Transmission in the sensorimotor synapses of the lumbar spinal cord in Rana ridibunda tadpoles

In experiments on preparations of isolated spinal cord of the tadpoles, intracellular studies have been made on the synaptic potentials evoked in the lumbar motoneurones during total activation of the fibers within the 9th dorsal root. It was shown that primary afferents form monosynaptic contacts with motoneurones at stages XIV-XXV. During larval development, the number of motor cells in which monosynaptic EPSPs are recorded increases, whereas the number of motoneurones with only polysynaptic reactions decreases. From the moment of formation of monosynaptic contacts, transmission in direct sensory-motor synapses is realised by a dual (electrical-chemical) mode. The data obtained are discussed in relation to the problem of evolution of synaptic transmission between heterotypic neurones in vertebrates.     

The ultrastructural characteristics of the motor neuron synaptic organization in the spinal cord of the frog Rana ridibunda

Electron microscopic studies on the spinal motor nuclei in amphibians indicate significant diversity in chemical synapses formed on motoneurones by axonal endings of supra- and intraspinal systems. High ultrastructural specialization was observed among axosomatic, axodendritic and axoaxonal synapses. Several types of axo-spine synapses and axodentritic synaptic complexes of the "glomerular" type were revealed. New data on ultrastructural peculiarities of chemical synapses presented in this paper, together with earlier detailed data on morphologically mixed and electrotonic synapses, increase our knowledge of evolutionary trends in synaptic organization of motoneurones in the spinal cord and suggest the existence of a complex mechanism of integration of synaptic influences in the spinal cord of lower vertebrates.     

Potentiation of postsynaptic potentials in motoneurons of the frog,Rana ridibunda, by gallamine

The potentiating effect of gallamine on postsynaptic potentials was studied intracellularly on the preparation of the isolated spinal cord of the frog,Rana ridibunda. The amplitude both of afferentPSP (stimulation of DR) and of descendingPSP (stimulation of LC and VC) increased to 200–600% of the initial value. The responses reached 20 mV, and a convulsive effect developed. Both early (mono- and disynaptic) and late components were considerably potentiated. New components, absent prior to the gallamine application, appeared. Responses to a shortterm application of glutamate were potentiated much less than PSP in the same cell. NMDAresponses were not potentiated. Brain treatment with EAA blockers showed that potentiation and convulsive effect might appear under conditions of blockage of NMDA receptors but not under conditions of blockage of non-NMDA receptors. Gallamine eliminated inhibition evoked by the agents that block EAA. After gallamine application the duration of antidromic and orthodromic AP did not change substantially, while after-depolarization increased considerably, which resulted in generation of 2-3 AP in response to a single stimulus. A competitive blocker of cholinergic transmission, d-tubocurarine, did not potentiate PSP but increased synaptic noise. Mechanisms of gallamine action are discussed.     

Ultrastructure of terminals of primary afferents in the ninth segment of the lumbar spinal cord of the tadpole Rana ridibunda

Studies have been made on the fine structure of HRP-labeled primary afferent endings in segment IX of the developing spinal cord during formation of direct sensory-motor contacts. It was found that single dorsal root collaterals reach the lateral motor nucleus and form synapses on motoneuronal bodies and their dendrites. In some of the synapses, two types of junctional specialization, i.e. active zone and gap junction, were observed.     

Participation of the GABA B receptors in presynaptic inhibition of the spinal cord descending projections fibers in the frog Rana ridibunda

(-)-Baclophen was found to depress in a dose-dependent and reversible way the excitatory post-synaptic potentials (EPSPS) of motor neurons and the ventral root potentials evoked by stimulation of fibres of the ipsi- and contralateral ventral columns. The (-)-baclophen depressing effect could be eliminated with saclophen. Picrotoxin eliminated the depressing effect of the GABA on the descending EPSPS. Depressing effects of (-)-baclophen and GABA upon the ventral root potentials were also shown. The data obtained in pharmacological analysis corroborate to a certain extent existence of the GABAB receptor presynaptic inhibition in descending fibres monosynaptically corrected with the spinal cord motor neurons in the frog Rana ridibunda.     

Characterization and localization of two forms of gonadotropin-releasing hormone (GnRH) in the spinal cord of the frog Rana ridibunda

Two molecular variants of gonadotropin-releasing hormone (GnRH) have been previously characterized in the brain of amphibians, i.e., mammalian GnRH (mGnRH) and chicken GnRH-II (cGnRH-II). The aim of the present study was to identify the molecular forms of gonadotropin-releasing hormone and to localize gonadotropin-releasing hormone-containing elements in the spinal cord of the frog Rana ridibunda using highly specific antisera against mGnRH and cGnRH-II. High-performance liquid chromatography (HPLC) analysis combined with radioimmunoassay (RIA) detection revealed that frog spinal cord extracts contained both mGnRH and cGnRH-II. Immunohistochemical labeling revealed that the frog spinal cord was devoid of GnRH-containing cell bodies. In contrast, numerous GnRH-immunoreactive fibers were observed throughout the entire length of the cord. mGnRH immunoreactivity was only detected in the rostral region of the cord and consisted of varicose processes located in the vicinity of the central canal. cGnRH-II-positive fibers were found throughout the spinal cord, the density of immunoreactive processes decreasing gradually toward the caudal region. Two main cGnRH-II-positive fiber tracts with a rostrocaudal orientation were observed: a relatively dense fiber bundle surrounding the central canal, and a more diffuse plexus in the white matter. In addition, short, varicose cGnRH-II-positive processes with a radial orientation were present throughout the gray matter. These fibers were particularly abundant ventromedially and formed a diffuse network that ramified laterally to end in the vicinity of motoneurons. Taken together, these data indicate that the frog spinal cord, like the frog brain, contains two forms of GnRH. The presence of numerous cGnRH-II-immunoreactive fibers in the ventral horn suggests that cGnRH-II may influence the activity of a subpopulation of motoneurons.     

Effect of light negative hydroaeroions on electromagnetic coupling of frog Rana ridibunda auricle

It is shown that negative hydroaeroions cause a decrease of the systole strength of freshly prepared auricle and activation of the tired auricle; a decrease of the amplitude and length of the heart trabecula action potentials due to depression of slow input Ca2+ current; shift of the threshold of activation of fast input Na+ current to low potentials; partly elimination of the rise of the auricle systole strength under novodrine. The latter effects were not observed under atropine and indomethacin.     

The heterogeneity of the excitatory synaptic inputs in the spinal motor neurons of the frog Rana ridibunda

The synaptic responses induced in motoneurones by the stimulations of the dorsal root (DR), single afferent fibres and reticular formation (RF) were intracellularly recorded in the isolated frog spinal cord. It was shown that argiopine (the selective blocker of glutamate receptors of non-NMDA type) in concentrations ranging from 3.10(-7) to 1.10(-5) M effectively suppressed the di- and polysynaptic, but not the monosynaptic components of EPSP's induced by DR stimulation. The initial reaction to argiopine consisted of the increase of this monosynaptic component of EPSP. In the same concentrations range, argiopine reduced both mono- and polysynaptic EPSP, evoked by RF stimulation. 2-amino-phosphonovaleric acid (1.10(-4) M) did not affect, whereas the kinurenate (1--2.10(-3) M) completely blocked the amplitude of all kinds of synaptic responses. The various effects of argiopine on the responses induced by microstimulation of presynaptic nerve terminals were observed. The data obtained speak in favour of heterogeneity of monosynaptic excitatory inputs in the motoneurones of frog spinal cord. Being the glutamatergic by nature, the inputs differ in the properties of postsynaptic receptors. All of these receptors concerning to non NMDA-type can be divided to argiopine-sensitive and argiopine-resistant. The first seem to be involved in the monosynaptic connections of RF and the second--in those of primary afferents with motoneurones.     

Characterization of the cDNA encoding proopiomelanocortin in the frog Rana ridibunda

In the amphibian pars intermedia, secretion of proopiomelanocortin (POMC)-derived peptides is controlled by multiple factors including classical neurotransmitters and neuropeptides. To pursue questions concerning the regulation of POMC gene expression in Rana ridibunda, we have isolated and characterized a full-length cDNA for frog POMC. A cDNA clone isolated from a frog pituitary library contains an open-reading frame of 780-bp that predicts a 260 amino acid POMC protein. The structure of frog POMC demonstrates considerable amino acid sequence similarity with POMC from other species. In particular, the sequence of alpha-melanotropin (alpha-MSH) is identical in frog and all mammalian species studied so far, while adrenocorticotropin (ACTH) and beta-endorphin exhibit 79% and 84% homology with their human counterpart. Frog POMC contains only one potential asparagine-linked N-glycosylation signal (Asn-Ser-Thr) within the gamma-MSH domain. The alpha-MSH sequence is C-terminally flanked by the Gly-Lys-Lys amidation signal while the joining peptide is not amidate.     

Ultrastructure and synaptic architecture of spinal motoneurons in the frog (Rana catesbeiana)

An electron-microscopic analysis of spinal motoneurons and their synapses was carried out in a frog (Rana catesbeiana). Six different types of boutons (S, F, M, P, C and GS) have been identified. Their distribution on spinal motoneuron somata and proximal dendrites is described. The mean linear percentage of the surface area covered by boutons is 26.1 +/- 1.9%. S-type boutons are preferentially concentrated on the soma and proximal dendrites. The relative number of S-type boutons (58.7%) was greater (p less than 0.01) than that of F-type boutons (41.3%). This is in contrast to mammalian spinal motoneurons where F-type boutons are much more numerous on the soma than S-type boutons. F-type boutons are randomly distributed and the average ratio of S:F-type boutons is 20:14 (S:F ratio = 1.4). In contrast, M-type boutons synapse exclusively on the distal part of the dorsal dendrites and are restricted to the intermediate zone or to the dorsal horn. P-type boutons form synapses upon the large M-type boutons. The polarity of these axoaxonic synapses is always from P to M. Similarities and differences between the synaptology of frog and mammalian spinal motoneurons are discussed.     

A study on cholesterol content in some tissues of the Saudi frog: Rana ridibunda.

1. Cholesterol content of selected tissues in the Saudi frog Rana ridibunda was determined. 2. Total cholesterol content was higher in tissues involved in water loss, ranging from 12.22 +/- 0.28 to 40.3 +/- 0.79 mg/g. 3. In tissues not involved in water loss, total cholesterol content ranged from 2.42 +/- 0.18 to 19.30 +/- 0.39 mg/g.     

Biochemical variability and genetic differentiation of marsh frog Rana ridibunda Pall. populations]

Marsh frog populations of Rana ridibunda s. lato from the Ukraine, Caucasus, Turkmenistan and Tadzhikistan have been analyzed genetically by 31 isozyme gene loci. The genetic distance between populations from Tadzhikistan on the one hand, and other regions, on the other hand is D-0.269, that assumes evolutionary divergence at the species level. The presence of fixed allelic substitutions between two populations from Tadzhikistan by five gene loci (Aat-S, Ldh-B, Alb, Es-5) supposes genetic heterogeneity of species in this region. Peculiarities of allozyme variation of this species are discussed.     

A light and electron microscopic study of the CB1 cannabinoid receptor in the primate spinal cord

The distribution of cannabinoid receptors was studied in the monkey spinal cord by immunocytochemistry and electron microscopy, using an antibody to the CB1 brain cannabinoid receptor. Large numbers of labelled neurons were observed in all portions of the grey matter of the spinal cord. These included small diameter 9–16µm neurons in the dorsal horn, larger (40–60µm) neurons in the intermediate grey, and very large (60–100µm), motor neurons in the ventral horn. Reaction product was observed in dendrites postsynaptic to unlabelled axon terminals. Since cannabinoid receptor activation decreases neuronal excitability by several mechanisms, including inhibition of voltage dependent calcium channels, the dense staining of CB1 in dorsal horn neurons suggests that CB1 could reduce calcium influx through such channels in these neurons. This, in turn, could decrease calcium-dependent changes in synaptic transmission and decrease sensitisation to nociceptive stimuli in these neurons. Similarly, the dense staining of CB1 in ventral horn cells suggests that cannabinoid receptors could limit calcium influx through voltage dependent calcium channels in these neurons, and could be significant in terms of neuroprotection to these neurons.     

Amplitude of miniature potentials of motor neurons in the frog Rana ridibunda

Miniature synaptic potentials have been recorded from motoneurones of the isolated spinal cord of the frog Rana ridibunda. In normal Ringer's solution, their frequency varied from 5 to 50/sec, whereas their amplitude reached 2-5 mV. Only 50-300 microV (rarely 0.5-1.5 mV) potentials persisted when TTX was added to Ringer's solution and/or Ca was replaced by Mn. However, in Ca-free solution, TTX in combination with Mn did not decrease the amplitude of miniature potentials, provided the initial values varied within 50-300 microV. Noise fluctuations did not exceed 40-50 microV, and the ratio of the number of miniature potentials of 50 microV to the number of 50 microV noise potentials was about 10:1. The observed miniature potentials with an amplitude of 50-100 microV coincide with the quantal units calculated by other authors from statistical analysis of the unitary EPSPs evoked by primary afferents or by ventrolateral tract fibers.