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.     

An ultrastructural study of nerve terminal degeneration in muscle spindles of the tenuissimus muscle of the cat

Summary Muscle spindles in the tenuissimus muscle of the cat were studied between 12 and 168 h after cutting or freezing the nerve to this muscle. Degenerative changes in sensory and motor nerve terminals on intrafusal muscle fibres were observed using the electron microscope. Comparisons were made with spindles from unoperated or sham-operated cats.The earliest degenerative changes were seen in sensory and motor terminals at 20–24 h after the lesion. No nerve endings were seen by 114 h after denervation. The most consistent initial signs of degeneration were: (1) the presence of abnormal mitochondria and dense bodies in sensory terminals, and (2) a decrease in the number and clumping of synaptic vesicles combined with an increase in glycogen and neurofilaments in motor endings. Intrafusal fibres participate in the removal of degenerating sensory endings. Schwann cells phagocytose degenerating motor terminals. The disappearance of nerve terminals precedes the complete degeneration of preterminal myelinated fibres within the muscle spindle.     

大鼠前庭脊髓束终末与脊髓小脑束起源细胞间的突触联系

目的　探讨前庭脊髓束终末与脊髓小脑束起源细胞间的突触联系。方法　采用HRP逆行追踪和顺行溃变结合电镜技术 ,对大鼠的前庭脊髓束终末与脊髓小脑束起源细胞之间的突触联系进行了研究。结果　发现溃变的前庭脊髓束轴突终末与HRP标记的脊髓小脑束神经元胞体之间形成的突触数量较少 ,其中轴 -树型突触数量多于轴 -体型突触。突触前终末含有丰富的清亮型扁平突触小泡 ,其次为清亮型圆形小泡。前庭脊髓束轴突溃变终末与脊髓边缘细胞形成的突触数量多于Clark s柱细胞形成的突触 ,并形成一些以标记树突为中心的突触复合体。结论　本研究从形态上证实前庭脊髓束终末直接与脊髓小脑束起源细胞发生突触联系 ,也表明前庭脊髓束对脊髓小脑束起源细胞向小脑的信息传递具有直接调控使用     

皮质—小脑前核—小脑通路—HRP法结合溃变电镜法研究

目的：观察大鼠小脑前核（桥核,下橄榄,外侧网状核,桥被盖网状核和旁正中网状核）中皮质纤维终末的溃变型及其与各核中的小脑投射神经元的突触联系。材料和方法：８只大鼠,用ＨＲＰ逆行示记法和溃变电镜技术。结果：（１）溃变型有三种,电子致密型,微丝增生型以及电子透明型。（２）电子致密型溃变有三种不同形态,即含圆形清亮型小泡,含多形清亮型小泡和混合型小泡的三种终末,其中绝大多数溃变终末含圆形亮型小泡。（４）溃     

The lateral cervical nucleus in the cat III. An electron microscopical study after transection of spinal afferents

Summary The ultrastructure of terminal degeneration within the lateral cervical nucleus (LCN) after transection of its spinal afferent fibers 2 days–2 years earlier is described. The degeneration after 2 days was of both the neurofilamentous and dense type. The highest number of degenerating terminals, about 15%, was found after 4–5 days. Then most of the degenerating boutons were of the dense type. The degenerating terminals had synaptic contact with cell bodies and dendrites of LCN-neurons. Removal of the degenerating boutons seemed to be effected by a phagocytic cell present in increased number compared to the normal LCN. In cases with long survival times an increase in the number of astroglial filaments was observed. In an animal where the spinal afferents to the LCN had been cut 2 years earlier a decrease in medium size of the neurons was observed. The amount of dendritic spines was also considerably smaller than normally.     

Gracile nucleus of streptozotocin-induced diabetic rats

Summary This study reports ultrastructural changes in the gracile nucleus of male Wistar rats after streptozotocin-induced diabetes. During the acute phase (3–7 days) degenerating electron-dense dendrites and axon terminals were dispersed in the neuropil. Degenerating dendrites were characterized by an electron-dense cytoplasm, swollen mitochondria, dilated endoplasmic reticulum and scattered ribosomes. Degenerating axon terminals were characterized by an electron-dense cytoplasm and clustering of small spherical agranular vesicles. Degenerating axon terminals may form part of a synaptic glomerulus with a central electron-dense dendrite, or they may form the central element of a synaptic glomerulus. These degenerating profiles were absent in the gracile nucleus of the 3 and 7 days insulin-treated post-streptozotocin rats. Macrophages were present in the neuropil and were in the process of engulfing neuronal elements. During the medium phase (1–6 months), most of the degenerating dendrites and axon terminals had been engulfed or removed by macrophages. During the late phase (9–12 months) a second wave of degeneration occurred in the gracile nucleus, similar to the acute phase. During the medium and late phases, dystrophic axonal profiles were also significantly increased in the rats after streptozotocin treatment.It is concluded that the ultrastructural changes observed in the gracile nucleus in the present study were the result of streptozotocin-induced diabetes rather than a toxic effect of streptozotocin, even in the acute phase.     

Connexions from large, ipsilateral hind limb muscle and skin afferents to the rostral main cuneate nucleus and to the nucleus X region in the cat.

1. Evidence is presented for an input from ipsilateral hind limb group I muscle afferents and low threshold cutaneous afferents, to cells in the rostral division of the main cuneate nucleus (rMCN) and in the region of the descending vestibular nucleus and the nucleus X of Brodal & Pompeiano (1957a), the (DV-X). 2. Thirteen group I-rMCN cells were recorded from. The functional properties of these cells were similar to those of nueleus Z (Landgren & Silfvenius, 1971; Johansson & Silfvenius, 1977a, b). The cells were monosynaptically linked to spinal dorsolateral fascicle (DLF) fibres. Nine cells projected to the contralateral thalamus, i.e. a second group I hind limb bulbothalamic tract is described. Ten cells were synaptically activated from the ipsilateral cerebellum from the anterior projection zone of the dorsal spinocerebellar tract (DSCT). Axon-collateral activation by DSCT fibres was established for two of these cells. They were both bulbothalamic relay cells. For the remaining eight cells, activated from the cerebellum, this was not proven. These cells could, however, either be linked to DSCT fibres or to short axon-collaterals of a cell body of unknown location. A projection from the rMCN to the cerebellum is described and agrees with recent anatomical findings. Two cells were not excited from the cerebellum. 3. Four rMCN cells were activated by cutaneous afferents with their secondary axons in the DLF. Suggestive evidence for a bulbothalamic cutaneous hind limb path via the rMCN is presented. Two cells were activated from the cerebellum, presumably via axon-collaterals of nonsegmental cells. 4. Eight group I-DV-X cells were recorded from. They were monosynaptically linked to spinal DLF fibres and resembled functionally the nucleus Z and rMCN cells when stimulated from the periphery. Two cells projected to the contralateral thalamus, and two others were synaptically excited. Seven cells were activated from the ipsilateral cerebellum. Two of them projected to the cerebellum, and three were synapitcally activated by axon-collaterals of an undefined non-segmental cell. 5. Two DV-X cells which were activated by cutaneous afferents possibly had their spinal fibres deep in the dorsal column. Both were activated from the cerebellum, one by collaterals of a spinal axon. The functional organization of the three juxtaposed medullary nuclei, Z, rMCN and DV-X is discussed.     

Effects of neonatal intraocular colchicine on synaptogenesis and on the retention of the ipsilateral retinofugal projection within the superior colliculus

Summary A normally transient ipsilateral retinofugal projection exists in the rat but is retained following eye removal because of the loss of competitive interaction between crossed and uncrossed fibers. To further explore this phenomenon, colchicine (10^–3M) was injected into the right eye of newborn albino rats to partially suppress axonal transport in optic fibers, alter the developmental time course of retinofugal synaptic terminals and determine if this would in turn extend the period of survival of the ipsilateral projection. Measurements of the number of fibers in the nerve were also made to insure that colchicine was not lethal to the retinofugal projection. Projections into the superior colliculus were demonstrated by anterograde movement of HRP from the left eye. TMB histochemistry revealed dense labeling of the contralateral retino-recipient layers at 5 dpn in untreated or saline-injected controls. The ipsilateral projection was seen as a lighter band of activity across the colliculus which was most concentrated in the antero-medial quadrant. This pathway was transient and degenerated by 10 dpn, except for a few antero-medial fibers. Animals treated with colchicine demonstrated a retention of this pathway through 20 dpn. A concomitant quantitative analysis of synaptic development within the superior colliculus revealed populations of boutons with round (R) and flattened (F) vesicles, as well as multiple junctional (MJ) and serial (S) complexes, most of which were specialized R boutons. The various synaptic categories displayed specific ratios unique to the different stages of maturation. Intraocular colchicine reduced the ratio of R, MJ and S boutons to F terminals between 5–15 dpn (P < 0.01). By 20 dpn, the proportions of MJ and S boutons remained depressed but the normal ratio of R to F boutons was restored. Areal determinations of each synaptic profile included in the counts revealed a significant reduction in the size of MJ synaptic profiles examined in colchicine-treated animals and this may have been reflected in the slight loss of tectal volume (6–9%). Removal of the left eye and assessment of degenerating boutons showed that the expanded ipsilateral projection was not sufficiently dense to produce such a restoration. It thus appears that colchicine delayed the growth of the R population, but the effect was reversible and this category of boutons continued to develop, albeit on a later time course. Continued depression of MJ and S boutons suggests that suppression of the rate and quantity of axonally-transported substances retards the final stages of tectal synaptic differentiation, reduces their competitive advantage and allows the retention of the ipsilateral optic projection.     

Neuron morphology and synaptic architecture in the medial superior olivary nucleus

Summary Dendritic arborization pattern, spatial and synaptic relations of various neuron types and the terminal distribution of afferent axons of various origin were studied in the medial superior olivary nucleus of the cat using Golgi, degeneration, electron microscope and horseradish peroxidase techniques. Three types of neurons clearly different in morphological features, distribution, neighbourhood relations, input and output characteristics were distinguished: (1) fusiform cells having specific dendritic orientations and arborization patterns and synaptic relations to various types of terminal axon arborizations (2) multipolar neurons with wavy dendrites bearing spine-like appendages, receiving relatively few synaptic contacts and having a locally arborizing axon, and (3) elongated marginal cells, largely restricted to the fibrous capsule of the nucleus. The fusiform and marginal neurons were identified by retrograde peroxidase labeling as the olivo-collicular projection cells.Ultrastructural analysis of normal and experimental material revealed the presence of four distinct kinds of axon terminals differing in size, synaptic vesicles type, relation to postsynaptic targets and in origin: (i) large terminals with multiple extended asymmetric synaptic membrane specializations and containing round, clear vesicles arise from the spherical cells of the ipsilateral anteroventral cochlear nucleus, (ii) most of the small axon terminal profiles — engaged in asymmetric synaptic contacts — originated from the trapezoid nucleus, (iii) terminal boutons containing pleomorphic vesicles belong to fibers descending from the ipsilateral multipolar neurons in the central nucleus of the inferior colliculus and from the nuclei of the lateral lemniscus while (iv) boutons containing exclusively ovoid vesicles and remaining intact after complete deafferentation of the nucleus were considered to be of local origin.     

Effects of intraneural injection of Ricinus communis agglutinin-60 into rat vagus nerve

Summary The dorsal motor nucleus (DMN) of the rat was studied at various survival periods following an intraneural injection ofRicinus communis agglutinin-60 (RCA-60) into the vagus nerve at the mid-cervical region. No obvious structural changes were noted in the DMN 2 and 4 days after the injection of RCA-60. At 5 and 6 days after the RCA-60 injection, the larger neurons (measuring 19 × 12 m) in the DMN underwent chromatolytic degeneration whereas the smaller ones (measuring 10 × 6 m), characterized by their infolded nuclei, remained unaffected. The majority of the degenerating DMN neurons became pale and crenated in outline. Other structural changes included swollen mitochondria with disrupted cristae and profiles of rough endoplasmic reticulum denuded of ribosome particles. A few of the degenerating neurons became extremely condensed and darkened. Axon terminals which showed synaptic contacts with these cells remained normal. Both pale and darkened degenerating dendrites, derived from the degenerating neurons, were present in the neuropil. In addition to these, degenerating axon terminals with clumping or swelling of synaptic vesicles were also present. They were presynaptic to dendrites of various sizes. Massive infiltration of mononuclear cells occurred in the DMN. These cells reached the DMN by diapedesis and were actively engaged in the phagocytosis of degenerating neuronal elements. While most of the invading cells transformed into active neuronal macrophages, some of them eventually died in the neuropil of the DMN.Light microscopic study by Fink-Heimer's method for degenerating fibres and terminals revealed their distribution to the DMN, nucleus of the tractus solitarius, nucleus commissuralis, dorsolateral and lateral part of the hypoglossal nucleus and the area postrema.It was concluded from this study that RCA-60, when injected into the cervical vagus was retrogradely transported to the cell body of the DMN neurons of the larger category. The selective destruction of the DMN neurons by RCA-60 elicited a massive infiltration of mononuclear cells which gave rise to the neural macrophages. The RCA-60 injected also killed the vagal sensory neurons as demonstrated by the numerous degenerating fibres and axon terminals in the DMN which would represent their central processes.     

The thalamic reticular nucleus of the adult rat: experimental anatomical studies

Summary The thalamic reticular nucleus (TRN) is a sheet-like nucleus partially enclosing the dorsolateral and anterior aspects of the thalamus and traversed by the thalamo-cortical and cortico-thalamic fibre systems. This paper describes the cellular and synaptic organization of the TRN in adult albino rats on the basis of LM and EM studies of normal animals and experimental animals with injections of horseradish peroxidase (HRP) and/or lesions in various parts of the brain. Particular attention was paid to the dorso-caudal part of the TRN, which establishes connections with visual centres.LM-HRP preparations show that the neurons of TRN project only to ipsilateral dorsal thalamus; no labelled cell bodies were found in TRN after injections into the cortex or any part of the brain stem caudal to the thalamus. Small injections into dorsal thalamus result in a small cluster of labelled neurons and an associated patch of terminal label in TRN. The dorso-caudal part of the nucleus projects to the dorsal lateral geniculate nucleus, the ventro-caudal part to the medial geniculate nucleus and a large part of the nucleus anterior to the areas associated with the geniculate nuclei projects to the ventrobasal nucleus. No evidence was found for a widespread distribution of reticulo-thalamic axons and the connections between TRN and the dorsal lateral geniculate nucleus and between TRN and the ventrobasal nucleus show a fine-grain topographical organization with more rostral and dorsal parts of TRN projecting to more rostral and dorsal parts of the dorsal lateral geniculate and ventrobasal nuclei.The neurons of TRN are variable in size (range of somal diametersc. 10–20 m), shape (cell bodies are most commonly ellipsoidal) and dendritic morphology (bitufted and bipolar arrangements most common), but no basis for subdividing them into more than one class was found with any of the techniques used. The cell body and dendrites are commonly aligned parallel to the surface of TRN and at right angles to the traversing fibre bundles. The dendrites do not branch extensively and are only moderately spinous. Long, hair-like spines corresponding to those described by Scheibel & Scheibel (1966) were not found: nor were dendritic bundles found to be as prominent in EM material as reported by these authors in LM-Golgi material. Plasma membranes of dendrites in small bundles and of contiguous somata were commonly in direct contact over large areas, but gap junctions between them were not seen.The neuropil of TRN is simple with three major axon terminal types.D-type terminals (about 56% of all terminals in visual TRN) have closely packed spherical synaptic vesicles (42 nm diameter);L-type terminals (about 31%) are paler, slightly larger and have less densely packed synaptic vesicles (46 nm diameter); both terminal types make Gray type 1 synaptic contacts on dendritic spines and dendritic shafts and rarely also on cell bodies and axon hillocks.F-type terminals (about 8%) contain flattened synaptic vesicles in a dark matrix and make Gray type 2 contacts with dendrites, cell bodies and axon hillocks. In visual TRN, D-type terminals (but not all) degenerate after ablation of ipsilateral visual cortex and L-type terminals (but not all) degenerate after lesion of ipsilateral dorsal lateral geniculate nucleus; the density of degenerating terminals is higher after cortical than after geniculate lesions. Indirect evidence suggests that F-type terminals may be (or may include) collaterals of reticulo-thalamic projection cells, but no evidence was found for a widespread or dense plexus of such collaterals.After injection of HRP into the dorsal lateral geniculate nucleus, labelled axon terminals in visual TRN (many clearly L-type) were found in synaptic contact with retrogradely labelled dendrites of reticulo-geniculate projection cells. When HRP injection was combined with ablation of ipsilateral visual cortex, degenerating axon terminals (most of them identifiable as D-type) were also found in synaptic contact with retrogradely-labelled dendrites of reticulo-geniculate projection cells.Thus, neurons of visual TRN in the rat receive monosynaptic, presumptively excitatory input from collaterals of cortico-geniculate and geniculo-cortical axons, and project in a topographically-organized manner to the ipsilateral dorsal lateral geniculate nucleus (where they make Gray type 2 GABAergic and presumptively inhibitory synaptic contacts chiefly with the dendrites of geniculo-cortical projection cells). A similar pattern of organization is seen in other parts of the TRN and these data are compatible with the view that the TRN (and the perigeniculate nucleus of the cat thalamus, which is similar in several respects to visual TRN) forms part of a negative feed-back system by which the activity of thalamo-cortical projection neurons is regulated.     

The development of the inferior olivary complex in preweanling opossums

Summary Pre- and postsynaptic elements within the developing inferior olive (IO) of both control and experimental opossums were examined via electron microscopy. Electron dense boutons identified di-/mesencephalic, cerebellar and spinal afferents within the IO of 8–71 day old animals, which survived 4–48 hours following either midbrain hemisections or spinal transections.During its initial stage of development (3–22 days) the neuropil of the IO is segregated into fields of small diameter neurites or flocculent profiles. Within the fields of flocculent profiles, synaptic interactions are established, which are both infrequent and immature. Although some flocculent profiles are presynaptic, most are postsynaptic and emanate from olivary somata and dendrites. Synaptic contacts also occur with olivary somata, dendritic shafts, spines and dendritic varicosities. Clear round vesicles (crv's; 40 m) predominate within all boutons, normal ones as well as those which degenerate after di-/mesencephalic, cerebellar and spinal lesions; however, larger (70 m) dense cored vesicles (dcv's) are occasionally observed within some boutons. Degenerating terminals from all three sources primarily contact flocculent profiles and dendritic shafts.As the opossum matures (42 days) dramatic increases occur in the number and complexity of both pre- and postsynaptic elements. Marked variations are observed in the matrix density of dendritic shafts. Although all terminal boutons predominantly contain crv's, the number of dcv's within the population of presynaptic elements increases markedly. Concurently, olivary neurons are profusely studded with spines. Simple dendritic spines and spiny appendages as well as dendritic shafts are the most frequent postsynaptic structures within the principal nucleus (PO). Olivary somata and their spines, however, are postsynaptic to degenerating de-/mesencephalic afferents within the PO. Flocculent profiles, which persist within the accessory nuclei, and dendritic shafts are postsynaptic to degenerating spinal boutons.By 70 days of age synaptic contacts appear more mature and more nearly approximate those seen in the adult (King 1980). Few somatic contacts, opaque dendrites, dendritic varicosities, and flocculent profiles are evident within the PO. Dendritic shafts and spines are the principal postsynaptic structures. Many di-/mesencephalic and cerebellar afferents synapse within maturing synaptic clusters on spines between which a rare gap junction is observed. Other di-/mesencephalic and cerebellar endings in the PO as well as spinal endings in the accessory nuclei are presynaptic to dendritic shafts and spines external to synaptic clusters. This predilection for contacting more specific loci on olivary neurons provides good evidence for synaptic remodeling.As the olivary nuclei develop further, the incidence of gap junctions increases and pleomorphic vesicles appear within boutons. The glial investment of neuronal elements, including synaptic clusters, also becomes more extensive.In conclusion, early di-/mesencephalic, cerebellar and spinal synaptic contacts appear qualitatively uniform in their synaptic features and postsynaptic interactions. As olivary development proceeds, however, the distinguishing synaptic features of the nuclear complex become more apparent. Synaptic remodeling occurs as some midbrain and cerebellar terminals are localized within synaptic clusters. The ultrastructural features characteristic of the adult IO are finally achieved by 80 days of age.This research was supported by N.I.H. Research Grant NS-08798     

Ultrastructural changes in the dorsal motor nucleus of monkey following bilateral cervical vagotomy

Summary The neurons of the dorsal motor nucleas (DMN) of the monkey (Macaca fascicularis) were of two main types: small (13 × 8 m) and medium-sized (20 × 13 m). The latter, which were the predominant form, contained a pale oval nucleus surrounded by organelle-rich cytoplasm. Between one and three long principal dendrites per section profile arose from eac¹ of the somata. Both axosomatic and axodendritic synapses were seen on these cells although the latter were more common.No structural changes were noted in the DMN 1–3 days after bilateral cervical vagotomy. Some of the dendrites of the medium-sized axotomized vagal neurons appeared darkened 5–10 days after the operation. With longer surviving intervals, i.e. 21 and 28 days after operation, darkened dendrites were more commonly seen and the cytoplasmic density of these dendrites was dramatically enhanced. Their mitochondria were pale and some of them also showed vesiculation. Both normal and degenerating axon terminals were seen to form synaptic contacts with the darkened dendrites. The degenerating axon terminals were characterized by the clumping of their round agranular vesicles. Both darkened dendrites and degenerating axon terminals were phagocytosed by hypertrophied astrocytes and activated microglial cells. Blood elements infiltrating into the DMN were a possible source for some of the neural macrophages.It was concluded from the present study that the dendrites of the vagal neurons were the first structures to degenerate in axotomy and these were subsequently removed by glial elements. Degenerating axon terminals on the darkened dendrites could represent endings of the central processes of peripheral vagal ganglion cells that had undergonetransganglionic degeneration after damage to their peripheral processes.     

Synaptic junctions between sympathetic axon terminals and pinealocytes in the monkey Macaca fascicularis

Summary The distribution of axon terminals in the pineal gland of monkeys was studied by electron microscopy. Numerous terminals bearing small pleomorphic agranular and dense-cored vesicles were localized in the perivascular space and among the pinealocytes in the parenchyma in normal monkeys. Following bilateral superior cervical ganglionectomy, they underwent degenerative changes, including the accumulation of glycogen masses, appearance of dense residual bodies and the displacement of synaptic vesicles. Some of these degenerating terminals showed synaptic contacts with the cell bodies of pinealocytes. At the synaptic junction the postsynaptic membrane was thickened asymmetrically. Examples of synaptic contacts were most frequently observed in 5 and 7 days postoperative animals. In the longer surviving (30 days) monkey, most of the axon terminals showed round agranular vesicles, and they were mainly presynaptic to the intrapineal ganglion cells with some of the pinealocytes. They remained structurally unchanged following the resection of both the superior cervical ganglia. A few axon terminals containing small dense-cored vesicles appeared to have survived the initial insult, but some of their vesicles appeared swollen 30 days after the operation. It is concluded from this study that some of the pinealocytes are under the influence by the postganglionic neurons in the superior cervical ganglia through direct synaptic contacts. The intrapineal ganglion cells are postsynaptic to fibres originating exclusively from the central nervous system. Some of these fibres, however, may be presynaptic directly to pinealocytes.     

A comparison of the mode of termination of the hippocampal and hypothalamic afferents to the septal nuclei as revealed by electron microscopy of degeneration

Summary Previous studies with the Nauta technique have established that fibres which originate in two important areas — the hippocampus and the hypothalamus — converge upon the cells of the septal nuclear complex. The purpose of this study was to investigate the anatomical basis of how the septal cells could differentiate between fibres from the two sources. Differences in the mode of termination of these two systems have been studied quantitatively at the electron microscope level by using the orthograde degeneration of terminals after lesions of the fimbria and the medial forebrain bundle. In the medial septal nucleus, the hippocampal fibres account for 35% of the terminals, and in the lateral septal nucleus, 43% of the terminals on the same side and a further 13% on the opposite side. These terminals are at least 98% axodendritic and 91% of them contain predominantly clear synaptic vesicles of 500 Å diameter. The hypothalamic fibres are the source of up to 19% of the axodendritic terminals in the medial septal nucleus, but considerably fewer in the lateral septal nucleus. In contrast to the hippocampal afferents, the hypothalamo-septal system has two characteristic features: firstly, the fibres give rise to up to 24% of the axosomatic terminals in the medial septal nucleus, and secondly, 63% of the terminals contain a population of vesicles with significantly higher proportions of dense centred vesicles of 800–1000 Å diameter.     

The termination of thalamo-cortical fibres in the visual cortex of the cat

Summary An electron microscopic study has been made of the site and mode of termination of thalamo-cortical fibres in area 17 of the visual cortex of the cat. Thalamo-cortical fibres had been selectively interrupted 4–5 days before perfusion of the brain. In agreement with previous studies, degenerating axon terminals were found in layer I, in the deep part of layer III, and in layer IV of the cortex. In addition, a few degenerating thalamo-cortical axon terminals were found making synapses upon spines and small dendrites in layer VI. Two examples were seen of degenerating axon terminals making axo-dendritic synapses upon dendrites in continuity with their cell bodies. These neurons were not large stellate cells and were probably pyramidal.     

Identification of dorsal root synaptic terminals on monkey ventral horn cells by electron microscopic autoradiography

Summary The projection of dorsal root fibres to the motor nucleus of the macaque monkey spinal cord has been examined utilizing light and electron microscopic autoradiography. Light microscopy demonstrates a very sparse labelling of primary afferent fibres in the ventral horn. Silver grains overlying radioactive sources are frequently clustered into small groups, often adjacent to dendritic profiles. Under the electron microscope, myelinated axons and a few large synaptic profiles containing rounded synaptic vesicles were overlain by numerous silver grains. These labelled profiles made synaptic contact with dendrites 1–3 m in diameter. The labelled profiles did not contact cell bodies or large proximal dendrites of ventral horn neurons. Frequently, small synaptic profiles containing flattened vesicles were presynaptic to the large labelled terminals and it is suggested that these axoaxonal synapses may mediate presynaptic inhibition of the primary afferent fibres. The relationship of the present findings to previously published physiological and anatomical studies is discussed.     

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.     

Mode of termination of primary vestibular fibres in the lateral vestibular nucleus an experimental electron microscopical study in the cat

Summary Following transection of the vestibular nerve in cats, the electron microscopical changes occurring in the lateral vestibular nucleus were studied after survival periods of 2–11 days. Material for study was taken from the rostroventral part of the nucleus of Deiters since this is known to receive the primary vestibular fibres.Degeneration of terminal boutons is evident two days after the lesion. Degenerating boutons show an increased electron optic density, mitochondrial changes and a loss of synaptic vesicles. They are often surrounded by a pericellular space filled with flocculent (probably protein) material. At three days and later this space is occupied by processes of astrocytes or of a type of phagocytic cells which surround or engulf the degenerating boutons. Nine to eleven days after the lesion almost all degenerating boutons have disappeared. There is evidence of phagocytosis of axons and myelin sheaths by astrocytes but mainly by phagocytes of yet undetermined origin. The filamentous type of bouton degeneration has not been observed.Degenerating boutons are found on neuronal perikarya and on proximal as well as on thin distal dendrites and on spines. They are common on small and medium-sized cells, but have also been seen on some giant cells. The degenerating boutons do not form series of synaptic complexes. Degenerating fibres and boutons have so far been found only ipsilateral to the lesion.The findings confirm and extend those made in corresponding experiments with silver impregnation procedures, but emphasize the limitations of the latter methods as regards conclusions concerning synaptic contacts.This investigation has been supported by Grant NB 02215-07 from the National Institute of Neurological Diseases and Blindness, US Public Health Service. This aid is gratefully acknowledged.A preliminary presentation of some of the findings described in this paper has been made by one of the authors (A. brodal) at a Ciba Foundation Symposium, Myotatic, Kinaesthetic and Vestibular Mechanisms, London, Sept. 27.–29., 1966.     

Fine structural survey of tyrosine hydroxylase immunoreactive terminals in the myenteric ganglion of the rat duodenum

We have examined whether the noradrenergic neurons have direct synaptic projections to the myenteric ganglion neurons of the duodenum and the ultrastructure of their terminals by using immunogold–silver labeling for tyrosine hydroxylase. In the neuropil of the myenteric ganglia, about half of the axon terminals contained round clear vesicles and the rest of them contained pleomorphic clear vesicles. The sizes of axon terminals contacting the dendrites as a whole were 1.62 ± 0.07 μm. All axon terminals formed asymmetric synaptic contacts with dendrites or somata. Immunohistochemical study revealed that the tyrosine hydroxylase-immunoreactive nerve terminals were distributed throughout the ganglia and contained exclusively pleomorphic clear synaptic vesicles (about 20–80 nm long). The tyrosine hydroxylase-immunoreactive terminals were generally large (1.99 ± 0.07 μm). A considerable number of the tyrosine hydroxylase-immunoreactive terminals made asymmetric synaptic contacts with small dendrites, spines or somata of the myenteric ganglion neurons. Serial ultrathin sections through the myenteric neurons revealed that about 16% of the total number of axosomatic terminals showed tyrosine hydroxylase immunoreactivity. These results indicated that the myenteric ganglion neurons of the duodenum receive direct synaptic projection of sympathetic noradrenergic neurons and that their terminals contain pleomorphic vesicles and form asymmetric synaptic contacts.