Proceedings of the National Academy of Sciences, 1988
The synaptic function of somatostatin-containing fibers in the nervous system is controversial. T... more The synaptic function of somatostatin-containing fibers in the nervous system is controversial. Therefore, we used a slice preparation of the rat brain stem to test the electrophysiological effects of prosomatostatin-derived peptides on neurons of the solitary tract complex, which contains an abundance of somatostatin-containing fibers and cell bodies. Superfusion of both somatostatin-14 and somatostatin-28 (the precursor for somatostatin-14), but not somatostatin-28-(1-12) or -(1-10), predominantly inhibited spontaneous spike and subthreshold (probably synaptic) activity. In intracellular recordings, somatostatin-14 and -28 hyperpolarized most neurons in association with a slight (10-35%) but reproducible decrease in input resistance. These hyperpolarizing responses were augmented in depolarized cells and persisted in cells in which spontaneous inhibitory postsynaptic potentials became depolarizing after Cl- injection. These data suggest that somatostatin receptors regulate a K+ co...
The Journal of neuroscience : the official journal of the Society for Neuroscience, 1991
Voltage-sensitive K+ channels were studied in rat cerebellar Purkinje neurons in culture using th... more Voltage-sensitive K+ channels were studied in rat cerebellar Purkinje neurons in culture using the single-channel recording technique. Recordings in the cell-attached and outside-out configuration revealed multiple voltage-sensitive K+ channel types in patches from both the somatic and the dendritic regions. K+ channel types were present in all patches studied. The same channel types were observed in somatic and dendritic recordings. Channel types were identified by reversal potential, single-channel conductance, voltage sensitivity, and patterns of activity. In cell-attached patches recorded under physiological conditions, 3 channel types were identified. Mean single-channel conductances were 92, 57, and 12 pS. All 3 channel types were activated by membrane depolarization. Similar channel types were identified in inside-out and outside-out patches recorded under physiological conditions. Two additional channel types were identified in the outside-out patches, with mean single-chann...
Glial fibrillary acidic protein- (GFAP) and calbindin D28k-immunoreactivity (IR) were investigate... more Glial fibrillary acidic protein- (GFAP) and calbindin D28k-immunoreactivity (IR) were investigated in the medial subdivision of the nucleus of the solitary tract (mNST) of prenatally X-irradiated rats. Pregnant rats were exposed to a single whole-body X-irradiation on day 11 or 16 of gestation at a dose of 1. 3 Gy. The offspring were killed at 7-14 days of age for the immunohistochemical observations. Rat pups showed strong GFAP-IR at the level rostral to the obex when receiving X-rays on day 11 of gestation, with hypertrophy of astrocyte cell bodies and cytoplasmic processes, but weak GFAP-IR when receiving X-rays on day 16 of gestation. Calbindin D28k-IR was stronger in the animals receiving X-rays on day 11 or 16 of gestation compared to that in the control animals. In the present study, the increase of GFAP- and calbindin D28k-IR cells in the mNST might indicate that adaptative mechanisms are taking place to preserve integrated nervous system function and possibly, to provide neuroprotection.
Pflügers Archiv European Journal of Physiology, 1986
Neurones within the ventral and ventrolateral nuclei of the solitary tract were analyzed under si... more Neurones within the ventral and ventrolateral nuclei of the solitary tract were analyzed under single-electrode current- and voltage-clamp conditions in rat brainstem slices. We present direct and indirect evidence for the existence of five different sorts of membrane currents: a tetrodotoxin-sensitive sodium current, a tetrodotoxin-resistant calcium current, a calcium-dependent potassium current, a non-inactivating potassium current which is inhibited by muscarine, an inactivating potassium current, which is inhibited by 4-aminopyridine. These membrane properties do not produce spontaneous bursting in these neurones. Assuming that neurones with such properties belong to the respiratory network, we discuss how conductances of this type may be involved in mechanisms regulating central respiratory activity.
Single channel activity of expiratory neurones was studied in outside-out recordings. Expiratory ... more Single channel activity of expiratory neurones was studied in outside-out recordings. Expiratory neurones were identified in the ventrolateral region of the in vitro isolated brain stem-spinal cord preparation of newborn rats in cell-attached and whole-cell configurations by their pattern of firing related to phrenic motor output. Three potassium (K+) channels of 10, 30 and 70 pS exhibited steady-state activity during long voltage commands (up to 5 min) and could be found associated together in the same patches. The 30pS channel showed voltage dependency, being most active at small depolarizations. The 70 pS channel showed little activity with < 1% of openings per sample time and 1 mM tetraethylammonium (TEA) sensitivity. At similar concentrations, the discharge of the phrenic nerve was also altered, as shown by the increase of the respiratory frequency and a tonic discharge. The association of these K+ channel types on the same patches may be specific of respiratory neurones and could contribute to their bursting activity.
Substance P (SP)-ergic neurons from 16/17 day-embryonic rat brain stem in primary culture were id... more Substance P (SP)-ergic neurons from 16/17 day-embryonic rat brain stem in primary culture were identified by immunocytochemistry using biotinylated avidin and phosphatase alkaline methods with affinity-purified anti-SP antibodies. An average of 84% of neurons contained SP from day 9 to day 21 after plating. These in vitro data show that SP-containing neurons develop in our culture conditions. SP may act as a maturation factor as well as a neurotransmitter.
In vitro, the respiratory activity in rodents is characterized by: (i) the rapidly peaking, slowl... more In vitro, the respiratory activity in rodents is characterized by: (i) the rapidly peaking, slowly decrementing pattern of spontaneous and rhythmic active phases recorded from the motor rootlets, and (ii) the specific location of their rhythmic generator in the ventrolateral medulla. The aim of the present study was to assess whether the trigeminal and facial motor rootlets still exhibit respiratory activity in the absence of peripheral and higher cerebral structures, and to compare the onset of their active phases with that of other respiratory rootlets, using the in vitro isolated brainstem--spinal cord preparation of the newborn mouse and rat. Spontaneous rhythmic activity was recorded from the trigeminal and facial rootlets. It was regular and synchronized bilaterally and ipsilaterally with the hypoglossal or cervical C1-C6 rootlets. Brainstem transection experiments demonstrated that for both the trigeminal and facial rootlets, the spontaneous rhythmic activity originates from the medulla, in a region consistent with the pre-Bötzinger complex and the rostral ventrolateral medulla. The pattern of the respiratory motor activity recorded from the trigeminal and facial rootlets was identical to the pattern recorded from the hypoglossal and cervical C1-C6 rootlets with rapidly peaking, slowly decrementing characteristics. The duration of the ascending part and the total duration of their active phases were similar. The onset of the active phases of the phrenic rootlets was delayed compared with that of the trigeminal, facial and hypoglossal rootlets. However, no difference in the onsets of the active phases of the cranial rootlets could be observed. Removal of the rostral pons suppressed the delay in onset of the active phases of the phrenic rootlets. Our findings show that: (i) rhythmic activities of the trigeminal and facial rootlets are preserved in absence of control by peripheral or high cerebral structures; (ii) the pattern and the location of the rhythmic generator for these activities are of the respiratory type; and (iii) the rostral pons is responsible for a delay in the onset of the active phases of the phrenic rootlets compared with that of the trigeminal, facial and hypoglossal rootlets.
The calcium sensitivity of a large conductance voltage-sensitive potassium channel found in cultu... more The calcium sensitivity of a large conductance voltage-sensitive potassium channel found in cultured rat cerebellar Purkinje neurons was studied in membrane patches from the somatic region of the cultured Purkinje neurons using single-channel recording techniques. The potassium channel had a conductance of approximately 94 picosiemens (pS) under physiologic ionic conditions and was active at depolarized membrane potentials. Activity due to this channel type was not observed when the saline at the internal surface of the membrane was calcium free. Low intracellular calcium concentration (10 nM) triggered channel activity at depolarized membrane potentials. Channel activity increased further with increasing intracellular calcium concentrations and was evident at more negative membrane potentials. The high sensitivity of this potassium channel type to intracellular calcium and its abundance in the Purkinje neuron membrane may reflect a prominent role in the control of action potential duration and interspike interval when the neurons are firing in a rapid, repetitive mode.
1. Respiratory neurons of mammals are rhythmically active because their membrane potential fluctu... more 1. Respiratory neurons of mammals are rhythmically active because their membrane potential fluctuates periodically over a voltage range of -70 to -55 mV. These respiratory drive potentials lead to periodic discharges of bursts of action potentials lasting for 1-2 s. The neuronal processes stabilizing this rhythmic activity involve excitatory and inhibitory synaptic processes that interact with specific membrane properties of the postsynaptic neurones. In the present experiments, performed on dorsal and ventral groups of respiratory neurones under in vivo and in vitro conditions, we verified the modulating feature of such intrinsic neuronal properties. 2. Intrinsic neuronal properties involve Ca2+ mechanisms that lead to intracellular Ca2+ accumulation, and consequently to activation of Ca(2+)-dependent K+ currents. 3. Blockade of intracellular Ca2+ accumulation significantly changed the amplitude and pattern of respiratory drive potentials, and blocked initial hyperpolarizing shifts...
Proceedings of the National Academy of Sciences, 1988
The synaptic function of somatostatin-containing fibers in the nervous system is controversial. T... more The synaptic function of somatostatin-containing fibers in the nervous system is controversial. Therefore, we used a slice preparation of the rat brain stem to test the electrophysiological effects of prosomatostatin-derived peptides on neurons of the solitary tract complex, which contains an abundance of somatostatin-containing fibers and cell bodies. Superfusion of both somatostatin-14 and somatostatin-28 (the precursor for somatostatin-14), but not somatostatin-28-(1-12) or -(1-10), predominantly inhibited spontaneous spike and subthreshold (probably synaptic) activity. In intracellular recordings, somatostatin-14 and -28 hyperpolarized most neurons in association with a slight (10-35%) but reproducible decrease in input resistance. These hyperpolarizing responses were augmented in depolarized cells and persisted in cells in which spontaneous inhibitory postsynaptic potentials became depolarizing after Cl- injection. These data suggest that somatostatin receptors regulate a K+ co...
The Journal of neuroscience : the official journal of the Society for Neuroscience, 1991
Voltage-sensitive K+ channels were studied in rat cerebellar Purkinje neurons in culture using th... more Voltage-sensitive K+ channels were studied in rat cerebellar Purkinje neurons in culture using the single-channel recording technique. Recordings in the cell-attached and outside-out configuration revealed multiple voltage-sensitive K+ channel types in patches from both the somatic and the dendritic regions. K+ channel types were present in all patches studied. The same channel types were observed in somatic and dendritic recordings. Channel types were identified by reversal potential, single-channel conductance, voltage sensitivity, and patterns of activity. In cell-attached patches recorded under physiological conditions, 3 channel types were identified. Mean single-channel conductances were 92, 57, and 12 pS. All 3 channel types were activated by membrane depolarization. Similar channel types were identified in inside-out and outside-out patches recorded under physiological conditions. Two additional channel types were identified in the outside-out patches, with mean single-chann...
Glial fibrillary acidic protein- (GFAP) and calbindin D28k-immunoreactivity (IR) were investigate... more Glial fibrillary acidic protein- (GFAP) and calbindin D28k-immunoreactivity (IR) were investigated in the medial subdivision of the nucleus of the solitary tract (mNST) of prenatally X-irradiated rats. Pregnant rats were exposed to a single whole-body X-irradiation on day 11 or 16 of gestation at a dose of 1. 3 Gy. The offspring were killed at 7-14 days of age for the immunohistochemical observations. Rat pups showed strong GFAP-IR at the level rostral to the obex when receiving X-rays on day 11 of gestation, with hypertrophy of astrocyte cell bodies and cytoplasmic processes, but weak GFAP-IR when receiving X-rays on day 16 of gestation. Calbindin D28k-IR was stronger in the animals receiving X-rays on day 11 or 16 of gestation compared to that in the control animals. In the present study, the increase of GFAP- and calbindin D28k-IR cells in the mNST might indicate that adaptative mechanisms are taking place to preserve integrated nervous system function and possibly, to provide neuroprotection.
Pflügers Archiv European Journal of Physiology, 1986
Neurones within the ventral and ventrolateral nuclei of the solitary tract were analyzed under si... more Neurones within the ventral and ventrolateral nuclei of the solitary tract were analyzed under single-electrode current- and voltage-clamp conditions in rat brainstem slices. We present direct and indirect evidence for the existence of five different sorts of membrane currents: a tetrodotoxin-sensitive sodium current, a tetrodotoxin-resistant calcium current, a calcium-dependent potassium current, a non-inactivating potassium current which is inhibited by muscarine, an inactivating potassium current, which is inhibited by 4-aminopyridine. These membrane properties do not produce spontaneous bursting in these neurones. Assuming that neurones with such properties belong to the respiratory network, we discuss how conductances of this type may be involved in mechanisms regulating central respiratory activity.
Single channel activity of expiratory neurones was studied in outside-out recordings. Expiratory ... more Single channel activity of expiratory neurones was studied in outside-out recordings. Expiratory neurones were identified in the ventrolateral region of the in vitro isolated brain stem-spinal cord preparation of newborn rats in cell-attached and whole-cell configurations by their pattern of firing related to phrenic motor output. Three potassium (K+) channels of 10, 30 and 70 pS exhibited steady-state activity during long voltage commands (up to 5 min) and could be found associated together in the same patches. The 30pS channel showed voltage dependency, being most active at small depolarizations. The 70 pS channel showed little activity with < 1% of openings per sample time and 1 mM tetraethylammonium (TEA) sensitivity. At similar concentrations, the discharge of the phrenic nerve was also altered, as shown by the increase of the respiratory frequency and a tonic discharge. The association of these K+ channel types on the same patches may be specific of respiratory neurones and could contribute to their bursting activity.
Substance P (SP)-ergic neurons from 16/17 day-embryonic rat brain stem in primary culture were id... more Substance P (SP)-ergic neurons from 16/17 day-embryonic rat brain stem in primary culture were identified by immunocytochemistry using biotinylated avidin and phosphatase alkaline methods with affinity-purified anti-SP antibodies. An average of 84% of neurons contained SP from day 9 to day 21 after plating. These in vitro data show that SP-containing neurons develop in our culture conditions. SP may act as a maturation factor as well as a neurotransmitter.
In vitro, the respiratory activity in rodents is characterized by: (i) the rapidly peaking, slowl... more In vitro, the respiratory activity in rodents is characterized by: (i) the rapidly peaking, slowly decrementing pattern of spontaneous and rhythmic active phases recorded from the motor rootlets, and (ii) the specific location of their rhythmic generator in the ventrolateral medulla. The aim of the present study was to assess whether the trigeminal and facial motor rootlets still exhibit respiratory activity in the absence of peripheral and higher cerebral structures, and to compare the onset of their active phases with that of other respiratory rootlets, using the in vitro isolated brainstem--spinal cord preparation of the newborn mouse and rat. Spontaneous rhythmic activity was recorded from the trigeminal and facial rootlets. It was regular and synchronized bilaterally and ipsilaterally with the hypoglossal or cervical C1-C6 rootlets. Brainstem transection experiments demonstrated that for both the trigeminal and facial rootlets, the spontaneous rhythmic activity originates from the medulla, in a region consistent with the pre-Bötzinger complex and the rostral ventrolateral medulla. The pattern of the respiratory motor activity recorded from the trigeminal and facial rootlets was identical to the pattern recorded from the hypoglossal and cervical C1-C6 rootlets with rapidly peaking, slowly decrementing characteristics. The duration of the ascending part and the total duration of their active phases were similar. The onset of the active phases of the phrenic rootlets was delayed compared with that of the trigeminal, facial and hypoglossal rootlets. However, no difference in the onsets of the active phases of the cranial rootlets could be observed. Removal of the rostral pons suppressed the delay in onset of the active phases of the phrenic rootlets. Our findings show that: (i) rhythmic activities of the trigeminal and facial rootlets are preserved in absence of control by peripheral or high cerebral structures; (ii) the pattern and the location of the rhythmic generator for these activities are of the respiratory type; and (iii) the rostral pons is responsible for a delay in the onset of the active phases of the phrenic rootlets compared with that of the trigeminal, facial and hypoglossal rootlets.
The calcium sensitivity of a large conductance voltage-sensitive potassium channel found in cultu... more The calcium sensitivity of a large conductance voltage-sensitive potassium channel found in cultured rat cerebellar Purkinje neurons was studied in membrane patches from the somatic region of the cultured Purkinje neurons using single-channel recording techniques. The potassium channel had a conductance of approximately 94 picosiemens (pS) under physiologic ionic conditions and was active at depolarized membrane potentials. Activity due to this channel type was not observed when the saline at the internal surface of the membrane was calcium free. Low intracellular calcium concentration (10 nM) triggered channel activity at depolarized membrane potentials. Channel activity increased further with increasing intracellular calcium concentrations and was evident at more negative membrane potentials. The high sensitivity of this potassium channel type to intracellular calcium and its abundance in the Purkinje neuron membrane may reflect a prominent role in the control of action potential duration and interspike interval when the neurons are firing in a rapid, repetitive mode.
1. Respiratory neurons of mammals are rhythmically active because their membrane potential fluctu... more 1. Respiratory neurons of mammals are rhythmically active because their membrane potential fluctuates periodically over a voltage range of -70 to -55 mV. These respiratory drive potentials lead to periodic discharges of bursts of action potentials lasting for 1-2 s. The neuronal processes stabilizing this rhythmic activity involve excitatory and inhibitory synaptic processes that interact with specific membrane properties of the postsynaptic neurones. In the present experiments, performed on dorsal and ventral groups of respiratory neurones under in vivo and in vitro conditions, we verified the modulating feature of such intrinsic neuronal properties. 2. Intrinsic neuronal properties involve Ca2+ mechanisms that lead to intracellular Ca2+ accumulation, and consequently to activation of Ca(2+)-dependent K+ currents. 3. Blockade of intracellular Ca2+ accumulation significantly changed the amplitude and pattern of respiratory drive potentials, and blocked initial hyperpolarizing shifts...
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