Morphine antinociceptive potency on chemical, mechanical, and thermal nociceptive tests in the rat.

Many tests are used to assess nociception in laboratory animals. The objective of this study was to compare morphine potency across tests. Rats were injected with saline or morphine (1-20 mg/kg SC), and nociception was assessed 15-20 min later. A consistent definition of antinociception-a change in response greater than 4 times the standard deviation above the mean for the saline-treated controls-was used to compare morphine potency on different tests. These data revealed 4 things. 1) Morphine potency was greatest on the paw pressure, hot plate, and tail withdrawal tests and lowest on the formalin test. 2) Stimulus intensity had no effect on morphine potency on the hot plate (ED50 = 4.5, 2.8, and 2.6 mg/kg for 49 degrees C, 52 degrees C, and 55 degrees C tests, respectively) or tail withdrawal tests (ED50 = 2.9 and 2.6 for 48 degrees C and 52 degrees C water, respectively). 3) Assessment of morphine potency using a within-subjects cumulative dosing procedure resulted in lower ED50 values compared to data collected using a between-subjects design (hot plate: 2.6 vs 4.9; tail withdrawal: 2.6-2.9 vs 5.7 mg/kg). 4) Adjusting the cutoff value from 4 to 5, 6, 7, and 8 standard deviations greater than the mean resulted in a progressive increase in ED50 values. These data demonstrate that morphine potency is dependent, in part, on the nociceptive test even when all other factors (eg, species, strain, age, gender, and cutoff value) are held constant. PERSPECTIVE: The ability of morphine to block nociception is influenced by many factors. The present study shows that the test used to assess nociception, but not the stimulus intensity, can have a dramatic effect on morphine potency. This finding shows that morphine potency varies depending on how pain is assessed.     

The induction of Fos-like immunoreactivity by noxious thermal, mechanical and chemical stimuli in the lumbar spinal cord of infant rats

The present study examined the maturation of nociceptive primary afferents using expression of Fos-like immunoreactivity in the second-order spinal cord neurons as an anatomical and functional marker. Pinch, immersion in hot water, or formalin injection applied to the hindpaw was used as the peripheral noxious stimulus in awake 0-, 1-, 2-, 3-, and 14-day-old rat pups. On the day of birth, all 3 stimuli elicited expression of the Fos protein in dorsal horn cells indicating that nociceptive primary afferents are functional at this age. The expression of the Fos protein was related to the intensity of stimulation since greater injection volumes of formalin or prolonged application of the thermal stimulus increased the number of stained nuclei. The number of stained nuclei was age dependent and older pups exhibited a greater number of stained nuclei. The results of this study are consistent with electrophysiological studies that have demonstrated that the primary nociceptive afferents continue to mature during the rats' postnatal life. Furthermore, the number of Fos immunoreactive neurons in immature rats are age and stimulus-intensity dependent.     

Trigeminal activation using chemical, electrical, and mechanical stimuli

Tactile, proprioceptive, and nociceptive information, including also chemosensory functions are expressed in the trigeminal nerve sensory response. To study differences in the processing of different stimulus qualities, we performed a study based on functional magnetic resonance imaging. The first trigeminal branch (ophthalmic nerve) was activated by (a) intranasal chemical stimulation with gaseous CO2 which produces stinging and burning sensations, but is virtually odorless, (b) painful, but not nociceptive specific cutaneous electrical stimulation, and (c) cutaneous mechanical stimulation using air puffs. Eighteen healthy subjects participated (eight men, 10 women, mean age 31 years). Painful stimuli produced patterns of activation similar to what has been reported for other noxious stimuli, namely activation in the primary and secondary somatosensory cortices, anterior cingulate cortex, insular cortex, and thalamus. In addition, analyses indicated intensity-related activation in the prefrontal cortex which was specifically involved in the evaluation of stimulus intensity. Importantly, the results also indicated similarities between activation patterns after intranasal chemosensory trigeminal stimulation and patterns usually found following intranasal odorous stimulation, indicating the intimate connection between these two systems in the processing of sensory information.     

The antinociceptive effects of stimulating the pretectal nucleus of the rat

Changes in the tail-flick latency to noxious heat were studied following electrical stimulation of the dorso-medial thalamus of the rat. Brief (15 sec), low intensity (35 microA) stimulation of the anterior pretectal nucleus caused no escape behavior or motor deficits but increased tail-flick latency for more than 45 min. Responses to non-noxious stimuli were enhanced but the animals were not hyperactive. The anterior pretectal nucleus does not receive retinal or accessory visual inputs like other parts of the pretectal complex but is known to receive axons from somatosensory cortex and project to the perirubral mesencephalic reticular formation and the periaqueductal gray (PAG). The antinociceptive effects of anterior pretectal stimulation were much longer lasting than those of PAG, less disrupting to motor performance and the stimulation was not aversive.     

Single trial analysis of evoked potentials to noxious thermal stimulation in man

Thermal (laser) evoked responses were obtained from 13 male volunteers. A single trial analysis technique with a latency adjusting adaptive filter was used to analyze evoked response amplitudes. Significant and substantial within-subject linear correlations were found between the magnitude (A) of the primary waveform (RMS μV of the P200-N300-P400 complex) and subjective pain response (R) as well as stimulus intensity (S). Since subjective pain response was strongly correlated with stimulus intensity, the partial correlation coefficients were calculated for R vs. A with S controlled, and S vs. A with R controlled, for each subject. The partial correlations revealed a much stronger relationship between subjective response and the evoked response amplitude, suggesting that the primary complex may measure neural events in the pain perception process rather than transduction and transmission of the stimulus event.     

Peripheral noxious stimulation reduces withdrawal threshold to mechanical stimuli after spinal cord injury: Role of tumor necrosis factor alpha and apoptosis

We previously showed that peripheral noxious input after spinal cord injury (SCI) inhibits beneficial spinal plasticity and impairs recovery of locomotor and bladder functions. These observations suggest that noxious input may similarly affect the development and maintenance of chronic neuropathic pain, an important consequence of SCI. In adult rats with a moderate contusion SCI, we investigated the effect of noxious tail stimulation, administered 1 day after SCI on mechanical withdrawal responses to von Frey stimuli from 1 to 28 days after treatment. In addition, because the proinflammatory cytokine tumor necrosis factor alpha (TNFα) is implicated in numerous injury-induced processes including pain hypersensitivity, we assessed the temporal and spatial expression of TNFα, TNF receptors, and several downstream signaling targets after stimulation. Our results showed that unlike sham surgery or SCI only, nociceptive stimulation after SCI induced mechanical sensitivity by 24 h. These behavioral changes were accompanied by increased expression of TNFα. Cellular assessments of downstream targets of TNFα revealed that nociceptive stimulation increased the expression of caspase 8 and the active subunit (12 kDa) of caspase 3, indicative of active apoptosis at a time point consistent with the onset of mechanical allodynia. In addition, immunohistochemical analysis revealed distinct morphological signs of apoptosis in neurons and microglia at 24 h after stimulation. Interestingly, expression of the inflammatory mediator NFκB was unaltered by nociceptive stimulation. These results suggest that noxious input caudal to the level of SCI can increase the onset and expression of behavioral responses indicative of pain, potentially involving TNFα signaling.     

GD3 synthase gene knockout mice exhibit thermal hyperalgesia and mechanical allodynia but decreased response to formalin-induced prolonged noxious stimulation

Gangliosides are a family of sialic acid-containing glycosphingolipids that are highly enriched in the mammalian nervous system. In particular, b- and c-series gangliosides, all of which contain alpha-2,8 sialic acids, have been considered to play important roles in adhesion, toxin-binding, neurite extension, cell growth and apoptosis. However, the neurobiological functions of these series of gangliosides remain largely unknown. To clarify the function of b- and c-series gangliosides in pain sensation in vivo, we generated mice in whom the gene for the alpha-2,8-sialyltransferase (GD3 synthase), which is responsible for the generation of all b-series gangliosides as well as c-series gangliosides, was disrupted. Compared to the wild-type mice, the mutant mice exhibited increased sensory responses to thermal and mechanical stimuli as measured by a hot plate test and von Frey test. In contrast, the mutant mice showed decreased responses during the late phase of the formalin test. Paw edema and Fos expression in the spinal cord after formalin injection were significantly decreased in the mutant mice compared to the wild-type mice. No significant differences in the conduction velocity of the sciatic nerve, and no apparent morphologic differences in the spinal cord and the sciatic nerve were detected between the wild-type and the mutant mice. These results suggested that b- and c-series gangliosides are critical in the development and/or maintenance of the sensory nervous system responsible for the transmission of acute pain sensation and pain modulation. Moreover, they play an important role in the development of hyperalgesia induced by inflammation.     

The use of thermal analysis in developing chemical tests of sterilization

A number of chemicals have been examined to find out whether they can be employed to test the temperature regimens of vapor and hot-air sterilization methods. Test kits for various temperature ranges and different sterilization modes have been selected. It is advisable that stains be used in tests (except the sulfur one) to control vapor (but not hot-air) sterilization.     

Sex differences in temporal summation of pain and aftersensations following repetitive noxious mechanical stimulation

The SF-36 is a well-validated health status instrument measuring eight different health concepts. One aim of this study was to compare health status as measured by SF-36 in subjects from the general population with no chronic pain (NCP), chronic regional pain (CRP), and chronic widespread pain (CWP). A second aim was to assess if SF-36 could reflect changes in pain status over time. A third aim was to study if health status at baseline, measured by SF-36, could predict pain status 3 years later. The study was designed as a 3-year follow up with a postal questionnaire, including the SF-36 health survey, to 2357 subjects from the general population aged 20-74 years. The results were controlled for age, sex, co-morbidity, and socio-economic status. At baseline, all eight health concepts of SF-36 discriminated between subgroups with NCP, CRP and CWP. Changes in SF-36 over the 3-year follow up time coincided with improvement or deterioration of pain status. Baseline SF-36 scores predicted pain outcome 3 years later. These results support that both physical and mental aspects of health status as measured by SF-36 are affected by the burden of musculoskeletal pain, are sensitive to changes in pain status, and also predict the further development of pain.     

Role of the paratrigeminal nucleus in nocifensive responses of rats to chemical, thermal and mechanical stimuli applied to the hind paw

Anatomical and immunohystochemical data suggest the paratrigeminal nucleus (Pa5) may play a role in nociceptive processing. The current study examines the influence of unilateral Pa5 lesion on nocifensive responses of conscious rats to noxious thermal (Hargreaves test), mechanical (electronic von Frey and Randall-Selitto tests), and chemical (formalin 2.5%; 50 microl) stimuli applied to the hind paw. Lesion of the Pa5 induced by ibotenic acid did not affect the latency for radiant heat-induced withdrawal of either paw. In contrast, the mean mechanical threshold for withdrawal of the contralateral (but not ipsilateral) paw in Pa5-lesioned rats was reduced by approximately 45% and 20%, in electronic von Frey and Randall-Selitto tests, respectively, when compared to sham-operated animals. Conversely, animals with Pa5 lesions injected with formalin in the contralateral paw spent less time engaged in focused (licking, biting or scratching the injected paw) and total nocifensive behavior (i.e., focused nocifensive behavior plus protection of the injected paw during movements) in both the first and second phases of the test [ approximately 50% inhibition of each parameter during first phase (0-5 min) and at 20, 25, and 30 min of second phase, relative to the sham-operated group], but the number of paw-jerks was unaffected. Pa5 lesion also delayed the onset of second phase focused pain induced by formalin in the ipsilateral paw. The results suggest that the Pa5 integrates the supraspinal pain control system and plays a differential modulatory role in the central processing of mechanical and chemical nociceptive information.     

The pattern of Fos expression in the spinal dorsal horn following acute noxious mechanical stimulation of bone

Little is known of the spinal mechanisms that mediate bone nociception. The aim of this study was to determine the pattern of neuronal activation in the spinal dorsal horn following acute noxious mechanical stimulation of bone. This was achieved by examining Fos expression in the spinal dorsal horn following acute, noxious mechanical stimulation of the rat tibia. Noxious mechanical stimuli were applied by bone drilling and raising tibial intra-osseous pressure. Control experiments consisted of surgery to expose the tibia. There was a significant increase in the number of Fos-like immunoreactive (Fos-LI) nuclei in the superficial, ipsilateral dorsal horn of animals in the bone drilling and pressure groups relative to animals of the control group at spinal cord segments L3 and L4 (P < 0.05). The number of Fos-LI nuclei in the deep dorsal horn was always lower than the number in the superficial dorsal horn (significant at L3 but not L4; P < 0.05). Whilst there appeared to be a small increase in the number of Fos-LI nuclei in the ipsilateral deep dorsal horn of bone drilling and pressure groups relative to the ipsilateral deep dorsal horn control group at both L3 and L4 segments, no significant effect was observed (P > 0.05). The present study implicates the superficial dorsal horn of the spinal cord as a region of interest in studies of acute bone pain, and highlights the notion that spinal mechanisms that mediate bone nociception may be different to those that mediate nociception of cutaneous and visceral origin.     

Nociceptor sensitization to mechanical and thermal stimuli in pig skin in vivo

Primary hyperalgesia to mechanical and thermal stimuli are major clinical symptoms of inflammatory pain and can be induced experimentally by ultraviolet‐B (UV‐B) irradiation in humans. We set‐up a pig model in order to have more options for pharmacological intervention on primary hyperalgesia. Pig skin was irradiated with a dose one‐ to threefold higher than the minimum erythema dose (MED) and investigated for mechanical and heat responsiveness 24 and 48h post UV‐B treatment. C‐fiber activation upon mechanical and thermal stimulation was assessed indirectly by extent of the axon reflex erythema (flare) measured by laser Doppler imaging. Mechanical stimulation with von Frey filaments (100mN) induced flare responses in UV‐B treated skin at 24 and 48h, but no effect was measured in normal untreated skin. Increased mechanical stimulation (600mN) elicited a small flare response in normal skin in an area of 1.8cm² on average that was extending about 2.5cm² in the UV‐B irradiated sites. Thermal stimuli provoked in normal pig skin flare areas of approximately 2cm² (45°C, 10s) and 4.5cm² (47°C, 10s) which increased to about 3.5cm² (45°C) and 5.5cm² (47°C) following UV‐B irradiation at 24 and 48h. No significant differences of mechanically or thermally induced hypersensitivity were seen between 24 and 48h after irradiation. We conclude that UV‐B induced mechanical and heat sensitization of primary afferent nociceptors can be assessed in pig skin, providing a new human‐like model of primary hyperalgesia. Sensitization of primarily mechano‐insensitive (silent) nociceptors, which are underlying the flare response in humans, most probably contributes to the observation presented here.     

Cerebral responses to noxious thermal stimulation in chronic low back pain patients and normal controls

Changes in regional cerebral blood flow (rCBF) have previously been demonstrated in a number of cortical and subcortical regions, including the cerebellum, midbrain, thalamus, lentiform nucleus, and the insula, prefrontal, anterior cingulate, and parietal cortices, in response to experimental noxious stimuli. Increased anterior cingulate responses in patients with chronic regional pain and depression to noxious stimulation distant from the site of clinical pain have been observed. We suggested that this may represent a generalized hyperattentional response to noxious stimuli and may apply to other types of chronic regional pain. Here these techniques are extended to a group of patients with nonspecific chronic low back pain. Thirty-two subjects, 16 chronic low back pain patients and 16 controls, were studied using positron emission tomography. Thermal stimuli, corresponding to the experience of hot, mild, and moderate pain, were delivered to the back of the subject's right hand using a thermal probe. Each subject had 12 measurements of rCBF, 4 for each stimulus. Correlation of rCBF with subjective pain experience revealed similar responses across groups in the cerebellum, midbrain (including the PAG), thalamus, insula, lentiform nucleus, and midcingulate (area 24') cortex. These regions represented the majority of activations for this study and those recorded by other imaging studies of pain. Although some small differences were observed between the groups these were not considered sufficient to suggest abnormal nociceptive processing in patients with nonspecific low back pain.     

Spinal cord monoamines modulate the antinociceptive effects of vaginal stimulation in rats

Perispinal administration (into the lumbar intrathecal space) of phentolamine (40 micrograms), an alpha-adrenergic receptor blocking agent, reduced the analgesic effect of vaginal stimulation by 39.7% (measured by tail flick latency) and 57.1% (measured by vocalization threshold) as compared to controls. Perispinal administration of methysergide (10 micrograms), a serotoninergic receptor blocking agent, reduced the analgesic effect of vaginal stimulation by 48.5% (measured by vocalization threshold), although it did not significantly affect the tail flick measure. In a separate experiment, vaginal stimulation activated the release of norepinephrine and serotonin into a superfusate of the spinal cord. During vaginal stimulation, levels of norepinephrine and serotonin increased about 2-fold above resting levels. These findings indicate that vaginal stimulation releases norepinephrine and serotonin into the spinal cord, thereby exerting an analgesic effect.     

The effects of thermal stimulation on clinical and experimental itch

In order to substantiate accidental observations on the influence of skin temperature on itch, and to elucidate a possible involvement of thermoreceptors in itch generation, the effects of thermostimulation on clinical and experimental itch were studied. Eighteen patients with atopic dermatitis rated the intensity of spontaneous itch on one of their forearms before, during, and after its immersion in a waterbath of either 10 degrees C or 45 degrees C. In 40 normal subjects itch was elicited by histamine topically applied to a 7 cm2 skin area of the volar forearm. Before and after histamine application thermal thresholds were recorded. Then the skin area was heated or cooled at a rate of 0.5 degrees C/sec and itch intensity was continuously rated. Cooling abolished itch in all patients and in most of the normal subjects. Heating produced less clear effects: in two-thirds of both patients and normal subjects itch disappeared or was reduced whereas in the others itch was aggravated. Usually after the end of thermostimulation the opposite changes in itch intensity occurred. In the normal subjects thermal thresholds were not significantly influenced by histamine. Over a certain temperature range itch and thermal sensations could coexist as separate modalities. The results indicate that changes in skin temperature have a marked influence on itch intensity. Whereas cooling seems to act directly on the sensory receptors mediating itch, warm stimuli could have a central inhibitory effect. A direct role of thermoreceptors in the generation of itch is improbable.     

Activation of anterior cingulate cortex produces inhibitory effects on noxious mechanical and electrical stimuli‐evoked responses in rat spinal WDR neurons

In present study, in vivo electrophysiological techniques were applied to examine the effects of anterior cingulate cortex (ACC) activation on mechanical and electrical stimuli‐evoked responses in rat spinal cord wide‐dynamic‐range (WDR) neurons. We found that bilateral ACC electrical stimulation (100 Hz, 20 V, 20 s) had different effects on neuronal responses to brush, pressure and pinch stimuli (10 s). The brush‐evoked neuronal responses at baseline, post 1 min and post 5 min were 60.8±15.0, 59.2±15.4 and 60.0±19.3 spikes/10 s, respectively (n =10, P >0.05 vs. baseline). The pressure‐evoked neuronal responses at baseline, post 1 min and post 5 min were 77.8±11.9, 38.0±7.8 and 45.8±7.6 spikes/10 s, respectively (n =10, P <0.05 vs. baseline). The pinch‐evoked neuronal responses at baseline, post 1 min and post 5 min were 137.6±16.7, 62.6±17.5 and 68.8±15.0 spikes/10 s, respectively (n =10, P <0.05 vs. baseline). Furthermore, ACC stimulation generated distinct effects on the different components of wind‐up response. The total numbers of late response (LR) and after‐discharge (AD), but not early response (ER), significantly decreased. Collectively, the present study demonstrated that short‐term ACC activation could generate long‐term inhibitory effects on the responses of WDR neurons to noxious mechanical (pressure and pinch) and electrical stimuli. The results indicated that ACC activation could negatively regulate noxious information ascending from spinal cord with long‐term effect, providing potential neuronal substrate for the modulation of ACC activation on nociception.     

Functional magnetic resonance imaging in rats subjected to intense electrical and noxious chemical stimulation of the forepaw

We examined whether cerebral activation to two different intense and painful stimuli could be detected using functional magnetic resonance imaging (fMRI) in alpha-chloralose anesthetized rats. Experiments were performed using a 9.4 T magnet and a surface coil centered over the forebrain. A set of gradient echo images were acquired and analyzed using our software based on fuzzy cluster analysis (EvIdent). Following the injection of 50 microl of formalin (5%) into the forepaw we observed a regional increase in signal intensity in the MR images in all animals. Anterior cingulate cortex, frontal cortex and sensory-motor cortex were some of the regions that activated frequently and often bilaterally. Surprisingly, activation appeared sequentially, often occurring first in either the right or the left hemisphere with a separation of seconds to minutes between peak activations. Morphine pre-treatment (1 mg/kg, i. v.) delayed and/or reduced the intensity of the activation resulting in a decrease in the overall response. Following episodes of intense electrical stimulation, produced by two brief stimulations (15 V, 0. 3 ms, 3 Hz) of the forepaw, activation was observed consistently in the sensory-motor cortex contralateral to the stimulation. Activation also occurred frequently in the anterior cingulate cortex, ipsilateral sensory-motor cortex and frontal cortical regions. All these regions of activation were markedly reduced during nitrous oxide inhalation. Treatment with morphine resulted in an inhibition of the activation response to electrical stimulation in most regions except for sensory-motor cortex. Thus, electrical and chemical noxious stimuli activated regions that are known to be involved in the central processing of pain and morphine modified the activation observed. fMRI combined with appropriate exploratory data analysis tools could provide an effective new tool with which to study novel analgesics and their effects on the CNS processing of pain in animal models.     

fMRI of pain processing in the brain: a within-animal comparative study of BOLD vs. CBV and noxious electrical vs. noxious mechanical stimulation in rat

This study aims to identify fMRI signatures of nociceptive processing in whole brain of anesthetized rats during noxious electrical stimulation (NES) and noxious mechanical stimulation (NMS) of paw. Activation patterns for NES were mapped with blood oxygen level dependent (BOLD) and cerebral blood volume (CBV) fMRI, respectively, to investigate the spatially-dependent hemodynamic responses during nociception processing. A systematic evaluation of fMRI responses to varying frequencies of electrical stimulus was carried out to optimize the NES protocol. Both BOLD and CBV fMRI showed widespread activations, but with different spatial characteristics. While BOLD and CBV showed well-localized activations in ipsilateral dorsal column nucleus, contralateral primary somatosensory cortex (S1), and bilateral caudate putamen (CPu), CBV fMRI showed additional bilateral activations in the regions of pons, midbrain and thalamus compared to BOLD fMRI. CBV fMRI that offers higher sensitivity compared to BOLD was then used to compare the nociception processing during NES and NMS in the same animal. The activations in most regions were similar. In the medulla, however, NES induced a robust activation in the ipsilateral dorsal column nucleus while NMS showed no activation. This study demonstrates that (1) the hemodynamic response to nociception is spatial-dependent; (2) the widespread activations during nociception in CBV fMRI are similar to what have been observed in ¹⁴C-2-deoxyglucose (2DG) autoradiography and PET; (3) the bilateral activations in the brain originate from the divergence of neural responses at supraspinal level; and (4) the similarity of activation patterns suggests that nociceptive processing in rats is similar during NES and NMS.     

An fMRI study of the anterior cingulate cortex and surrounding medial wall activations evoked by noxious cutaneous heat and cold stimuli

The anterior cingulate cortex (ACC) and adjacent regions in the medial wall have been implicated in sensory, motor and cognitive processes, including pain. Our previous functional magnetic resonance imaging (fMRI) studies have demonstrated pain-related activation of the posterior portion of the ACC during transcutaneous electrical nerve stimulation (TENS) and variable patterns of cortical activation with innocuous and noxious thermal stimuli in individual subjects. The present study represents the companion paper to our recent study of pain- and thermal-related cortical activations with the aim to use fMRI to delineate the activations in the ACC and surrounding regions of the medial wall during application of innocuous and noxious thermal stimuli as well as during performance of a motor task in individual subjects. Ten normal subjects were imaged on a conventional 1.5 T GE 'echospeed' system. Functional images were obtained from sagittal sections through each hemisphere centered at approximately 3-5 and 7-9 mm from midline. Each subject was imaged during innocuous (cool, warm) and noxious thermal (cold, hot) stimulation of the thenar eminence, and execution of a motor (sequential finger-thumb opposition) task. Task-related activations were mostly confined to contralateral and medial ipsilateral images. Although the present results demonstrate intersubject variability in the task-related activations, some general modality-specific patterns were apparent: (i) innocuous thermal-related activations were located mainly in the anterior ACC; (ii) noxious thermal-related activations were primarily located in the anterior ACC, the ventral portion of the posterior ACC, and the supplementary motor area (SMA); (iii) motor-related activations were primarily located in the SMA and dorsal portion of the posterior ACC. These results indicate that specific spatial patterns of activation exist within the ACC and surrounding regions of the medial wall for innocuous and noxious thermal stimuli, and that noxious thermal- and motor-related activations appear to be segregated within the ACC. Therefore, we propose a segregation of the ACC into an anterior non-specific attention/arousal system and a posterior pain system.     

Assessment of brain responses to innocuous and noxious electrical forepaw stimulation in mice using BOLD fMRI

Functional magnetic resonance imaging (fMRI) using the blood oxygen level-dependent (BOLD) contrast was used to study sensory processing in the brain of isoflurane-anesthetized mice. The use of a cryogenic surface coil in a small animal 9.4T system provided the sensitivity required for detection and quantitative analysis of hemodynamic changes caused by neural activity in the mouse brain in response to electrical forepaw stimulation at different amplitudes. A gradient echo-echo planar imaging (GE-EPI) sequence was used to acquire five coronal brain slices of 0.5 mm thickness. BOLD signal changes were observed in primary and secondary somatosensory cortices, the thalamus and the insular cortex, important regions involved in sensory and nociceptive processing. Activation was observed consistently bilateral despite unilateral stimulation of the forepaw. The temporal BOLD profile was segregated into two signal components with different temporal characteristics. The maximum BOLD amplitude of both signal components correlated strongly with the stimulation amplitude. Analysis of the dynamic behavior of the somatosensory ‘fast’ BOLD component revealed a decreasing signal decay rate constant k_off with increasing maximum BOLD amplitude (and stimulation amplitude). This study demonstrates the feasibility of a robust BOLD fMRI protocol to study nociceptive processing in isoflurane-anesthetized mice. The reliability of the method allows for detailed analysis of the temporal BOLD profile and for investigation of somatosensory and noxious signal processing in the brain, which is attractive for characterizing genetically engineered mouse models.