Ultrastructural identification of degenerating boutons of monosynaptic pathways to the lumbosacral segments in the cat after spinal hemisection

Summary Electrophysiological results suggest that the supra-spinal monosynaptic pathways to the lumbosacral motoneurones in cats are located in the vestibulospinal and reticulospinal tracts. In the present study these pathways were interrupted in the upper thoracic region by hemisection of the spinal cord. The boutons undergoing degeneration were then studied both qualitatively and quantitatively by means of serial and single sections of selected areas of the ventral horn in spinal cord segments L 6, L 7 and S 1. The degenerating boutons were distributed over the cell soma and larger dendrites (but not the most proximal portions) of the motoneurones. Two days postoperatively boutons showing signs of degeneration were readily identified in the latero-ventral motoneurone nucleus and Rexed's lamina VIII. These boutons which were swollen had a greatly increased number of neurofilaments and a decreased number of synaptic vesicles. Five days postoperatively these boutons which were now shrunken, contained swollen mitochondria and a dense, granular material. Between 5 and 8 days post-operatively the boutons were invaded by glial cell processes and digested. A small percentage of the F- and S-type boutons (approximately 0.5 and 3.0% respectively) were identified as forming the terminals of the experimentally interrupted pathways. This study did not exclude the possibility that a proportion of these boutons were the terminals of the propriospinal, monosynaptic fibers derived from the cervical region of the cord.On sabbatical leave from the Department of Zoology, University of Melbourne     

Projections from the lateral vestibular nucleus to the spinal cord in the mouse

The present study investigated the projections from the lateral vestibular nucleus (LVe) to the spinal cord using retrograde and anterograde tracers. Retrogradely labeled neurons were found after fluoro-gold injections into both the cervical and lumbar cord, with a smaller number of labeled neurons seen after lumbar cord injections. Labeled neurons in the LVe were found in clusters at caudal levels of the nucleus, and a small gap separated these clusters from labeled neurons in the spinal vestibular nucleus (SpVe). In the anterograde study, BDA-labeled fiber tracts were found in both the ventral and ventrolateral funiculi on the ipsilateral side. These fibers terminated in laminae 6–9. Some fibers were continuous with boutons in contact with motor neurons in both the medial and lateral motor neuron columns. In the lumbar and sacral segments, some collaterals from the ipsilateral vestibulospinal tracts were found on the contralateral side, and these fibers mainly terminated in laminae 6–8. The present study reveals for the first time the fiber terminations of the lateral vestibular nucleus in the mouse spinal cord and therefore enhances future functional studies of the vestibulospinal system.     

Ultrastructure of supraspinal dorsal root projections in the toad. II. The cerebellar granular layer

Summary Following section of the left dorsal roots, degenerating fibres and boutons were observed in the granular layer of the ipsilateral cerebellum. The degenerating terminals were identified as largeen passant varicosities of mossy fibres contacting the dendrites of presumptive granule cells. They contained round synaptic vesicles and neurofilaments and established Gray type I contacts. The terminals initially underwent filamentous degeneration with neurofilamentous hypertrophy, swollen mitochondria and loss of synaptic vesicles. At later survival times (6–30 days) they acquired an electron-dense appearance due to an increase and clumping of the filamentous component.After injection of horseradish peroxidase into the left cerebellum, all ipsilateral spinal ganglia showed a few (2–3%) labelled cells, indicating that a primary afferent contribution to this pathway originated from each segment of the spinal cord.     

Ipsilateral, ventral corticospinal tract of the adult rat: ultrastructure, myelination and synaptic connections

The corticospinal tract (CST) of the rat is a widely used model system in developmental, physiological, and regeneration studies. The CST of the rat consists of a main tract, that runs in the dorsomedial funiculus and several minor components. We have shown earlier that one of the minor components, the ipsilateral, ventral CST, projects all the way down the spinal cord in the adult rat and single fibers form large terminal arbors in their spinal target areas. Here we investigated its ultrastructure and compared it to that of CST fibers of the main tract. By the use of anterograde axonal tracing with biotin dextran-amine (BDA) and pre-embedding avidin-peroxidase histochemistry we investigated axon diameters and myelination using electron microscopy. Ipsilateral, ventral CST fibers were found to run in the ventral funiculus close to the midline. They were intermingled with heavily myelinated large diameter axons, presumably reticulospinal, vestibulospinal, or tectospinal fibers. Ipsilateral, ventral CST fibers were of small diameter (0.68 m, ±0.04) and about [frac34] of them were moderately myelinated (9.64 ± 0.7 layers of myelin). Co-localization of a rhodamine-dextrane anterograde tracer with the presynaptic marker synaptophysin using confocal microscopy and electron microscopy revealed varicosities on terminal arborisations to be presynaptic boutons and clearly demonstrated contacts to neurons in intermediate laminae of the spinal cord at lumbar spinal levels. This study extends our earlier work indicating that the ipsilateral, ventral CST component of the adult rat is a morphologically complete CST component and may perform similar functions to the main CST component.     

Electron microscopic investigation of the vestibular projection to the cat trochlear nuclei

Ultrastructural degeneration studies were carried out on the cat trochlear nucleus following lesion of the vestibulo-trochlear pathway in order to characterize the location and type of presynaptic endings involved in this pathway. Four types of boutons are found in the normal trochlear nucleus. Types I and II are large and demonstrate typical en passant profiles with small diameter synaptic vesicles (35 and 40 nm). These terminals are characterized by the absence of neurofilaments in the Type II endings. Types III and IV are smaller boutons, located more axondendritically, and contain larger diameter synaptic vesicles (45 nm). Type V terminals contain large, granulated vesicles and occur only rarely. Following the interruption of the ascending projection from the ipsilateral superior and medial vestibular nuclei by parasagittal medullary lesions, degeneration of Type II boutons was commonly encountered in the ipsilateral trochlear nucleus. Predominantly Type III degeneration was found in the contralateral trochlear nucleus. Electrical stimulation of the vestibular nerve showed that these lesions resulted in (1) a complete loss of inhibition in the ipsilateral trochlear nucleus and (2) a significant (75-90%) reduction in the contralateral excitatory pathway to the trochlear nucleus. Midline sagittal lesions in the floor of the fourth ventricle interrupting the decussating fiber projection from the bilateral medial vestibular nuclei resulted in selective degeneration of only Type III boutons in both trochlear nuclei. We conclude that inhibitory vestibular neurons eminating from the superior vestibular nucleus terminate on trochlear motoneurons with Type II boutons and excitatory vestibular neurons from the contralateral medial vestibular nucleus end on trochlear motoneurons with Type III boutons.     

Light microscopic and ultrastructural localization of immunoreactive substance P in the dorsal horn of monkey spinal cord

Light- and electron-microscopic localization of substance P in the monkey spinal cord was studied by the peroxidase anti-peroxidase technique with the particular aim of examining types of interactions made by substance P-positive boutons with other neuronal elements in the dorsal horn. By light-microscopy dense labeling for immunoreactive substance P was found in laminae I, II (outer zone) and V (lateral region), consistent with findings in other mammalian species. By electron-microscopy, substance P-positive staining was mostly in unmyelinated and in some thinly myelinated small diameter fibers. Substance P-positive terminals contained both large granular vesicles (80-120 nm diameter), which were filled with reaction product, and clear round vesicles (40-60 nm). Substance P-positive large granular vesicles were sometimes observed near presynaptic sites and in contact with dense projection there. Immunoreactive substance P boutons were small to large in size (1-4 micron), formed synapses with somata and large dendrites and were the central axons of synaptic glomeruli where they were in synaptic contact with numerous small dendrites and spines. Substance P-labeled axons frequently formed synapses with dorsal horn neurons which were also postsynaptic to other types of axons. Substance P-positive profiles participated in numerous puncta adhaerentia with unlabeled cell bodies, dendrites and axons. Only rarely, some suggestive evidence was obtained indicating that axons might synapse onto substance P-containing boutons. Biochemical analysis of monkey spinal cord tissue extracts, undertaken to characterize more precisely the immunoreactive substances, indicated that only substance P and its oxide derivative were detected with the antiserum used in the immunocytochemistry. These morphological findings show that substance P is contained within a class of axon terminals, many of which have been shown previously in the monkey to originate from the dorsal root. The results suggest that modulation of substance P primary afferents terminating in the outer dorsal laminae of the monkey spinal cord occurs in part via axonal inputs onto dorsal horn neurons postsynaptic to the primary afferent.     

Terminations of reticulospinal fibers originating from the gigantocellular reticular formation in the mouse spinal cord

The present study investigated the projections of the gigantocellular reticular nucleus (Gi) and its neighbors—the dorsal paragigantocellular reticular nucleus (DPGi), the alpha/ventral part of the gigantocellular reticular nucleus (GiA/V), and the lateral paragigantocellular reticular nucleus (LPGi)—to the mouse spinal cord by injecting the anterograde tracer biotinylated dextran amine (BDA) into the Gi, DPGi, GiA/GiV, and LPGi. The Gi projected to the entire spinal cord bilaterally with an ipsilateral predominance. Its fibers traveled in both the ventral and lateral funiculi with a greater presence in the ventral funiculus. As the fibers descended in the spinal cord, their density in the lateral funiculus increased. The terminals were present mainly in laminae 7–10 with a dorsolateral expansion caudally. In the lumbar and sacral cord, a considerable number of terminals were also present in laminae 5 and 6. Contralateral fibers shared a similar pattern to their ipsilateral counterparts and some fibers were seen to cross the midline. Fibers arising from the DPGi were similarly distributed in the spinal cord except that there was no dorsolateral expansion in the lumbar and sacral segments and there were fewer fiber terminals. Fibers arising from GiA/V predominantly traveled in the ventral and lateral funiculi ipsilaterally. Ipsilaterally, the density of fibers in the ventral funiculus decreased along the rostrocaudal axis, whereas the density of fibers in the lateral funiculus increased. They terminate mainly in the medial ventral horn and lamina 10 with a smaller number of fibers in the dorsal horn. Fibers arising from the LPGi traveled in both the ventral and lateral funiculi and the density of these fibers in the ventral and lateral funiculi decreased dramatically in the lumbar and sacral segments. Their terminals were present in the ventral horn with a large portion of them terminating in the motor neuron columns. The present study is the first demonstration of the termination pattern of fibers arising from the Gi, DPGi, GiA/GiV, and LPGi in the mouse spinal cord. It provides an anatomical foundation for those who are conducting spinal cord injury and locomotion related research.     

Spinal projections to the lateral reticular nucleus in the rat

Summary The synaptic relationships and the distribution of the afferent terminals of the spinal pathway to the lateral reticular nucleus (LRN) of the rat were examined following induced degeneration. After high cervical hemisections, the spino-LRN projection was first examined with the Fink-Heimer silver impregnation method. Degeneration was confined primarily to the ipsilateral LRN and all three divisions of the nucleus were involved. Maximum degeneration was observed in the caudal regions of the parvocellular division. The magnocellular division, except for the extreme dorsomedial area, showed substantial degeneration as well. The subtrigeminal division throughout its entire length contained only sparse degeneration.Electron microscopic examination following spinal cord lesions revealed both round and pleomorphic-vesicle terminals in various stages of electron dense degeneration. The majority of the degenerating terminals were of the round-vesicle variety. Both types of terminals contacting somata were also observed to degenerate but their number was small in comparison to those on dendritic profiles. Terminals in synaptic contact with two dendritic profiles were also observed to degenerate. Some of the large terminals belonging to synaptic configurations (glomeruli) underwent degeneration and were therefore of spinal origin as well.     

鸡脊髓上行投射在脑干的分布

本文用Nauta法追踪鸡在颈髓半横切后脊髓上行的溃变纤维,观察的结果可知,鸟类动物脊髓有上行纤维直接向丘脑投射,主要终止于丘脑的背外侧核(DL)及背内侧核(DM)的腹侧份。根据比较解剖学的结果认为该区域相当于哺乳类动物的板内核群,是旧脊丘系的一部分,可能与动物的原始感觉有关。脊髓还发出纤维向下丘脑的内细胞层(SI)和外细胞层(SE)投射,该联系可能与动物痛的情绪反应有关。此外,在中脑的顶盖、中央灰质及网状结构中也有溃变标记。在延、桥脑的下橄榄(IO)、迷走背核(NX)、孤束核(S)、外侧网状核(RL)、小细胞网状核(Rpc)及巨细胞网状核(Rgc)等都有溃变标记,通过对这些通路及终末前溃变分布的研究,进一步探讨了哺乳类动物中枢神经系统中在种族发育上部分属于比较古老的结构。     

Ultrastructure of degenerating cerebellothalamic terminals in the ventral medial nucleus of the cat

Summary Terminal degeneration of cerebellar afferents in the ventral medial thalamic nucleus (VM) was studied in cats at the ultrastructural level after uni- or bilateral lesions in the brachium conjunctivum (BC). To achieve discrete lesions within the BC, a new very accurate stereotaxic technique was used. Numerous large terminals belonging to a population of so-called LR boutons were observed degenerating in the VM. The boutons displayed a wide variety of degenerative changes. Some revealed the features of the classical neurofilamentous type of degeneration. Others, although containing a slightly increased number of neurofilaments, featured much more prominently large numbers of coated vesicle shells and heavy accumulations of a flocculent electrondense material. Degeneration in a third group of boutons similar to some extent to the light type of degeneration was characterized by tight clumping of enormously swollen or distorted synaptic vesicles within a light matrix. At later stages, however, all these boutons were believed to become shrunken and electron-dense since intermediate stages between the light- and dark-appearing boutons were observed. The degenerating cerebellar boutons formed asymmetrical synaptic contacts. Groups of 3 or 4 boutons terminated upon dendrites of projection neurons synapsing more frequently on spines than on dendritic stems. The synaptic contacts between cerebellar boutons and the vesicle-containing dendrites of local circuit neurons were encountered as often if not more than the contacts on projection neuron dendrites. Triads consisting of cerebellar boutons and dendrites of both types of neurons were observed very regularly. This synaptic arrangement provides the anatomical basis for the modification of cerebellar input in the VM by interneurons.     

Unilateral retrogasserian rhizotomy causes contralateral degeneration in spinal trigeminal nuclei of cats: an ultrastructural study

We are studying the response to injury within the brainstem trigeminal nucleus following trigeminal nerve lesions. We have previously shown with light microscopy and reduced silver stains that unilateral retrogasserian rhizotomy results not only in massive degeneration throughout the ipsilateral spinal trigeminal nucleus; in addition, degeneration is seen in a ventral position at the periobex region (involving caudal pars interpolaris and rostral pars caudalis) in the contralateral spinal trigeminal nucleus. In the present study, we have used electron microscopy to identify the source of the degenerating elements seen bilaterally after unilateral retrogasserian rhizotomy in eight adult felines with survival times ranging from 3 to 20 days. At short survival times (3–7 days) degenerating terminals with round synaptic vesicles (R terminals) and type 1, asymmetric contacts predominate bilaterally, while fewer degenerating terminals with flattened synaptic vesicles (F terminals) and type 2, symmetric contacts are seen. At longer survival times more F terminal degeneration is seen, especially on the contralateral side. Postsynaptic sites and dendrites show minimal alterations. These findings suggest that the degenerating R terminals seen on the contralateral side originate from primary afferents while the degenerating F terminals seen on the contralateral side originate from intrinsic sources involving a crossed internuclear pathway. In addition, the finding of degenerating F terminals may represent a novel form of selective transynaptic change of intrinsic neurons, associated with minimal dendritic or somatic alterations.     

Pallidosubthalamic projection in the Cat

Summary The degenerative changes within the cat's subthalamic nucleus (Sth) following lesions of the external pallidum were studied by electron microscopy.Four to five days following pallidal lesions a great number of terminals undergoing degenerative changes were encountered in the ipsilateral Sth. The contralateral Sth was free of degeneration. The degenerating terminals show predominantly the light degenerative type, less frequently the dark degenerative pattern, and occasionally exhibit signs of filamentous hyperplasia. The degenerated boutons usually insert on perikarya of the large Sth neurons, on proximal dendrites, and more rarely contact dendritic spines. They were observed neither to perform synaptic contacts with the perikarya of the small Sth neurons nor with other vesicle-containing profiles. On the basis of the ultrastructural aspect of the degenerating terminals, they were identified as F1 terminals, discriminated in a previous study (Romansky et al., 1978). The normal appearance, the synaptic relationships, and the degenerative features of the F1 terminals in the Sth closely resemble the entopeduncular terminals in the thalamus described by Rinvik and Grofová (1974a), and Grofová and Rinvik (1974).The possible contribution of the interrupted passing fibers to the observed degeneration is discussed. The present findings corroborate the relevant morphological, physiological, neurochemical, and neuropharmacological data in the literature.     

Inhibitory zinc-enriched terminals in mouse spinal cord

The ultrastructural localization of zinc transporter-3, glutamate decarboxylase and zinc ions in zinc-enriched terminals in the mouse spinal cord was studied by zinc transporter-3 and glutamate decarboxylase immunohistochemistry and zinc selenium autometallography, respectively.The distribution of zinc selenium autometallographic silver grains, and zinc transporter-3 and glutamate decarboxylase immunohistochemical puncta in both ventral and dorsal horns as seen in the light microscope corresponded to their presence in the synaptic vesicles of zinc-enriched terminals at ultrastructural levels. The densest populations of zinc-enriched terminals were seen in dorsal horn laminae I, III and IV, whereas the deeper laminae V and VI contained fewer terminals. At ultrastructural levels, zinc-enriched terminals primarily formed symmetrical synapses on perikarya and dendrites. Only relatively few asymmetrical synapses were observed on zinc-enriched terminals. In general, the biggest zinc-enriched terminals contacted neuronal somata and large dendritic elements, while medium-sized and small terminals made contacts on small dendrites. The ventral horn was primarily populated by big and medium-sized zinc-enriched terminals, whereas the dorsal horn was dominated by medium-sized and small zinc-enriched terminals.The presence of boutons with flat synaptic vesicles with zinc ions and symmetric synaptic contacts suggests the presence of inhibitory zinc-enriched terminals in the mammalian spinal cord, and this was confirmed by the finding that zinc ions and glutamate decarboxylase are co-localized in these terminals. The pattern of zinc-enriched boutons in both dorsal and ventral horns is compatible with evidence suggesting that zinc may be involved in both sensory transmission and motor control.     

An electron microscopic analysis of pyramidal tract terminations in the spinal cord of the cat

Summary The ultrastructure of the external basilar region (EBR) of the spinal cord has been investigated in normal cats and after ablation of the contralateral sensorimotor cortex. This region (lateral part of lamina V of Rexed) is the main site of termination of descending pyramidal fibres.The EBR contains neurons with light and dark cytoplasm and correspondingly light and dark dendrites. Axon terminals in the EBR can be divided into four types on the basis of the following structural criteria: (1) spheroid synapt vesicles, (2) flattened synaptic vesicles, (3) spheroid clear vesicles and large dense-core vesicles, (4) electron-dense plasma matrix and numerous spheroid vesicles. Types (1) and (2) prevail, forming mainly axondendritic contacts, generally with thin protrusions and spine-like appendages of the dendrites. A few larger synaptic arrangements of glomerular structure were observed.Signs of terminal degeneration can be recognized in the EBR as early as 30 hours after cortical ablation. The structural elements of these terminals are fused into a compact electron-dense material corresponding to the known dark type of degeneration. Three to five days after the lesions a second set of degenerating terminals become recognizable, in which the lysis of cytoplasmic elements is prevalent. This slower modus of degeneration might be distinguished as the light type. Degenerating terminals are located mainly on dendrites of the EBR neurons and are of small size. The shape of the synaptic vesicles can be recognized in some terminals undergoing degeneration according to the light type; they are, as a rule, of flattened type. Synaptic terminals do not degenerate in the glomerular complexes.     

Reticulospinal and vestibulospinal pathways in the snake Python regius

In the present HRP study extensive reticulospinal projections and more modestly developed vestibulospinal pathways have been demonstrated in the snake Python regius. The funicular trajectories of the main reticulospinal pathways have been shown: via the lateral funiculus pass spinal projections of the nucleus reticularis superior pars lateralis, the nucleus reticularis inferior and nucleus raphes inferior; via the ventral funiculus fibers arising in the nucleus reticularis superior and nucleus reticularis medius. Spinal projections of the locus coeruleus and subcoeruleus area reach their targets via both the lateral and ventral funiculi. Two vestibulospinal pathways have been demonstrated: an ipsilateral tractus vestibulospinalis lateralis arising in the ventrolateral vestibular nucleus, and a contralateral tractus vestibulospinalis medialis from the descending and ventromedial vestibular nuclei. After HRP gel implants into the vestibular nuclear complex direct vestibulocollic projections to motoneurons in the rostral spinal cord were observed. Spinal projections from the ventral part of the nucleus reticularis inferior and the descending and ventromedial vestibular nuclei are mainly aimed at the thin "neck area" (approximately the first 50 spinal segments). This area is extensively used in such acts as orientation and prey-catching, requiring a rather delicate brain stem control.     

The central cervical nucleus in the cat. IV. Afferent fiber connections

Summary Afferent input to the central cervical nucleus (CCN) in the C1–4 segments was studied with degeneration methods after sectioning of dorsal roots (DRs) or lesions in the spinal cord or brain stem. Degeneration in the CCN was heavy after sectioning of the DRs C1–4, moderate after sectioning DRs C5–8, and scanty after sectioning DRs T1–4. One to 2 days after sectioning of the C2 dorsal root the resulting degeneration had a granular appearance. At 4 days after the operation coarser argyrophilic fragments appeared, and this type of degeneration dominated at longer postoperative intervals. Degeneration in the ipsilateral CCN was found after lesions of the ventral and lateral funiculi of the thoracic cord. No degeneration was found after lesions of the dorsal funiculus caudal to T4 or after lesions in the ventral and lateral funiculi of the lumbar cord. Degeneration in the ipsilateral CCN was found after lesions in the brain stem in cases with lesions involving the medial and caudal part of the medulla. These afferents may run in the medial longitudinal fascicle (MLF).Supported by the Swedish Medical Research Council, project no. 553     

猫皮质-后索核-脊髓通路的实验形态学研究

用Nauta法和HRP法研究了35只猫的皮质后索核投射和后索核脊髓投射。结果表明,投向后索核的皮质纤维起自后乙状回第Ⅴ层锥体细胞,该回的内侧面和十字沟上岸投向薄束核,背外侧面外2/3投向楔束核,内1/3投向二核。皮质纤维下行于锥体束内,大部分经锥体交叉达对侧后索核,少量不交叉的纤维至同侧。后索核的中间腹侧部是皮质纤维分布的中心。投向脊髓灰质的后索核纤维主要发自核中间腹侧部的大、中型三角、多角和梭形细胞。薄束核投向下段脊髓灰质,楔束核投向上段脊髓灰质。后索核纤绀经后角尖周围的白质下行,主要终止于同侧脊髓灰质Ⅳ、Ⅴ层。皮质纤维在后索核中的终止部位与后索核脊髓纤维的起源区基本一致,据此试将两者归为一个投射系统,即皮质-后索核-脊髓通路加以描述。     

大鼠脊神经后根切断后脊髓和背根神经节CGRP的表达变化

为观察大鼠脊神经后根切断后相应背根神经节(DRG)和脊髓节段CGRP的表达变化,本研究采用25只健康成年SD大鼠随机分为正常对照组、假手术对照组和L4、5后根切断后3d、7d和14d组(n=5),用免疫组织化学方法结合图像分析技术检测各组相应DRG和脊髓节段内CGRP的表达变化。结果如下:后根切断后3d、7d和14d伤侧DRG内CGRP表达较对照组和对侧明显增强;后根切断后3d脊髓后角CGRP免疫阳性纤维减少,7d、14d时进一步减少;后根切断后3d脊髓前角运动神经元内CGRP表达增加,免疫阳性细胞数增多,7d和14d时表达进一步增强。以上结果提示,脊神经后根切断后DRG和脊髓CGRP表达变化呈现一定的时空模式,可能参与了神经损伤后的再生过程。     

Immunocytochemical localization of substance P in the rat spinal cord with special reference to fibers within the ventral column of the rostral lumbar segments

The distribution of substance P (SP) in the rat spinal cord was investigated by peroxidase-anti-peroxidase (PAP) immunocytochemistry. A dense network of SP-immunoreactive fibers and terminals was detected in the ventral column of the rostral lumbar cord with a different density and extent from the other segmental levels. These fibers and terminals were accumulated within and around the centromedial nucleus (CM) of the L1 and L2 segments, with some bundles of immunoreactive fibers between the CM and other areas; i.e. laminae V and X and the contralateral CM. They formed a dense network, such as in arborization of immunoreactive fibers and terminals on the transverse plane and in a comb-shaped structure on the horizontal plane. The origin of the SP in this network was examined. It was determined that neither a total transection of spinal cord at a low thoracic level or mid-lumbar level, nor at an ipsilateral or bilateral section of the 3-5 dorsal roots, containing L1 and L2 roots, induced any visible changes in the SP staining pattern. An intrathecal injection of colchicine revealed the presence of SP-immunoreactive neurons in the dorsal horn and intermediate gray matter at the spinal cord including the rostral lumbar cord. The present findings suggested that the majority of SP immunoreactivities in the above network are derived from local circuit interneurons of the spinal cord.     

Establishment of synaptic connections between explants of embryonic neural tissue in culture: experimental ultrastructural studies

Summary The development of synaptic interconnections between co-cultured explants of central and peripheral nervous tissue from chick embryos has been investigated by light and electron microscopy. Two sets of co-cultured explants were used: (a) dorsal root ganglion (DRG) and spinal cord and (b) retina and tectum. Both sets of co-cultured explants became linked by bundles of fibres but the most consistent results were obtained with the DRG-spinal cord explants. Thus axons from the DRG extended large distances across the culture substrate to reach and enter mainly the dorsal horn region of the spinal cord explants. In contrast retina-tectum links were less frequently established and were less extensive, possibly because there are fewer cells in retinal explants capable of establishing contacts in tectal explants than there are cells in DRG explants capable of establishing contacts in the spinal cord. In order to distinguish between synapses involving only neuronal elements within an expiant and those involving ingrowing fibres, fibre bundles linking adjacent explants were transected and the preparations fixed two to six hours later. Electron microscope study of such cultures revealed degenerating neurites and terminals in the spinal cord explants receiving DRG fibres but none in the corresponding DRG explants. Retinal explants contain numerous synapses of many types but degenerating terminals could not be found within the retinal explants after nerve fibre transections. Degenerating neurites and terminals were found within tectal explants but they were fewer and more difficult to locate than those found within spinal cord explants. The reasons for such differences are discussed.