鼻科学

20050021胚胎型神经细胞黏附分子在人嗅黏 膜内的发育表达/任翔…//临床耳鼻咽喉科杂 志.2004,18(5).259～260 目的:探讨胚胎发育期人嗅黏膜内胚胎型神经 细胞黏附分子(ENCAM)的表达变化规律及生理 功能。方法:使用免疫组织化学方法对人胚胎发育 不同时期嗅黏膜内ENCAM的表达情况进行检 测。结果:ENCAM的表达开始于胚胎发育的第 l4周,阳性反应位于嗅神经元细胞胞体、树突、轴突 及固有层内的嗅神经纤维束;随胚胎发育阳性细胞 数目逐渐增多,嗅神经纤维束染色逐渐增强;第28 周达峰值后逐渐下降。在发育不同时期,ENCAM 阳性细胞均位…     

神经特异性烯醇化酶及嗅标记蛋白在不同胎龄胎儿嗅黏膜中的表达

目的 探讨神经特异性烯醇化酶(neuron—specific enolase，NSE)及嗅标记蛋白(olfactory marker protein，OMP)在不同胎龄胎儿嗅黏膜中的表达。方法 以免疫组化方法检测NSE及OMP在12、16、20、24，28和34周6例不同胎龄胎儿嗅黏膜中的表达。结果 NSE免疫阳性反应在孕12～34周的胎儿嗅黏膜中均有表达，各胎龄胎儿嗅黏膜切片中均可见大量阳性着色的双极嗅神经细胞。孕12周时，阳性细胞数量很多，排列紧密，胞体多位于嗅上皮中下部且阳性嗅神经上皮占据胎儿鼻腔上2／3的黏膜，随着孕龄的增大，阳性细胞逐渐成多层次排列，所占鼻腔黏膜面积却逐渐减小，至孕34周时，仅局限于鼻腔上1／3黏膜：0MP免疫反应在孕12周胎儿鼻腔上2／3黏膜切片中仅发现少量阳性着色的双极神经细胞，明显少于同龄胎儿NSE阳性反应细胞。随着胎龄的增加，0MP阳性细胞逐渐增多，且胞体多位于嗅上皮中上部，但其数量仍相对少于同龄NSE阳性细胞。结论 人胚胎12周时嗅黏膜中已有大量的嗅神经细胞，其中少数嗅神经细胞已开始发育成熟。随着胎龄的增大，嗅上皮面积逐渐缩小，但成熟的嗅神经细胞逐渐增多，胚胎发育后期的胎儿嗅化学感受器发育已趋于成熟。     

神经特异性烯醇化酶及嗅标记蛋白在不同胎龄胎儿嗅黏膜中的表达

目的　探讨神经特异性烯醇化酶 (neuron specificenolase ,NSE)及嗅标记蛋白 (olfactorymarkerprotein ,OMP)在不同胎龄胎儿嗅黏膜中的表达。方法　以免疫组化方法检测NSE及OMP在1 2、1 6、2 0、2 4、2 8和 34周 6例不同胎龄胎儿嗅黏膜中的表达。结果　NSE免疫阳性反应在孕 1 2～ 34周的胎儿嗅黏膜中均有表达 ,各胎龄胎儿嗅黏膜切片中均可见大量阳性着色的双极嗅神经细胞。孕1 2周时 ,阳性细胞数量很多 ,排列紧密 ,胞体多位于嗅上皮中下部且阳性嗅神经上皮占据胎儿鼻腔上2 /3的黏膜 ,随着孕龄的增大 ,阳性细胞逐渐成多层次排列 ,所占鼻腔黏膜面积却逐渐减小 ,至孕 34周时 ,仅局限于鼻腔上 1 /3黏膜。OMP免疫反应在孕 1 2周胎儿鼻腔上 2 /3黏膜切片中仅发现少量阳性着色的双极神经细胞 ,明显少于同龄胎儿NSE阳性反应细胞。随着胎龄的增加 ,OMP阳性细胞逐渐增多 ,且胞体多位于嗅上皮中上部 ,但其数量仍相对少于同龄NSE阳性细胞。结论　人胚胎 1 2周时嗅黏膜中已有大量的嗅神经细胞 ,其中少数嗅神经细胞已开始发育成熟。随着胎龄的增大 ,嗅上皮面积逐渐缩小 ,但成熟的嗅神经细胞逐渐增多 ,胚胎发育后期的胎儿嗅化学感受器发育已趋于成熟     

神经元核心抗原在小鼠嗅球和嗅上皮发育中的表达

目的：探讨神经元核心抗原（NeuN）在嗅球和嗅上皮发育过程中的表达特性和意义。方法：在小鼠胚胎发育9．5、11．5、14．5、17．5d,出生当天和3个月成年个体的头部切片标本中,以免疫荧光染色方法检测NeuN的表达。结果：刚出生小鼠嗅球内层状结构尚不明显,NeuN表达于嗅球周边区域。在3个月大成年小鼠,嗅球的层状结构已清晰可见,NeuN表达阳性的成熟神经元主要聚集于接近嗅球中心的颗粒细胞层。位于嗅上皮的双极嗅感觉神经元在小鼠胚胎和成年个体中均未见NeuN表达。结论：NeuN通常被认为表达于几乎所有部位的成熟神经元。但在嗅球和嗅上皮中,NeuN只表达于成熟嗅球中的颗粒细胞层。小鼠出生到发育为成熟个体的过程中,NeuN表达阳性的成熟神经细胞由周边逐渐向嗅球中心迁移,可能与气味模式的形成有关。     

外伤性嗅觉障碍大鼠嗅黏膜的组织学变化

目的 构建大鼠外伤性嗅觉障碍模型,观察不同时间点嗅黏膜组织学变化.方法 神经切断组(40只)和对照组(20只)大鼠均在显微镜下暴露左侧嗅球,沿筛板切断神经组切断大鼠左侧嗅神经.采用嗅觉诱发电位(olfactory evoked potentials,OEPs)和神经示踪验证造模效果.术后1天、5天、2周、3周、6周处理大鼠,处理前1天每组各取2只大鼠经鼻腔滴注辣根过氧化物酶(horseradish peroxidase,HRP).嗅黏膜及嗅球冰冻切片后观察嗅上皮的厚度、细胞数量的变化以及嗅神经的连续性,并且行免疫组化观察嗅上皮中的新生嗅感觉神经元(olfactory receptor neurons,ORNs).结果OEPs及神经示踪证实手术方法可以完全切断嗅神经.术后1天,切断侧黏膜中ORNs出现凋亡,两组大鼠双侧嗅上皮厚度和细胞数量比值无明显变化.5天时切断侧嗅上皮中细胞数量减少,上皮厚度变薄,嗅球中无HRP标记纤维.术后2、3周大鼠嗅球中出现蓝色标记,嗅上皮厚度和细胞数量逐渐增加,但仍然与对照组有差异,此时嗅上皮中出现大量的新生ORNs.经过6周的恢复,嗅上皮厚度及细胞数量基本恢复至对照组水平,嗅上皮中有较多的新生ORNs,其轴突与嗅球重新建立神经联系,但是上皮中仍然有一定数量的凋亡细胞.结论 嗅神经切断术可以作为制作外伤性嗅觉障碍的可靠方法;由于ORNs具有再生能力,大鼠嗅神经切断后嗅黏膜在一定时间内基本恢复至正常水平.     

大鼠嗅感觉上皮发育及嗅细胞凋亡的研究

目的 探讨神经特异性烯醇酶（NSE）、嗅细胞标记蛋白（OMP）及细胞凋亡在不同胎龄大鼠嗅黏膜中的表达。方法 免疫组化方法检测NSE及OMP在不同孕期大鼠嗅上皮中的表达及其规律。TUNEL方法检测不同孕期大鼠嗅上皮中凋亡细胞并计算细胞凋亡指数（AI）。结果 E13d鼻腔黏膜中即有NSE阳性表达,细胞数量多。E13d嗅上皮中未见OMP阳性表达细胞。在E14d,嗅上皮中出现嗅OMP阳性细胞,数量少,随胎龄增加阳性细胞数量增多,至17d达高峰并逐渐趋于稳定。E13～E15d细胞凋亡数量较稳定,至E16d凋亡细胞数量明显增加,达到高峰,E18～E21d凋亡细胞数量逐渐减少渐趋于稳定。E16dAI与其他d数差异有统计学意义。结论 E14d嗅黏膜中已有发育成熟的嗅细胞,数量少,胚胎发育后期大鼠嗅化学感受器发育已趋于成熟。在嗅上皮发育过程中存在细胞凋亡,E16d嗅细胞凋亡出现一高峰,细胞凋亡在嗅觉发育过程中起重要作用。     

鼻科学

2仪M侧)19神经特异性烯醇化酶及嗅标记蛋白在不同胎龄胎儿嗅私膜中的表达/史丽…//中华耳鼻咽喉科杂志一2(X)3,38(3)一180一282 目的:探讨神经特异性烯醇化酶(NSE)及嗅标记蛋白(OMP)在不同胎龄胎儿嗅勃膜中的表达(方法:以免疫组化方法检测NSE及OMP在12、16、20、24、28和34周6例不同胎龄胎儿嗅豁膜中的表达。结果:NSE免疫阳性反应在孕12一34周的胎儿嗅薪膜中均有表达,各胎龄胎儿嗅勃膜切片中均可见大量阳性着色的双极嗅神经细胞。孕]2周时,阳性细胞数量很多,排列紧密,胞体多位于嗅上皮中下部且阳性嗅神经上皮占据胎儿鼻腔上2/3的茹膜…     

嗅神经离断后大鼠嗅黏膜气味受体的表达变化

明显减少,差异具有统计学意义(t=63.960,P=0.000).随着术后恢复时间的延长,嗅神经离断侧再生嗅黏膜表达Olr1493基因受体的细胞逐渐增多,至第6周时基本和对照侧一致,分别为(417.8±32.4)个和(445.3±10.0)个,差异无统计学意义(t=2.600,P=0.060).结论 大鼠嗅神经离断后再生嗅感觉神经元及其受体数量可以恢复至正常水平,表达位置亦未发生变化.     

放疗后嗅粘膜超微结构观察

嗅区粘膜放疗后可引起迟发性嗅觉障碍，为了解其机制，对放疗后嗅粘膜用透射电镜观察其超微结构改变，发现其上皮层变性嗅神经原增多，固有层内纤维母细胞增多，胞内含有有大量粗面内质网和线粒体，在纤维母细胞在周围大有量整齐排列的胶原纤维，放疗后嗅粘膜的纤维化是嗅觉障碍的原因之一。     

放疗后嗅粘膜超微结构观察

嗅区粘膜放疗后可引起迟发性嗅觉障碍，为了解其机制，对放疗后嗅粘膜用透射电镜观察其超微结构改变。发现其上皮层变性嗅神经原增多；固有层内纤维母细胞增多，胞浆内含有大量粗面内质网和线粒体，在纤维母细胞周围有大量整齐排列的胶原纤维。放疗后嗅粘膜的纤维化是嗅觉障碍的原因之一。     

Expression of neuron-specific enolase and olfactory marker protein in the developing olfactory mucosa of human fetuses]

OBJECTIVE: To study the expression of neuron-specific enolase (NSE) and olfactory marker protein (OMP) in the developing olfactory mucosa of human fetuses. METHOD: The expression of NSE and OMP in the olfactory mucosa of 6 human fetuses (12, 16, 20, 24, 28 and 34 weeks) was studied using the technique of immunohistochemistry. RESULTS: NSE immunological positive reactions were seen in all 6 fetal mucosa from gestational 12 (G12) to G34, with plenty of positive-stained dual-pole neuron cells. At G12, the positive cells aligned tightly, the cell bodies were localized in the lower portion of olfactory epithelium and the positive-stained area occupied upper 2/3 of fetal nasal mucosa. With the development, the positive cells gradually became multilayer, but the density and the relative area of positive-cells reduced. At G34, the positive cells were located only in upper 1/3 of nasal mucosa. OMP-positive reactions were localized in a few dual-pole neurons at G12, the number was much less than NSE-positive cells in the same fetus. With the development, the OMP-positive cells gradually increased with most of the cell bodies located in the upper portion of epithelium, but number still relatively less than the NSE-positive cells at the same age. CONCLUSION: At G12, there were lots of olfactory neuron in the olfactory mucosa and only a few olfactory neurons had became mature. With the development, the olfactory epithelial area reduced but the number of mature olfactory neurons increased. At the last trimester, fetal olfactory sensor was almost matured.     

Immunohistochemical study of the human olfactory system]

Taurine (2-aminoethane sulfonic acid) and carnosine (beta-alanyl-L-histidine) are found in large quantities in the olfactory epithelium and bulb. Taurine is a structurally simple amino acid, and has been reported to have several putative roles, such as neurotransmitter, neuromodulator, neurogrowth factor and to function in membrane stabilization. Carnosine, on the other hand, has been suggested as a putative neurotransmitter in the olfactory system. We have succeeded in visualizing taurine- and carnosine-like immunoreactivities (LI) in the human olfactory mucosa, and also carnosine-LI in the human olfactory bulb. For this investigation, we collected specimens of the human olfactory bulb by autopsy and from the olfactory mucosa by biopsy, and compared localization of taurine- and carnosine-LI in several cases. By means of biopsy using Nakano's forceps, samples of olfactory mucosa were obtained from 5 cases: a 17 year old female, 23 year old male, 46 year old male, 47 year old male, and a 57 year old male. The olfactory bulb of a 1 month old male was collected at autopsy. These specimens were processed for immunohistochemical study according to the peroxidase-antiperoxidase (PAP) method. In the olfactory epithelium, taurine-LI was demonstrated in some primary olfactory neurons, and in basal cells. Carnosine-LI was observed only in primary olfactory neurons, i.e., dendrites, vesicles and axonal bundles of olfactory receptor cells, but not in basal cells. In the olfactory bulb, the olfactory nerve layer and the glomerular layer showed carnosine-LI positive reactions. Therefore, taurine and carnosine may possibly coexist in some olfactory neurons. Olfactory receptor cells are classified as sensory neurons.(ABSTRACT TRUNCATED AT 250 WORDS)     

Upregulation of neural growth-associated protein and neural cell adhesion molecule in mouse olfactory epithelium and axons after unilateral removal of the olfactory bulb

The expression of synaptosomal-associated protein (SNAP-25), neural growth-associated protein (GAP-43) and neural cell adhesion molecule (NCAM) were studied in mouse olfactory cells and axons for 2 weeks following unilateral bulbectomy. The olfactory cells and axons in the control olfactory epithelium were positive for SNAP-25 but levels decreased in the atrophic olfactory epithelium 3 days after bulbectomy. There was no expression of SNAP-25 in the olfactory epithelium on the bulbectomy side 7 days after bulbectomy, indicating that this protein may be a good marker for the degeneration of olfactory cells. The expression of NCAM was still found in the atrophic olfactory epithelium at 7 days after bulbectomy, while the expression of NCAM in the olfactory epithelium of the bulbectomy side was stronger than that on the control side at 14 days after bulbectomy. The expression of GAP-43 in the olfactory axonal bundles of the bulbectomy side at 3 and 4 days after bulbectomy was stronger than that on the control side. These results suggest that upregulation of NCAM may be related to the regeneration of the olfactory cells, with upregulation of GAP-43 probably playing a role in axonal regeneration after bulbectomy. Received: 6 January 1998 / Accepted: 22 April 1998     

Developmental changes in expression of a calcium-binding protein (spot 35-calbindin) in the Nervus terminalis and the vomeronasal and olfactory receptor cells.

The detailed localization of spot 35-calbindin and its ontogenic change were studied in Nervus terminalis, the vomeronasal organ and the olfactory epithelium of the rat by immunohistochemistry. At the embryonic days 12 and 13 (E 12-13), calbindin-immunoreactive cells were found in the outermost layer of the presumptive olfactory bulb and within the olfactory placode. At E 14 to the postnatal day 1 (P 1), intense calbindin-immunoreactivity was localized in ganglionated fiber bundles of Nervus terminalis coursing through the mesenchymal spaces on both sides of the nasal septum. Nervus terminalis decreased the immunoreactivity abruptly after P 1 and it showed no distinct immunoreactivity for calbindin at P 7 and thereafter. On the other hand, numerous receptor cells in the olfactory epithelium and the thicker vomeronasal epithelium exhibited weak to moderate immunoreactivity for calbindin at E 18-P 1. Their immunoreactivity decreased in intensity progressively after P 7 and no distinct immunoreactivity for calbindin was detected in most of the receptor cells, whereas moderate immunoreactivity was detected in most of the vomeronasal and olfactory nerves at P 28 and P 63.     

嗅球摘除后嗅觉神经元凋亡的实验研究

目的研究凋亡是否参与嗅上皮正常生理更替和嗅球摘除后嗅觉神经元死亡而后再生的过程,探讨凋亡与神经元再生的关系。方法应用原位末端标记法和透射电镜观察正常成年大鼠以及摘除嗅球后大鼠嗅上皮16、32、48 h和3、7、30 d时凋亡的出现和改变情况。结果正常成年大鼠嗅上皮中有末端脱氧核苷酸转移酶介导的脱氧三磷酸尿苷缺口末端标记(TUNEL)的阳性细胞(1.93±0.31)个/200μm。摘除嗅球后TUNEL阳性细胞数增加,32 h达峰值(90.9±18.03)个/200μm,以后迅速下降,维持于低水平。透射电镜下见嗅球摘除术后嗅觉神经元出现胞质浓缩、胞核染色质边聚等细胞凋亡的特征性超微结构改变。尚可见少量胞质内出现自噬泡以及除线粒体以外的细胞器扩张,但胞核正常的嗅觉神经元。结论凋亡参与成年大鼠嗅觉神经元生理性更替以及实验性嗅觉神经元死亡和再生的过程。此外,尚存在自噬型和胞质型神经元死亡。     

Electron microscopic study on the esophageal mucosa in human fetuses

Morphogenetic studies of the esophageal mucosa in human fetuses have been few and there is only one report at the ultrastructural level. We thus studied the esophageal mucosa in human fetuses (at the gestational ages from the 7th to 21st week) by scanning (SEM) as well as transmission electron microscopy (TEM). Our results and the review of the literature lead to the following conclusions: 1) Primary cilia were seen in the 7th and 8th week of gestation. 2) Ciliated cells appeared around the 8th week of gestation. They increased in number according to the fetal ages, but gradually decreased after the 14th week of gestation. Their degenerative process at the ultrastructural level was similar to that reported previously. 3) The stratified squamous epithelia appeared at the 14th week of gestation, but the squamous cells remained immature even at the 21st week. 4) Glycogen granules in non-ciliated cells decreased as the differentiation of the cells proceeded, suggesting that they provide an energy source necessary for the cell differentiation.     

Olfactory secretion and sodium, potassium-adenosine triphosphatase: regulation by corticosteroids

OBJECTIVES: To investigate the cellular distribution and relative intensity of the immunoreactivity associated with the expression of sodium, potassium-adenosine triphosphatase (Na, K-ATPase) in cells of the olfactory mucosa. Second, changes in the activity of this enzyme in the olfactory mucosa are correlated with changes in the circulating corticosteroid aldosterone. METHODS: Combination of immunohistochemical and biochemical techniques were employed to examine the olfactory Na, K-ATPase. RESULTS: Within the olfactory epithelium, the Na, K-ATPase immunoreactivity was greatest at the supranuclear region of sustentacular cells and/or dendrites of olfactory receptor neurons (ORNs). Cell bodies of ORNs demonstrated moderate immunoreactivity, whereas the duct cells of Bowman's gland exhibited moderate to intense immunoreactivity. Acinar cells of the Bowman's gland were the most intensely stained components of the lamina propria, exhibiting strong immunoreactivity at the basolateral plasma membrane domains of the acinar cells and less within the cytoplasm. Binding of ouabain, a specific inhibitor of Na, K-ATPase, was significantly elevated for aldosterone-injected versus sham-injected controls. CONCLUSION: These results suggest that olfactory Na, K-ATPase is regulated by the systemic corticosteroid aldosterone. The results are consistent with the hypothesis that corticosteroids regulate olfactory secretion.