Search PubMed⌕ Search

Biomedical subjects

L Urban

Publications and source records attributed to L Urban.

83 records · Page 5Linked to original sources
PubMed1989

Cholinergic effects on spinal dorsal horn neurons in vitro: an intracellular study.

The cholinoceptive properties of dorsal horn neurons (lamina III-V) were investigated by means of intracellular recordings from the rat isolated spinal cord slice preparation. In half of the neurons investigated, acetylcholine (ACh) evoked a dose-dependent slow depolarization and increase in excitability; hyperpolarization was observed in 10% of neurons. Acetyl-beta-methylcholine (MCh) similarly depolarized 39% and hyperpolarized 25% of neurons tested; depolarization was also observed following bethanechol. Responses to the muscarinic agonists were abolished by atropine (10(-5) M). Nicotine depolarized 84% of tested neurons; dihydro-beta-erythroidine (5 x 10(-5) M) and (+)-tubocurarine (10(-6) M) antagonized this depolarization. ACh-, MCh- and nicotine-induced depolarizations, associated with changes in input resistance, were maintained in the presence of tetrodotoxin (10(-6) M). Substance P, as well as repetitive electrical stimulation of the dorsal root, also evoked depolarization in ACh-sensitive neurons. Atropine, but not (+)-tubocurarine, diminished responses to both substance P and dorsal root stimulation. These results indicate that dorsal horn neurons are ACh-sensitive and possess both muscarinic and nicotinic receptors. In addition, the parallel sensitivity of neurons to muscarinic agonists, substance P and dorsal root stimulation, as well as the parallel antagonistic effect of atropine, are supportive of a common ionic mechanism underlying the activation of muscarinic and substance P receptors.

Action Potentials↗
PubMed1989

Postischemic synaptic physiology in area CA1 of the gerbil hippocampus studied in vitro.

After transient forebrain ischemia in the Mongolian gerbil, CA1b hippocampal pyramidal cells degenerate during a period of 2-4 d. We tested the hypothesis that this delayed neuronal death is preceded by excessive synaptic excitation. Hippocampal slices were prepared from gerbils that had been subjected to a 5 min occlusion of both common carotid arteries. Input/output curves demonstrated enhancement of the initial slope of the Schaffer collateral-commissural focally recorded EPSP at all stimulus currents between 5 and 10 hr after the ischemic insult. The duration of the focally recorded EPSP also increased. At the same time, the excitability of the CA1b pyramidal cells decreased. Thus, the EPSP brought fewer pyramidal cells to threshold than the same size EPSP in control slices. During the first 14 hr after ischemia, the antidromic population spike remained unaffected. By 24 hr after ischemia, however, the focally recorded EPSP and both orthodromic and antidromic population spikes were markedly depressed, and they declined further over the next 2 d. No recovery was detected. In the same slices, transient ischemia only mildly and reversibly affected the response of dentate granule cells to perforant path stimulation and did not affect their response to antidromic stimulation. Hippocampal slices adjacent to those used for electrophysiological recording were analyzed histologically. Examination of somatic argyrophilia confirmed that CA1b pyramidal cells suffered delayed neuronal death, whereas dentate granule cells remained intact. Pyramidal cell argyrophilia was, however, not detected until 2 d after these neurons had become virtually inexcitable. We conclude that CA1b pyramidal cells begin to lose electrophysiological function well before definite morphological signs of degeneration become visible. The observation of enhanced excitatory transmission 5-10 hr after reperfusion is consistent with the idea that delayed ischemic neuronal death results, at least in part, from excessive excitation.

Action Potentials↗
PubMed1988

An intracellular analysis of amino acid induced excitations of deep dorsal horn neurones in the rat spinal cord slice.

The rat spinal cord slice preparation has been used to investigate the sensitivity of deep dorsal horn neurones to the excitatory amino acids N-methyl-D-aspartate (NMDA), quisqualate and L-glutamate. Intracellular recordings were made from 44 neurones in laminae III-VI of 14- to 16-day rats. Superfusion of quisqualate (30 microM) excited all neurones, NMDA (50 microM) excited 72% and L-glutamate (0.5-1 mM) 63% of the neurones. Depolarizations were retained after tetrodotoxin but with a reduced amplitude. The NMDA antagonist D-aminophosphonovalerate (D-APV, 10 microM) reduced NMDA and L-glutamate depolarizations by 66% and by 40%, respectively, while the quisqualate responses were enhanced by 27%. Dorsal root stimulation elicited two main patterns of activity; short-latency single/double spikes followed by subthreshold excitatory postsynaptic potentials (EPSPs) or a burst of spikes rising from a long duration composite EPSP. D-APV reduced the long-latency components of the first type and reduced the amplitude and duration of the composite EPSP of the second. These results support a specialized role for NMDA receptors in synaptic transmission in the dorsal horn.

2-Amino-5-phosphonovalerate↗
PubMed1988

The responses recorded in vitro of deep dorsal horn neurons to direct and orthodromic stimulation in the young rat spinal cord.

The electrophysiological properties of 87 neurons in the deep dorsal horn (laminae III-VI) of the rat spinal cord have been investigated in vitro. Two preparations have been used; the transverse spinal cord slice preparation from the third or fourth lumbar segments of 14-16-day-old rats (71 cells) and a hemisected lumbar spinal cord preparation from 10-12-day-old rats (16 cells). The input impedances (range 11-128 M omega), membrane potentials (-67 +/- 8 mV S.D.), action potential amplitude (77 +/- 11.8 mV) duration (1.4 +/- 0.5 ms) and afterpotentials, were effectively identical in the neurons recorded from the two preparations. Neurons in both preparations when activated with long-duration (1-2 s) outward current pulses showed a single steady-state firing range with little adaptation of firing frequency or action potential amplitude. This pattern of responses was unaffected by changing the membrane potential. Orthodromic synaptic activity could be elicited in the neurons by stimulating either the small dorsal root remnants in the slice or the dorsal roots in the hemisected spinal cord. The responses evoked by single stimuli of increasing intensity varied in different neurons in both preparations. The commonest response (32/62) consisted of a short-latency, short-duration composite excitatory postsynaptic potential which generated one or two spikes with no further spiking activity at longer latency when the stimulus intensity was increased beyond threshold. In 20 neurons, graded stimulation produced a graded response with recruitment, at high intensities, of a discharge of action potentials lasting several hundred milliseconds. A small number of cells (4) responded to the single stimulus with a train of action potentials lasting several seconds. Stimulating adjacent dorsal roots in the hemisected cord preparation could evoke quite different responses from the neurons. The heterogeneity of the types of orthodromic responses obtained in both preparations, in spite of the almost uniform intrinsic membrane properties, is likely to reflect differences in the strength, location and type of afferent and interneuronal input to different dorsal horn cells.

Action Potentials↗
PubMed1987

Distribution, origin and sensitivity to capsaicin of primary afferent substance P-immunoreactive nerves in the heart.

This report is intended as an overview of the distribution, origin and sensitivity to capsaicin of substance P-immunoreactive (SP-I) primary afferent cardiac nerves. Immunohistochemical and physiological methods were employed to compare the presence and density of these nerve fibers in the guinea pig and rat hearts. SP-I fibers are numerous in the guinea pig heart including the parietal pericardium, atria, ventricles, valves, coronary arteries and around intrinsic cardiac ganglion cells. The rat heart contains few SP-I fibers. Vagotomy does not influence the number of intensity of immunoreactive fibers in the guinea pig heart. By stimulating the atrium or ventricle and recording from the second or third thoracic dorsal roots Ad1, Ad2 and C fibers were demonstrated in the atria, but only Ad fibers in the guinea pig ventricle; in addition, only Ad fibers were recorded from the vagus nerves. Only Ad1 fibers were demonstrated in the rat heart. Treatment with capsaicin depletes the SP-I and decreases the conduction velocity of C-fibers and some Ad2 fibers in the guinea pig heart. We suggest that SP-I primary afferent nerve fibers are unmyelinated (C-type) or small myelinated (Ad2-type) nerves in the guinea pig heart and that their cell bodies of origin are predominantly in dorsal root ganglia.

Afferent Pathways↗
PubMed1986

Effects of polyclonal and monoclonal antibodies to substance P on slow excitatory transmission in rat spinal dorsal horn.

The effects of bath perfusion of polyclonal and monoclonal antibodies to substance P (SP) on slow excitatory transmission in rat dorsal horn have been investigated by intracellular recording in the immature rat spinal cord slice preparation. Both polyclonal and monoclonal antibodies to SP produced a significant decrease in the amplitude and the duration of the slow depolarization generated in dorsal horn neurons by high intensity, repetitive dorsal root stimulation or exogenous SP application. The effect of endogenous SP, or SP-related peptide, released during dorsal root stimulation appears likely since bath perfusion of a slice with a normal rabbit serum, or affinity chromatography preadsorbed SP antiserum, or non-specific IgG, or 5-hydroxytryptamine antiserum had no similar depressant effect. These results, if taken together with other experimental evidence, suggest that SP, or SP-like peptide, is in some way involved in a generation of the dorsal root-evoked slow depolarization. In addition, a novel approach is presented for using polyclonal and monoclonal antibodies to SP as pharmacological antagonists. Use of a specific characterized monoclonal antibody for the detection of physiological and pharmacological effects of putative peptide transmitters in vitro opens new avenues for further investigations.

Animals↗
PubMed1985

Origin of small primary afferent substance P-immunoreactive nerve fibers in the guinea-pig heart.

Primary afferent and substance P (SP)-immunoreactive nerve fibers of the guinea pig and rat heart were investigated by physiological and immunohistochemical methods. Immunohistochemistry revealed abundant SP-positive fibers in the guinea pig atria, with fewer in the ventricles. Only an occasional fiber was seen in the rat atrium or ventricle. Sectioning the vagus nerve did not noticeably influence the supply of SP-immunoreactive nerve fibers in the guinea pig heart. When the atria or ventricles were stimulated, afferent nerve fiber activity was recorded from the second and third thoracic dorsal roots. In guinea pig atria 3 types of fibers were identified on the basis of conduction velocities: A delta 1, A delta 2 and C fibers. Only A delta fibers were identified in the ventricle. By vagal recordings, A delta fibers were demonstrated but a C fiber response could not be shown in this nerve. SP-immunoreactivity in primary afferent fibers was depleted by the neurotoxin capsaicin. Capsaicin treatment also caused a reduction in the conduction velocity of small diameter myelinated A delta 2 (by 29%) and unmyelinated C fibers (by 46%). In the rat heart, evidence for A delta 2 or C fibers was not found. These results indicate that primary afferent and SP-immunoreactive fibers are numerous in guinea pig heart, but few in the rat. It is concluded that most of these fibers have their cell bodies of origin in the dorsal root ganglia.

Animals↗
PubMed1981

Peripheral reinnervation patterns and dorsal root ganglion cell topography in skin-grafted Rana pipiens frogs.

Behavioral responses to mechanical stimulation of the skin were observed in unoperated, 180 degrees skin-rotated and sham-grafted Rana pipiens frogs before and after crushing various cutaneous nerve bundles. In the two control groups, wiping responses directed towards the animal's dorsum were mediated solely via dorsomedial (DM) and dorsolateral (DL) nerve trunks. In skin-rotated frogs, DM and DL nerve trunks were responsible for almost all of the misdirected responses elicited from dorsal cutaneous areas. Six frogs (2 sham- and 4 skin-rotated) possessed some areas of plical skin from which responses could only have been mediated via more ventrally located nerve trunks. However, such dorsal expansions of ventral receptive fields never included the midline skin areas from which misdirected responses had been elicited in the intact animal. The topographic arrangement of cells within the DRG appears to be weakly polarized in cobalt-stained sections, but with large individual variations in size and observed number of neuronal profiles. For the skin-rotated group the ratio of cobalt-filled profiles in the dorsal as compared with the ventral half of the ganglion was 1.6, which did not differ significantly from the control value of 1.9. Taken as a whole, the results of the present study indicate that selective (re)innervation of displaced skin by the original nerve fibers cannot explain the origin of misdirected responses in Rana pipiens.

Animals↗
PubMedDate not supplied

Effects of substance P analogues on spinal dorsal horn neurons.

The effects of iontophoretically applied (D-Pro2, D-Phe7, D-Trp9)-SP and (D-Pro2, D-Trp7,9)-SP on the spontaneous and evoked activity of functionally identified cat spinal dorsal horn neurons have been investigated in vivo by means of extracellular single unit recording technique. In addition, the rat spinal cord slice preparation has been used to study the actions of (D-Pro2, D-Trp7,9)-SP and (D-Arg1, D-Pro2, D-Trp7,9, Leu11)-SP on the resting membrane potential of dorsal horn neurons and also on their responses to dorsal root stimulation and exogenous SP application. We have observed that both (D-Pro2, D-Phe7, D-Trp9)-SP and (D-Pro2, D-Trp7,9)-SP produced an excitation of about 15% of all neurons tested and had a weak antagonistic effect against SP in the cat spinal cord. (D-Pro2, D-Trp7,9)-SP suppressed the SP-induced excitation in 63% of examined cells. In addition, depression of the glutamate-induced excitation and spontaneous activity was evident in 10% and 19% of the cat dorsal horn neurons tested, respectively. In the spinal cord slice preparation (D-Arg1, D-Pro2, D-Trp7,9, Leu11)-SP proved to be a more potent antagonist of the SP-induced depolarization and the dorsal root-elicited slow depolarization, if compared with (D-Pro2, D-Trp7,9)-SP.

Animals↗
PubMedDate not supplied

Correlation between function and structure in "epileptic" human hippocampal tissue maintained in vitro.

Intracellular and extracellular recordings were obtained from the CA1 region and gyrus dentatus of human hippocampi that had been surgically removed for treatment of intractable seizures. Pathologic diagnoses were obtained, and electrophysiologic results were correlated with degree of principal cell loss (sclerosis) in the regions studied. Extracellular field potentials were absent or smaller in amplitude, with abnormal characteristics, in sclerotic regions. In all regions, regardless of the degree of sclerosis, individual pyramidal or granule cells could be penetrated. These neurons showed apparently normal electrophysiologic characteristics and intact excitatory synaptic input. No signs of hyperexcitability were observed.

Adolescent↗