Projections of afferent ventral root fibers on dorsal horn neurons in the cat spinal cord

Mechanisms of the effect of stimulation of afferent fibers in ventral roots on dorsal horn interneurons were investigated in experiments on anesthetized cats. Dorsal horn interneurons on which such fibers project were shown to exist. In particular, some dorsal horn interneurons can exert an inhibitory influence on effects of dorsal root fiber activation.Institute of Physiology, Academy of Sciences of the Kazakh SSR, Alma-Ata. Translated from Neirofiziologiya, Vol. 17, No. 3, pp. 300–305, May–June, 1985.     

Fos protein expression in the medulla oblongata and changes in size of spinal lateral horn neurons after 4-wk simulated weightlessness in rats

Responses of the neurons in medulla oblongata and C8-T1 spinal cord lateral horn of rats induced by simulated weightlessness were investigated using anti-Fos protein and anti-tyrosine hydroxylase (TH) double staining immunohistochemical methods, and Nissl-staining technique respectively. After four weeks of tail-suspension, many Fos-like positive neurons were localized in the medullary visceral zone (MVZ), predominantly in the nucleus of tractus solitarii and ventrolateral medulla, and some of them showed TH-like immunoreactivity. Sizes of the cell bodies of the lateral horn neurons in C8-T1 segment were significantly increased in 4-wk tail-suspended rats (P<0.05) as compared with that in controls. The results suggest that the neurons in MVZ and the spinal lateral horn may be involved in the adaptation of central cardiovascular regulation during weightlessness.     

Structure and localization of sympathetic neurons in the cat spinal cord

By the method of retrograde axonal transport of horseradish peroxidase (HP) structure and localization of sympathetic neurons sending axons to the cranial cervical ganglion (CCG) have been revealed ipsilaterally in the ventral horns and in 4 nuclei of the spinal cord: nucl ILp, nucl. ILf. nucl. IC, nucl. ICpe. Orientation of the neurons, their number, structure of the nuclei formed by them, degree of the CCG efferentation by the preganglionic fibres, which run from various nuclei, are different. In nucl. ILf two types of neurons have been revealed-triangle and spindle-shaped, they always orienting by their long axis in mediolateral direction. The greatest amount of HP-positive neurons are found in nucl. ILp. They form a well distinquished compact nucleus in the lateral horns. HP-labelled neurons in nucl. ILp are found at the level of segments T1-T8 with their maximal amount at the level of segments T1-T3. HP-positive neurons are detected in nucl. ILf beginning from the segment C8 up to the middle of T4, in nucl. IC-from the segment C8 up to T6, in nucl. ICpe-from the segment C8 up to T5, in the ventral horns-from the segment T1 up to T5. In rostocaudal direction from the segment C8 up to T8 the number of HP-positive neurons is decreasing, but the part of nucl. ILp neurons in the CCG efferentation, comparing to the neurons in other sympathetic structures of the spinal cord, is increasing.     

Development of catecholaminergic neurons in the human medulla oblongata

Distribution and maturation of catecholaminergic (CA) neurons have been studied by tyrosine hydroxylase immunohistochemistry in the medulla oblongata of human fetuses aged 14.5-25 weeks of gestation. Already at 14.5 weeks, CA neurons were observed in two longitudinally oriented cell clusters, one located ventrolaterally in the area of the lateral reticular and ambiguous nuclei, the other one dorsomedially forming 4 groups related to the dorsal vagal nucleus, the commissural nucleus of the vagus, the nucleus of the tractus solitarius and the area postrema. CA neurons in the area postrema were often found close to blood vessels. Scattered intermediate CA neurons were seen between these two larger clusters. CA neurons still appeared immature exhibiting bipolar morphology with only one or two short stout processes, which hardly branched. At 21 weeks, CA neurons occupied essentially the same location, but had a more mature morphology. Though still bipolar in shape, they had thinner and much longer processes which frequently branched. Both in the ventrolateral and the dorsomedial cell clusters, these processes were frequently lying close to blood vessels. At 25 weeks, CA cells had matured into multipolar neurons with long thin processes forming fine fiber networks in the ventrolateral medulla as well as around and within the dorsal vagal and solitarius nuclei. Only at this stage, a distinct CA fiber tract was seen located in the region of the tractus solitarius. Our results indicate that CA neurons in the human medulla, which are presumably involved in the control of ventilation and blood pressure, though generated rather early during development, mature relatively late.     

Changes in gas composition of expired air during electric stimulation of the ventral medulla oblongata]

The experiments on urethane-anesthetized cats with the electrically stimulated ventral brain stem revealed that caudal ventral medulla at the depth of 1500 microns possesses structures whose electrical activation increases the level of carbon dioxide in arterial blood and in the end portion of expirate, on the one hand, and decreases the oxygen content in expirate and arterial blood, on the other hand.     

Synaptic responses of medulla oblongata neurons to stimulation of the spinal stepping atria in the cat

Synaptic responses of medulla oblongata (bulbar) neurons to microstimulation of stepping points in the spinal dorsolateral funiculi were recorded in decrebrate cats. Upon stimulation of the stepping point both in the ipsi- and contralateral funiculi, 40% of the neurons generated synaptic responses; the remaining cells responded to stimulation of only one stepping point. A part of the bulbar neurons responds to stimulation of stepping points both at the C₂ and Th₁₂ level. The latent periods of the synaptic responses of the bulbar neurons to stimulation of the stepping point at the C₂ level were in the 2–10 msec range. The data obtained indicate that bulbar neurons, along with propriospinal neurons, can participate in evoking locomotion upon stimulating the stepping point.Institute of Information Transmission Problems, Academy of Sciences of the USSR, Moscow. M. V. Lomonosov State University, Moscow. Translated from Neirofiziologiya, Vol. 23, No. 3, pp. 328–333, May–June, 1991.     

The renin-angiotensin system and noradrenaline release in the hypothalamus and medulla oblongata

The effect of angiotensin II (AII) and 48 h bilateral nephrectomy on the 3H-norepinephrine (3H-NE) and 3H-NE metabolites release in vitro was studied in slices of male Wistar rat hypothalamus and medulla oblongata. The total 3H outflow of radioactivity was higher in AII exposed tissues than in nephrectomized ones of both organs. The 3H-NE and 3H-NE metabolites remanent radioactivity in the tissues increased in both the soluble cytoplasmatic fractions and the granular vesicle ones, in the two organs from the nephrectomized rats. The ratio between granular and cytoplasmatic NE and granular and cytoplasmatic radioactive metabolites was not noticeably altered in any of the groups. The release of 3H-NE caused by AII and the opposite effect by nephrectomy, agree with the inverse relationship demonstrated between endogenous NE content in the central nervous system and AII plasmatic levels. AII might act on presynaptic NE receptors of the cellular membrane. The relationship between the renin-AII system and the central nervous system catecholamines could be involved in the control of development and maintenance of the renal arterial hypertension.     

Glycine immunoreactive neurons in the medulla oblongata in cats

Using a highly specific antiserum to Glycine and a very sensitive immunohistochemical technique with streptavidin-HRP, we visualized for the first time a considerable number of glycine immunoreactive cell bodies and fibers in the cat medulla oblongata. These results suggest that glycine may play an essential role in nearly all the physiological functions involving the medulla oblongata, including the muscular atonia occurring during paradoxical sleep.     

Ultrastructural localization of substance P immunoreactivity in the ventral horn of the rat spinal cord

At the light microscope level, the minute concentrations of substance P (SP) in rat spinal ventral horn can be visualized best by amplification with the double bridge PAP method of Vacca et al. (1975; 1980) in 5 microns paraffin tissue sections. Morphologically, the immunoreactive sites resemble punctate bodies. They occur in close apposition with the large ventral horn cells and their associated neuronal processes. By the Sternberger PAP procedure, we now describe these punctate bodies at the electron microscope level. Ultrastructurally, they appear as tiny boutons (terminal and preterminal) and small unmyelinated processes. The boutons and processes typically contain one to several immunolabeled dense core vesicles among many immunolabeled clear vesicles. They range in size near the limit of resolution of the light microscope (LM), thereby justifying further the use of LM amplification staining by the double bridge method. The immunolabeled boutons often synapse with large smooth dendrites (which may originate from motoneurons) by asymmetrical or symmetrical synaptic densities. Their synaptic densities appear immunostained as well. The data support the view that the electrophysiological action of SP in the ventral horn occurs in part by synaptic action along the processes of the ventral horn cells. Other mechanisms of action are considered for the peptide as well. Additional types of membrane specializations (synaptoid junctions) and SP neural circuits are described below.     

Connections of neurons in the lumbar ventral horn of spinal cord are altered after long-standing limb loss in a macaque monkey

Explanations for the massive reorganization in primary motor cortex, M1, after limb amputation typically focus on processes that occur in cortex. Few have investigated whether changes in more peripheral parts of the pathway might also play a role in the reorganization. In the present study, we examined the integrity and connectivity of the spinal cord motoneurons in a macaque monkey (Macaca mulatta) that lost a hindlimb as a result of accidental injury more than 3.5 years earlier. To label motoneurons, multiple small injections of a neuroanatomical tracer were placed in the muscles of the hip just adjacent to the stump of the amputated leg, and in matched locations in the opposite side for control purposes. Injections of a second tracer were made in the intact foot. In the ventral horn that related to the intact hindlimb, motoneurons labeled by the hip injections were concentrated rostral and ventromedial to those labeled by the foot injections. Hip injections on the side of the amputation labeled neurons that were located well beyond the normal territory for motoneurons related to the hip and into the zone normally occupied by neurons projecting to the foot. Labeled motoneurons innervating the intact limb were significantly larger than neurons on the side of the amputation (x = 2410 and 2061 microm(2), respectively). The findings suggest that many neurons survived the long-standing amputation, and made new connections with remaining intact muscles. These new patterns of connectivity likely contribute to the reorganization of motor cortex in amputees, and perhaps to abnormal behaviors like those reported by human amputees.     

Intracellular pH regulation in ventral horn neurones cultured from embryonic rat spinal cord

Intracellular pH was measured with the pH-sensitive fluorescent probe BCECF in spinal cord neurones cultured from rat embryos. At an external pH of 7.3, the average steady-state pHi was 7.18 +/- 0.03 (SEM, n = 97) and 7.02 +/- 0.01 (n = 221) in HEPES-buffered and in bicarbonate-buffered medium, respectively. In both external media, pHi was strongly dependent on external pH (pHe). In HEPES-buffered medium, pHi recovery following an acid load induced by transient application of ammonium required external Na+ and was inhibited by amiloride, indicating the presence of a Na+/H+ exchange. Na(+)- and HCO3(-)-dependent, DIDS-sensitive alkalinizing mechanisms also contributed to pHi regulation in CO2/bicarbonate-buffered medium. The presence of an electrogenic Na(+)-HCO3- cotransporter was confirmed by the alkalinizing effect of KCl application. The fact that pHi is lower in CO2/bicarbonate- than in HEPES-buffered medium and the alkalinization observed upon suppression of external Cl- suggest that the acidifying Cl-/HCO3- transporter plays an important role in defining pHi.     

Intracellular pH regulation in ventral horn neurones cultured from embryonic rat spinal cord

Summary

We have isolated and characterized a cDNA from the marine sponge Geodia cydonlum coding for a new member of the tyrosine protein kinase (TK) family. The cDNA encodes a protein of M_r = 68 710, termed GCTK, which is homologous to class II receptor tyrosine kinases (RTKs). GCTK contains conserved amino acids (aa) characteristic of all protein kinases, and the sequences DLATRN and PIRWMATE which are highly specific for TKs. Furthermore, the sequence N-L-Y-x(3)-Y-Y-R Is highly homologous to the sequence D-[LIV]-Y-x(3)-Y-Y-R found only in class II RTKs. The sponge TK, when compared with mammalian class II RTKs, shows maximum 31% homology in the TK domain indicating that this the oldest member of class II RTK started to diverge from the common ancestral protein kinase 650 million years ago. Using GCTK as a probe we identified three mRNA signals ranging from 2μ6 to 0μ6 kb. Kinase activity was localized only in the cell membranes from G. cydonium (M_r = 65 000), and was not detected in the cytosol of this organism. Antibodies raised against a synthetic peptide, corresponding to the aa residues within the catalytic domain of the sponge TK, recognized strongly two proteins of M_r = 65 000; these proteins, present in membrane fractions, also bound to the anti-phosphotyrosine antibody. These data suggest that the TK cloned from the sponge is a membrane-associated 65 kDa protein. Moreover these results demonstrate that RTKs are present from the lowest group of multicellular eukaryotes, sponges, to mammals, and may suggest that RTKs are involved in a signal transduction pathway.     

Intracerebral connections after administration of kainate into the caudal part of ventral sections of the medulla oblongata]

It was found that 1 day after unilateral destruction with kainate of neurons in the caudal ventral medulla unanesthetized animals showed disorders of arterial pressure control with a prevalence of hypertensive response. In 7 days after operation there was a decrease in the rate of neuronal uptake of serotonin and an increase in choline-, dopamine-, noradrenaline-, and glycinergic mediation in the basal hypothalamus and rostral ventral medulla, which characterize the functional state of synaptic formations in the brain stem structures connected to neurons in the caudal ventral medulla and involved in cardiovascular regulation.     

Nitroxidergic neurons in nuclei of rat and human medulla oblongata

Four types of neurons distinguished by NADPH-diaphorase reaction have been identified in seven nuclei of human and rat medulla oblongata. It was found that nitrogen oxide (NO)-positive neurons had similar distributions; neurons with high NO-synthase activity predominated in vasomotor nuclei and, in sensitive nuclei, most neurons had low NO-syntase activity.     

Effect of mediodorsal thalamic nucleus on the respiratory neurons from medulla oblongata and the rat respiration during hypoxia]

In the normal as well as in the oxygen deficiency conditions the research has been conducted to study the influence of associative mediodorsal (MD) nucleus of thalamus on impulsive activity of respiratory neurons of medulla oblongata of respiration. In conditions of normal atmospheric pressure, before the uplift of the animals, the electrical stimulation of MD of nucleus of thalamus has had mainly inhibiting influence. In the initial phase, on 4-5 thousand meter altitude, activation of frequent discharge of neurons occurred, the respiration has become frequent as well. In this situation the inhibiting influence of stimulation of MD nucleus of thalamus was more accentuated than in conditions of normoxia. In the second phase, 7.5-8 thousand meters, the opposite occurred, i.e. reduction of respiratory center activity of medulla oblongata and thalamus. In this difficult conditions of hypoxia, a reduction of impulsive activity of neurons has been observed; the respiration was becoming slower and surface. Meanwhile, the inhibiting influence of thalamus was not significant.