The Golgi silver impregnation technique gives detailed information on neuronal morphology of the few neurons it labels, whereas the majority remain unstained. In contrast, the Nissl staining technique allows for consistent labeling of the whole neuronal population but gives very limited information on neuronal morphology. Most studies characterizing neuronal cell types in the context of their distribution within the tissue slice tend to use the Golgi silver impregnation technique for neuronal morphology followed by deimpregnation as a prerequisite for showing that neuron's histological location by subsequent Nissl staining. Here, we describe a rapid method combining Golgi silver impregnation with cresyl violet staining that provides a useful and simple approach to combining cellular morphology with cytoarchitecture without the need for deimpregnating the tissue. Our method allowed us to identify neurons of the facial nucleus and the supratrigeminal nucleus, as well as assessing cellular distribution within layers of the dorsal cochlear nucleus. With this method, we also have been able to directly compare morphological characteristics of neuronal somata at the dorsal cochlear nucleus when labeled with cresyl violet with those obtained with the Golgi method, and we found that cresyl violet–labeled cell bodies appear smaller at high cellular densities. Our observation suggests that cresyl violet staining is inadequate to quantify differences in soma sizes.

Retrograde transport of horseradish peroxidase, applied to cut peripheral nerves, was used to determine the rostrocaudal distribution of motoneurones supplying different branches of the ventral ramus for a single mid‐ or caudal thoracic segment in the cat. The motoneurones occupied a length of spinal cord equal to the segmental length but displaced rostrally from the segment as defined by the dorsal roots, with the number of motoneurones per unit length of cord higher in the rostral part of a segment (close to the entry of the most rostral dorsal root) than in the caudal part. The cross‐sectional area of the ventral horn showed a rostrocaudal variation that closely paralleled the motoneurone distribution. The ratio between the number of motoneurones per unit length in the caudal and rostral regions of a segment (0.70) was similar to the ratio previously reported for the strength of functional projections of expiratory bulbospinal neurones (0.63). This is consistent with the motoneurones being the main targets of the bulbospinal neurones. J. Comp. Neurol. 472:281–291, 2004. © 2004 Wiley‐Liss, Inc.

Peripheral input convergence on trigeminal premotor neurons in the vicinity of trigeminal motor nucleus has been investigated. Thirty neurons were identified by their antidromic responses to microstimulation of the masseteric subnucleus of trigeminal motor nucleus (NVmot-mass). Peripheral receptive fields were found in the buccal mucosae, periodontal ligaments, palate, tongue and vibrissae for 16 neurons located in the intertrigeminal area (NVint), supratrigeminal area (NVs), main sensory trigeminal nucleus (NVsnpr) and subnucleus gamma of the oral nucleus of the spinal trigeminal tract (NVspo-gamma). Eleven neurons in the NVint, NVs and NVspo-gamma responded to passive jaw opening: nine neurons were activated and two were inhibited. None of the neurons responded to both the orofacial mechanical stimulation and passive jaw opening. Forty-six percent of neurons (13 out of 28 tested) received inputs from the inferior alveolar nerve (IAN) and 53% of neurons (8 out of 15 tested) received inputs from the infraorbital nerve (ION). Out of 15 neurons tested for inputs from the IAN and ION, 7 neurons in the NVsnpr and NVspo-gamma received input from both. Sixteen percent of neurons (4 out of 25) received inputs from the masseteric nerve (MassN). None of the neurons with inputs from IAN and/or ION also received inputs from the MassN. We suggest that trigeminal premotor interneurons with projections to the NVmot-mass fall into two broad categories, those with inputs from the IAN and/or ION and those with inputs from the MassN, possibly muscle spindle afferents, and no neuron receiving inputs from both.

The location of motoneurones of digastricus pars occipitomandibularis (DPO) muscle (or retractor mandible) of the rabbit was investigated using the technique of retrograde transport of intramuscularly applied horseradish peroxidase (HRP). Our aim was to establish whether the "digastric" classification of DPO is also reflected in the central distribution of its motoneurones within cranial nerve nuclei known to innervate conventional digastric muscles. Our observations suggest that DPO shares motor innervation with jaw-closing muscles and not jaw-opening muscles. Consequently, the anatomical classification of rabbit DPO is in question.

Physiological recordings made from neurones of the caudal medulla in the vicinity of the obex have revealed evidence of sensitivity to passive stretches applied to jaw muscles. All stretch-sensitive neurones discharged during passive muscle lengthening but fell silent on shortening. Whilst some stretch-sensitive neurones showed responses to tactile palpation of jaw muscles, none responded to pressure applied to the teeth or electrical stimulation of the chorda-lingual nerve. Response characteristics of these stretch-sensitive neurones were reminiscent of those seen in jaw muscle spindle afferents. We suggest that these neurones were most probably axons of jaw muscle spindle afferents running in the tract of Probst.

Interneurones are no longer considered to merely relay signals from single modalities or change their sign. Instead, it has become clear, that in addition to subserving local reflex arcs, they also participate in many complex functions including central motor pattern generation. In this article. I review the literature on inputs that converge on trigeminal premotor interneurones and show that they originate from a wide range of modalities, including the masticatory central pattern generator (CPG). Using a "black box" systems analysis approach, I suggest that the masticatory CPG ma!: give out both rhythmic and tonic drive potentials during fictive jaw movements. I also confirm previous findings by others, namely, that when fictive movements are evoked by stimulation of different cortical sites, the firing characteristics of the interneurone under investigation can also change. These observations indicate that the masticatory CPG is capable of outputting different drive potentials to the same interneurone. It remains unclear, however, whether these interneurones merely relay these outputs of the masticatory CPG without modifying them or they themselves constitute the final part of the masticatory CPG.

Single jaw-muscle spindle afferent axons were characterized physiologically and intracellularly stained to determine whether particular physiological types of spindle afferent show distinctive morphologies. Microelectrodes filled with either horseradish peroxidase (HRP) or biotinamide (Neurobiotin) were advanced into the mesencephalic trigeminal nucleus (Vme) in anesthetized rats. Intracellular recordings then were characterized by their response: to palpation of the jaw muscles; when pressure was applied to the teeth and during passive ramp and hold and sinusoidal jaw movement. Seventy-one afferents were characterized physiologically and injected with HRP; an additional 61 afferents were typed and injected with biotinamide. The response of 43 stained neurons was recorded in the presence of suxamethonium. The major projection areas of these afferents were the: trigeminal motor nucleus (Vmo); region dorsal to Vmo; reticular formation, spinal trigeminal nucleus, superior cerebellar peduncle and Vme. One afferent type was modulated strongly during stretching of the jaw-elevator muscles. Based on their high sensitivity during stretching of the jaw muscles and/or their silencing during the release phase of muscle stretch, these afferents were classified as primary-like spindle afferents. These afferents projected most strongly to Vmo. A second type of afferent was modulated only modestly during stretching of the jaw-elevator muscles. These tonic afferents were classified as secondary-like spindle afferents because of their low dynamic sensitivity during ramp muscle stretch and their continued discharge during the release phase of muscle stretch. Secondary-like afferents projected most strongly to the region dorsal to Vmo. Boutons (n = 3,834) from 11 afferents were studied in detail. Secondary-like afferents had statistically larger boutons within Vmo. In both secondary- and primary-like spindle afferents, only a small number of boutons were associated closely with the somata and proximal dendrites of trigeminal motoneurons. In these cases, however, two to five boutons appeared to contact individual motoneurons, implying multiple monosynaptic inputs to a selective subset of jaw-elevator motoneurons. Some "giant" boutons were present dorsal to Vmo and in Vme. These results demonstrate that dynamically sensitive and nondynamically sensitive jaw-elevator muscle spindle afferents project preferentially to different regions. Primary-like spindle afferents are capable of providing feedback related to the dynamic phases of muscle stretch and project most heavily to Vmo. Secondary-like spindle afferents can transmit a feedback signal associated with muscle length and project most strongly to the supratrigeminal region. Both types of afferent have projections caudal to Vmo that may serve longer latency jaw-muscle stretch reflexes and/or the projection of proprioceptive information to the thalamus and cerebellum.

The effects of electrical stimulation within the midbrain on fusimotor output to the jaw elevator muscles were studied in anaesthetized cats. Muscle spindle afferents recorded in the mesencephalic trigeminal nucleus were categorised as primary or secondary by their responses to succinylcholine during sinusoidal or rampand-hold stretches. Changes in their stretch responses during midbrain stimulation were then assessed by changes in bias and in dynamic sensitivity. Problems were encountered in interpreting changes in sine wave stretch responses of primary afferents, in some of which a very small change in firing pattern produced large changes in estimates of the reponse amplitude. Sine wave testing also sometimes over-estimated static effects and under-estimated dynamic effects relative to ramp responses. On other occasions a small amount of static fusimotor activity caused a marked increase in sine response amplitude, which could be wrongly interpreted as a dynamic effect. Consequently, ramp responses only were used for diagnosing fusimotor changes. The most effective region for producing pure dynamic fusimotor excitation was directly rostral to the red nucleus, extending dorsally and ventrally approximately in the course of the retroflex bundle. Stimulation of regions caudal and dorso-caudal to the red nucleus, previously designated as the mesencephalic area for dynamic fusimotor control of leg muscles, gave static or mixed static and dynamic effects on jaw spindles. The use of midbrain stimulation to identify fusimotor neurones of jaw muscles as static or dynamic would be most reliable with stimulation just rostral to the red nucleus and would require spindle afferent behaviour to be monitored at the same time with ramp stretches.

In anaesthetized cats, it was possible to enhance the dynamic sensitivity to stretch of jaw muscle spindle primary afferents by electrical stimulation in the midbrain. This effect was taken to be due to excitation of dynamic fusimotor neurones and was used to identify fusimotor fibres as static or dynamic in recordings from the masseter nerve. During reflex movements, some fusimotor fibres fired tonically, while others were markedly modulated with increases in firing corresponding approximately to muscle shortening. The modulated group had a significantly higher mean resting firing frequency than the tonic group. Midbrain stimulation at points shown to produce dynamic effects on muscle spindle primary afferents increased the discharge frequency of the tonic fusimotor units, but did not significantly affect the modulated fusimotor units. We conclude that the fusimotor fibres in the masseter nerve which fire tonically during reflex jaw movements are probably of the dynamic type, while those which show strongly modulated increases in firing are probably static.