GAP-43免疫反应神经纤维在小鼠脾脏的分布及其性质的研究

用免疫组化方法研究了GAP－43免疫反应阳性神经纤维在成年小鼠脾脏的分布及其性质。结果显示，GAP43样免疫反应阳性纤维不仅广泛分布于脾内血管周围，还存在于白髓、红髓的淋巴细胞之间。应用6-羟多巴（6－OHDA）损毁儿茶酚胺能神经后，TH、NPY免疫组化染色不能显示任何阳性神经纤维，而GAP－43免疫反应阳性神经纤维尚有部分存留．它们多位于较大的血管周围，较直且平滑、膨体少。此结果表明，成熟小鼠脾内绝大部分单胺能神经纤维和少部分非单胺能神经纤维含有GAP－43，是具有可塑性的神经纤维。     

日本猴坐骨神经皮支和肌支中NPY和VIP免疫反应阳性纤维的分布和…

结扎猴坐骨神经皮支和肌支近侧端，观察其中ＮＰＹ和ＶＩＰ免疫反应产物蓄积状况以及其所支配的肌肉和皮肤中ＮＰＹ，ＶＩＰ免疫反应阳性纤维的分布。结扎神经干７ｄ后，发现皮支的结扎近心端含有ＮＰＹ和ＶＩＰ；而肌支的结扎近心端只含ＮＰＹ，无ＶＩＰ免疫反应阳生产物。在皮肤，大量ＶＩＰ免疫反应阳性神经纤维分布在汁腺周围，含ＮＰＹ听神经纤维分小血管周围。肌肉组织中有害富的ＮＰＹ免疫反应阳性神经纤维沿血管分布，但几乎     

不同年龄大鼠脑内血管TH能神经分布的观察──免疫组织化学法研究

应用ＡＢＣ法，以酪氨酸羟化酶作为标记物，对幼年、成年、老年Ｗｉｓｔａｒ大鼠（每组各１０只）脑实质内血管的免疫阳性神经纤维的分布类型、纤维密度等进行了观察。结果是：在三组不同年龄大鼠脑皮质血管壁上均出现免疫反应阳性纤维。幼年鼠脑皮质血管神经纤维分布类型有环行型（占５８．１４％）和纵行型（占４１．８６％），纤维密度较小；成年鼠分布类型主要有网状型（占８９．１２％），其次为环行型（占８．３４％），少数为纵行型，纤维密度较大；老年鼠分布类型大多数为环行型（７６．０３％），其余为网状型（占２３．９７％），纤维密度明显较成年鼠稀疏。本文讨论了脑内血管神经纤维的分布类型、纤维密度与脑内微循环的神经源性调节的关系。     

日本猴坐骨神经皮支和肌支中NPY和VIP免疫反应阳性纤维的分布和起源的比较研究

结扎猴坐骨神经皮支和肌支近侧端，观察其中ＮＰＹ和ＶＩＰ免疫反应产物蓄积状况以及其所支配的肌肉和皮肤中ＮＰＹ、ＶＩＰ免疫反应阳性纤维的分布。结扎神经干７ｄ后，发现皮支的结扎近心端含有ＮＰＹ和ＶＩＰ；而肌支的结扎近心端只含ＮＰＹ，无ＶＩＰ免疫反应阳性产物。在皮肤，大量ＶＩＰ免疫反应阳性神经纤维分布在汗腺周围，含ＮＰＹ的神经纤维分布在小血管周围。肌肉组织中有丰富的ＮＰＹ免疫反应阳性神经纤维沿血管分布，但几乎见不到ＶＩＰ阳性神经纤维。用ＴｒｕｅＢｌｎｅ（ＴＢ）逆行追踪结合荧光免疫组化技术研究猴坐骨神经皮支和肌支含肽能神经的起源，发现标记物给予腓肠内侧皮神经末端时，于Ｌ７和Ｓ１椎旁交感节出现ＴＢ与ＶＩＰ、ＴＢ与ＮＰＹ双标细胞；而支配腓肠肌的肌支只在Ｌ６、Ｌ７发现ＴＢ和ＮＰＹ双标细胞而含ＴＢ的细胞与ＶＩＰ免疫反应阳性细胞完全不重叠。本研究证明日本猴坐骨神经的皮支和肌支中ＮＰＹ和ＶＩＰ免疫反应阳性神经的分布和起源有明显的差异。     

成年和发育中大鼠胼胝体轴突含生长抑素的免疫组织化学证据

本实验用免疫组织化学ＡＢＣ法发现大鼠胼胝体内存在生长抑素免疫反应阳性轴突．此阳性轴突可见于生后不同发育阶段及成年大鼠的胼胝体内．出生时胼胝体内此轴突的数量很少；生后１周内其密度逐渐增高，至第２周初期达到高峰，此时胼胝体内可见大量生长抑素阳性轴突．然而，至第２周末，生长抑素阳性胶胝体轴突的密度已下降至接近成年时的水平，即仅有少量此纤维存在于胼胝体内。这些结果提示在大鼠早期生后发育过程中许多生长抑素阳性胼胝体轴突是暂时性的．结合本文作者等以往对成年和发育中大鼠和猫的与胼胝体联系的神经介质的研究结果，可以认为胼胝体中的活性物质是多样的，在哺乳动物大脑皮质发育中可能起着不同的作用。     

大鼠生后脑下垂体前叶CGRP免疫反应阳性神经纤维的发育

应用免疫组织化学方法观察了大鼠生后其脑下垂体前叶 CGRP免疫反应阳性神经纤维的发育变化。 SD大鼠按生后天数分为 P1～ 2、P8、P15、P30、P45等五个实验组。结果证明 ,P1～ 2 d,从周围脑膜中来的 CGRP阳性神经纤维进入前叶浅表 ,形态较短直 ,前叶中央部未见阳性纤维。P8d,CGRP阳性纤维的数量较 P1～ 2 d者显著增多 ,CGRP阳性纤维细而弯曲 ,富含膨体 ,呈网状或簇状分布于前叶背侧中央部及吻侧部。P15 d,CGRP阳性纤维可分为粗直类纤维和弯曲的膨体型细纤维 ,前者分布于前叶大部分 ,后者主要呈网状或簇状分布于前叶吻端近腹侧部及背尾部。 P30 d,CGRP阳性纤维也分为粗直类纤维和弯曲的膨体型细纤维 ,粗直类纤维分布于整个垂体前叶 ;弯曲的膨体型细纤维较 P15 d者分布广泛。P45 d CGRP阳性纤维类型和分布接近成年大鼠。本研究结果表明 ,大鼠垂体前叶的 CGRP阳性神经纤维随出生后年龄的增长 ,其类型、数量、分布逐渐由少到多 ,由分布于浅表到广泛分布于整个垂体前叶 ,呈现一种动态变化 ,说明出生后大鼠垂体前叶中 CGRP阳性纤维发育迅速     

神经肤Y免疫反应阳性神经纤维在大鼠脾脏内的分布

用ABC免疫组织化学法结合葡萄糖氧化酶-DAB-硫酸镍铵(GDN)显色技术,研究神经肽Y(NPY)免疫反应阳性神经纤维在大鼠脾脏内的分布。结果表明,在脾脏内有较丰富的NPY免疫反应(NPY-IR)阳性神经纤维,它们多呈串珠状,主要伴动脉及其分支走行,也见于脾被膜的结缔组织内,白髓、红髓和边缘区等部位的淋巴组织中,以及脾血窦周围。脾内NPY免疫反应阳性神经纤维与血管和淋巴细胞的关系密切,提示它们对脾淋巴细胞的发育和功能可能有调节作用。NPY可能直接作用于淋巴细胞或通过调节脾的血液循环间接地发挥作用。     

哺乳动物垂体后叶比较化学神经解剖学 Ⅱ.NT和CCK免疫阳性神经纤维在猴、狗、大鼠垂体后叶的比较分布

本文报道了神经紧张素和胆囊收缩素免疫阳性神经纤维不仅存在于大鼠垂体后叶，也存在于猴和狗的垂体后叶.神经紧张素免疫阳性神经纤维在3种动物垂体后叶均为少量稀疏分布，狗和大鼠稍密集，猴最少;猴和狗垂体柄及其邻近的神经叶相对较多，大鼠垂体后叶形成弥散的阳性纤维网，局部可见岛状增密的纤维簇。胆羹收缩素阳性纤维以大鼠垂体后叶密度为最大，狗次之，猴最少;大鼠垂体后叶的中等密度的胆囊收缩素阳性纤维网，吻侧部多于尾侧部，周边区比中央区稍密;猴、狗垂体后叶胆囊收缩素阳性纤维为散在分布。本文还注意到一些神经紧张素和胆翼收缩素免疫阳性神经纤维盘绕于血管分布的现象，其功能意义尚不清楚。     

P物质免疫反应阳性神经纤维在雌性大鼠生殖器官的分布

目的和方法:实验采用亲和免疫组化SABC法对雌性大鼠动情间期P物质的正常分布进行了观察.结果:P物质免疫反应阳性纤维在子宫角与阴道分布最为丰富,宫颈和宫体分布中等,卵巢内免疫阳性神经纤维较少,且免疫阳性神经纤维在子宫角有从卵巢端向阴道端递减的趋势,在各管状生殖道,免疫阳性神经纤维以内膜和外膜分布较多.结论:分析研究雌性大鼠生殖器官内P物质免疫反应阳性神经纤维的分布,提示P物质可能与卵泡发育、性激素的产生、血管及非血管平滑肌的运动调节有关.     

哺乳动物垂体后叶比较化学神经解剖学Ⅱ.NT和CCK免疫阳性神经纤维在？…

本文报道了神经紧张素和胆囊收缩素免疫阳性神经纤维不仅存在于大鼠垂体后叶，也存在于猴和狗的垂体后叶。神经紧张素免疫阳性神经纤维在３种动物垂体后叶均为少量稀疏分布，狗和大鼠稍密集，猴最少；猴和狗垂体柄及其邻近的神经叶相对较多，大鼠垂体后叶形成弥散的阳性纤维网，局部可见岛状增密的纤维簇。胆囊收缩素阳性纤维以大鼠垂体后叶密度为最大，狗次之，猴最少；大鼠垂体后叶的中等密度的胆囊收缩素阳性纤维网，吻侧部多于尾     

Development of growth-associated protein-43 and neurotrophin receptor immunoreactivities in the rat urinary bladder

Little is known about how the prenatal innervation of the urinary bladder develops. The objective of this study was to define the timing and pattern of innervation of the developing rat bladder using growth-associated protein-43 (GAP-43) immunohistochemistry. In addition, we examined the development of two different classes of transmembrane receptor proteins, the Trk family of tyrosine kinases and neurotrophin receptor p75 immunoreactivities. GAP-43-positive nerves were first detected in the bladder at embryonic day (E) 16. They were growing rapidly to the bladder dome and covered the whole bladder by E18. The density of GAP-43-containing nerves increased in the muscle layer by postnatal day (P) 0. Both Trk receptor-positive and p75 neurotrophin receptor-positive fibers were also first seen at E16. The number of Trk immunoreactive nerves reached a peak at E18, and then decreased over the following days. In contrast, p75-labeled fibers were abundant at E18-P14. There were few GAP-43 or neurotrophin receptor-positive fibers in the adult. GAP-43 immunohistochemistry provided us with a picture of innervation in the developing rat bladder. Furthermore, the demonstration of neurotrophin receptors positive fibers in the prenatal and early postnatal bladders suggests that neurotrophins may contribute to the development of the peripheral nervous system in the urinary bladder.     

Immunohistochemical distribution of growth-associated protein 43 (GAP-43) in developing rat nasoincisor papilla

We employed immunohistochemistry of growth-associated protein 43 (GAP-43) to trace the early development of gustatory nerves and alpha-gustducin to demonstrate mature taste buds in the rat nasoincisor papilla (NP). The sequential changes of gustatory structures revealed eight characteristic stages. One, at embryonic day 16 (E16), GAP-43-immunoreactive (IR) nerve fibers were observed in close relation with presumptive taste buds in the lateral apical epithelium on each side of NP; meanwhile, no immunoreactivity could be observed in the papillary epithelium. Two, at E17, fine GAP-43-IR nerve fibers first invaded the apical epithelium of the papilla. Three, at E19, GAP-43-IR nerve fibers were extensive in apical epithelium and colonized in immature taste buds. Four, at E20, GAP-43-IR nerve fibers were first observed in ductal epithelium (lining the medial wall of nasoincisor ducts). Five, at postnatal day 1 (P1), immunoreactive nerve fibers first coincided with immature taste buds in the ductal epithelium. Six, at P3, alpha-gustducin-IR cells identical for mature taste buds were simultaneously demonstrated in both apical and ductal epithelium. Seven, at P14, progressive taste bud proliferation and maturation as well as neural invasion were demonstrated in all regions of the epithelium. Eight, during advanced stage in adult animals, extensive innervation was traced especially in close relation with taste buds. The sequential topographic patterns of NP gustatory structures seem very specific as compared to those in other locations of mammalian gustatory system. The present study reveals that gustatory nerves preceded the development of taste buds. However, further investigations are required to examine such a characteristic model for the neurogenic theory of taste induction.     

自体移植脾组织内GAP-43神经的生成机制

为探索自体移植脾组织内GAP-43神经的再生机制,将Wistar大鼠42只随机分为实验组和对照组。实验组切除脾脏以后,切取1／2脾脏,切成1 mm×1 mm×1 mm大小的组织块植入大网膜内,术后7、14、30、60、90、120、180 d取脾组织标本通过原位杂交检测GAP-43 mRNA、NGF mRNA和TrkA mRNA,同时进行免疫组化染色观察GAP-43神经。对照组手术松动脾脏,在术后相同时间点取脾组织作对照观察。结果显示:术后30 d检测到移植脾组织内有GAP-43 mRNA、NGF mRNA和TrkA mRNA表达,90 d达高峰后开始下降;术后60 d明显可见GAP~43染色阳性神经纤维,90 d密度最大,主要存在于血管周围,以后无明显改变。结果提示:自体移植脾组织内GAP-43神经再生与内源性NGF和TrkA表达密切相关。     

Sympathetic innervation of fetal mouse thymus

This study was carried out to investigate the presence of sympathetic nerve fibers in the developing thymus. Thymic rudiments from 13 days of gestation up to birth and adult thymuses were examined by histofluorescence microscopy to detect the presence of these nerve fibers during ontogeny. Sympathetic nerve fibers were first visible around day 17 of gestation and increased in density and distribution by the time of birth and in the adult thymus. The appearance of immunoreactive cells about the same time as the presence of nerve fibers in the thymic rudiments has been discussed in light of a regulatory role for the sympathetic neuronal input on the maturation of thymocytes during ontogeny.     

Developmental regulation of parasympathetic nerve density by sympathetic innervation in the tarsal smooth muscle of the rat.

The developmental influence of sympathetic innervation on parasympathetic nerve density was investigated in the tarsal smooth muscle of the rat. Specificity of acetylcholinesterase staining as a marker for parasympathetic innervation was first determined by acute selective denervations. Excision of the ipsilateral superior cervical ganglion caused a 39% reduction in the density of acetylcholinesterase-positive nerves seven days later, indicating that sympathetic nerves contribute to cholinesterase-positive tarsal muscle innervation. Excision of the pterygopalatine ganglion concurrent with superior cervical ganglionectomy caused a virtually complete disappearance of acetylcholinesterase-positive innervation within seven days, indicating that non-sympathetic cholinesterase-positive fibers derive from the pterygopalatine ganglion and are presumed to be parasympathetic. Analysis of the control population indicated that parasympathetic nerve density did not vary significantly between males and females, between the superior and inferior muscles, or in rats studied at four and 12 months of age. The influence of sympathetic innervation on parasympathetic nerve density during postnatal development was examined by conducting surgical sympathectomies on postnatal day 5 and quantifying acetylcholinesterase-positive nerve density at four months of age. Neonatal sympathectomy caused a 46% reduction in cholinesterase-positive nerve density beyond that which occurred in acutely sympathectomized adult controls. It is concluded that sympathetic innervation is required for developing parasympathetic nerves to attain their normal density within the rat tarsal muscle. This finding is consistent with the idea that sympathetic nerves can exert positive effects on parasympathetic nerve outgrowth during development.     

Growth-associated protein-43 is elevated in the injured rat sciatic nerve after low power laser irradiation

Low power laser irradiation (LPLI) has been used in the treatment of peripheral nerve injury. In this study, we verified its therapeutic effect on neuronal regeneration by finding elevated immunoreactivities (IRs) of growth-associated protein-43 (GAP-43), which is up-regulated during neuronal regeneration. Twenty Sprague-Dawley rats received a standardized crush injury of the sciatic nerve, mimicking the clinical situations accompanying partial axonotmesis. The injured nerve received calculated LPLI therapy immediately after injury and for 4 consecutive days thereafter. The walking movements of the animals were scored using the sciatic functional index (SFI). In the laser treated rats, the SFI level was higher in the laser treated animals at 3-4 weeks while the SFIs of the laser treated and untreated rats reached normal levels at 5 weeks after surgery. In immunocytochemical study, although GAP-43 IRs increased both in the untreated control and the LPLI treated groups after injury, the number of GAP-43 IR nerve fibers was much more increased in the LPLI group than those in the control group. The elevated numbers of GAP-43 IR nerve fibers reached a peak 3 weeks after injury, and then declined in both the untreated control and the LPLI groups at 5 weeks, with no differences in the numbers of GAP-43 IR nerve fibers of the two groups at this stage. This immunocytochemical study using GAP-43 antibody study shows for the first time that LPLI has an effect on the early stages of the nerve recovery process following sciatic nerve injury.     

Developmental changes in pulpal sensory innervation of rat incisors and molars shown on a single injection of the fluorescent dye AM1-43

Developmental changes in pulpal innervation of rat incisors and molars were examined using the fluorescent styryl dye AM1-43, which labels sensory cells and nerves in vivo. From 2 to 40 days after birth, the animals were injected once with a small amount of AM1-43 solution (2 microg/g bodyweight). One day after the injection, the animals were killed and examined. In 3-day-old rats, neither incisors nor molars were innervated. In 7-day-old rats, the pulp of incisors and molars was innervated as indicated by fine intensely stained varicose sensory fibers and thicker intensely stained fibers running mostly along the blood vessels. In 15-, 27-, and 41-day-old rats, sensory nerve fibers neither passed through the odontoblast layer nor penetrated into the dentin in incisors, whereas the sensory nerve fibers penetrated into the coronal dentin through the odontoblast layers in molars. These results suggest that innervation of dental pulp is composed of two phases: (i) linear penetration of nerve fibers along blood vessels into the pulp space; and (ii) sprouting and extension of nerve fibers into coronal dentin. Innervation of the incisor pulp may stop at the first phase.     

The rat renal nerves during development

The prenatal and postnatal development of the innervation of the rat kidney has been investigated using immunocytochemical methods. The efferent innervation was studied using dopamine-beta-hydroxylase and neuropeptide Y antibodies. Calcitonin gene related peptide and substance P antibodies were used to investigate the afferent innervation. Kidneys from embryos of 14 to 20 days, from newborn rats, and from animals of 4, 10, 12, 21, 38, 60, and 90 days of age were studied. Slices of whole kidneys were analyzed, and frozen sections were used to investigate the location of the nerves in more detail. Both afferent and efferent nerves are observed inside the kidney by embryonic day 16. At birth, the afferent nerves are found (1) forming a rich plexus in the renal pelvis; (2) associated with the renal vasculature as far as the interlobular arteries (cortical radial arteries) and (3) in the corticomedullary connective tissue. The efferent innervation appears, at birth, to extend to the interlobular arteries and to the afferent arterioles of the perihilar juxtamedullary nephrons. The efferent innervation increases rapidly during the following days, and by postnatal day 21 a distribution of the innervation similar to that of the adult is observed. While the afferent innervation reaches the major target regions of the kidney by birth, the efferent does most of its expansion into the kidney postnatally. Afferent and efferent fibers are found, extrarenally and intrarenally, in the same nerve bundles. This proximity between afferent and efferent fibers may represent anatomical bases for their interaction in the adult as well as during development.Supported by U.S. Public Health Service Grant Rol 18340 from the National Institute of Health     

GAP-43 in the axons of mammalian CNS neurons regenerating into peripheral nerve grafts

Summary Although mature mammalian CNS neurons do not normally regenerate axons after injury, it is well established that they will regrow axons over long distances into peripheral nerve implants. We have autografted segments of sciatic nerve into the brains of adult albino rats and have used light and electron microscopic immunocytochemistry to examine the distribution of the growth associated protein GAP-43 in and around the graft in the first two weeks following implantation. GAP-43 was present, 3–14 days after grafting, in small non-myelinated axonal sprouts in the brain parenchyma around the proximal tip of the graft. At 11–14 days after implantation similar sprouts within the graft itself were GAP-43 immunoreactive. The sprouts were either naked or associated with other cell processes (chiefly of Schwann cells; to a lesser extent of astrocytes). We also show that small numbers of neuronal perikarya around the tip of the graft become GAP-43 immunoreactive 11–14 days after implantation. Thus mature mammalian CNS neurons regenerating axons into a PNS graft display a marked increase in their content of GAP-43. In addition, we report that small plaques of GAP-43 reaction product are sometimes present on the plasma membranes of Schwann cells or astrocytes adjacent to immunoreactive axons, and that narrow sheet-like or filopodial processes of astrocytes, Schwann cells and possibly other non-neuronal cell types, may contain small amounts of GAP-43.