Accessory oculomotor nuclei of man. 1. The nucleus of Darkschewitsch: a Nissl and Golgi study

A morphoquantitative study was carried out to provide detailed information regarding the cytoarchitecture and neuronal morphology of the nucleus of Darkschewitsch (ND) of man. The neuronal population showed heterogeneity of shape and size of the nerve cell bodies. Small and medium-sized neurons appeared scattered in a wide neuropil. In the Golgi material, two types of neurons were identified: multipolar and fusiform cells. Multipolar cells, which were the most numerous (77%), had 3-5 dendrites giving off primary bifurcations at a short distance from the nerve cell body. Sometimes dendrites and axons were seen to spread outside the ND. The fusiform cells had 1-2 dendrites emerging from the opposite poles of the elongated nerve cell bodies. The dendrites tended to run unbranched for long distances in the section plane before dichotomizing. The dendrites and axons of the fusiform cells always lay inside the ND. The cytoarchitectural features of the ND corresponded to the characters of the reticular formation so that the ND of man could be considered to be a typical reticular nucleus inside the central gray matter. The prevailing presence of multipolar neurons whose processes often spread outside the ND could suggest that the ND is a mainly projective nucleus.     

The interstitial nucleus of Cajal of the cat. II. Effects of kainic acid lesion on vertical optokinetic nystagmus and after-nystagmus

The effects of bilateral lesions of the interstitial nucleus of Cajal (INC) by ka?nic acid on vertical optokinetic nystagmus (OKN) and after-nystagmus (OKAN) were studied in four cats: in three cats, in the acute stage from 1 to 60 days after the lesions; in the fourth cat, they were studied 3 years after the lesions were made. Histological control of lesions showed that the whole INC was bilaterally destroyed in two cats of the acute group and only the upper part of INC in the third cat. In the chronic cat, the density of cell bodies in both INC was lower than normal. In the acute group, the cats exhibited a spontaneous downward eye drift in light and in darkness. During an upward optokinetic stimulation, the effect of INC lesions was dramatic: upward slow phases and downward quick phases of OKN were abolished. Sixty days post lesions, small upward slow eye movements were again observed. During a downward optokinetic stimulation, the defect was much less; in particular, after a slight impairment of downward slow phases, during the first days post lesions, they recovered quickly. The secondary optokinetic after nystagmus (OKAN II) ensuing a downward OKN was cancelled and did not reappear 60 days post lesions. In the chronic stage, three years after the lesions, during an upward optokinetic stimulation, the cat showed upward slow phases with velocities close to normal. However, upward slow phases were curved: the velocity at the end of the slow phases was lower than at the beginning. After an upward OKN (the direction of slow phases gives the direction of the OKN and OKAN), the ensuing OKAN was present but abnormal.     

Plasticity of interstitial cells of Cajal: a study of mouse colon

Ablation of the myenteric plexus in mouse colon with the detergent benzalkonium chloride (BAC) is followed by considerable recovery of the nerves, indicating that this plexus is capable of regeneration and has plasticity. Interstitial cells of Cajal (ICC) are closely associated with enteric nerves, and the acquisition and maintenance of their adult phenotype are nerve-dependent. Little is known about the regenerative processes of ICC or about the possible dependence of these processes on neurons. To address these questions, we ablated the myenteric plexus in the mouse colon with BAC and followed changes in the adjacent ICC (ICC-MP) from day 2 to day 70 after treatment, by using c-kit-immunohistochemistry and electron microscopy. In the untreated area, c-kit-positive cells and ICC-MP with normal ultrastructural features were always present. The region partially affected by BAC contained some c-kit-positive cells, and either normal or vacuolated ICC-MP were observed by electron microscopy. Moreover, at days 60–70, ICC-MP with particularly extended rough endoplasmic reticulum were present in this area. In the treated area, either denervated or reinnervated, c-kit-positive cells were always absent. By day 14 after BAC treatment, nerve fibers had started to grow back into the treated region and, in the reinnervated area, cells with fibroblast-like features appeared and were seen to contact both nerve endings and smooth muscle cells and to acquire some typical ICC features. Thus, ICC are vulnerable to external insult but appear to have some ability to regenerate.This work was supported by the US-Israel Binational Science Foundation (BSF, 98-00185; to M.H.) and University funds “quota di ateneo ex 60%” (M.-S. F.-P.).     

The interstitial nucleus of Cajal of the cat. I. Neurons activity related to vertical eye movements

Interstitial nucleus of Cajal (INC) neurons activity was studied during vertical optokinetic nystagmus (OKN) and after-nystagmus (OKAN) in awake cats lying on their right side. The activity of one hundred neurons was recorded in the left INC and analysed in relation with the vertical component of OKN and OKAN. The activity of 27 neurons was correlated either to eye position or to both eye velocity and eye position; 18 of these neurons were recorded in their on-direction and their off-direction. The analysis of the 18 neurons showed that the activity of 8 of them was correlated to eye position in the on-direction and in the off-direction and the correlation to eye position was higher than to eye velocity; these neurons are considered as position neurons. Seven other neurons had a higher correlation to eye position that to eye velocity in the on-direction and this relation reversed in the off-direction, these neurons are considered as position-velocity neurons. Thirty two burst-neurons were activated only during quick phases of OKN and OKAN and they were silent during slow phases and periods of fixation. Nine burst neurons had an upward on-direction and 23 neurons a downward on-direction. The eye velocity-average burst frequency (ABF) and quick phase duration-burst duration relationships had low correlations and suggested that INC burst neurons were excitatory premotor neurons. Statistical analysis showed that downward on-direction burst neurons had a higher ABF that upward on-direction burst neurons. Moreover, during OKN and OKAN, the velocity sensitivity of INC burst neurons was the same. The activity of the remaining neurons (41 neurons) was not quantitatively correlated to vertical and horizontal eye movements; they were classified as irregular tonic neurons. This study shows that INC neurons carry an eye position signal which was never reported before. This supports the results of INC lesion studies which showed that INC is involved in the vertical velocity to position integration. Moreover, there is an up versus down asymmetry in the frequency of INC burst neurons.     

The organization of the reptilian brainstem reticular formation: A comparative study using Nissl and Golgi techniques

The brainstem reticular formation has been studied in 16 genera representing 11 families of reptiles. Measurements of Nissl-stained reticular neurons revealed that they are distributed along a continuum, ranging in length from 10 μm to 95 μm. Reticular neurons in crocodilians and snakes tend to be larger than those found in lizards and turtles. Golgi studies revealed that reticular neurons posess long, rectilinear, sparsely branching dendrites. Small reticular neurons ( > 31 μm length) possess fusiform or triangular somata which bear two or three primary dendrites. These dendrites have a somewhat simpler ramification pattern when compared with those of large reticular neurons (< 30 μm length). Large reticular neurons generally possess perikarya which are triangular or polygonal in shape. The somata of large reticular neurons bear an average of four primary dendrites. The dendrites of reptilian reticular neurons ramify predominantly in the transverse plane and are devoid of spines or excrescences. The dendritic ramification patterns observed in the various repitilian reticular nuclei were correlated with known input and output connections of these nuclei. Nissl and Golgi techniques were used to divide the reticular formation into seven nuclei. A nucleus reticularis inferior (RI) is found in the myelencephalon, a reticularis medius (RM) in the caudal two-thirds of the metencephalon, and a reticularis superior (RS) in the rostral metencephalon and caudal mesencephalon. Reticularis inferior can be subdivided into a dorsal portion (RID) and a ventral portion (RIV). All reptilian groups possess RID and RM but RIV is lacking in turtles. Reticularis superior can be subdivided into a large-celled lateral portion (RSL) and a small-celled medial portion (RSM). All reptilian groups possess RSM and RSL, but RSL is quite variable in appearance, being best developed in snakes and crocodilians. The myelencephalic raphe nucleus is also quite variable in its morphology among the different reptilian families. A seventh reticular nucleus, reticularis ventrolateralis (RVL), is found only in snakes and in teiid lizards. It was noted that the reticular formation is simpler (fewer numbers of nuclei) in the representatives of older reptilian lineages and more complex (greater numbers of nuclei) in the more modern lineages. Certain reticular nuclei are present or more extensive in those families which have prominent axial musculature.     

A rapid method combining Golgi and Nissl staining to study neuronal morphology and cytoarchitecture.

The Golgi silver impregnation technique gives detailed information on neuronal morphology of the few neurons it labels, whereas the majority remain unstained. In contrast, the Nissl staining technique allows for consistent labeling of the whole neuronal population but gives very limited information on neuronal morphology. Most studies characterizing neuronal cell types in the context of their distribution within the tissue slice tend to use the Golgi silver impregnation technique for neuronal morphology followed by deimpregnation as a prerequisite for showing that neuron's histological location by subsequent Nissl staining. Here, we describe a rapid method combining Golgi silver impregnation with cresyl violet staining that provides a useful and simple approach to combining cellular morphology with cytoarchitecture without the need for deimpregnating the tissue. Our method allowed us to identify neurons of the facial nucleus and the supratrigeminal nucleus, as well as assessing cellular distribution within layers of the dorsal cochlear nucleus. With this method, we also have been able to directly compare morphological characteristics of neuronal somata at the dorsal cochlear nucleus when labeled with cresyl violet with those obtained with the Golgi method, and we found that cresyl violet-labeled cell bodies appear smaller at high cellular densities. Our observation suggests that cresyl violet staining is inadequate to quantify differences in soma sizes.     

The Nissl substance of living and fixed spinal ganglion cells. II. An ultraviolet absorption study

Living chick spinal ganglion neurons grown for 19 to 25 days in vitro were photographed with a color-translating ultraviolet microscope (UV-91) at 265, 287, and 310 mmicro. This instrument was unique in permitting rapid accumulation of ultraviolet information with minimal damage to the cell. In the photographs taken at 265 mmicro of the living neurons, discrete ultraviolet-absorbing cytoplasmic masses were observed which were found to be virtually unchanged in appearance after formalin fixation. These were identical with the Nissl bodies of the same cells seen after staining with basic dyes. The correlation of ultraviolet absorption, ribonuclease extraction, and staining experiments with acid and basic dyes confirmed the ribonucleoprotein nature of these Nissl bodies in the living and fixed cells. No change in distribution or concentration of ultraviolet-absorbing substance was observed in the first 12 ultraviolet photographs of a neuron, and it is concluded that the cells had not been subjected to significant ultraviolet damage during the period of photography. On the basis of these observations, as well as previous findings with phase contrast microscopy, it is concluded that Nissl bodies preexist in the living neuron as discrete aggregates containing high concentrations of nucleoprotein.     

Physiology and pathophysiology of the interstitial cell of Cajal: from bench to bedside. I. Functional development and plasticity of interstitial cells of Cajal networks

Interstitial cells of Cajal (ICC) are the pacemaker cells in gastrointestinal (GI) muscles. They also mediate or transduce inputs from enteric motor nerves to the smooth muscle syncytium. What is known about functional roles of ICC comes from developmental studies based on the discovery that ICC express c-kit. Functional development of ICC networks depends on signaling via the Kit receptor pathway. Immunohistochemical studies using Kit antibodies have expanded our knowledge about the ICC phenotype, the structure of ICC networks, the interactions of ICC with other cells within the tunica muscularis, and the loss of ICC in some motility disorders. Manipulating Kit signaling with reagents to block the receptor or downstream signaling pathways or by using mutant mice in which Kit or its ligand, stem cell factor, are defective has allowed novel studies of the development of these cells within the tunica muscularis and also allowed the study of specific functions of different classes of ICC in several regions of the GI tract. This article examines the role of ICC in GI motility, focusing on the functional development and maintenance of ICC networks in the GI tract and the phenotypic changes that can occur when the Kit signaling pathway is disrupted.     

Demonstration of a sparse direct projection from the interstitial nucleus of Cajal to the cerebellum in the cat

In the cat the possible contribution of cerebellar afferents from the accessory oculomotor nuclei was investigated by means of retrograde transport of the wheat germ agglutinin-horseradish peroxidase complex. A sparse bilateral projection from the interstitial nucleus of Cajal was demonstrated. The fibres reach cerebellar cortical as well as nuclear regions. No retrogradely labelled cells were found in the nucleus of Darkschewitsch and the nucleus of the posterior commissure.     

Antidromic and synaptic potentials evoked in neurons of the lateral vestibular nucleus by stimulation of the interstitial nucleus of Cajal and Darkshevich's nucleus in cats

Effects of stimulation of the interstitial nucleus of Cajal and Darkshevich's nucleus on unit activity in the lateral vestibular nucleus of Dieters were investigated in cats anesthetized with pentobarbital. Stimulation of the above-mentioned structures was shown to lead to antidromic and orthodromic activation of Dieters neurons. Axon collateral of vestibular neurons, ascending to the above-mentioned brain-stem structures were discovered electrophysiologically. Stimulation of the nuclei of Cajal and Darkshevich was shown to evoke mono- and polysynaptic EPSPs and IPSPs in neurons of Deiters nucleus. Convergence of influences from both nuclei on the neurons studied was demonstrated. The particular features and functional role of the influences observed are discussed.L. A. Orbeli Institute of Physiology, Academy of Sciences of the Armenian SSR, Erevan. Translated from Neirofiziologiya, Vol. 16, No. 6, pp. 822–829, November–December, 1984.     

Inherent rhythmcity and interstitial cells of Cajal in a frog vein

Interstitial cells of Cajal are responsible for rhythmic contractions of the musculature of the gastrointestinal tract and blood vessels. The existence of these cells and spontaneous rhythmicity were noticed in amphibian vein and the findings are reported in this paper. The postcaval vein was identified in the frog, Rana tigrina and was perfused with amphibian Ringer solution after isolation. Contractile activity was recorded through a tension transducer connected to a polygraph. The isolated postcaval vein showed spontaneous rhythmic activity. Addition of cold Ringer solution decreased, while warm Ringer increased, the rate of contraction. Adrenaline caused inhibition of rhythmic activity at a dosage that increased the rate of isolated sinus venosus. Sections of the postcaval vein, when stained supravitally with methylene blue, showed the presence of interstitial cells of Cajal. Photic stimulation of the vein in the presence of methylene blue led to a significant decrease in the rate of spontaneous beating of the vein. These findings indicate that the postcaval vein of frog is capable of inherent rhythmcity, which is dependent on the interstitial cells of Cajal but is independent of the sinus venosus.     

Reciprocal connections between the red nucleus and the trigeminal nuclei: a retrograde and anterograde tracing study.

An anterograde biocytin and a retrograde WGA-colloidal gold study in the rat can provide information about reciprocal communication pathways between the red nucleus and the trigeminal sensory complex. No terminals were found within the trigeminal motor nucleus, in contrast with the facial motor nucleus. A dense terminal field was observed in the parvicellular reticular formation ventrally to the trigeminal motor nucleus. The parvicellular area may be important for the control of jaw movements by rubrotrigeminal inputs. On the other hand, the contralateral rostral parvicellular part of the red nucleus receives terminals from the same zone in the rostral part of the trigeminal sensory complex, where retrogradely labelled neurones were found after tracer injections into the red nucleus. Such relationships could be part of a control loop for somatosensory information from the orofacial area.     

The neuronal cell types in the arcuate nucleus in normal and monosodium glutamate treated rats: a Golgi study

Two histochemical different populations of neurons migrate from the median eminence (ME) into the arcuate nucleus (ARC) at puberty in rats. Therefore the cellular composition of the ARC was studied in normal and monosodium glutamate (MSG) treated female rats at 22, 35, 55 days after birth and in mature ones. Golgi-Cox and mainly Rio-Hortega techniques with Nissl counterstaining were applied. In the mature arcuate nucleus several types of Golgi impregnated cells can be discerned. Neonatal MSG-treatment exclusively destructs the arcuate dopaminergic cells. From the Golgi-preparations it appears that in these animals one type is indeed damaged in or at the ARC.     

The pacemaker activity of interstitial cells of Cajal and gastric electrical activity

Interstitial cells of Cajal (ICC) are the pacemaker cells in the gut. They have special properties that make them unique in their ability to generate and propagate slow waves in gastrointestinal muscles. The electrical slow wave activity determines the characteristic frequency of phasic contractions of the stomach, intestine and colon. Slow waves also determine the direction and velocity of propagation of peristaltic activity, in concert with the enteric nervous system. Characterization of receptors and ion channels in the ICC membrane is under way, and manipulation of slow wave activity markedly alters the movement of contents through the gut. Gastric myoelectrical slow wave activity produced by pacemaker cells (ICC) can be reflected by electrogastrography (EGG). Electrogastrography is a perspective non-invasive method that can detect gastric dysrhythmias associated with symptoms of nausea or delayed gastric emptying.     

Physiology and pathophysiology of the interstitial cells of Cajal: from bench to bedside. IV. Genetic and animal models of GI motility disorders caused by loss of interstitial cells of Cajal

Several human motility disorders have been shown to be associated with loss or defects in interstitial cells of Cajal (ICC) networks. Because tissue samples for these studies were taken from patients with well-advanced motility problems, it is difficult to determine whether the loss of ICC is a cause or a consequence of the disease process. To establish the cause-and-effect relationship of ICC loss in motility disorders, it may be feasible to use animal models in which ICC are lost as motility dysfunction develops. Several models with defects in ICC networks have been developed, and these include animals with defects in the Kit signaling pathway (e.g., white-spotting mutants that have defects in Kit receptors; steel mutants that have mutations in stem cell factor, the ligand for Kit; and animals that are chronically treated with reagents that block Kit or downstream signaling proteins). ICC do not die when Kit signaling is blocked, rather, they redifferentiate into a smooth muscle-like phenotype. Diabetic animals (NOD/LtJ mice), animals with chronic bowel obstruction, and inflammatory bowel models also have defects in ICC networks that have been associated with motility disorders. By studying these models with molecular and genomic techniques it may be possible to determine the signals that cause loss of ICC and find ways of restoring ICC to dysfunctional tissues. This article discusses recent progress in the utilization of animal models to study the consequences of losing ICC on the development of motility disorders.     

Golgi study of the anterior dorsal ventricular ridge in a lizard. II. Neuronal cytodifferentiation

The development of the morphologic features of neurons in the anterior dorsal ventricular ridge (ADVR) has been followed in Golgi preparations from the lizard Gallotia galloti between embryonic stage 32 and post-eclosion stages of specimens 3.6–4.5 cm in length. The differentiation sequence of multipolar and bitufted neurons was established. Dendritic growth cones are present after stage 34. Filiform dendritic processes are replaced later on by spines. Clusters of neurons first appear at stage 39 in the periventricular zone, the cells becoming Golgi-impregnated in pairs. After hatching, the number of impregnated cells per cluster increases.