PubMed Health⌕ Search

Biomedical subjects

J Lipski

Publications and source records attributed to J Lipski.

At least 37 records · Page 2Linked to original sources
PubMed1998

Whole cell patch-clamp study of putative vasomotor neurons isolated from the rostral ventrolateral medulla.

A distinct subpopulation of neurons in the rostral and ventrolateral part of the medulla oblongata (RVL) plays a key role in controlling sympathetic vasomotor tone. To characterize these neurons under conditions in which all cell-to-cell interactions are eliminated, RVL neurons were acutely dissociated from 13- to 19-day old rats. Cells projecting to the upper thoracic segments were retrogradely labeled with fluorescent beads. Fifty-two percent (17/33) of examined spinally projecting neurons were catecholaminergic, as demonstrated by single-cell reverse transcription-polymerase chain reaction or immunocytochemistry. No spontaneous (capacitive) spikes were revealed in the tight seal cell-attached configuration. Whole cell recordings were made from 54 spinally projecting neurons using Cs(+)- or K(+)-containing pipettes. No spontaneous firing was observed in current-clamp mode with K(+)-based pipettes (membrane potential, -61.5 +/- 2.3 mV). Step depolarizations (300- or 400-ms pulses, up to 100 pA) evoked regular firing or one to four spikes. Several voltage-gated currents, resembling the transient and persistent Na+, delayed rectifier and low- and high-threshold Ca2+, were revealed in voltage-clamp mode. These results show that isolated spinally projecting RVL neurons display no pacemaker-like activity. Because data from the literature indicate that these neurons are capable of generating such activity under different experimental conditions, the factors responsible for different behavior need to be determined. Dissociated RVL neurons provide a useful new model for studying biophysical and other properties of neurons involved in blood pressure control.

Animals↗
PubMed1997

P2 receptor excitation of rodent hypoglossal motoneuron activity in vitro and in vivo: a molecular physiological analysis.

The role of P2 receptors in controlling hypoglossal motoneuron (XII MN) output was examined (1) electrophysiologically, via application of ATP to the hypoglossal nucleus of rhythmically active mouse medullary slices and anesthetized adult rats; (2) immunohistochemically, using an antiserum against the P2X2 receptor subunit; and (3) using PCR to identify expression of P2X2 receptor subunits in micropunches of tissue taken from the XII motor nucleus. Application of ATP to the hypoglossal nucleus of mouse medullary slices and anesthetized rats produced a suramin-sensitive excitation of hypoglossal nerve activity. Additional in vitro effects included potentiation of inspiratory hypoglossal nerve output via a suramin- and pyridoxal-phosphate-6-azophenyl-2',4'-disulphonic acid (PPADS)-sensitive mechanism, XII MN depolarization via activation of a suramin-sensitive inward current, decreased neuronal input resistance, and a slow-onset theophylline-sensitive reduction of inspiratory output likely resulting from hydrolysis of extracellular ATP to adenosine and activation of P1 receptors. Immunohistochemically, P2X2 receptors were detected in inspiratory XII MNs that were labeled with Lucifer yellow. These data, combined with identification of mRNA for three P2X2 receptor subunit isoforms within the hypoglossal nucleus (two of which have not been localized previously in brain) and the previous demonstration that P2X receptors are ubiquitously expressed in cranial and spinal motoneuron pools, support not only a role of P2 receptors in modulating inspiratory hypoglossal activity but a general role of P2 receptors in modulating motor outflow from the CNS.

Adenosine↗
PubMed1997

O2-sensitive K+ current in undifferentiated and NGF-treated PC12 cell variants.

It has been reported that cultured PC12 cells can be used as a model for studying mechanisms of O2 sensitivity, previously examined in peripheral chemoreceptors and some neurons. This study compared the hypoxic depression of K+ currents in two PC12 variants, before and after differentiation into neurone-like cells induced by nerve growth factor (NGF). The results show that interaction of O2 and K+ channels is strongly-voltage dependent in the PC12/TM but not the PC12/ES subline. In PC12/TM cells an effect of hypoxia on the K+ current was appreciable only at moderately depolarized voltages, with a loss of sensitivity at +40 to +50 mV. NGF-induced transformation did not affect the responses seen in undifferentiated cells. These results emphasize the importance of screening PC12 cells before selecting a variant for studying O2 sensitivity. In view of evidence cited in the literature that hypoxia may effect membrane channels directly, further molecular and biophysical studies of the differences among PC12 variants are required.

Animals↗
PubMed1996

Properties of presympathetic neurones in the rostral ventrolateral medulla in the rat: an intracellular study "in vivo'.

1. Intracellular recordings were made in pentobarbitone-anaesthetized rats from sixty-eight neurones located in the rostral ventrolateral medulla (RVLM), which responded with inhibition (latency, 33.6 +/- 9.3 ms) after stimulation of the aortic depressor nerve with short bursts of pulses. This inhibition was due to chloride- and voltage-dependent IPSPs. 2. Seventeen neurones could be excited antidromically after stimulation in the T2 spinal segment (conduction velocity 1.9-8.5 m.s-1) and were classified as RVLM presympathetic vasomotor neurones. 3. "Spontaneously' active neurones (n = 29) displayed a largely irregular pattern of firing, with no clear relationship between the level of the membrane potential and cycles of phrenic nerve activity at end-tidal CO2 < 5.0%. Cardiac cycle-related shifts of the membrane potential were not considered indicative of baroreceptor input as they could be due to movement artifacts. 4. All neurones displayed large synaptic activity (EPSPs and IPSPs, peak-to-peak amplitude > 5.0 mV). The depolarizing IPSPs observed during injection of chloride and/or negative current consisted of a phasic and a tonic component. 5. The on-going activity of these neurones resulted from synaptic inputs, with individual action potentials usually preceded by identifiable fast EPSPs. 6. No evidence was found for the presence of gradual depolarizations (autodepolarizations) between individual action potentials, and therefore under these experimental conditions the activity of RVLM presympathetic neurones did not depend on intrinsic pacemaker properties. 7. These results are consistent with the "network' hypothesis for the generation of sympathetic vasomotor tone.

Animals↗
PubMed1996

Pre-sympathetic neurones in the rostral ventrolateral medulla of the rat: electrophysiology, morphology and relationship to adjacent neuronal groups.

The activity of preganglionic sympathetic neurones largely depends on synaptic excitation from antecedent reticulospinal neurones located in the rostral ventrolateral medulla (RVLM). Our study, conducted in anaesthetized rats, showed that all RVLM pre-sympathetic neurones display a substantial synaptic noise and their action potentials are usually preceded by fast EPSPs. No evidence was found for presence of gradual depolarizations (autodepolarizations) between individual spikes. Therefore our results are consistent with the "network" hypothesis for the generation of sympathetic vasomotor tone. Axons of some pre-sympathetic neurones intracellularly labelled with Neurobiotin or Lucifer Yellow had collaterals arborizing in several medullary regions. Thus these neurones have synaptic inputs not only to preganglionic sympathetic neurones, but also to other, yet unidentified cells in the brainstem. Finally, our results show that anatomically adjacent RVLM pre-sympathetic and Bötzinger respiratory neurones from two functionally distinct neuronal subpopulations, and that some pre-sympathetic neurones have an adrenergic phenotype.

Animals↗
PubMed1995

A comparative study of pre-sympathetic and Bötzinger neurons in the rostral ventrolateral medulla (RVLM) of the rat.

The aim of this study was to investigate the degree of functional and anatomical overlap between two major neuronal subpopulations in the rostral ventrolateral medulla: pre-sympathetic (sympathoexcitatory) neurons, and expiratory neurons of the Bötzinger complex. Extracellular recordings were made with dye-filled microelectrodes in pentobarbital anesthetized, paralyzed and artificially ventilated adult Wistar rats. Tests applied included stimulation of baroreceptor afferents, activation of peripheral chemoreceptors and lung stretch receptors, changes in central respiratory drive with hyper- or hypoventilation, nociceptive stimulation, and antidromic stimulation from the T2 segment of the spinal cord or medulla oblongata at obex level. The two groups of neurons showed different patterns of spontaneous activity and generally different responses to these stimuli. The recording positions showed some overlap, but the majority of Bötzinger neurons were dorsolateral to pre-sympathetic neurons. There was a large overlap between the location of pre-sympathetic neurons and the lateral part of the C1 adrenergic group, but only a small overlap between these adrenergic neurons and Bötzinger neurons. These results indicate that the anatomically adjacent pre-sympathetic and Bötzinger expiratory neurons form two functionally distinct neuronal subpopulations.

Animals↗
PubMed1995

Barosensitive neurons in the rostral ventrolateral medulla of the rat in vivo: morphological properties and relationship to C1 adrenergic neurons.

The aim of this study, conducted in anaesthetized rats, was to examine the morphology of barosensitive neurons in the rostral ventrolateral medulla and their immunoreactivity for a catecholamine synthesizing enzyme, tyrosine hydroxylase. Thirty neurons displaying inhibitory postsynaptic potentials following stimulation of the aortic depressor nerve were intracellularly labelled with Lucifer Yellow or Neurobiotin. Some of these neurons could be excited antidromically from the second thoracic segment of the spinal cord, with conduction velocities of spinal axons ranging from 1.9 to 7.2 m/s. The filled somas were found immediately caudal to the facial nucleus and ventral or ventromedial to compact formation of the nucleus ambiguus. Some dendrites reached the ventral medullary surface. Axons usually projected dorsomedially and then made a sharp rostral and/or caudal turn. The caudally projecting axon could, in some cases, be followed to the first cervical segment of the spinal cord. Seven cells issued fine axon collaterals on the ipsilateral side. These were identified mainly in two areas: in the rostral ventrolateral medulla (or immediately dorsomedial to that region), and within the dorsal vagal complex. Seven of 27 examined cells (26%) were tyrosine hydroxylase-immunoreactive and were classified as C1 adrenergic neurons. No clear relationship was found between the presence or absence of adrenergic phenotype and the morphology of filled cells. However, the amplitude of aortic nerve-evoked inhibitory postsynaptic potentials was significantly larger in tyrosine hydroxylase-positive neurons. Possible reasons for the low percentage of barosensitive cells with tyrosine hydroxylase immunoreactivity found in this study, in comparison with previously published estimates, are discussed. This is the first study describing the morphology of neurons in this part of the medulla identified as barosensitive in vivo, and directly demonstrating adrenergic phenotype in a subset of these neurons.

Adrenergic Fibers↗
PubMed1994

Neurone-specific enolase persists in some hippocampal cells 24 hours after an NMDA injection.

Neurone-specific enolase immunoreactivity (NSE-IR) was examined in the hippocampal dentate gyrus 5 h, 24 h and 6 days following local injection of N-methyl-D-aspartate (NMDA), and was compared with Nissl substance and acid fuchsin staining. Five and 24 hours post-injection extracellular NSE-IR was increased, and the number of NSE-IR neurones in the granule cell layer was decreased. However, at 24 h post-injection only, 10-15% of the granule cells were strongly NSE-IR, in contrast to the Nissl/acid fuchsin staining which showed a homogeneous population of degenerating neurones. By 6 days there were no viable neurones in the lesion area stained by either method. These results show that neuronal degeneration after an excitotoxic insult occurs in a non-uniform way in an apparently homogeneous group of central neurones.

Animals↗
PubMed1994

Morphological study of long axonal projections of ventral medullary inspiratory neurons in the rat.

The aim of this study was to examine medullary and spinal axonal projections of inspiratory bulbospinal neurons of the rostral ventral respiratory group (VRG) in the rat. A direct visualization of long (9.8-33 mm) axonal branches, including those projecting to the contralateral side of the medulla oblongata and the spinal cord, was possible due to intracellular labeling with neurobiotin and long survival times (up to 22 h) after injections. Seven of the nine labeled neurons had bilateral descending axons, which were located in discrete regions of the spinal white matter; ipsilateral axons in the lateral and dorsolateral funiculus, contralateral in the ventral and ventromedial funiculus. The collaterals issued by these axons at the mid-cervical level formed close appositions with dendrites of phrenic motoneurons, which had also been labeled with neurobiotin. None of these collaterals crossed the midline. The significance of this finding is discussed in relation to the crossed-phrenic phenomenon. Additional spinal collaterals were identified in the C1 and T1 segments. Within the medulla, collaterals with multiple varicosities were identified in the lateral tegmental field and in the dorsomedial medulla (in the hypoglossal nucleus and in the nucleus of the solitary tract). These results demonstrate that inspiratory VRG neurons in the rat have some features which have not been previously described in the cat, including frequent bilateral spinal projection and projection to the nucleus of the solitary tract. In addition, this study shows that intracellular labeling with neurobiotin offers an effective way of tracing long axonal projections, supplementing results previously obtainable only with antidromic mapping, and providing morphological details which could not be observed in previous studies using labeling with horseradish peroxidase.

Animals↗
PubMed1994

Electrophysiological study of dorsal respiratory neurons in the medulla oblongata of the rat.

There has been controversy whether the dorsal respiratory group (DRG), identified in the cat and several other species as a concentration of mainly inspiratory neurons located in the ventrolateral subnucleus of the solitary tract, also exists in the rat. The aim of this study was to re-examine this question by systematically exploring this region with extracellular microelectrodes, in anesthetized and artificially ventilated rats. One-hundred and forty-two units were recorded which fired in phase with central respiratory cycles (determined by recording from the phrenic nerve) and/or lung inflations. One-hundred and nineteen recordings were thought to be from neuronal cell bodies (confirmed in some cases by excitatory responses to microelectrophoretic administration of DL-homocysteic acid), while the remaining 23 were from lung vagal afferents. Most neurons in the former group (87/119) were inspiratory. Out of 96 neurons tested for spinal projections only 14 (12 inspiratory, 2 expiratory) responded antidromically following stimulation at C3 segment. These results confirm the existence of the DRG in the rat and demonstrate that neurons located in this region have firing patterns generally similar to those previously described in the cat. The main difference is the relative paucity in the rat of neurons projecting spinally below the C2 level, which indicates that most DRG neurons in this species do not project directly to phrenic and intercostal motoneurons, but to other, as yet unidentified, neuronal groups within the brainstem or upper cervical segments.

Animals↗
PubMed1994

Projections from inspiratory neurons of the ventral respiratory group to the subretrofacial nucleus of the cat.

Arterial blood pressure and the activity of many sympathetic nerves are known to be affected by changes in central respiratory activity. The central neurons responsible for this respiratory modulation are unknown. In the present study we have labelled inspiratory neurons (n = 24) in the rostral ventral respiratory group and Bötzinger complex in the medulla oblongata of the cat using intracellular injection of biocytin. The filled neurons were examined to see if they had axonal projections to the subretrofacial nucleus, an important brainstem nucleus in the tonic and reflex control of blood pressure. The subretrofacial nucleus was identified histologically as a cluster of neurons in the rostral ventrolateral medulla, some of which are tyrosine hydroxylase immunoreactive. Varicose axons arising from labelled inspiratory neurons were mostly found dorsal to this cluster, within the area corresponding to the Bötzinger complex. A small number of axon varicosities were seen in the subretrofacial nucleus. The results suggest that a part of the respiratory modulation of sympathetic nerve activity may be due to a direct synaptic input from inspiratory neurons of the ventral respiratory group to neurons of the subretrofacial nucleus.

Animals↗
PubMed1994

An electrophysiological and morphological study of esophageal motoneurons in rats.

Previous anatomic studies have shown that motoneurons supplying the striated musculature of the esophagus form a tightly grouped cluster in the rostral portion of the nucleus ambiguus, known as the compact formation. This study, conducted in anesthetized rats, presents the first in vivo intracellular and extracellular recordings from this group of motoneurons, which were identified by antidromic stimulation directly from the esophagus (latency 7-68 ms). The motoneurons were silent at rest, and those impaled intracellularly (n = 44) showed no respiratory modulation of their membrane potential. Intracellular labelling with Lucifer Yellow (n = 3) or Neurobiotin (n = 15) revealed multipolar somas with longitudinally oriented dendritic trees mainly confined to the compact formation. No axon collaterals were found. When swallowing-like activity was induced by muscarine applied to the dorsal medullary surface, the motoneurons displayed bursting activity, with the majority of bursts occurring during expiration. These results show that antidromic stimulation of esophageal motoneurons with an electrode inserted into the esophagus provides a simple way of identifying these motoneurons. In the absence of pharmacological stimulation, these motoneurons receive no respiratory-modulated synaptic input, in contrast to adjacent motoneurons in the semicompact formation (supplying the upper airways), which are known to display respiratory activity. However, some synchronization of respiratory and swallowing-like activity was observed after pharmacological activation of the swallowing pattern generator in the dorsal medulla.

Animals↗
PubMed1993

Expiratory neurons of the Bötzinger Complex in the rat: a morphological study following intracellular labeling with biocytin.

The term "Bötzinger Complex" (BOT) refers to a distinct group of neurons, located near the rostral portion of the nucleus ambiguus, which are known to play an important role in the control of respiratory movements. Previous studies conducted in cats have demonstrated that most of these neurons are active during expiration, exerting a monosynaptic inhibitory action on several subpopulations of inspiratory neurons in the medulla and spinal cord. The aim of this study was to examine morphological properties and possible synaptic targets of BOT neurons in the rat. Forty-one expiratory neurons were labeled intracellularly with biocytin; 12 were interneurons (BOT neurons) and 29 were motoneurons. The latter could not be antidromically activated following stimulation of the superior laryngeal or vagal nerves. BOT neurons showed extensive axonal arborisations in the ipsilateral medulla, with some projections to the contralateral side. Bouton-like axon varicosities mainly clustered in two areas: near the parent cell bodies, and in the area corresponding to the rostral part of the ventral respiratory group (VRG). In five pairs of labeled neurons, each consisting of one BOT neuron and one inspiratory neuron in the rostral VRG, no appositions were identified at the light microscopic level between axons of BOT neurons and dendrites or cell bodies of inspiratory neurons. These results demonstrate that some features of BOT expiratory neurons in the rat are similar to those previously described in cats. The differences include their more ventral location in relation to the compact formation of nucleus ambiguus (retrofacial nucleus), and the relative paucity in the rat of neurons displaying an augmenting pattern of activity and of neurons with spinally projecting axons. In addition, we were unable to find morphological evidence for contacts between labeled BOT neurons and ipsilateral inspiratory neurons near the obex level, a finding not consistent with previous electrophysiological studies in the cat in which such synaptic connections have been identified.

Animals↗
PubMed1993

Upper cervical inspiratory neurons in the rat: an electrophysiological and morphological study.

Upper cervical inspiratory neurons from a distinct neuronal column located near the lateral edge of the intermediate grey matter in the rostral spinal segments. Previous studies conducted in cats have demonstrated synaptic inputs to these neurons from several respiratory-related regions of the medulla, and long descending axonal projections mainly towards the motoneurons supplying the intercostal muscles. The aim of this study was to examine the electrophysiological and morphological properties of this propriospinal system in the rat. Extracellular recordings were made from 127 cervical inspiratory units, mainly in the C1 and C2 segments. Eighty-two percent could be antidromically activated from the C7/C8 border. No evidence of monosynaptic connection was obtained by cross-correlating the activity of some of these units with the discharge of the phrenic nerve. Intracellular recordings were made from seven neurons, three of which were labelled with biotinamide (neurobiotin). Long "survival times" after intracellular injections (up to 23 h) resulted in staining of axons for long distances, at least to the C5 segment. Each of the three labelled axons issued only one short collateral which arborized in the region of the phrenic nucleus. These results demonstrate that upper cervical inspiratory neurons in the rat have features similar to those previously described in the cat, including only a limited projection to the phrenic nucleus. In addition, this study provides the first morphological identification of these neurons.

Animals↗
PubMed1992

Projections from the commissural subnucleus of the nucleus of the solitary tract: an anterograde tracing study in the cat.

The commissural subnucleus (COM) of the nucleus of the solitary tract (NTS) is known to receive primary afferents from the lungs and other viscera innervated by the vagus nerve, and thus to participate in central autonomic and respiratory control. The aim of the present study was to identify the areas of terminal arborizations of COM neurons in order to examine brainstem sites which may be involved in reflex responses mediated by these neurons. The projections were studied in cats, using biocytin as an anterograde tracer. Labeled fibers and terminal boutons were visualized by horseradish-peroxidase histochemistry, 2-3 days after microinjection of the tracers into the COM 1-2 mm caudal to the obex. Labeled axons were examined in the brainstem from the rostral pons to the caudal medulla and were found bilaterally, with an ipsilateral predominance, mainly in the following regions: (1) The dorsolateral rostral pons. Terminal boutons were observed in the lateral and medial parabrachial nuclei, Kölliker-Fuse nucleus, and around the mesencephalic trigeminal tract. This area corresponds to the pontine respiratory group also known as the "pneumotaxic center." (2) The pontine area dorsolateral to the superior olivary nucleus. This region contains the A5 noradrenergic cell group; (3) Near the ventral surface, below the facial nucleus. This area overlaps with the 'retrotrapezoid nucleus.' (4) Respiration-related areas of the medulla, including the dorsal and ventral respiratory groups, and the Bötzinger complex. (5) The dorsal motor nucleus of the vagus. These results suggest that the COM is involved in reflex arcs, which have both respiratory functions and autonomic functions. The pathway to the dorsolateral pons, which has been identified in our recent electrophysiological study is likely to play a role in mediating respiratory responses from pulmonary rapidly adapting receptors. Other pathways may represent additional projections from second-order neurons receiving input from this group of lung receptors, or projections from as yet unidentified neurons that relay information from different afferents terminating in the COM.

Animals↗
PubMed1992

5-Hydroxytryptophan (5-HTP) augments spontaneous and evoked phrenic motoneuron discharge in spinalized rats.

Experiments on anesthetized, spinalized rats were conducted to determine the effects of systemic 5-hydroxytryptophan (5-HTP) administration on: (1) spontaneous phrenic nerve activity and (2) evoked phrenic responses to short latency, non-serotonergic synaptic inputs elicited by electrical stimulation of lateral funiculus. 5-HTP augmented spontaneous phrenic activity and allowed expression of a second, longer latency evoked response. Both effects were antagonized by methysergide. Our results suggest that spinal serotonin increases the efficacy of synaptic inputs to phrenic motoneurons.

5-Hydroxytryptophan↗