NITRIC OXIDE (NO), CITRULLINE - NO CYCLE ENZYMES,GLUTAMINE SYNTHETASE AND OXIDATIVE STRESS IN ANOXIA (HYPOBARIC HYPOXIA) AND REPERFUSION IN RAT BRAIN

Nitric oxide is postulated to be involved in the pathophysiology of neurological disorders due to hypoxia/ anoxia in brain due to increased release of glutamate and activation of N-methyl-D-aspartate receptors. Reactive oxygen species have been implicated in pathophysiology of many neurological disorders and in brain function. To understand their role in anoxia (hypobaric hypoxia) and reperfusion (reoxygenation), the nitric oxide synthase, argininosuccinate synthetase, argininosuccinate lyase, glutamine synthetase and arginase activities along with the concentration of nitrate /nitrite, thiobarbituric acid reactive substances and total antioxidant status were estimated in cerebral cortex, cerebellum and brain stem of rats subjected to anoxia and reperfusion. The results of this study clearly demonstrated the increased production of nitric oxide by increased activity of nitric oxide synthase. The increased activities of argininosuccinate synthetase and argininosuccinate lyase suggest the increased and effective recycling of citrulline to arginine in anoxia, making nitric oxide production more effective and contributing to its toxic effects. The decreased activity of glutamine synthetase may favor the prolonged availability of glutamic acid causing excitotoxicity leading to neuronal damage in anoxia. The increased formation of thiobarbituric acid reactive substances and decreased total antioxidant status indicate the presence of oxidative stress in anoxia and reperfusion. The increased arginase and sustained decrease of GS activity in reperfusion group likely to be protective.     

Calretinin- and parvalbumin-immunoreactive neurons in the rat main olfactory bulb do not express NADPH-diaphorase activity

The presence of nitric oxide synthase (NOS) in neuronal elements expressing the calcium-binding proteins calretinin (CR) and parvalbumin (PV) was studied in the rat main olfactory bulb. CR and PV were detected by using immunocytochemistry and the nitric oxide (NO) -synthesizing cells were identified by means of the reduced nicotinamide adenine dinucleotide phosphate diaphorase (NADPH-diaphorase) direct histochemical method. The possible coexistence of NADPH-diaphorase and each calcium-binding protein marker was determined by sequential histochemical-immunohistochemical double-labeling of the same sections. Specific neuronal populations were positive for these three markers. A subpopulation of olfactory fibers and olfactory glomeruli were positive for either NADPH-diaphorase or CR. In the most superficial layers, groups of juxtaglomerular cells, superficial short-axon cells and Van Gehuchten cells demonstrated staining for all three markers. In the deep regions, abundant granule cells were NADPH-diaphorase- and CR-positive and a few were PV-immunoreactive. Scarce deep short-axon cells demonstrated either CR-, PV-, or NADPH-diaphorase staining. Among all these labeled elements, no neuron expressing CR or PV colocalized NADPH-diaphorase staining. The present data contribute to a more detailed classification of the chemically- and morphologically-defined neuronal types in the rodent olfactory bulb. The neurochemical differences support the existence of physiologically distinct groups within morphologically homogeneous populations. Each of these groups would be involved in different modulatory mechanisms of the olfactory information. In addition, the absence of CR and PV in neuronal groups displaying NADPH-diaphorase, which moreover are calmodulin-negative, indicate that the regulation of NOS activity in calmodulin-negative neurons of the rat olfactory bulb is not mediated by CR or PV.     

Projections of two subclasses of vomeronasal nerve fibers to the accessory olfactory bulb in the rabbit

The organization of the projections of subclasses of vomeronasal nerve fibers to the accessory olfactory bulb was analysed using monoclonal antibodies generated against a homogenate of the rabbit olfactory bulb. Monoclonal antibody R2D5 labels all the somata of vomeronasal receptor cells in the vomeronasal organ as well as all their axons (vomeronasal nerve fibers). Another monoclonal antibody (R4B12), which has been shown to selectively bind and thus identify a subclass of olfactory nerve fibers, also labels a subclass of vomeronasal nerve fibers. The R4B12-positive subclass of vomeronasal nerve fibers project to the glomeruli in the rostrolateral part of the accessory olfactory bulb. The third monoclonal antibody (R5A10) recognizes a complementary subclass of vomeronasal nerve fibers projecting to the glomeruli in the caudomedial part of the accessory bulb. In contrast to the clearly segregated terminations in the accessory bulb, the two subclasses of vomeronasal nerve fibers are intermingled with each other in the vomeronasal nerve bundles. Retrograde labeling of vomeronasal receptor cell somata following injection of horseradish peroxidase within the rostrolateral (R4B12-positive) part of the accessory bulb indicates that vomeronasal receptor cells of this subtype are widely distributed in the vomeronasal sensory epithelium. These results demonstrate the heterogeneity of vomeronasal receptor cells and the specificity of projections arising from subclasses of vomeronasal nerve fibers to the accessory olfactory bulb.     

Distribution of NADPH diaphorase-containing neurons in the pigeon central nervous system

The aim of the present study was to determine the distribution of nitric oxide-synthesizing neurons in the pigeon brain and spinal cord. Tissue sections were stained for reduced nicotinamide adenine dinucleotide phosphate-diaphorase (NADPH-d). In the telencephalon, intensely stained neurons with dendrites extending distally were seen in most regions. The ectostriatum was characterized by intensely and diffusely stained neuropil. In the diencephalon, intensely positive neurons were seen in the lateral hypothalamic region and lateral mammillary nucleus. In the mesencephalon, intensely stained, multipolar neurons were abundantly scattered in the central gray, nucleus intercollicularis, reticular formation, nucleus tegmenti pedunculo-pontinus, pars compacta, area ventralis of Tsai, and ansa lenticularis. In the rhombencephalon, positively-stained neurons were found in the pontine nuclei and reticular formation. The cerebellar cortex, except for Purkinje cells, was a preferential region for NADPH-d activity. Positive end-bulbs made contact on somata in the nucleus magnocellularis cochlearis. In the spinal cord, NADPH-d positive neurons were seen in layer II and the marginal nucleus. Our results demonstrated that the distribution of NADPH-d-containing neurons in the pigeon brain and spinal cord is more complex than in other avian species. Our findings indicate that NADPH-d-containing neurons are present in several sensory pathways, including olfactory, visual, auditory, and somatosensory tracts, although some nuclei in each system did not show NADPH-d activity. The wide distribution of NADPH-d activity in the pigeon CNS suggests that nitric oxide modulates sensory transmission in avian central nervous system.     

Immunohistochemical localization of argininosuccinate synthetase in the rat brain in relation to nitric oxide synthase-containing neurons.

The distribution of the urea cycle enzyme, argininosuccinate synthetase, in the rat brain was determined using immunohistochemistry. This enzyme participates in the only known metabolic pathway for citrulline, its condensation with aspartate to form argininosuccinate, which can then be cleaved to fumarate and arginine. It may thus provide a mechanism to recycle citrulline, formed in the nervous system via nitric oxide synthase activity, back to the nitric oxide precursor, L-arginine. Argininosuccinate synthetase immunoreactivity was detected in discrete populations of neurons throughout the brain. Double-staining with nicotinamide adenine dinucleotide phosphate (reduced form)-diaphorase histochemistry for the localization of nitric oxide synthase demonstrated that argininosuccinate synthetase coexists with nitric oxide synthase in some brain regions. However, many neurons were found that contained one of these two enzymes, but not the other. Thus some nitric oxide synthase-containing neurons appear able to recycle citrulline via argininosuccinate, while others do not. Additional roles for argininosuccinate synthetase in the brain are discussed.     

急性缺氧小鼠海马CAl区NOS活力和nNOS蛋白表达的时程变化

目的:观察急性缺氧小鼠海马CAl区一氧化氮合酶(NOS)和神经元型一氧化氮合酶(nNOS) 阳性神经元的时程变化,探讨NO在脑缺氧中的作用并为抗脑缺氧提供依据。方法:复制小鼠急性缺氧模型,采用NADPH-d组织化学和nNOS免疫组织化学方法,研究急性缺氧后不同时程点小鼠海马CAl区NADPH-d 和nNOS阳性神经元数量的变化。结果:与正常对照组相比较,急性缺氧后0.5h组小鼠海马CAl区NADPH-d 和nNOS阳性神经元的数量无明显变化,差异无显著性(P>0.05),3h、6h和12h组逐渐增多并于12h升高达到最高峰,差异有显著性(P<0.05),而于24h后开始降低,48h恢复正常。结论:急性缺氧后早期海马CAl区NOS和nNOS水平明显增多,NO在缺氧所致早期脑损伤中起重要作用。     

Distribution of NADPH-diaphorase and nitric oxide synthase-containing neurons in the intramural ganglia of guinea pig urinary bladder

The cell population and distribution of NADPH-diaphorase positive and NOS immunoreactive intramural ganglion cells were examined on stretched whole-mount preparations of the guinea pig urinary bladder which was divided into 3 regions: base, body and dome. The results showed that the highest frequency both of NADPH-d and NOS positive neurons was observed in the bladder base. Cell counts in the whole bladder showed that the number of NADPH-d positive neurons was much more than that of NOS immunoreactive neurons. Using neuron specific enolase (NSE) positive neurons as a reference (100%), NADPH-d positive neurons accounted for 84% while NOS immunoreactive neurons only made up 45% of the total neuronal population. These results, along with previous studies on the function of nitric oxide, suggest that nitric oxide may be involved in the relaxation activity in the bladder base during micturition. The significant difference in the number of NADPH-d positive and NOS immunoreactive neurons suggests that the localisation of one enzyme does not necessarily reflect the presence of the other.     

大鼠下丘脑一氧化氮合酶（NOS）阳性神经元的分布

观察大鼠下丘脑各核团NOS阳性神经元的分布。采用还原型尼克酰胺腺嘌呤二核苷酸脱氢酶（NADPH－d）法，结果显示，大量NOS阳性神经元见于下丘脑外侧区、视上核（SO）和室旁核（Pa）；出现较多NOS阳性神经元的部位是视前大细胞核，见到少量NOS阳性神经元的部位是室周核、视前内侧区、视前外侧区和下丘脑前区。结论：NOS阳性神经元分布于下丘脑的许多核团。     

Neuronal nitric oxide synthase (nNOS) immunoreactivity in the olfactory system of a cartilaginous fish

Nitric oxide is a regulative molecule with important roles in the olfactory system of vertebrates. Chondrichtyans have a key position in vertebrate evolution and nothing is known about nitric oxide in their olfactory system. Aim of this work was to investigate the neuronal nitric oxide synthase (nNOS) immunoreactivity in the olfactory system of the shark Scyliorhinus canicula. Because nitric oxide is often related to GABA in the olfactory system, also the distribution of GABA and its synthesis enzyme GAD has been investigated. In the olfactory epithelium scattered cells in the basal and medial zone of the epithelium thickness presented nNOS-like immunoreactivity. In the olfactory bulb the nNOS-like immunoreactivity has been highlighted in nerve fibers around some blood vessels and in scattered GABAergic granule cells. The presence of nNOS in the olfactory system of S. canicula is overall lesser than that described in other vertebrates, even if nitric oxide probably keeps some essential functions.     

Heterogeneity of nitric oxide synthase-containing neurons in the mouse main olfactory bulb

The distribution and structural features of nitric oxide [corrected] synthase (NOS) containing intrinsic neurons were studied in the mouse main olfactory bulb (MOB). NOS positive neurons were heterogeneous, including some subpopulations of periglomerular cells, granule cells, interneurons in the external plexiform layer, superficial and deep short-axon cells and stellate cells. NOS positive periglomerular cells were frequently calretinin immunoreactive and, although rarely, calbindin positive. Importantly, some middle and external tufted cells were also confirmed to be NOS positive, some of which were also cholecystokinin (CCK) positive. Retrograde tracer experiments showed that some NOS positive tufted cells, which were also CCK positive, constitute the intrabulbar association system and the projection system to the olfactory tubercle. In addition, another particular subpopulation of NOS positive neurons with no or little CCK immunoreactivity appeared to project to areas covering the dorsal endopiriform nucleus, claustrum and insular cortex. Furthermore, diverse types of neurons other than mitral/tufted cells were also suggested to be projection neurons of the MOB. The present study revealed the diversity of NOS positive neurons in the mouse MOB and further revealed that they were different from those reported previously in the rat MOB in structural and chemical properties.     

Induction of inducible nitric oxide synthase expression in activated microglia following domoic acid (DA)-induced neurotoxicity in the rat hippocampus

Neuronal degeneration followed by glial activation (microglia and astrocytes) and nitric oxide synthase (NOS) expression in the hippocampus was investigated at 3 months after domoic acid (DA) administration and compared with DA treated rats at 5 days time interval which was reported earlier. Massive degeneration with complete absence of neurons in the hippocampal CA1 and CA3 regions and hypertrophied microglial cells showing intense immunoreaction with the antibody OX-42 was observed at 3 months after DA administration. Sparsely distributed OX-42 positive microglial cells were observed in the hippocampus of control rats at 3 months after saline treatment No apparent changes could be observed in the immunoreactivity of GFAP at 3 months after saline and DA administration. Neuronal nitric oxide synthase immunoreactive neurons were completely absent in the hippocampus at 3 months after DA administration. In contrast, nicotinamide adenine dinucleotide phosphate diaphorase (NADPH-d) histochemical analysis revealed absence of NADPH-d reactivity in the neurons, but positive reactivity in the microglial cells of CA1-CA3 regions in the hippocampus after DA treatment. Double immunofluorescense revealed co-expression of inducible nitric oxide synthase with immunoreactive OX-42 positive microglial cells in the hippocampal subfields at 3 months after DA administration. The microglia-produced NO appears to be a secondary phenomenon in the prolonged inflammatory process following DA-induced neuronal degeneration.     

Localization of C-PON immunoreactivity in the rat main olfactory bulb. Demonstration that the population of neurons containing endogenous C-PON display NADPH-diaphorase activity

The presence of the neuropeptide C-terminal flanking peptide of neuropeptide-Y, C-PON, has been investigated in the main olfactory bulb of the rat using conventional fluorescence and peroxidase-antiperoxidase immunocytochemical techniques. The distribution of immunoreactive structures to C-PON was examined in both horizontal and coronal sections. Endogenous C-PON was localized within two types of short-axon cells including (1) superficial short-axon cells in the glomerular layer and (2) deep short-axon cells lying in the deepest portion of the granule cell layer and in the adjacent white matter. In addition, varicose immunoreactive processes were detected in all layers, although they were more numerous in the deepest portion of the granule cell layer. Immunoreactive cell bodies and processes were also observed in the nucleus olfactorius anterior and in the intrabulbar portion of the anterior commissure. Nevertheless, immunoreactive structures were not localized in the lateral olfactory tract. The indirect immunofluorescence technique to detect endogenous C-PON in combination with the enzyme histochemical demonstration of NADPH-diaphorase activity, in single sections, showed that the NADPH-diaphorase procedure is a reliable marker for these C-PON positive cells. Also, indirectly, that, in the rat main olfactory bulb, C-PON and neuropeptide-Y are contained in the same cell types. Many glomeruli were stained following the NADPH-diaphorase procedure, but they were not C-PON immunoreactives. Results of this study provide evidence suggesting that C-PON may influence polysynaptically the function of mitral cells and, therefore, the olfactory bulb output.     

Nitric oxide synthase protein levels, not the mRNA, are downregulated in olfactory bulb interneurons of reeler mice

Homozygous mutations in the Reelin gene result in severe disruption of brain development. The histogenesis of layered regions, like the neocortex, hippocampus and the cerebellum, is most notably affected in mouse reeler mutants and similar traits are also present in mice lacking molecular components of the Reelin signalling pathway. Moreover, there is evidence for an additional role of Reelin in sustaining synaptic plasticity in adult networks. Nitric oxide is an important gaseous messenger that can modulate neuronal plasticity both in developing and mature synaptic networks and has been shown to facilitate synaptic changes in the hippocampus, cerebellum and olfactory bulb. We studied the distribution and content of neuronal nitric oxide synthase in the olfactory bulbs of reeler and wildtype mice. Immunocytochemistry reveals that Reelin and neuronal nitric oxide synthase containing interneurons are two distinct, non overlapping cell populations of the olfactory bulb. We show by in situ hybridization that both nitrergic and Reelin expressing cells represent only a subset of olfactory bulb GABAergic neurons. Immunoblots show that neuronal nitric oxide synthase protein content is decreased by two thirds in reeler mice causing a detectable loss of immunolabelled cells throughout the olfactory bulb of this strain. However, neuronal nitric oxide synthase mRNA levels, essayed by quantitative real-time RT-PCR, are unaffected in the reeler olfactory bulb. Thus, disruption of the Reelin signalling pathway may modify the turnover of neuronal nitric oxide synthase in the olfactory bulb and possibly affects nitric oxide functions in reeler mice.     

3H-thymidine-radiographic studies of neurogenesis in the rat olfactory bulb

Neurogenesis in the rat olfactory bulb was examined with 3H-thymidine-radiography. For the animals in the prenatal groups, the initial 3H-thymidine exposures were separated by 24 h; they were the offspring of pregnant females given two injections on consecutive embryonic (E) days (E12-E13, E13-E14, . . . E21-E22). For the animals in the postnatal (P) groups, the initial 3H-thymidine injections were separated by 48 h, each group receiving either four (PO-P3, P2-P4, . . . P6-P9) or two (P8-P9, P10-P11, . . . P20-P21) consecutive daily injections. On P60, the percentage of labeled cells and the proportion of cells added during either 24 h or 48 h periods were quantified at several anatomical levels for each neuronal population in the main olfactory bulb (mitral cells, tufted cells, granule cells, interneurons in the external plexiform layer, periglomerular granule cells) and accessory olfactory bulb (output neurons, granule cells, periglomerular granule cells). The total time span of neurogenesis extends from E12 to beyond P20. Output neurons are prenatally generated over 5-9 day periods (with most neurogenesis occurring over 2-4 days) in a strict sequential order beginning with the accessory bulb output neurons (E13-E14) and ending with the interstitial tufted cells lying between the glomeruli in the main bulb (E20-E22). These data are correlated with the main and accessory bulb projection fields in the amygdala and with the chronology of amygdala neurogenesis. With the exception of the granule cells in the accessory bulb (88% generated between E15-E22), the rest of the interneuronal populations are generated postnatally and nearly simultaneously. While most neurons (75-80%) originate during the first three weeks of life, all interneuronal populations, including accessory bulb granule cells, show some neurogenesis beyond P20. Injections of 3H-thymidine in juvenile and adult rats indicates neurogenesis up to P60 in the accessory bulb and up to P180 in the main bulb, especially in the main bulb granule cell population. There is circumstantial evidence for turnover of main bulb granule cells during adult life.     

肝性脑病大鼠海马CA3区神经元形态和一氧化氮合酶表达的变化

目的观察肝性脑病模型组和正常对照组大鼠脑海马CA3区神经元的变化及一氧化氮合酶(NOS)的表达;探讨海马CA3区神经元的形态学改变及一氧化氮(NO)在肝性脑病发病机制中的作用。方法雄性大鼠50只,实验开始前所有动物均进行莫里斯水迷宫测试,之后将动物分为对照组和实验组。9周后建立CCL4肝性脑病模型,分别取两组大鼠海马组织进行尼氏染色及烟酰胺腺嘌呤二核苷酸-黄递酶(NADPH-d),染色。结果尼氏染色发现,实验组大鼠海马神经元数目减少、染色较浅,胞质内尼氏体减少或消失;NADPH-d染色发现,实验组可见粗大轴突着色,树突联系广泛;对照组则少有粗大轴突着色,树突间联系不如实验组广泛。实验组NOS阳性神经元染色较对照组深,为紫蓝或深蓝色(强阳性及阳性),且阳性神经元数目较多;而对照组染色浅淡,呈浅蓝或与背景同色,为弱阳性。结论肝性脑病时海马受到损伤,NO可能介导了神经元的损伤并参与了肝硬化和肝性脑病的发病,血氨升高是肝性脑病(HE)致病因素之一。     

PKC抑制剂灯盏花素乙对甲醛炎性痛大鼠脊髓NOS表达及NO含量的影响

目的：观察蛋白激酶C（PKC）选择性抑制剂灯盏花素乙（CH）对甲醛炎性痛时大鼠自发痛反应、脊髓一氧化氮合酶（NOS）表达和一氧化氮（NO）含量的影响，探讨炎性痛时脊髓内NO产生是否受PKC调控。 方法： 采用右后掌足底注射甲醛复制炎性痛模型；计数缩足反射次数反映自发痛程度；应用NADPH-d组织化学法测定脊髓NOS表达；硝酸/亚硝酸还原法测定脊髓腰膨大部位NO2-/NO3-含量。 结果： 甲醛炎性痛大鼠L5脊髓后角浅层和中央管周围灰质NADPH-d阳性细胞的数目、阳性细胞胞体及纤维的染色深度均明显大于正常对照组，脊髓腰膨大部位 NO2-/NO3-含量明显增高。预先鞘内给予CH，可明显抑制甲醛炎性痛诱导的大鼠第二相自发痛反应以及脊髓NOS表达和NO2-/NO3-的含量。 结论： 炎性痛时，脊髓伤害性感受神经元内PKC激活可以促进NOS表达和NO的产生。     

Distribution and expression pattern of the nitrergic system in the cerebellum of the sheep

The nitrergic system produces nitric oxide as an atypical neurotransmitter in the nervous system. Nitric oxide is produced from l-arginine through specific enzymes known as nitric oxide synthases. Of these, the more abundant form in neurons is the constitutive neuronal nitric oxide synthase, although the inducible isoform can be expressed as well, especially following stress or other injuries. The excessive formation of nitric oxide results in protein nitration, particularly at tyrosine residues, thus the presence of nitrotyrosine can be used as a marker of nitric oxide production. In previous studies we have shown the distribution of the components of the nitrergic system in the cerebellum of rodents, where neuronal nitric oxide synthase immunoreactivity was present in stellate and basket cells, and occasionally in granule cells. Here, we present evidence that in the sheep, as a model of larger mammals, most cerebellar neurons display an intense immunostaining for neuronal nitric oxide synthase, including unipolar brush cells, and Lugaro and Golgi neurons, which are not immunoreactive in rodents. In addition, weak immunoreactivity for inducible nitric oxide synthase and nitrotyrosine was found in particular cell types, indicating a basal expression for these markers. Our results suggest a larger dependence on the nitrergic system for the cerebella of larger mammals. Since this increase happens in both activating and inhibitory neurons of the cerebellar circuitry, we propose that in these animals there is a higher steady-state regulation of the cerebellum based on nitric oxide.     

A Golgi study on the olfactory bulb in the lamprey, Lampetra japonica

The intrinsic organization of the olfactory bulb in the lamprey was studied using the rapid Golgi method. Although not as discrete as in many vertebrates, a laminar organization was recognized. From the periphery inward, the following layers were discernible: the layer of the olfactory fibers, the olfactory glomeruli with the mitral cells, the granule cells, and the ependymal cells. Just beneath the surface of the olfactory bulb, the olfactory fibers extended over the entire bulb forming a dense fiber plexus terminating in the olfactory glomeruli which were arranged in one to two layers internally to the layer of the olfactory fibers. The mitral cells formed no discrete layer and were located mainly around the olfactory glomeruli. The mitral cells in the lamprey were lacking in secondary dendrites, but had two or more primary dendrites which terminated in the olfactory glomeruli. The axons of the mitral cells proceeded inwardly and accumulated diffusely in the granule cell layer which occupied a wide area internally to the layer of the olfactory glomeruli with the mitral cells. The granule cell layer was composed of densely packed small spindle or fusiform axonless cells, the processes of which extended superficially to be distributed in the olfactory glomeruli. At the deepest region of the bulb was a layer of the ependymal cells lining the surface of the olfactory ventricle. The external and internal plexiform layers were not evident. Thus, while the major constituents of the olfactory bulb of the vertebrate could be identified in that of the lamprey, the general laminar organization seemed indiscrete.     

大鼠端脑内一氧化氮合酶阳性神经元的发育

目的研究大鼠胚胎时期及生后早期一氧化氮合酶（NOS）阳性神经元在端脑的分布,探讨一氧化氮（NO）在脑发育过程中的作用。方法应用还原型尼克酰胺腺嘌呤二核苷酸磷酸脱氢酶（NADPH-d）组织化学方法观察孕14d起至生后14d大鼠端脑内NOS阳性神经元的形态和分布。结果孕14d没有观察到阳性神经元。孕15d纹状体腹外侧已有NOS阳性表达。孕17d在大脑皮质、梨状皮质观察到NOS阳性神经元,但胞体小,树突短,且分支少。随着年龄的增长神经元的胞体数目增多、染色增强或维持一定的水平。到孕20d,NOS阳性神经元分布广泛,梨状皮质、纹状体腹外侧及终纹床核均有大量NOS阳性神经元,其胞体明显增大,树突分支复杂化,长度增加。在生后,除上述脑区的阳性神经元进一步发育分化,大脑皮质和纹状体的NOS阳性纤维相互编织成疏密不等的纤维网外,在胼胝体、海马也观察到NOS阳性神经元。到生后14d,NOS阳性神经元的分布模式总体上已与成年大鼠相似。结论NOS阳性神经元在端脑独特的表达模式提示NO在脑发育和成熟过程中扮演重要角色。