were prospectively included, and their serum NSE levels were measured. We investigated serum NSE levels in patients with TGA. In addition, we analyzed the differences in clinical characteristics between patients with elevated and normal serum NSE levels. Of the 48 patients with TGA, 16 patients (33.3%) had elevated serum NSE levels (25.0 ± 11.5 ng/mL), whereas 32 patients (66.7%) showed normal serum NSE levels (12.8 ± 2.1 ng/mL). The patients with elevated serum NSE levels exhibited higher levels of cognitive impairment than those with normal serum NSE levels (4/16 vs. 1/32, p = 0.036). The serum NSE levels showed a relatively high discrimination (AUC 0.684) between patients with and without cognitive impairment, with 80.0% sensitivity and 74.4% specificity at a cut-off value 17.3 ng/mL. A third of all patients with TGA carry elevated serum NSE levels, which suggests that the neuronal cell dysfunction could be associated with TGA pathogenesis. In addition, it might be correlated with cognitive impairment.     

Serum and CSF neuron-specific enolase in patients with West syndrome

OBJECTIVE: To determine whether frequent seizures and/or hypsarrhythmia may cause neuronal injury in West syndrome. BACKGROUND: West syndrome is an age-related epileptic syndrome of infancy characterized by clusters of epileptic spasms, a peculiar interictal EEG pattern of hypsarrhythmia, and mental deterioration. Recent clinical studies demonstrated that serum and CSF neuron-specific enolase (NSE)-a marker of neuronal injury-were increased after status epilepticus. METHODS: The authors examined serum and CSF NSE levels in 18 newly diagnosed infants (8.4 +/- 2.2 months) with West syndrome (3 cryptogenic, 15 symptomatic). In patients who showed complete resolution of spasms and disappearance of hypsarrhythmia (responders), additional serum NSE levels were determined several weeks after cessation of seizures. Serum NSE levels were obtained from 28 age-matched infants with normal neurologic development (control group), and 10 infants with an acute neurologic insult. RESULTS: There were no significant differences (p > 0.05) in serum NSE levels between the group with West syndrome (12.9 +/- 3.4 ng/mL) and the control group (13.2 +/- 3.1 ng/mL). The serum NSE value in the group with an acute insult (100.3 +/- 67.4 ng/mL) was significantly higher (p < 0.0001) than that for the West syndrome and the control groups. The mean +/- SD CSF NSE level was 7.3 +/- 3.6 ng/mL, which is similar to the reported CSF NSE levels of Japanese infants without neurologic disease. Thirteen responders showed no significant (p > 0.05) change in serum NSE after cessation of epileptic spasms. CONCLUSION: Normal serum and CSF neuron-specific enolase levels provided no evidence that seizures and/or hypsarrhythmia induced neuronal injury in West syndrome.     

癫痫患者血清和脑脊液神经元特异性烯醇化酶的测定

目的 探讨癫痫发作对脑神经元的损伤。方法 应用酶联免疫反应法动态测定癫痫发作后患者血清和脑脊液（CSF）中神经元特异性烯醇化酶（NSE）的含量。结果 癫痫组血清和CSF中NSE含量在发作后明显升高，血清NSE水平在发作后第1天最高，1周左右开始下降，2周左右降至正常；抽搐发作及频繁发作时，血清和CSF中NSE升高更明显。结论 癫痫发作引起血清和CSF中NSE水平的升高；癫痫发作导致神经元损伤，抽搐发作及频繁发作时神经元损伤更严重。     

癫(癎)患者血清、脑脊液神经元特异性烯醇化酶的测定及意义

目的探讨癫癎患者癎性发作后对神经元和血脑屏障的损伤。方法采用酶联免疫法测定癫癎患者在癎性发作后2d内血清和脑脊液(CSF)中神经元特异性烯醇化酶(NSE)的含量,与非神经系统疾病的神经症对照组对比分析。结果患者组CSF中NSE较对照组升高,差异有统计学意义;但血清中NSE含量与对照组差异无统计学意义。结论癫癎患者癫癎发作后脑脊液中NSE升高,提示癎性发作对中枢神经的神经元有损伤;而血中NSE正常,提示血脑屏障正常。     

血清神经元特异性烯醇化酶与急性脑出血关系的研究

目的探讨血清神经元特异性烯醇化酶（NSE）与急性脑出血的关系。方法采用酶联免疫吸附法对40例急性脑出血患者血清NSE水平进行动态检测,分析其与脑出血量、神经功能缺损程度评分（NDS）、脑出血部位的关系,并与正常对照组比较。结果脑出血组患者在发病48h内血清NSE水平即明显升高,3-5d达到峰值,6-9d和10-14d时仍明显高于正常对照组（P〈0.01-0.001）;脑出血患者血清NSE水平与出血量及NDS呈正相关（r=0.421,P〈0.01;r=0.415,P〈0.01）;与脑出血部位无明显关系（P〉0.05）。结论急性脑出血患者血清NSE水平明显升高,并且与脑出血量和病情有关。     

脑出血急性期神经元特异性烯醇化酶的变化及临床研究

目的　探讨脑出血急性期血清神经元特异性烯醇化酶 (NSE)浓度变化及其与神经功能缺损和预后之间的关系。方法　用酶联免疫分析法分析测定 4 0例脑出血患者和 2 0例健康人血清 NSE浓度。神经功能缺损评定按 SSS标准 ,出血量以入院时 CT结果计算。结果　脑出血患者血清 NSE浓度明显高于正常对照组 (P<0 .0 5 ) ,与出血量呈明显正相关 (P<0 .0 5 ) ,与神经功能缺损程度呈正相关 (P<0 .0 5 ) ,与预后呈负相关 (P<0 .0 5 )。结论　脑出血急性期检测血清 NSE浓度 ,有助于判断神经功能缺损程度及预后。     

Prognostic value of serum neuron-specific enolase levels after head injury

Abstract

Neuron-specific enolase (NSf) is a dimeric cytoplasmic enzyme detected in high levels in neurons and acts in the glycolytic pathway. It is known that there is a quantitative relationship between the concentration of serum NSf and the degree of cell damage in the central nervous system. We examined serum levels of NSf by enzyme immunoassay in 89 patients with head injury and aimed to evaluate its relationship with neurological status and prognosis of the patients. [Neural Res 1998; 20: 418–420]     

颅脑损伤后血清神经元特异性烯醇化酶变化

神经元特异性烯醇化酶(neuron-specific enolase,NSE)是神经元损伤的敏感标志.血清与脑脊液NSE水平与脑损伤的严重性与预后的关系较密切[1].为探讨颅脑损伤患者血清NSE水平的变化规律与颅脑损伤程度及临床意义,我们检测了123例颅脑损伤患者血清NSE水平.报告如下: