The effects of temperature and synaptic blockade in vitro on neurons of the horizontal limb of the diagonal band of Broca in the rat basal forebrain

We studied the thermosensitivity of neurons in the rat horizontal limb of the diagonal band of Broca (HDB) in vitro under normal conditions and under conditions of a low calcium/high magnesium synaptic blockade (SB). Of 52 HDB neurons tested, 34 neurons (65%) were warm-sensitive (WS), three neurons (6%) were cold-sensitive (CS), 11 neurons (21%) were temperature-insensitive (TI) and four additional neurons (8%) were both warm- and cold-sensitive (WS/CS). Of 34 neurons tested for thermosensitivity under SB, 11 were WS, 4 were CS and 19 were TI. Nearly half (48%) of the WS neurons maintained warm sensitivity under SB, 43% became TI and 9% became CS. Baseline firing rates of neurons significantly decreased during SB and then increased during recovery from SB. In addition, a distinct anatomical distribution of thermosensitive neurons was found in the HDB. The most ventral aspect of the HDB (interaural +0.9-1.3 mm) had proportionally fewer temperature sensitive neurons (65% vs. 88%) than areas more dorsal (interaural +1.3-1.7 mm), and only one of seven ventral HDB neurons (14%) remained thermosensitive during SB. In the dorsal HDB, 65% of the neurons maintained thermosensitivity during SB. These results demonstrate that the HDB contains inherently thermosensitive neurons, and that a difference in thermal characteristics exists between the ventral and dorsal HDB neurons.     

Depression of agonistic behavior after diagonal band of Broca lesions in rats.

28 male Sprague-Dawley rats received either lesions of the diagonal band of Broca (DBB) or sham DBB lesions followed by septal lesions, or septal lesions followed by DBB lesions. Shock-elicited fighting was diminished significantly by DBB lesions. Septal lesions elevated the DBB lesion-induced depression transiently. It is concluded that the DBB lesions interrupted a fiber system which either carries a facilitatory effect upon agonistic behavior from the amygdala to the dorsal forebrain or participates in the response to nociaceptive stimulation. (19 ref) (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Ionic mechanisms of action of neurotensin in acutely dissociated neurons from the diagonal band of Broca of the rat

Whole cell recordings were performed on acutely dissociated neurons from the horizontal limb of the diagonal band of Broca (hDBB) from rats to elucidate the ionic mechanisms of action of neurotensin. Neurotensin caused a decrease in whole cell voltage-activated outward currents and failed to elicit a response when Ca2+ influx was blocked by changing the external solution to the one containing 0 mM Ca2+ and 50 microM Cd2+, suggesting the involvement of Ca2+-dependent conductances. Charybdotoxin, a specific blocker of voltage-sensitive calcium-activated K+ channels (IC), caused a decrease in outward currents comparable with that caused by blocking calcium influx and occluded the neurotensin-induced decrease in outward currents. Similarly, 50 microM tetraethylammonium ions also blocked the neurotensin response. Also neurotensin reduced whole cell barium currents (IBa) and calcium currents (ICa). Amiloride and omega-conotoxin GVIA, but not nimodipine, were able to eliminate the neurotensin-induced decrease in IBa. Thus T- and N- but not L-type calcium channels are subject to modulation by neurotensin, and this may account for its effects on IC. The predicted changes in action potential as a result of the blockade of currents through calcium channels culminating into changes in IC were confirmed in the bridge current-clamp recordings. Specifically, neurotensin application led to depolarization of the resting membrane potential, broadening of spike and a decrease in afterhyperpolarization and accommodation. These alterations in action potential characteristics that resulted in increased firing rate and excitability of the hDBB neurons also were produced by application of charybdotoxin. Neurotensin effects on these properties were occluded by 2 - [(1 - 7 - chloro - 4 - quinolinyl) - 5 - (2, 6 - di - methoxyphenyl) pyrazol-3-yl) carbonylamino] tricyclo (3.3.1.1.)decan-2-carboxylic acid, a nonpeptide high-affinity neurotensin receptor antagonist. Neurotensin blockade of IC, possibly through ICa, is a potential physiological mechanism whereby this peptide may evoke alterations in the cortical arousal, sleep-wake cycle, and theta rhythm.     

Inhibitory effects of berberine on voltage- and calcium-activated potassium currents in human myeloma cells

The effects of berberine, an isoquinoline alkaloid, were investigated in human myeloma cells. In cells with intracellular Ca2+ concentration ([Ca2+]i) = 10 nM, the depolarizing square pulses from -80 mV elicited an instantaneous outward current with an inactivation. This outward current was voltage dependent, activating at -30 mV and showed inactivation with repetitive depolarization, and was hence believed to be n type voltage-activated K+ current (IK(V)). Berberine (30 microM) produced a prolongation in the recovery of IK(V) inactivation. In cells with [Ca2+]i = 1 microM, berberine also inhibited A23187-induced IK(Ca). Berberine (1-300 microM) caused the inhibition of IK(V) and IK(Ca) in the concentration-dependent manners. The IC50 values of berberine-induced inhibition of IK(V) and IK(Ca) were approximately 15 microM and 50 microM, respectively. In inside-out configurations, berberine inside the pipette suppressed the activity of K(Ca) channels without changing the single channel conductance. Berberine also inhibited the proliferation of this cell line and the IC50 value of berberine-induced inhibition of cell proliferation was 5 microM. Thus, the cytotoxic effect of berberine in cancer cells may be partially explained by its direct blockade of these K+ channels.     

Learning and memory of odor-reward association: Selective impairment following horizontal diagonal band lesions.

Rats with horizontal diagonal band of Broca (HDBB) lesions were trained in a successive-cue olfactory discrimination using different intertrial intervals (ITIs). They learned the paradigm of the task, but substantial impairment in performance of odor–reward associations was observed when the ITIs were longer than 15 sec. They performed as well as control animals with short ITIs when they were trained previously with long ITIs, but forgetting appeared very soon thereafter (24 hrs later). The finding is that the HDBB is an essential relay between the hippocampal system and the olfactory cortex. The HDBB allows associative memory storage when a limited-duration short-memory system located elsewhere is overlooked. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Metabotropic glutamate receptor inhibition of visceral afferent potassium currents

The effects of metabotropic glutamate receptor activation (mGluR) on voltage-gated potassium currents have been characterized in visceral sensory afferent neurons. L-Glutamate is known to be a primary neurotransmitter in visceral afferents which terminate at the level of the nucleus of the solitary tract (NTS). Synaptic communication between these afferents and the NTS has been shown to involve both postsynaptic ionotropic and presynaptic metabotropic glutamate receptor activation. The purpose of the present study was to determine the effects of mGluR activation on voltage-gated potassium currents in visceral sensory neurons. Application of mGluR agonist t-ACPD inhibited both the peak and the steady state voltage-gated potassium current in 39 out of 56 visceral afferent neurons tested (70%) by 22.0 +/- 3 and 22.8 +/- 2%, respectively. Voltage and pharmacological protocols were utilized to isolate the potassium current affected by mGluR activation. Increasing the holding potential from -100 mV to -30 mV only partially attenuated the inhibitory effects of t-ACPD (decreased effect by 11%), suggesting that t-ACPD modulates both a voltage insensitive and a voltage-sensitive potassium current. In addition, 4-aminopyridine (5 microM) was applied to eliminate the 4-AP sensitive transient current. Also, this protocol only partially attenuated the inhibitory effects of t-ACPD (decreased effect by 6.3%), suggesting that mGluR activation inhibits both a 4-AP-sensitive and 4-AP-insensitive potassium current in visceral afferent neurons. Results from this study suggest that mGluRs may regulate visceral sensory afferent neuronal activity through inhibition of voltage-gated potassium channels.     

P2 receptor modulation of voltage-gated potassium currents in Brown adipocytes

Using patch voltage-clamp techniques, we find there are two components to the voltage-gated potassium current (IKv) in rat brown adipocytes. The components differ in their gating and responses to purinergic stimulation, but not their pharmacology. IKv-A recovers from inactivation at physiological membrane potentials, while IKv-B inactivation recovers at more negative potentials. Both currents are >90% blocked by similar concentrations of quinine and tetraethylammonium, but not by beta-dendrotoxin, charybdotoxin, or apamin. The two current components are differentially modulated by extracellular ATP. ATP shifts the voltage dependence of IKv-A inactivation negative by 38 +/- 5 mV (n = 35, +/-SEM) and shifts activation by -14 +/- 2 mV in whole-cell experiments. ATP did not affect the steady state inactivation voltage dependence of IKv-B, but did apparently convert IKv-A into IKv-B. The pharmacology of the inactivation shift is consistent with mediation by a P2 purinergic receptor. Purinergic stimulation of perforated-patch clamped cells causes hyperpolarizing shifts in the window current of IKv-A by shifting inactivation -18 +/- 4 mV and activation -7 +/- 2 mV (n = 16). Since perforated-patch recordings will most closely resemble in vivo cell responses, this ATP-induced shift in the window current may facilitate IKv activation when the cell depolarizes. IKv activity is necessary for the proliferation and differentiation of brown adipocytes in culture (Pappone, P.A., and S.I. Ortiz-Miranda. 1993. Am. J. Physiol. 264:C1014-C1019) so purinergic modulation of IKv may be important in altering adipocyte growth and development.     

Vasopressin receptors modulate the pharmacological phenotypes of Cushing's syndrome

We have examined the expression profiles of the different vasopressin receptors (V1, V2, V3) that can be expressed in the three different types of tumors associated with Cushing's syndrome. V3 (V1b) receptor cDNA was cloned from a pituitary tumor responsible for Cushing's disease. We show that it is overexpressed in these tumors and can respond to DD-AVP. High expression of the V3 receptor on highly differentiated, ACTH-secreting, bronchial carcinoid tumors explain why these non-pituitary tumors occasionally respond to vasopressin, mimicking a "pituitary-like" behavior. A retrospective analysis showed that vasopressin induced an ACTH-independent cortisol rise in 27% of the adrenocortical tumors responsible for Cushing's syndrome. V1 mRNA was detected in normal adrenal cortex and in all tumors. Adenomas had significantly higher levels than carcinomas. V1 mRNA levels were higher in responders than in non-responders. One adenoma which had a brisk cortisol response in vivo, also had in vitro cortisol responses that were inhibited by a specific V1 antagonist. In situ hybridization showed the presence of V1 mRNA in the normal human adrenal cortex where the signal predominated in the compact cells of the zona reticularis. A positive signal was also present in the tumors with high V1 mRNA levels determined by RT-PCR; its distribution pattern was heterogeneous and showed preferential association with compact cells. High-and not ectopic-expression of the V1 receptor occurs in a minority of adrenal cortical tumors which become directly responsive to vasopressin stimulation.     

Different effects on learning ability after injection of the cholinergic immunotoxin ME20.4IgG-saporin into the diagonal band of Broca, basal nucleus of Meynert, or both in monkeys.

Immunotoxic lesions of the diagonal band of Broca (VDB) in monkeys disrupted cholinergic input to the hippocampus, producing impaired learning of visuospatial conditional discriminations but not simple visual discriminations. Immunotoxic lesions of the basal nucleus of Meynert (NBM) deprived the cortex of most of its cholinergic input, producing impaired learning of simple visual discriminations but not visuospatial conditional discriminations. Combined lesions of the NBM?+?VDB resulted in impaired learning of both types of task. The impairment after NBM lesions ameliorated with time but could be reinstated by a low dose of the glutamate blocking drug MK801, which, at this dose, did not impair simple visual discrimination learning in normal monkeys. The cholinergic projections from the NBM and VDB may sustain the function of the glutamatergic pyramidal cell pathways within the cortex and hippocampus, respectively. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Selectivity of linopirdine (DuP 996), a neurotransmitter release enhancer, in blocking voltage-dependent and calcium-activated potassium currents in hippocampal neurons

Linopirdine [DuP 996, 3, 3-bis(4-pyridinylmethyl)-1-phenylindolin-2-one], a putative cognition enhancing drug, increases acetylcholine release in rat brain tissue and improves performance in animal models of learning and memory. The mechanism whereby linopirdine enhances acetylcholine release has been proposed to involve inhibition of the M-type K+ current (IM). Our study examines the selectivity of linopirdine for IM by determining its effects on other ionic currents present in rat hippocampal CA1 neurons using patch clamp techniques. Linopirdine was found to block voltage-gated, calcium-activated and leak K+ currents in a dose-dependent manner. Of the seven currents measured, linopirdine was most selective for IM with an IC50 of 2.4 +/- 0.4 microM, followed by IC (measured as a medium afterhyperpolarization tail current, ImAHP) with an IC50 of 16.3 +/- 2.4 microM. Both IM and IC were completely suppressed by linopirdine. At a concentration of 100 microM, linopirdine weakly inhibited the K+ leak current, IL, the transient outward current, IA, the delayed rectifier, IK, and the slow component of IAHP, by 28 +/- 8, 37 +/- 10, 36 +/- 9 and 52 +/- 10 percent, respectively. The mixed Na+/K+ inward rectifying current, IQ, was essentially unaffected by linopirdine (IC50 >300 microM). These results indicate that linopirdine selectively blocks IM at concentrations 相似文献   

The supramammillary nucleus innervates cholinergic and GABAergic neurons in the medial septum-diagonal band of Broca complex

In the present study, the connectivity between two subcortical nuclei involved in hippocampal theta activity, the supramammillary nucleus and the medial septum-diagonal band of Broca complex, was examined. Targets of the supramammillary afferents in the medial septum-diagonal band of Broca complex were identified by combining anterograde transport of Phaseolus vulgaris leucoagglutinin with immunostaining for putative postsynaptic neurons, i.e. for parvalbumin and choline acetyltransferase that are known to label the GABAergic and cholinergic neurons, respectively, of the medial septum-diagonal band of Broca complex. Double retrograde transport experiments using the tracers horseradish peroxidase and wheat germ agglutinin-conjugated colloidal gold were employed to identify supramammillary neurons that project both to the hippocampus and the medial septum-diagonal band of Broca complex. Phaseolus vulgaris leucoagglutinin injections into the supramammillary nucleus of the rat resulted in dense fibre and terminal labelling in the medial septum-diagonal band of Broca complex. Labelled terminals formed asymmetric synapses mainly on distal dendrites of medial septal neurons. Proximal dendrites and somata were rarely contacted. The supramammillary afferents showed no target selectivity for a particular cell type; they innervated both cholinergic and GABAergic cells. Occasionally, perisomatic, basket-like terminals of supramammillary origin were found around parvalbumin-containing neurons. Double-retrograde experiments revealed that at least 25% of the supramammillo-hippocampal cells also projected to the medial septum-diagonal band of Broca. These data suggest that the nucleus, known to modulate the hippocampal electrical activity directly by the supramammillo-hippocampal pathway, also has the potential for an indirect action via the innervation of both the GABAergic and cholinergic septohippocampal neurons. This dual modulation may originate, at least in part, from the same population of supramammillary neurons.     

A remark on the high-conductance calcium-activated potassium channel in human endothelial cells

Iron plays a central role in the pathogenesis of Mycobacterium tuberculosis, the principal causative agent of tuberculosis. To learn more about iron acquisition by this bacterium, its iron regulated proteins (IRPs) were investigated. Seven IRPs were identified - three increased by high iron concentrations, and four by low iron concentrations. The smallest protein induced by low iron, Irp10, is tightly iron regulated as it is virtually absent in bacteria cultured in the presence of high iron concentrations. The gene (irpA ) encoding this protein and an adjacent open reading frame, mtaA, were cloned and sequenced. The protein encoded by mtaA (Mta72) has striking homology to metal transporting P-type ATPases. This study suggests that Irp10 and Mta72 function as a two-component metal transport system in M. tuberculosis.     

Gender difference in the cycle length-dependent QT and potassium currents in rabbits

Women are known to have a longer electrocardiographic Q-T than men, which may contribute to their being at greater risk of developing drug-induced polymorphic ventricular arrhythmias. However, little is known about the underlying mechanisms. In the present study, we evaluated potential gender differences in Q-T interval in isolated perfused rabbit hearts using the Langendorff technique and evaluated the density of outward potassium currents in single ventricular myocytes using the whole-cell patch-clamp technique. We found that female hearts demonstrated a greater Q-T lengthening (delta Q-T%) upon an increase in cycle length (CL), resulting in a significantly longer Q-T (301 +/- 4.8 ms, CL = 2.3 s) at a long CL in female hearts compared with male hearts (267 +/- 4.0 ms, P < .01). Ventricular myocytes isolated from female hearts showed a smaller IK(tail) and peak IKI outward current density. A 50% reduction in extracellular K+ and Mg++ shifted the I-V relationship of IKI and Ito and reduced their amplitude. However, neither the I-V relationship of IKr nor the gender difference in the Q-T-CL relationship was significantly altered. We conclude that 1) female rabbit ventricular myocytes have significantly lower IKr and IKl outward current densities than do male cells, which may contribute to the gender difference in Q-T, and 2) a lower base-line IKr density may contribute to the steeper Q-T-CL relationship in female hearts.     

Specific residues in the Pbx homeodomain differentially modulate the DNA-binding activity of Hox and Engrailed proteins

Two classes of homeodomain proteins, Hox and Engrailed, have been shown to act in concert with the atypical homeodomain proteins Pbx and extradenticle. We now show that specific residues located within the Pbx homeodomain are essential for cooperative DNA binding with Hox and Engrailed gene products. Within the N-terminal region of the Pbx homeodomain, we have identified a residue that is required for cooperative DNA binding with three Hox gene products but not for cooperativity with Engrailed-2 (En-2). Furthermore, there are similarities between heterodimeric interactions involving the yeast mating type proteins MATa1 and MATalpha2 and those that allow the formation of Pbx/Hox and Pbx/En-2 heterodimers. Specifically, residues located in the a1 homeodomain that were previously shown to form a hydrophobic pocket allowing the alpha2 C-terminal tail to bind, are also required for Pbx/Hox and Pbx/En-2 cooperativity. Furthermore, we show that three residues located in the turn between helix 1 and helix 2, characteristic of many atypical homeodomain proteins, are required for cooperative DNA binding involving both Hox and En-2. Replacement of the three residues located in the turn between helix 1 and helix 2 of the Pbx homeodomain with those of the atypical homeodomain proteins controlling cell fate in the basidiomycete Ustilago maydis, bE5 and bE6, allows cooperative DNA binding with three Hox members but abolishes interactions with En-2. The data suggest that the molecular mechanism of homeodomain protein interactions that control cell fate in Saccharomyces cerevisiae and in the basidiomycetes may well be conserved in part in multicellular organisms.     

Cardiopulmonary actions of muscarinic receptor subtypes in the newborn dog

We addressed the role of muscarinic receptor subtypes in neurally mediated bronchoconstriction in vivo and airway smooth muscle contraction in vitro in the newborn dog. The in vivo dose-response effects of "selective" muscarinic antagonists on changes in lung resistance (RL) and heart rate (HR) in response to electrical stimulation of the vagus nerves were obtained in four groups of newborns. Each group was exposed to a different muscarinic antagonist: M1-selective pirenzepine (pir), M2-selective AF-DX 116 (11-[2-[(diethylamino)methyl]-1-piperidinyl]acetyl-5,11-dihydro-6H-pyrid o- [2,3-b]-[1,4]-benzodiazepine-6-one), M3-selective p-F-HHSiD (p-fluoro-hexahydro-sila-difenidol), and nonselective atropine (atr). In vitro concentration-response effects of pir and AF-DX 116 were obtained for neurally induced contractions of tracheal smooth muscle, elicited by electrical field stimulation. In a separate series of experiments we measured the bronchoconstrictor response to the muscarinic agonist acetylcholine delivered by right heart injection. Muscarinic antagonists reduced RL and HR responses to vagal stimulation in a dose-dependant fashion; however, ED50 values and selectivity for airway and cardiac responses (HR/RL ED50 ratio) were significantly different between antagonists. The rank order of potencies for inhibition of the increase in RL was atr > pir, M1 > p-F-HHSiD, M3 > AF-DX 116, M2, while that for HR was atr > AF-DX 116 > pir > p-F-HHSiD. AF-DX 116 preferentially inhibited the HR response, as reflected by the lowest HR/RL ED50 ratio (p < 0.001). The remaining antagonists preferentially inhibited RL, with the highest HR/RL ED50 ratio seen for p-F-HHSiD. These data suggest that muscarinic receptor subtypes are differentiated at birth and mediate cardiac and airway responses to vagal stimulation. We did not find autoinhibitory actions of airway M2 receptors on either the in vivo bronchoconstrictor response or the in vitro contractile response to electrical field stimulation. This suggests that neonatal airway M2 receptors, but not myocardial M2 receptors, are reduced in number or weakly coupled to muscarinic signal transduction mechanisms. Direct activation of airway smooth muscle by acetylcholine caused dose-dependent increases in RL that reached a plateau at approximately 200% at 100 micrograms, similar to values reported for vagal stimulation.