Axonal projection of the medullary expiratory neurons in the feline thoracic spinal cord.

Expiratory neurons in the caudal ventral respiratory group extend descending axons to the lumbar and sacral spinal cord, and they possess axon collaterals, the distribution of which has been well-documented. Likewise, these expiratory neurons extend axons to the thoracic spinal cord and innervate thoracic expiratory motoneurons. These axons also give rise to collaterals, and their distribution may influence the strength of synaptic connectivity between the axons and the thoracic expiratory motoneurons. We investigated the distribution of axon collaterals in the thoracic spinal cord using a microstimulation technique. This study was performed on cats; one cat was used to make an anatomical atlas and six were used in the experiment. Extracellular spikes of expiratory neurons were recorded in artificially ventilated cats. The thoracic spinal gray matter was microstimulated from dorsal to ventral sites at 100-µm intervals using a glass-insulated tungsten microelectrode with a current of 150-250 µA. The stimulation tracks were made at 1 mm intervals along the spinal cord in segments Th9 to Th13, and the effective stimulating sites of antidromic activation in axon collaterals were systematically mapped. The effective stimulating sites in the contralateral thoracic spinal cord with expiratory neurons in the caudal ventral respiratory group (cVRG) occupied 14.4% of the total length of the thoracic spinal cord examined. The mean percentage of effective stimulating tracks per unit was 18.6 ± 4.4%. The distribution of axon collaterals of expiratory neurons in the feline thoracic spinal cord indeed resembled that reported in the upper lumbar spinal cord. We propose that a single medullary expiratory neuron exerts excitatory effects across multiple segments of the thoracic spinal cord via its collaterals.

Morphological analysis of neck muscle nerves and neurons in cats.

Previous studies have failed to show morphological differences between neck muscle alpha and gamma motor fibers or alpha and gamma motoneurons. The present study aimed to investigate the morphological features of neck muscle motor nerves and motoneurons in cats. To determine the morphological features of peripheral motor fibers, the value of the outer contours of each fiber was converted into a perfect circle after ganglionectomy to remove sensory fibers, and the fiber diameters were calculated based on their circumferences. The sizes of neck motor fibers in the peripheral nerves had an evident bimodal distribution into small and large fiber groups, as depicted in histograms. The sizes of small and large motor fibers ranged from 2 to 12 µm and from 12 to 40 µm, respectively. The small fiber group is likely to correspond to gamma motor fibers and the large fiber group to alpha motor fibers. The morphological features of neck muscle motoneurons sectioned in the horizontal plane were examined using the horseradish peroxidase (HRP) retrograde labeling technique. The diameters of the biventer cervicis and complexus motoneurons had bimodal distributions. The inflection point between the small and large diameter population was 28 µm for the biventer cervicis and 26 µm for the complexus. We also observed that larger neurons displayed more dendrites. In conclusion, we could identify morphological differences likely to correlate with alpha and gamma motoneurons in both neck muscle peripheral nerves and neck motoneurons.

Properties of Renshaw-like cells excited by recurrent collaterals of pudendal motoneurons in the cat.

Although anatomical studies have indicated pudendal motoneurons to give off recurrent collaterals, they are not considered to make synapses onto interneurons, such as Renshaw cells, and rather terminate their own signals. No study till date has examined interneurons being driven by recurrent collaterals of pudendal motoneurons. Here, we aimed to investigate the existence of Renshaw cells driven by pudendal motoneurons along with the recurrent inhibition of the latter. Extracellular recordings were obtained from the ventral horn of the sacral spinal cord of anesthetized cats. Dorsal roots were sectioned, and motor axons were electrically stimulated. Renshaw-like cells driven by recurrent collaterals, with high-frequency firings at short latency discharge, were observed around Onuf's nucleus. However, the recurrent inhibitory post-synaptic potentials were not recorded by intracellular recordings from the pudendal motoneurons. In summary, we found Renshaw-like cells driven by pudendal motoneurons, but we could not identify the synaptic connection of these neurons.

Firing properties of medullary expiratory neurons during fictive straining in cats.

Expiratory (E) neurons in the caudal nucleus retroambigualis extend descending spinal axons to the lumbar and sacral spinal cord. Discharge rates of single E neurons were recorded to examine differences in activity of E neurons projecting to the lumbar or sacral spinal cord during fictive straining induced by distention of the colon with a balloon. Firing frequencies of E neurons with descending axons in the thoracic and lumbar spinal cord increased during the repetitive rise of rectum pressure, whereas those of E neurons with descending axons in the sacral spinal cord decreased. E neurons with descending axons in the thoracic/lumbar and sacral spinal cord exhibit different firing characteristics during the repetitive rise of rectum pressure when straining during defecation. The activity of abdominal nerves during fictive straining is in phase with changes in rectum pressure, but out of phase with the activity of E neurons.

Electrophysiological properties of Ia excitation and recurrent inhibition in cat abdominal motoneurons.

Ia excitation and recurrent inhibition are basic neuronal circuits in motor control in hind limb. Renshaw cells receive synaptic inputs from axon collaterals of motoneurons and inhibit motoneurons and Ia inhibitory interneurons. It is important to know properties of Ia excitation and recurrent inhibition of trunk muscle such as abdominal muscles. The abdominal muscles have many roles and change those roles for different kind of functions. Intracellular recordings were obtained from the abdominal motoneurons of the upper lumbar segments in cats anesthetized. First, dorsal roots were left intact, and sensory and motor axons were electrically stimulated. Ia excitatory post-synaptic potentials were elicited in five of eight motoneurons at same segment stimulated. Second, dorsal roots were sectioned, and motor axons were electrically stimulated. Recurrent inhibitory post-synaptic potentials were elicited in one of 11 abdominal motoneurons. Renshaw cells extracellularly fired high-frequency bursts at short latency and at same segment stimulated.

Effect of streptozotocin-induced diabetes on motoneurons and muscle spindles in rats.

This study examined the alterations in the number and size of motoneurons innervating the medial gastrocnemius (MG) and biceps femoris (BF) motor nuclei in diabetic rats (12 or 22 weeks after injection of streptozotocin) and age-matched controls using retrograde labeling technique. Additionally, morphological alterations of muscle spindles in BF and MG muscles were tested. Significantly fewer labeled MG motoneurons were found in 12- and 22-week diabetic rats as compared with age-matched control animals. In contrast, the number of BF motoneurons was preserved in each group. Compared to control animals, the ratio of larger motoneurons of MG and BF muscle were decreased at 12 weeks, and smaller MG motoneurons were drastically decreased at 22 weeks. Moreover, MG muscle spindle showed reduction of its number and increase of intrafusal muscle fibers; however, BF muscle spindles showed little or no difference from control animals. We conclude that there is an early loss of alpha motoneurons for both MG and BF muscles followed by a later loss of gamma motoneurons in MG muscle in diabetic animals. Moreover, loss of gamma motoneuron might induce atrophy of MG muscle spindles.

Discharge patterns of abdominal and pudendal nerves during induced defecation in anesthetized cats.

Defecation is thought to be achieved not only by contraction of the colon, but also by a rise in intra-abdominal pressure. In this study we recorded the discharges of nerves innervating the abdominal (Abd) muscles, diaphragm, external anal sphincter (EAS) muscle and pelvic floor (PF) muscles during induced defecation evoked by distention of an expellable balloon to reveal defecation-related muscle activities. The discharges of the Abd muscle and phrenic (Phr) nerves increased when rectal pressure increased. The discharges of the EAS and PF nerves usually increased in proportion to the pressure in the rectum and maintained a constant activity level, although some trials showed inhibition. The results suggest that activities of these muscles increase the intra-abdominal pressure.

The size of motoneurons of the gastrocnemius muscle in rats with diabetes.

Alterations in the number and size of motoneurons were studied in the medial gastrocnemius (MG) motor nucleus of diabetic rats (12 or 22 weeks after injection of storeptozotocin) and age-matched controls. Each group contained 6 animals. MG motoneurons were retrogradely labeled by dextran-fluorescein and the number and size of cell bodies were examined. Significantly fewer labeled MG motoneurons were found in the 22-week diabetic rats as compared with age-matched control animals. The mean soma diameter of MG motoneurons was significantly smaller in the 12- and 22-week diabetic animals. Furthermore the soma size for 22-week diabetic animals was smaller than for 12-week diabetic animals. The distribution of average soma diameters in the MG nucleus of control animals was bimodal; cells with larger average diameter were presumed to be alpha-motoneurons and those with smaller diameters were presumed to be gamma. Compared to control animals, the number of smaller MG motoneurons was reduced in 12 week diabetic animals. By 22 weeks, diabetic animals had no small MG motoneurons and the size distribution became unimodal. We conclude that there is a significant decrease in the absolute number and size of MG motoneurons in diabetic rats, with the possibility that the decrease occurred predominantly among the smaller gamma-motoneurons.

The diaphragmatic activities during trunk movements.

In order to investigate how the diaphragm and trunk muscles are recruited during various voluntary movements, we recorded EMG signals and video images simultaneously and analyzed EMGs of the diaphragm and trunk muscles during the voluntary movements that required trunk muscles. During trunk movements, the duration of the diaphragmatic activity became irregular and the intensity of the activity increased. Further analysis revealed that the diaphragmatic activities were consisted of two components, such as respiratory and non-respiratory activities during voluntary movements. Our results led to the idea that the diaphragmatic activities may be controlled from different control mechanisms of central nervous system.

Activity patterns of the diaphragm during voluntary movements in awake cats.

The diaphragm is an important inspiratory muscle, and is also known to participate in the postural function. However, the activity of the diaphragm during voluntary movements has not been fully investigated in awake animals. In order to investigate the diaphragmatic activity during voluntary movements such as extending or rotating their body, we analyzed the electromyogram (EMG) of the diaphragm and trunk muscles in the cat using a technique for simultaneous recordings of EMG signals and video images. Periodic respiratory discharges occurred in the left and right costal diaphragm when the cat kept still. However, once the cat moved, their periodicity and/or synchrony were sometimes buried by non-respiratory activity. Such non-periodic diaphragmatic activities during voluntary movements are considered as the combination of respiratory activity and non-respiratory activity. Most of the diaphragmatic activities started shortly after the initiation of standing-up movements and occurred after the onset of trunk muscle activities. Those activities were more active compared to the normal respiratory activity. During rotation movements, left and right diaphragmatic activities showed asymmetrical discharge patterns and higher discharges than those during the resting situation. This asymmetrical activity may be caused by taking different lengths of each side of the diaphragm and trunk muscles. During reaching movements, the diaphragmatic activity occurred prior to or with the onset of trunk muscle activities. It is likely that diaphragmatic activities during reaching movements and standing-up movements may have been controlled by some different control mechanisms of the central nervous system. This study will suggest that the diaphragmatic activity is regulated not only by the respiratory center but also by inputs from the center for voluntary movements and/or sensory reflex pathways under the awake condition.

Morphological study of external oblique motor nerves and nuclei in cats.

In order to clarify the morphological features of peripheral motor nerves and motoneurons that innervate trunk muscles, the size distribution of external oblique (EO) peripheral motor fibers and motoneurons of the thoracic and the lumbar segments were examined. Histograms of the size distribution of EO motor fibers in peripheral nerves after ganglionectomy clearly had a bimodal distribution of small fiber groups and large fiber groups. It is very likely that small fiber groups correspond to gamma motor fibers and large fiber groups to alpha motor fibers. Gamma and alpha motor fiber groups were separated at 8-14 microm. The average diameter of the gamma and alpha motor fibers were different in each segment. The ratio of gamma and alpha motor fibers was approximately 1:2.0 in the thoracic segments and from 1:1.8 to 1:0.9 in the lumbar segments. Horseradish peroxidase was applied to the central stump of EO nerves, and the size distribution of EO motoneuron cell bodies in the thoracic and the lumbar spinal cords was examined. The size distribution of motoneuron cell bodies was bimodal in one cat (small and large motoneurons) and unimodal in three cats. When the ratio of small motor fibers to large motor fibers in peripheral nerves was applied to that of small motoneurons to large motoneurons, the separation of small and large motoneurons was approximately 40 microm. These results suggest that the morphological characteristics in peripheral nerves of trunk muscles are not reflected in motoneurons.

Morphological analysis of the external anal sphincter motor nerve and its motoneurons in the cat.

To investigate the spinal neural circuitry that controls the tonus of the external anal sphincter (EAS) in the cat, the size distribution of EAS motor fibers and their motoneurons (MN) was examined, and the presence of muscle spindles in EAS musculature was also tested for. The size distribution of EAS motor fibers was examined after degeneration of afferent fibers and that of their MN was measured, after labeling the cells with horseradish peroxidase. Both distributions were unimodal, thereby demonstrating the difficulty of distinguishing between alpha and potential gamma MN; but muscle spindles were not found in the musculature. Mechanisms underlying the spinally controlled tonus of the EAS remain unclear, including the nature and role of spinal reflexes. It is argued that non-spindle sensory receptors in the anal canal may provide the sensory component of a reflex circuit that contributes to this tonus.

Analysis of activity of motor units in the biceps brachii muscle after intercostal-musculocutaneous nerve transfer.

We examined respiratory activity of motor units (MUs) in the internal intercostal nerves (IICNs)-transferred biceps brachii muscle (IC-biceps) in cats. MUs of IC-biceps showed respiratory discharges in inspiratory and expiratory phases, and these were enhanced by CO2 inhalation. Narrowing the airway also enhanced inspiratory and expiratory MUs activity. A mechanical load to the thorax immediately enhanced inspiratory MUs activity and weakened expiratory MUs activity. We analyzed the cross-correlation of MUs activity in interchondral muscle and IC-biceps to characterize the respiratory spinal descending inputs to motoneurons. We confirmed the short-term synchronization from interchondral muscles indicating divergence of a single respiratory presynaptic axon to thoracic motoneurons, but could not find synchronization from IC-biceps. The motor axonal conduction velocity (axonal CV) of IC-biceps MUs was lower than that of interchondral muscles. There was no correlation between the respiratory recruitment order of IC-biceps MUs and their axonal CV. These results indicate that IC-biceps shows the respiratory activities and afferent inputs from intercostal muscle spindles in the neighboring segments remain influential on activity of IC-biceps. In addition, the short-term synchronization from IC-biceps could not be found, suggesting that the intercostal nerve transfer alters the respiratory spinal descending inputs to thoracic motoneurons.