Increase in the discharge of muscle spindles during diaphragm fatigue.

The discharge of 18 single unit spindles located in the right crural diaphragm was recorded during rhythmic diaphragmatic contractions before the onset of and during fatigue. Spindle discharge was significantly greater (P<0.05) during rhythmic exercise when the diaphragm was fatigued than spindle discharge during rhythmic exercise when the diaphragm was not fatigued. The increase in spindle discharge during diaphragmatic fatigue is inconsistent with the notion that spindles comprise the afferent arm of a fatigue-induced inhibitory reflex originating in the diaphragm.     

Fatigue-induced changes in group IV muscle afferent activity: differences between high- and low-frequency electrically induced fatigues

Recordings of group IV afferent activity of tibialis anterior muscle were performed in paralysed rabbits during runs of electrically induced fatigue produced by direct muscle stimulation at a high (100 Hz, high-frequency fatigue HFF) or a low rate (10 Hz, low-frequency fatigue LFF). In addition to analysis of afferent nerve action potentials, muscle force and compound muscle action potentials (M waves) elicited by direct muscle stimulation with single shocks were recorded. Changes in M wave configuration were used as an index of the altered propagation of membrane potentials and the associated efflux of potassium from muscle fibers. The data show that increased group IV afferent activity occurred during LFF as well as HFF trials and developed parallel with force failure. Enhanced afferent activity was significantly higher during LFF (maximal Δf_impulses=249±35%) than HFF (147±45%). No correlation was obtained between the responses of group IV afferents to LFF or to pressure exerted on tibialis anterior muscle. On the other hand, decreased M wave amplitude was minimal with LFF while it was pronounced with HFF. Close correlations were found between fatigue-induced activation of group IV afferents and decreases in force or M wave amplitude, but their strength was significantly higher with LFF compared to HFF. Thus, electrically induced fatigue activates group IV muscle afferents with a prominent effect of low-frequency stimulation. The mechanism of muscle afferent stimulation does not seem to be due to the sole increase in extracellular potassium concentration, but also by the efflux of muscle metabolites, present during fatiguing contractions at low rate of stimulation.     

Somato-vesical reflexes in chronic spinal cats

The effects of afferent volleys in hindlimb cutaneous and muscle nerves on vesical tone and contractility and on the discharges in pelvic nerves to the bladder were measured in anesthetized CNS-intact and 2-19 months chronic spinal cats. In chronic spinal cats volleys in group III and IV fibers increased the tone of the quiet, empty bladder (excitatory somato-vesical reflex). The same volleys inhibited the slow, large, rhythmic micturition contractions of the expanded bladder (inhibitory somato-vesical reflex). In CNS intact cats single or short tetanic volleys induced a reflex discharge in pelvic vesical nerve branches with 3 distinct components. These reflexes could be observed during micturition contractions, not markedly between the contractions or when the bladder was empty and quiet. The latencies of the 3 components were 90, 320 and 770 ms, respectively. The two early components (AI- and A2-reflex) were evoked by volleys in group II and III hindlimb afferents. The late component (C-reflex) was induced by group IV volleys. In chronic spinal cats a group II and III-induced A-reflex (latency 90 ms) and a group IV-induced C-reflex (latency 340 ms) were observed. The central pathways and the physiological significance of the various somato-vesical reflexes are discussed.     

Spindle gain increase during muscle unit fatigue

In anaesthetized cats, medial gastrocnemius motor units (MUs) were stimulated with random sequences (mean rates between 6 and 12 pps) of electrical pulses delivered to their axons in small ventral root filaments. Muscle tension was recorded under isometric conditions, and spike trains of muscle spindle afferents were recorded from small dorsal root filaments during prolonged MU activation. Time-domain (PSTH) and frequency-domain (gain) computations were performed to study the effects of fatiguing muscle unit contractions on the signal transmission from skeletomotor efferents to spindle afferents. In the course of muscle unit fatigue, during which the gain of the force-producing sub-system decreased, the gain of the sub-system transforming force to afferent discharge increased so that the overall gain between skeletomotor efferents and spindle afferents remained relatively high. This could be a mechanism that preserves a high quality of afferent information on MU contractions.     

Morphology of central terminations of intra-axonally stained, low-threshold mechanoreceptive primary afferent fibers from oral mucosa and periodontium in the rat

Horseradish peroxidase (HRP) was injected intra-axonally into functionally identified primary afferent fibers within the rat spinal trigeminal tract in order to study the morphology of their central terminations. They were physiologically determined to be large, myelinated afferent fibers from periodontium or oral mucosa by means of electrical and mechanical stimulation of their receptive fields. Twenty-eight axons that innervated the periodontium of incisors and 21 axons that innervated the oral mucosa were stained for distances of 2-5 mm from the injection sites at the levels of the main sensory nucleus (Vms), spinal trigeminal nucleus and rostral cervical spinal cord. The collaterals of these primary afferent fibers formed terminal arbors in the medial or dorsomedial part of the Vms, and the oral and interpolar spinal trigeminal nuclei (Vo and Vi). In the caudal spinal trigeminal nucleus (Vc), the collaterals of one half of the periodontium afferent fibers terminated mainly in lamina V at the rostral and middle levels of Vc. On the other hand, the collaterals of the other half of the periodontium afferent fibers terminated mainly in lamina IV at the rostral level of Vc, and rostrally these terminal areas shifted to the most medial part of Vi. The collaterals of mucosa afferent fibers terminated in lamina V at the rostral level of Vc, and these terminal areas shifted gradually to laminae III and IV as the parent axons traveled more caudally. These shifts were staggered rostrocaudally according to the rostrocaudal locations of the receptive fields. The density of collaterals of periodontium afferent fibers in Vi was significantly larger than that of mucosa afferent fibers. The average size of the varicosities of periodontium afferent fibers was significantly larger than those of mucosa afferent fibers in Vo, Vi and Vc. The average number of varicosities belonging to single collaterals of slowly-adapting periodontium afferent fibers in Vi were significantly larger than those in Vo. In Vi, the average number of varicosities of single collaterals of slowly-adapting periodontium afferent fibers were significantly larger than those of rapidly-adapting periodontium afferent fibers.     

Electrophysiological study of diaphragmatic myoclonus.

This is the first reported detailed electrophysiological study of diaphragmatic myoclonus. An 86 year old woman had rapid, intermittent epigastric pulsations. Neurological examination and imaging studies of the brain and spinal cord were normal. Needle EMG showed rhythmic contractions of the diaphragm and external intercostal muscles at 4 to 5 Hz. These contractions were often associated with suppression of normal breathing and were capable of maintaining adequate ventilation. Both diaphragms were involved but showed considerable variability in their relative latencies. Automated interference pattern analysis suggested a change in recruitment order, with selective activation of large phrenic motoneurons. The supraspinal mechanisms mediating diaphragmatic myoclonus are different from that of voluntary and involuntary rhythmic breathing, and seem to be unrelated to palatal myoclonus. The generator source is likely related to respiratory centres in the rostral medulla.     

Group III and IV muscle afferent discharge patterns after repeated lengthening and shortening actions

The purpose of this study was to test the hypothesis that group III and IV muscle afferent activity would differ after concentric‐ and eccentric‐type fatiguing tasks. Tibialis anterior afferent activities from adult rats were measured in three conditions: before and after a rest period (C), and after concentric (CC) or eccentric (EC) exercise. Specific activators were used to elicit increases in afferent discharge rates, i.e., electrically induced fatigue (EIF), or potassium chloride (KCl) and lactic acid (LA) injections. After the rest period (POST‐condition), the control group displayed a pattern of response to stimuli similar to that obtained in baseline condition (PRE‐condition). However, responses were significantly different in the exercise groups: afferent responses were blunted in the CC group and were almost suppressed in the EC group. These results demonstrate that the type of muscular contraction involved in the fatiguing task can affect group III and IV afferent fiber activity differently and, potentially, can differentially affect the regulation of central motor command. Muscle Nerve, 2009     

Modifications of afferent activities from Tibialis anterior muscle in rat by tendon vibrations, increase of interstitial potassium or lactate concentration and electrically-induced fatigue

Although previous experiments with a partially similar objective have been described in dogs, cats and rabbits, the purpose of this study was to identify and characterize mechanosensitive and chemosensitive muscle afferents in the anaesthetized rat since it is a widely used laboratory animal. The peroneal nerve innervating the tibialis anterior muscle was studied. Measurement of conduction velocities from compound action nerve potentials evoked by peripheral nerve stimulation allowed identification of group I-II (10.79+/-1.02 m/s), group III (2.96+/-0.58 m/s) and group IV (0.46+/-0.07 m/s) afferent fibers. Computation of the different compound potential areas showed that afferents I and II arising from spindles and tendon organs represented 9.65+/-2.2%, whereas afferents III and IV arising from free nerve endings in muscle represented 90.35+/-2.2% (III, 46.66+/-2.71% and IV, 43.69+/-2.52%). Action potentials were recorded from teased nerve filaments. Mechanical tendon vibrations (10 to 90 Hz) were used to activate mechanoreceptors. Peak increase in afferent discharge (fimpulses) was measured at 50 Hz (n = 12/19 units) or 70 Hz (n = 7/19 units). Intra-arterial bolus injections of different concentrations of potassium chloride (KCl: 1 to 20 mM) or lactic acid (LA: 0.5 to 3 mM) elicited marked activation of III and IV afferents (n = 124). Enhancement of fimpulses was not proportional to the increase in [KCl] or [LA]. Activation of afferents plateaued when [KCl] was equal or greater than 5 mM while fimpulses peaked, then decreased, when [LA] was 1 mM. Muscle fatigue induced by direct electrical muscle stimulation (EIF) markedly activated group III-IV (n = 17/18) afferents (176.9+/-29.7% of control) which persisted for the 3 minutes of recovery from fatigue. Maximal fimpulses increases in response to LA (+67%) and KCl (+46.9%) injections and to EIF (+76.9%) were similar. This procedure for characterizing the functional properties of sensory nerve endings in a skeletal muscle may be used to assess further changes in sensory muscle paths in experimental rodent pathophysiological systems.     

Stimulation of the MLR inhibits the discharge of dorsal horn neurons responsive to muscular contraction

We found that electrical stimulation of the mesencephalic locomotor region (MLR) inhibited the discharge of deep dorsal horn neurons receiving group III afferent input from the triceps surae muscles. In contrast, contraction of these muscles induced by electrical stimulation of the tibial nerve activated these dorsal horn neurons. Our findings show that descending central motor commands can inhibit dorsal horn interneurons receiving input from group III afferents during exercise.     

Discharge rate and reflex responses of fusimotor neurons during muscle ischemia

Reflex effects of muscle ischemia on fusimotor neurons were investigated in decerebrate cats. Nerve impulses of single fusimotor neurons were recorded from filaments cut out of otherwise intact nerves to triceps surae muscles. Muscle ischemia was induced by clamping a. and v. femoralis. Fusimotor discharge rate was counted before, during, and after ischemia of both resting and contracting muscles. Muscle contractions were induced either by direct electrical stimulation or reflexly by muscle stretch. No changes were found in either the mean discharge rate or in the reflex response of fusimotor neurons to muscle stretches and contractions during muscle ischemia. It could be concluded that group III and IV muscle afferent fibers, responding to contractions of the ischemic muscle, have no reflex effects on fusimotor neurons. It also does not seem probable that fusimotor neurons contribute to ischemic muscular cramps.     

Effects of neonatal capsaicin deafferentation on neuromuscular adjustments, performance, and afferent activities from adult tibialis anterior muscle during exercise

To investigate the role played by muscle afferents in the sensorimotor loops, we measured the effects of capsaicin injection in newborns on the mechano- and metabosensitive discharges and the running performance at adulthood. Female Sprague Dawley rats received a subcutaneous injection of either 50 mg/kg capsaicin or solvent (10% ethanol, 10% Tween 80 in 0.9% saline) during their second day of life. These two groups were compared with a control, untreated group. Four months later, treadmill running performance and muscle afferent (mechanosensitive and metabosensitive) activities from the tibialis anterior muscle were measured. The capsaicin-treated group demonstrated a reduced maximal exercise capacity (time to exhaustion) and a reduced response of muscle metabosensitive fibers (group III and IV nerve endings) to the examined stimuli (arterial KCl and lactic acid injections, electrically induced fatigue) compared with the sham-injected solvent and control groups. Group IV afferent responses were absent in the compound nerve action potentials evoked by peripheral nerve stimulation. The response to mechanosensitive fibers to tendon vibration was also affected in the capsaicin group compared with the control and sham-injected groups, which presented a bimodal response corresponding to the activation of muscle spindles and Golgi tendon organs. Finally, measurements of the force developed by the tibialis anterior muscle from the beginning to the end of a 3-min muscle stimulation revealed a more significant fall in the capsaicin group compared with the others. The present experiments reveal that the pharmacological alteration of muscular metabosensitive afferent resulted in drastic changes in the neuromuscular sensory encoding and in the central neural network that could accelerate failure of the task during fatigue.     

Crossed and uncrossed projections to cat sacrocaudal spinal cord: I. Axons from cutaneous receptors

Intra-axonal recording and horseradish peroxidase staining techniques were used to map terminal fields of primary afferent fibers from cutaneous receptors within the cat sacrocaudal spinal cord. It was hypothesized that projection patterns of cutaneous afferent fibers mirror the known somatotopic organization of sacrocaudal dorsal horn cells. Forty-three primary afferent fibers, innervating either slowly adapting type I receptors, hair follicles, or slowly adapting type II receptors, all on the tail, were recovered. All collaterals (N = 372) branched from parent axons in the dorsal columns. Most collaterals coursed rostromedially to the ipsilateral gray matter, penetrated the medial dorsal horn, and arborized within laminae III, IV, and to a lesser extent, V. Ipsilateral projections to dorsal horn were as follows: axons with dorsal or dorsolateral receptive fields (RFs; n = 20) to the lateral portion, axons with lateral RFs (n = 4) to the central portion, and axons with ventral or ventro-lateral RFs (n = 19) to the medial portion. Most axons (16 of 20) with dorsal or dorsolateral RFs also had contralateral projections to lateral dorsal horn and most axons (15 of 19) with ventral or ventrolateral RFs also had contralateral projections to medial dorsal horn. No axons with lateral RFs had crossed projections. These data represent the first complete mapping of the somatotopic organization of primary afferent fiber projection patterns to a spinal cord level. The findings demonstrate that ipsilateral projection patterns of sacrocaudal primary afferent fibers are in register with the somatotopic organization of the dorsal horn. Our earlier suggestion that crossed projections of primary afferent fibers give rise to crossed components of dorsal horn RFs spanning the midline is supported by these results.     

Central projection of unmyelinated (C) primary afferent fibers from gastrocnemius muscle in the guinea pig

We have demonstrated the central projections of muscle C or group IV afferent fibers in the guinea pig by tracing arborizations in the spinal cord. C afferent fibers from the gastrocnemius muscle (GCM) were electrophysiologically identified by conduction velocity (less than 1 m/second). A single neuron in the lumbar 5 dorsal root ganglion (L5 DRG) was intracellularly labeled with Phaseolus vulgaris leucoagglutinin (PHA-L). After iontophoretic injection of PHA-L, we processed the lumbar cord and L5 DRG for PHA-L immunohistochemistry. Six muscle C afferent fibers from 40 animals were labeled, and whole trajectories were recovered. Labeled fibers were reconstructed by tracing of the arbor in serial parasagittal sections. The GCM C afferents projected rostrocaudally for two or three segments and ran at the surface of the dorsal funiculus, giving off collaterals into laminae I and II and sometimes into parts of lamina III. We determined, based on the branching pattern and form of the terminal plexus, that the branching of muscle C afferent fibers showed an intermediate pattern that fell morphologically between the terminal patterns of somatic and visceral afferents. The numbers and sizes of fiber swellings and terminal swellings were measured on all collateral branches. We found that the area of distribution of the terminal swellings of muscle C afferent fibers is larger than that of somatic terminals but that the density of terminal swellings in the terminal area was lower than that of the somatic terminals.     

Origin and distribution of phrenic primary afferent nerve fibers in the spinal cord of the adult rat

Previous studies from this laboratory have localized and morphologically characterized phrenic motor neurons in the rat spinal cord at light and electron microscopic levels. The present investigation used a modification of the TMB method for the retrograde transport of horseradish peroxidase (HRP) to describe at light microscope levels the origin and distribution of phrenic primary afferent axons in the adult rat spinal cord. Dry HRP crystals were applied to the central stump of the transected phrenic nerve in the neck to label spinal ganglion cell bodies and thus determine the levels of origin of afferent axons in the phrenic nerve. Camera lucida drawings were then made from serial sections through the appropriate spinal cord levels to determine the specific distribution of transganglionically labeled phrenic central axonal processes within the spinal cord. HRP-labeled phrenic primary neurons were observed in the C3 to C7 spinal ganglia. The camera lucida studies indicated that the transganglionically labeled central processes of phrenic primary afferent axons distributed into the dorsal horn at the C4 and C5 levels of the spinal cord. Furthermore, central processes distributing to C5 were more numerous than those that distributed to C4. Afferent axons were never seen in the dorsal horn at C3, C6, or C7. As spinal ganglion cells were labeled at C3 above and C6 and C7 below, it follows that central processes of phrenic afferent fibers descend and ascend in the dorsal columns of the spinal cord before distributing into the dorsal horn. Specifically, the labeled primary afferent axons and their collateral branches were found in the fasciculus cuneatus, and in laminae I, II, III, and IV of the dorsomedial aspect of the dorsal horn. The function of these central axonal processes is unknown, but based on a comparison of our morphologic data with previous physiological and anatomical studies, we suggest that phrenic afferent fibers may arise from proprioceptors (muscle spindles and Golgi tendon organs), nociceptors, or rapidly adapting mechanoreceptors (Pacinian corpuscles) within the diaphragm.     

Alterations in primary afferent input to substantia gelatinosa of adult rat spinal cord after neonatal capsaicin treatment

Primary afferent fibers are divided into three main subgroups: Abeta-, Adelta-, and C-fibers. Morphological studies have demonstrated that neonatal capsaicin treatment (NCT) depletes C-fiber inputs in the spinal dorsal horn; the electrophysiological features of the NCT-induced changes, however, remain unclear. This issue was addressed by performing whole-cell voltage-clamp recordings from substantia gelatinosa (SG) neurons in dorsal root-attached spinal cord slices. When estimated from excitatory postsynaptic currents (EPSCs) evoked by stimulating primary afferent fibers, 24 (49%) of 49 neurons examined exhibited C-fiber EPSCs that were either monosynaptic (n = 15) or polysynaptic (n = 9) in origin; only two of the neurons had Abeta-fiber responses. In NCT rats, however, SG neurons exhibiting C-fiber-mediated EPSCs decreased to 7% (3 of 41 neurons tested), whereas Abeta-fiber EPSCs were observed in 21 (51%) of the neurons, and 14 (67%) of them exhibited monosynaptic ones. There was no change in the cell proportion having Adelta-fiber innervation after NCT. Our electrophysiological data suggest that NCT diminishes primary afferent C-fiber inputs while enhancing Abeta-fiber direct innervation in the SG in adulthood.     

Ultrastructural three-dimensional reconstruction of group III and group IV sensory nerve endings ("free nerve endings") in the knee joint capsule of the cat: evidence for multiple receptive sites

The noncorpuscular endings ("free nerve endings") of thinly myelinated group III and nonmyelinated group IV afferent nerve fibers have been examined in the knee joint capsule of sympathectomized cats by transmission electron microscopy and three-dimensional reconstruction of series of semi- and ultrathin sections. The sensory ending is the most distal part of a group III or IV nerve fiber that consists only of the sensory axon and associated Schwann cells but lacks a myelin sheath and is not surrounded by perineurium. The sensory axon divides into several branches and forms a terminal tree. The branches run either as single fibers or within small Remak bundles in parallel to sensory axons of other endings; they spread along vessel walls and also extend into dense connective tissue. Each sensory axon consists of a series of spindle-shaped thick segments ("beads") connected by waist-like thin segments. Thus all axons of sensory endings have a string-of-beads appearance, which resembles that of efferent sympathetic nerve fibers. The beads of the sensory axon and the end bulb at its tip show the same ultrastructural features which are characteristic of receptive sites: an accumulation of mitochondria and glycogen particles and various vesicles in the axoplasm and "bare" areas of axolemma that are not covered by Schwann cell processes. Group III and group IV sensory endings differ in the length of their branches (up to 200 microM in group III vs. more than 300 microM in group IV), number of beads per 100 microM axon length (about seven vs. nine or ten), mean diameter of axons (0.9-1.5 microM vs. 0.3-0.6 microM), and the presence of a neurofilament core consisting of bundles of parallel microfilaments only in group III. In conclusion, we propose that the sensory part of noncorpuscular "free nerve endings" is formed by the entire terminal tree of group III or group IV nerve fibers and that the beads in the course of the sensory axon represent multiple receptive sites.     

Unitary discharges in dorsal and ventral roots after the administration of 4-aminopyridine in the cat

The administration of 4-aminopyridine (4-AP) in decerebrate paralyzed cats induces centrifugal rhythmic discharges in both ventral and dorsal roots. This study describes the mode of discharge of individual primary afferents as well as some ventral root fibers. Several patterns of antidromic discharge have been observed in primary afferents after the administration of 4-AP. A large proportion of the units (n = 96; 53%) showed rhythmic bursts of discharge related (n = 41) or not (n = 55) to the ongoing rhythmic activity in the peripheral nerves. Other units (n = 86; 47%) discharged either tonically, sporadically or had no antidromic activity at all. The conduction velocity of the non-bursting units was significantly higher (89.7 +/- 18.4 m/s) than that of the bursting units (70.6 +/- 15.4 m/s; P less than 0.01). Ventral roots showed rhythmic activity although less intense than that of the dorsal roots. As in dorsal roots, some fibers showed a rhythmical pattern of discharge related to the mass activity recorded from whole dorsal roots or peripheral nerves, while other units were not related. It is concluded that bursting activity which occurs in peripheral nerves after the administration of 4-AP is mainly the result of the antidromic activation of medium to small size primary afferent fibers.     

Pain from excitation of identified muscle nociceptors in humans

The technique of intraneural microstimulation (INMS) combined with microneurography was used to excite and to record impulse activity in identified afferent peroneal nerve fibers from skeletal muscle of human volunteers. Microelectrode position was minutely adjusted within the impaled nerve fascicle until a reproducible sensation of deep pain projected to the limb was obtained during INMS. During INMS trains of 5–10 s in duration and at threshold for sensation, volunteers perceived a well defined area of deep pain projected to muscle. Psychophysical judgements of the magnitude of pain increased with increasing rates of INMS between 5 and 25 Hz. Also, the area of the painful projected field (PF) evoked during trains of INMS of various duration but constant intensity and rate typically expanded with duration of INMS. The intraneural microelectrode was alternatively used to record neural activity originating from primary muscle afferents. Eight slowly adapting units with moderate to high mechanical threshold were identified by applying pressure within or adjacent to the painful PF. Conduction velocities ranged from 0.9 to 6.0 m/s, and fibers were classed as Group III or Group IV. Capsaicin (0.01%) injected into the RF of two slowly conducting muscle afferents (one Group III and one Group IV) produced spontaneous discharge of each fiber and caused intense cramping pain, suggesting that the units recorded were nociceptive. Our results endorse the concept that the primary sensory apparatus that encodes the sensation of cramping muscle pain in humans is served by mechanical nociceptors with slowly conducting nerve fibers. Results also reveal that muscle pain can be precisely localized, although the human cortical function of locognosia for muscle pain becomes blunted as a function of duration of the stimulus.     

Modulation of the spinal excitability by muscle metabosensitive afferent fibers

The aim of this study was to identify the effect of chemical activation of muscle metabosensitive afferent fibers from groups III and IV on Hoffmann (H‐) reflex modulation in the vastus medialis muscle. The experiment was conducted in rats and was divided into two experiments. The first experiment consisted of recording the metabosensitive afferent activity from femoral nerve in rats in response to KCl intraarterial injections in nontreated adults and adults treated neonatally with capsaicin. Thus, the dose‐response curve was determined. The second experiment consisted of eliciting the H‐ and M‐waves before and after KCl injection in nontreated adult animals and those treated neonatally with capsaicin. Thus, the H_max/M_max ratio was measured. Results indicated that, 1) in nontreated animals, afferent fibers peak discharge was found after 10 mM KCl injection; 2) no significant increase in afferent discharge rate was found in capsaicin‐treated animal after KCl injections, confirming that capsaicin is an excitotoxic agent that had destroyed the thin metabosensitive nerve fibers; 3) in nontreated animals, H_max/M_max ratio was significantly attenuated after a 10 mM KCl injection activating metabosensitive afferent fibers; and 4) in capsaicin‐treated animals, no significant change in H_max/M_max ratio was observed after the KCl injection. These results reinforce the hypothesis that the spinal reflex response was influenced by metabosensitive muscle fibers and provide direct evidence that activation of these fibers could partially explain the reported decrease in H‐reflex when metabolites are released in muscle. © 2010 Wiley‐Liss, Inc.     

Firing patterns of pre-Bötzinger and Bötzinger neurons during hypocapnia in the adult rat

Controversy exists about how a coordinated respiratory rhythm is generated in the brainstem. Some authors suggest that neurons in the pre-B?tzinger complex are key to initiation of all types of breathing. While, on the other hand, it has been reported that some pre-B?tzinger neurons fail to maintain a rhythmic discharge in phase with phrenic nerve discharge during mechanical hyperventilation. Extracellular recordings were made from respiratory units in the pre-B?tzinger and B?tzinger complexes of 13 anaesthetised, paralysed and vagotomised rats. Central respiratory activity was monitored from the C5 phrenic nerve. During mechanical hyperventilation, several changes were observed in the phrenic neurogram. Firstly, the frequency and amplitude of integrated phrenic nerve discharge were reduced and reversibly stopped. Secondly, the patterned discharges changed from an augmenting to a variety of non-augmenting patterns in 53 of 60 cases. In some cases (n=9) we observed that the pattern appeared to have two components, an early short duration discharge followed by a longer duration discharge. Respiratory units also started to show different firing patterns during mechanical hyperventilation. In general, they were divided into those units that fired tonically (n=28) and units that became silent (n=32), before phrenic nerve discharge ceased coincidently with complete apnoea. Of particular interest were those expiratory-inspiratory units in the pre-B?tzinger complex (n=8) that narrowed their firing period towards late expiration and early inspiration during mechanical hyperventilation. Given their firing features, it is possible that these expiratory-inspiratory units may participate in generation of the early inspiratory component of phrenic nerve discharge.