Escape Threshold Research Articles

Ca 2+/calmodulin-protein kinase IIα in the trigeminal subnucleus caudalis contributes to neuropathic pain following inferior alveolar nerve transection

Calcium-calmodulin protein kinase IIα (CaMKIIα) is mainly found in brain cells, and the mRNA concentrates highly in the postsynaptic density. CaMKIIα is an effector of calcium and calmodulin mediated functions, and the phosphorylated CaMKIIα (pCaMKIIα) activates glutamate receptors, such as the AMPA receptor, and enhances its function. In the present study, we examined whether CaMKIIα in trigeminal brainstem neurons contributed to the neuropathic pain induced by inferior alveolar nerve (IAN) transection. Using immunohistochemistry and in situ hybridization, we found that the expression of CaMKIIα and pCaMKIIα increased in the trigeminal subnucleus caudalis (Vc) after IAN transection. The significant increase in the protein of CaMKIIα peaked at 30 min after IAN transection, and the mRNA of CaMKIIα increased from 2 to 14 days. Double immunofluorescent staining for CaMKIIα and MAP2, a marker of dendrite, revealed a significant increase in the overlapping area at 30 min after injury. This suggests that CaMKIIα protein is synthesized from the local mRNA pool in the dendrite 30 min after IAN transection and may quickly transmit information after nerve injury. In the behavioral test in which the escape threshold from mechanical stimulation to the lateral face was measured, intrathecal administration of KN-93, a CaMKII inhibitor, for 7 days significantly inhibited mechano-allodynia induced by IAN transection, as compared with administration of a control peptide. These data suggest that CaMKIIα in the trigeminal subnucleus caudalis may be involved in neuropathic pain caused by IAN transection.

SHORT COMMUNICATIONS: Increases in extracellular levels of 5‐HT and dopamine in the basolateral, but not in the central, nucleus of amygdala induced by aversive stimulation of the inferior colliculus

Consistent evidence has shown that dopamine release in the prefrontal cortex is increased by electrical stimulation of the inferior colliculus (IC) as unconditioned stimulus. Recent reports have also demonstrated that inactivation of the basolateral nucleus of the amygdala (BLA) with muscimol enhances the behavioural consequences of the aversive stimulation of the IC and reduces the dopamine release in the prefrontal cortex. Moreover, neurotoxic lesions of the BLA enhance whereas those of the central nucleus of the amygdala (CeA) reduce the aversiveness of the electrical stimulation of the IC. Based on these findings the present study examined the effects of the electrical stimulation of the IC on the extracellular levels of serotonin and dopamine in the BLA and CeA. To this end, rats implanted with a stimulation electrode in the IC also bore a microdialysis probe in the BLA or CeA for determination of the release of dopamine and serotonin. IC electrical stimulation at the freezing and escape thresholds increased the levels of serotonin ( approximately 70%) and dopamine ( approximately 60%) in the BLA related to the basal values. Similarly, the metabolites DOPAC and 5-HIAA increased in a parallel fashion in BLA. No significant changes could be detected in these biogenic amines and metabolites in CeA following IC aversive stimulation. These findings point to a differential role of serotonergic and dopaminergic mechanisms of the BLA and CeA in the setting up of adaptive responses to fear states generated at the inferior colliculus level.

Temporomandibular Joint Inflammation Potentiates the Excitability of Trigeminal Root Ganglion Neurons Innervating the Facial Skin in Rats

The aim of this study was to test the hypothesis that temporomandibular joint (TMJ) inflammation alters the excitability of trigeminal root ganglion (TRG) neurons innervating the facial skin, by using behavioral, electrophysiological, molecular, and immunohistochemical approaches. Complete Freund's adjuvant (CFA) was injected into the rat TMJ to produce inflammation. The threshold for escape from mechanical stimulation applied to the orofacial area in TMJ-inflamed rats was significantly lower than that in naïve rats. The TRG neurons innervating the inflamed TMJ were labeled by 2% Fluorogold (FG) injection into the TMJ. The number of FG-labeled substance P (SP)-immunoreactive neurons in the inflamed rats was significantly increased compared with that in the naïve rats. On the other hand, medium- and large-diameter TRG neurons (>30 microm) innervating the facial skin were labeled by FG injection into the facial skin. In the FG-labeled cutaneous TRG neurons, the occurrence of SP (100 nM) induced membrane depolarization in inflamed rats (medium: 73.3%, large : 85.7%) was larger than that in the naïve rats (medium: 29.4%, large : 0%). In addition, SP application significantly increased the firing rate evoked by depolarizing pulses in the neurons of inflamed rats compared with those of naïve rats. Quantitative single-cell RT-PCR analysis showed the increased expression of mRNA for the NK1 receptor in FG-labeled TRG neurons in inflamed rats compared with that in naive rats. The numbers of SP and NK1 receptors/neurofilament 200 positive immunoreactive TRG neurons innervating the facial skin (FG-labeled) in the inflamed rats were significantly increased compared with those seen in naïve rats. These results suggest that TMJ inflammation can alter the excitability of medium- and large-diameter TRG neurons innervating the facial skin and that an increase in SP/NK1 receptors in their soma may contribute to the mechanism underlying the trigeminal inflammatory allodynia in the TMJ disorder.

Early Painful Diabetic Neuropathy Is Associated with Differential Changes in Tetrodotoxin-sensitive and -resistant Sodium Channels in Dorsal Root Ganglion Neurons in the Rat

Diabetic neuropathy is a common form of peripheral neuropathy, yet the mechanisms responsible for pain in this disease are poorly understood. Alterations in the expression and function of voltage-gated tetrodotoxin-resistant (TTX-R) sodium channels have been implicated in animal models of neuropathic pain, including models of diabetic neuropathy. We investigated the expression and function of TTX-sensitive (TTX-S) and TTX-R sodium channels in dorsal root ganglion (DRG) neurons and the responses to thermal hyperalgesia and mechanical allodynia in streptozotocin-treated rats between 4-8 weeks after onset of diabetes. Diabetic rats demonstrated a significant reduction in the threshold for escape from innocuous mechanical pressure (allodynia) and a reduction in the latency to withdrawal from a noxious thermal stimulus (hyperalgesia). Both TTX-S and TTX-R sodium currents increased significantly in small DRG neurons isolated from diabetic rats. The voltage-dependent activation and steady-state inactivation curves for these currents were shifted negatively. TTX-S currents induced by fast or slow voltage ramps increased markedly in neurons from diabetic rats. Immunoblots and immunofluorescence staining demonstrated significant increases in the expression of Na(v)1.3 (TTX-S) and Na(v) 1.7 (TTX-S) and decreases in the expression of Na(v) 1.6 (TTX-S) and Na(v)1.8 (TTX-R) in diabetic rats. The level of serine/threonine phosphorylation of Na(v) 1.6 and In Na(v)1.8 increased in response to diabetes. addition, increased tyrosine phosphorylation of Na(v)1.6 and Na(v)1.7 was observed in DRGs from diabetic rats. These results suggest that both TTX-S and TTX-R sodium channels play important roles and that differential phosphorylation of sodium channels involving both serine/threonine and tyrosine sites contributes to painful diabetic neuropathy.

Fos-like immunoreactive neurons following electrical stimulation of the dorsal periaqueductal gray at freezing and escape thresholds

Electrical stimulation of the dorsal regions of the periaqueductal gray (PAG) leads to defensive reactions characterized as freezing and escape responses. Until recently it was thought that this freezing behavior could be due to the recruitment of neural circuits in the ventrolateral periaqueductal gray (vlPAG), while escape would be mediated by other pathways. Nowadays, this view has been changing mainly because of evidence that freezing and escape behaviors thus elicited are not altered after lesions of the vlPAG. It has been suggested that there are at least two pathways for periaqueductal gray-mediated defensive responses, one involving the hypothalamus and the cuneiform nucleus (CnF) which mediates responses to immediate danger and another one involving the amygdala and vlPAG which mediates cue-elicited responses, either learned or innate. To examine this issue further we measured Fos protein expression in brain areas activated by electrical stimulation of the dorsolateral PAG (dlPAG) at the freezing and escape thresholds. The data obtained showed that freezing-provoking stimulation caused increases in Fos expression in the dorsomedial PAG (dmPAG), while escape-provoking stimulation led to increases at both dmPAG and dlPAG. Surprisingly, neither escape- nor freezing-provoking stimulations altered Fos expression in the central nucleus of amygdala (CeA). Escape-provoking stimulation caused increased Fos expression in the ventromedial hypothalamus (VMH), dorsal premammilary nucleus (PMd) and in the cuneiform nucleus. Significant increases in Fos labeling were found in the dmPAG and PMd following freezing-provoking stimulation. Therefore, the present data support the notion of a neural segregation for defensive behaviors in the dorsal columns of PAG, with increased Fos expression in the dmPAG following freezing, while dlPAG is affected by both freezing and escape responses. dlPAG, CnF, VMH and PMd are part of a brain aversion network activated by fear unconditioned stimuli. The present data also suggests that the defensive responses generated at the dlPAG level do not recruit the neural circuits of the vlPAG and CeA usually activated by conditioned fear stimuli.

A new model of experimental parotitis in rats and its implication for trigeminal nociception.

A rat model of chronic parotitis was developed following a direct injection of Complete Freund's adjuvant (CFA) into the unilateral parotid gland via the parotid duct without skin incision. The nocifensive behavior, plasma extravasation in the parotid gland, and trigeminal Fos protein expression, a marker of neuronal activation, were analyzed in this model and compared to that of the saline-injected rats. A significant reduction of the escape threshold to mechanical stimulation of the lateral face on the ipsilateral side to the CFA injection was observed at 1-6 days after CFA injection as compared to that of the pre-CFA control ( P<0.01). The lateral face region contralateral to the CFA injection also showed mechanical hyperalgesia at 1-6 days after injection ( P<0.05). The plasma extravasation was significantly increased in the parotid gland ipsilateral to CFA injection as compared to that of the parotid gland with saline injection at 3 days after injection as shown by Evans' blue dye extravasation ( P<0.05). Bilateral expression of Fos protein-like immunoreactive cells was observed in the transition zone between the trigeminal spinal nucleus interpolaris (Vi) and caudalis (Vc) and paratrigeminal nucleus (Pa5). On the other hand, a significant unilateral expression of Fos protein-positive cells was observed on the ipsilateral side of the upper cervical (C2) dorsal horn ( P<0.05). This model of parotitis can be used to study trigeminal pain mechanisms associated with sialadenitis. A unique feature of this preparation is that the inflammation was limited to the parotid gland after intraductal injection of CFA, allowing analysis of peripheral input from a defined orofacial region. The model will be useful in developing new strategies to treat chronic orofacial pain.

Noise-induced escape through a chaotic saddle: lowering of the activation energy

The noise-induced escape mechanism is investigated for a prototype time-discrete nonequilibrium system describing a laser in a ring cavity with optical feedback, the Ikeda map. For certain parameter values a chaotic saddle is embedded in the basin of attraction of the metastable state. This results in a lowering of the escape threshold for the activation energy, which is established by employing the theory of quasipotentials. Our findings are explained by the computation of the most probable exit path (MPEP), which lies partly on the chaotic saddle. The enhancement of escape turns out to be maximal if the MPEP lies entirely on the chaotic saddle.

Contrasting magmas in metapelitic and metapsammitic migmatites in the Cooma Complex, Australia

Metapelite-derived migmatites (“bedded migmatites”) formed in the low-pressure/high-temperature (LPHT) Cooma Complex, southeastern Australia, contain magma (neosome and leucosome) confined to the metapelitic beds in which they were generated. The metapsammitic beds were more ductile than the metapelitic beds (and the metapelitic parts of graded beds), which underwent fracture and boudinage, thereby providing space for the magma, though some also occurs in axial surface folia. Transitions from bedded to stromatic migmatites can be seen, but the magma mainly remained in the metapelites, even in the most strongly deformed stromatic migmatites. This, together with boudinage and transposition of the leucosome, as well as microstructural evidence of quartz recrystallization, suggest that much or most of the stromatic layering was formed by solid-state deformation. In contrast, magmas (neosomes) formed by partial melting of feldspathic metapsammites at Cooma moved out of their parent rocks, and coalesced into veins and small intrusions of diatexite, because (1) the host rocks deformed more homogeneously, and no interboudin space was made for the melts, and (2) the melt escape threshold was exceeded, probably with the assistance of deformation. Metapsammite melting occurred after solidification of the metapelite-derived magma, and the mobile metapsammite-derived magma (diatexite) disrupted and incorporated fragments of the metapelitic migmatites. The metapsammite-derived magma, together with this solid metapelitic material, locally coalesced into bodies closely resembling the Cooma Granodiorite.

Effects of naloxone on rewarding and aversive brain sites

The effects of naloxone on the rewarding and aversive properties of brain stimulation derived from the ventral tegmental area and the nucleus reticularis gigantocellularis, respectively, were assessed in rats, based on the following measures—the current threshold for latency to escape aversive nucleus reticularis gigantocellularis stimulation, the frequency threshold for rewarding ventral tegmental area stimulation, and the frequency threshold for self-stimulation obtained from delivery of concurrent ventral tegmental area and nucleus reticularis gigantocellularis stimulation, before and after three systemic doses of naloxone (0, 10, and 20mg/kg); in the latter case, the stimulation trains were interdigitated with an interpulse interval of 2ms. Initially, thresholds for concurrent stimulation were elevated relative to the values obtained for ventral tegmental area stimulation alone, returning to baseline values only when the nucleus reticularis gigantocellularis stimulation no longer induced escape. After each pairing of the two sites, the current threshold for escape gradually increased until the maximum value administered, 700μA, at which point aversive responses were no longer observed. This required very few pairings, between one and five trials across animals. Drug tests were then begun and produced a significant dose-response threshold increase across animals, without reinstating the latency to escape nucleus reticularis gigantocellularis stimulation. These findings are discussed in terms of a dissociation between the analgesic and rewarding properties of ventral tegmental area stimulation.

Apomorphine enhances conditioned responses induced by aversive stimulation of the inferior colliculus.

Consistent evidence has shown that learning may be produced in paradigms using electrical stimulation of the inferior colliculus (IC) as unconditioned stimulus (UCS). Recent reports have also demonstrated that aversive stimulation of the IC, at the escape threshold, enhances dopamine (DA) release in the prefrontal cortex. The purpose of the present study was to determine whether dopaminergic mechanisms are involved in the Pavlovian conditioning and latent inhibition using IC stimulation as UCS and light as conditioned stimulus (CS). Rats were placed inside a shuttle box and subjected to a two-way avoidance paradigm. IC aversive electrical stimulation was used as UCS and shuttle box illumination as CS. The rats quickly learned to avoid or interrupt the IC stimulation. Apomorphine injections produced a dose-dependent increase in the number of avoidance responses. On the other hand, chlorpromazine administration promoted a dose-dependent reduction of the avoidance responses. Previous injections of chlorpromazine inhibited the effects of apomorphine. Also, previous exposure to unreinforced light weakened the strength of the conditioning. Apomorphine blocked this latent inhibition effect, which was antagonized by previous injections of chlorpromazine. These findings bring evidence for the involvement of DA in the setting up of adaptive responses to aversive states generated at the IC level, which may underlie stressful situations present in anxiety.

Photodetachment of He- in the vicinity of the two-electron escape threshold.

Recording the yield of He(1snl(3)L) Rydberg states for n=11-14, we measure the photodetachment cross sections of metastable He-(1s2s2p(4)P(o)) ions in the vicinity of the two-electron escape threshold. We observe a large number of double Rydberg He- quartet state resonances and report energies and widths of intrashell states in the n=13-15 manifolds. Sharp thresholds are measured at He((3)P(o)) and He((3)D(e)) Rydberg states with preference for population of the former, whereas the He((3)S(e)) states are not populated, in agreement with qualitative theoretical arguments.

A model for the conservation management of a ‘secondary’ prey: sooty shearwater ( Puffinus griseus) colonies on mainland New Zealand as a case study

New Zealand's native birds constitute the supplementary or ‘secondary’ prey of introduced mammalian predators. Predation on secondary prey is inversely density dependent. Small populations will therefore be at greater risk of extinction than large, which escape due to the buffering effect of their size on predation impact. A matrix model of a mainland sooty shearwater ( Puffinus griseus) population predicted a mean growth rate of 1.044 (0.967–1.130) with no predation. Survival of adults attending the colony was the most influential population parameter. With predation effects included, colony growth was sensitive to both management intensity and colony size. Small colonies required the removal of almost all predation risk to survive, although the level of management required for growth decreased with increasing colony size. Management intensity may be relaxed as a colony grows and an ‘escape threshold' colony size is eventually reached at which growth continues without predator management.

Induction of Fos protein-like immunoreactivity in the trigeminal spinal nucleus caudalis and upper cervical cord following noxious and non-noxious mechanical stimulation of the whisker pad of the rat with an inferior alveolar nerve transection

After transection of the inferior alveolar nerve (IAN: the third branch of the trigeminal nerve), the whisker pad area, which is innervated by the second branch of the trigeminal nerve, showed hypersensitivity to mechanical stimulation. Two days after IAN transection, the threshold intensity for escape behavior to mechanical stimulation of the ipsilateral whisker pad area was less than 1.0 g, a sign of allodynia, and returned to the preoperative level (preoperative threshold: 52.0 g) at 32 days after surgery. This decrement of escape threshold lasted for more than 3 weeks. The whisker pad area contralateral to the IAN transection also showed a decrease in escape threshold to non-noxious mechanical stimulation as compared with sham-operated rats. However, the change in threshold intensity for the side contralateral to transection was not as pronounced as that on the ipsilateral side. Fos protein-like immunoreactive (LI) cells were observed in the superficial laminae but not dominant in deeper laminae of the trigeminal spinal nucleus caudalis (Vc) and the first segment of the spinal cord (C1) after non-noxious mechanical stimulation of the whisker pad area in the rats with IAN transection. Fos protein-LI cells were expressed bilaterally in the Vc and C1, but were more numerous on the ipsilateral side to transection than on the contralateral side. The largest number of Fos protein-LI cells was observed at 2400 μm caudal from the trigeminal subnucleus interporalis (Vi)–Vc border both in ipsilateral and contralateral sides. The number of Fos protein-LI cells increased after application of 1, 4, and 16 g stimuli as compared to rats without mechanical stimulation. Furthermore, an extensively greater number of Fos protein-LI cells were expressed both in superficial and deep laminae of the bilateral Vc and C1 of the spinal cord after subcutaneous injection of mustard oil into the whisker pad. Fos protein expression after mustard oil injection was much stronger than that observed after any mechanical stimulation in the rats with IAN transection. These data suggest that the change in the numbers and spatial arrangement of nociceptive neurons in the Vc and C1 after IAN transection reflect the development of mechanical hyperalgesia in the area adjacent to the IAN innervated region.

Alteration of medullary dorsal horn neuronal activity following inferior alveolar nerve transection in rats.

The effects of inferior alveolar nerve (IAN) transection on escape behavior and MDH neuronal activity to noxious and nonnoxious stimulation of the face were precisely analyzed. Relative thresholds for escape from mechanical stimulation applied to the whisker pad area ipsilateral to the transection were significantly lower than that for the contralateral and sham-operated whisker pad until 28 days after the transection, then returned to the preoperative level at 40 days after transection. A total of 540 neurons were recorded from the medullary dorsal horn (MDH) of the nontreated naive rats [low-threshold mechanoreceptive (LTM), 27; wide dynamic range (WDR), 31; nociceptive specific (NS), 11] and sham-operated rats with skin incision (LTM, 34; WDR, 30; NS, 23) and from the ipsilateral (LTM, 82; WDR, 82; NS, 31) and contralateral MDH relative to the IAN transection (LTM, 77; WDR, 82; NS, 33). The electrophysiological properties of these neurons were precisely analyzed. Background activity of WDR neurons on the ipsilateral side relative to the transection was significantly increased at 2-14 days after the operation as compared with that of naive rats. Innocuous and noxious mechanical-evoked responses of LTM and WDR neurons were significantly enhanced at 2-14 days after IAN transection. The mean area of the receptive fields of WDR neurons was significantly larger on the ipsilateral MDH at 2-7 days after transection than that of naive rats. We could not observe any modulation of thermal responses of WDR and NS neurons following IAN transection. Also, no MDH neurons were significantly affected in the rats with sham operations. The present findings suggest that the increment of neuronal activity of WDR neurons in the MDH following IAN transection may play an important role in the development of the mechano-allodynia induced in the area adjacent to the area innervated by the injured nerve.

Neuroanatomical and psychopharmacological evidence for interaction between opioid and GABAergic neural pathways in the modulation of fear and defense elicited by electrical and chemical stimulation of the deep layers of the superior colliculus and dorsal periaqueductal gray matter

The effects of central administration of opioid antagonists on the aversive responses elicited by electrical (at the freezing and escape thresholds) or chemical stimulation (crossings, rearings, turnings and jumps, induced by microinjections of bicuculline) of the midbrain tectum were determined. Central microinjections of naloxone and naltrexone in the mesencephalic tectum caused a significant increase in the freezing and escape thresholds elicited by electrical midbrain tectum stimulation. Furthermore, both opioid antagonists caused a significant decrease in the mean incidence of aversive behavioral responses induced by microinjections of bicuculline in the deep layers of the superior colliculus (DLSC) and in dorsal aspects of the periaqueductal gray matter (DPAG), as compared with controls. These findings suggest an opioid modulation of the GABAergic inhibitory inputs controlling the aversive behavior elicited by midbrain tectum stimulation. In fact, immunohistochemical evidence suggests that the dorsal mesencephalon is rich in β-endorphin-containing neurons and fibers with varicosities. Iontophoretical microinjections of the neurotracer biodextran in the substantia nigra, pars reticulata (SNpr), show nigro-tectal pathways connecting SNpr with the same neural substrate of the DPAG rich in neuronal cells immunoreactive for opioid peptides. Labeled neurons of the DLSC and periaqueductal gray matter send inputs with varsicosities to ipsi- and contralateral DPAG and ipsilateral SNpr. These findings, in addition to the psychopharmacological evidence for the interaction between opioid and GABAergic mechanisms, offer a neuroanatomical basis of a possible presynaptic opioid inhibition of GABAergic nigro-tectal neurons modulating the fear in aversive structures of the cranial mesencephalon, in a short link, and maybe through a major neural circuit, also in GABA-containing perikarya of nigro-tectal neurons.

Factors that influence the persistence of stimulation-induced aversion

Brain stimulation reward in certain regions has been shown to produce analgesia to externally applied painful stimuli. In the present experiments, we studied how electrical self-stimulation of the dorsal raphe (DR) nucleus modifies the aversive effects of electrical stimulation of the nucleus reticularis gigantocellularis (Gi) or of the dorsal tegmentum (DTg). In the first study, the threshold for latency to escape aversive Gi stimulation was tracked before and after exposure to rewarding DR stimulation. Only a few sessions of DR self-stimulation were required to produce a complete and long-lasting inhibition of Gi aversion. In the second study, the aversion induced by DTg stimulation rapidly disappeared following a few test sessions at that site. Unlike our previous experience with Gi aversion that required either pairing with rewarding lateral hypothalamic (LH) or ventral tegmental area (VTA) pulses in order to increase the threshold for latency to escape Gi aversion, in this study, simply brief experience with rewarding DR stimulation in unpaired trials was sufficient to entirely suppress Gi-induced aversion. Even more surprising was the finding that unlike the Gi, aversion obtained from activation of the DTg does not persist, its threshold for escape quickly increases, and within a few sessions is no longer evident. One interpretation of these findings is that the aversion mechanisms associated with the Gi and DTg are differentially susceptible to analgesic processes.

Conditioned antinociception and freezing using electrical stimulation of the dorsal periaqueductal gray or inferior colliculus as unconditioned stimulus are differentially regulated by 5-HT2A receptors in rats.

Electrical stimulation of the dorsal periaqueductal gray matter (dPAG) and the inferior colliculus (IC) has been used as an aversive unconditioned stimulus. However, studies on the behavioral, sensorial and autonomic components of the conditioned fear elaborated in the midbrain tectum are lacking. This study was undertaken to investigate the nature of the aversiveness of stimulation of the dPAG and IC as well as the modulation by 5-HT mechanisms of the fear conditioned responses to these stimulations. Animals chronically implanted with an electrode glued to a guide cannula into the dPAG or the IC were submitted to one, two or three sessions of conditioning. Each session consisted of ten pairings of the light in a distinctive chamber (CS) with the electrical stimulation of one of these regions at the escape threshold determined previously. Control groups were submitted to the same procedure, except for the conditioning sessions in which the conditioned stimuli were presented alone in one case and performed in a different context in the other. On the next day, each animal was exposed only to the CS (testing) and the duration of freezing, number of rearings, grooming, bouts of micturition and fecal boli were recorded for 5 min. Before and after the testing session, the animals were submitted to the tail-flick test. The data showed that the conditioning with electrical stimulation of the dPAG and the IC caused significant increases in the time of freezing, defecation and micturition, and significant reductions in the number of rearings and grooming. On the other hand, only the conditioning with electrical stimulation of the dPAG produced significant conditioned antinociception. Microinjections of methysergide, a non-specific antagonist of 5-HT receptors, or ketanserin, an antagonist of 5-HT2A receptors, into the dPAG before testing significantly inhibited the antinociception without affecting any of the behavioral or autonomic conditioned responses. 1) Conditioned freezing may be produced using the electrical stimulation of the dPAG or IC as unconditioned stimuli, 2) only the pairing of CS plus dPAG but not with IC stimulation, produces significant conditioned antinociception, 3) blockade of 5-HT2A receptors inhibits conditioned antinociception but not the conditioned defensive behavior using the electrical stimulation of the dPAG as unconditioned stimulus.

Viral escape at the molecular level explained by quantitative T-cell receptor/peptide/MHC interactions and the crystal structure of a peptide/MHC complex

Viral escape, first characterized for the lymphocytic choriomeningitis virus (LCMV) in a mouse transgenic for the P14 T cell-receptor (TCR), can be due to mutations in T-cell epitopes. We have measured the affinity between the H-2D b containing the wild-type and two of its “viral escape” epitopes, as well as other altered peptide ligands (APL), by using BIACORE analysis, and solved the crystal structure of H-2D b in complex with the wild-type peptide at 2.75 Å resolution. We show that viral escape is due to a 50 to 100-fold reduction in the level of affinity between the P14 TCR and the binary complexes of the MHC molecule with the different peptides. Structurally, one of the mutations alters a TCR contact residue, while the effect of the other on the binding of the TCR must be indirect through structural rearrangements. The former is a null ligand, while the latter still leads to some central tolerance. This work defines the structural and energetic threshold for viral escape.

Continuous vs. discontinuous melt segregation in migmatites: insights from a cellular automaton model

A cellular automaton model is presented that simulates melt extraction from migmatites. The varying parameters are the relative amount of melt, thresholds for melt connection leading to movement and melt escape, and type of deformation (pure and/or simple shear). It is shown that when the melt percentage is about half that of the melt escape threshold, the competition between production and deformation‐assisted extraction results in a discontinuous rate of melt extraction. The process can be compared to stick‐slip motion during frictional experiments. At higher melt fraction, it is continuously extracted. Deformation assists significantly in segregating melt, particularly when a pure shear (compaction) component is added to simple shear.

Electrical stimulation of the midbrain tectum enhances dopamine release in the frontal cortex

One widely used animal model of anxiety is the electrical stimulation of a given structure supposed to be involved in the neural circuitry underlying emotional behavior. Indeed, electrical stimulation of midbrain structures with substrates for the processing of fear-like responses, such as the dorsal periaqueductal gray matter (DPAG) or the inferior colliculus (IC), produces behavioral, sensorial and autonomic responses very similar to the defense reactions observed in environmental threatening situations. It has also been proposed that the required level of integration of all these components of the defense reaction needs an integrative process situated at higher brain level, as the prefrontal cortex. As a matter of fact, substantial cortical inputs to the midbrain tectum have already been found. In view of this evidence, it seems important to know whether animals stimulated in the midbrain tectum would present neurochemical changes in the prefrontal cortex. To this end, we examined the temporal course of the effects of the electrical stimulation of the DPAG and IC on the dopamine (DA) release in the prefrontal cortex. Electrical stimulation of these structures was performed at the alertness (control) and escape thresholds. Electrical stimulation of the inferior colliculus at the escape threshold produced a long-lasting increase in the levels of corticofrontal dopamine in relation to these measurements in the control group. No significant changes in extracellular DA release in this cortical area could be observed following DPAG electrical stimulation. These findings bring evidence for the involvement of dopamine of the frontal cortex in the setting up of adaptive responses to stressful situations generated at the inferior colliculus level.