Number and distribution of stapedius motoneurons in cats

Cell bodies of stapedius motoneurons were identified by retrograde transport of horseradish peroxidase (HRP) following injections into the stapedius muscle. Large injections were made in an attempt to label all stapedius motoneurons. To control for labeling of non-stapedial neurons resulting from spread of HRP, we determined the locations of brainstem neurons labeled by HRP applied to the facial nerve, the chorda tympani nerve, the auricular branch of the vagus nerve, the tensor tympani muscle, and the cochlea. In three cats analyzed in detail, 1,133-1,178 neurons projecting to the stapedius muscle were identified. Arguments are given which suggest that in these three cats all stapedius motoneurons were labeled. The labeled stapedius neurons may all be motoneurons because they all stain positively for acetylcholinesterase and have medium-coarse Nissl bodies. Most stapedius motoneurons were located around the motor nucleus of the facial nerve. Staphedius motoneurons were also found near the descending limb of the facial-nerve root, in the peri-olivary neuropil, and in the reticular formation with the ascending fibers of the facial-nerve root.     

Intracellularly labeled stapedius-motoneuron cell bodies in the cat are spatially organized according to their physiologic responses

This study examines whether the locations of stapedius-motoneuron cell bodies are correlated with their responses to sound. Single-unit recordings and injections of horseradish peroxidase were made in axons of stapedius motoneurons in the fascicles which run from the facial nerve to the stapedius muscle in the cat. Single units were characterized physiologically by their responses to ipsilateral, contralateral, and binaural sounds. Labeled cell bodies (N = 28) were found in all of the brainstem regions previously identified as containing stapedius motoneurons. Motoneurons characterized as having similar response properties had cell bodies in relatively circumscribed locations. Most (eight of 12) motoneurons excited by sound in either ear had cell bodies in a narrow band around the facial nucleus. Most (seven of eight) motoneurons excited by ipsilateral but not contralateral sound had cell bodies in the cleft between the superior olivary complex and the facial nucleus. All four motoneurons excited by contralateral but not ipsilateral sound had cell bodies located ventromedial to the facial nucleus. The three motoneurons excited only by binaural sound had cell bodies located dorsal to the superior olivary complex. (Two of these were also in the cleft between the superior olivary complex and the facial nucleus.) The cell body of the one motoneuron showing activity in the absence of sound stimulation was located dorsolateral to the facial nucleus. These results show that the cell bodies of stapedius motoneurons with similar electrophysiologic properties tend to have similar locations in the brainstem. The results are consistent with the idea that the stapedius-motoneuron pool is divided into subgroups that are spatially segregated in terms of their patterns of input from the two ears.     

Brainstem facial-motor pathways from two distinct groups of stapedius motoneurons in the cat

To determine the brainstem origins and axonal routes of stapedius motoneurons, we labeled motoneurons by injecting cat stapedius muscles with horseradish peroxidase. Some injections were made in normal cats and some in cats in which the middle segment of the internal facial genu had been cut. By tracing labeled axons and by comparing the locations of labeled cell bodies in normal and lesioned cats, we divided stapedius motoneurons into two groups: "perifacial" and "accessory." Perifacial stapedius motoneurons have cell bodies located around the motor nucleus of the facial nerve and axons which follow the classical course of facial motor axons through the internal genu of the facial nerve. Accessory stapedius motoneurons have cell bodies near the descending facial motor root and axons which ascend to the rostral tip of the internal facial genu, abruptly reverse direction, and then join the descending facial motor root. The sharply hooked course of axons of accessory stapedius motoneurons is similar to the course of axons from other accessory nuclei of cranial nerves V-VII. Our present results, with those of McCue and Guinan (J. Neurophysiol. 60:1160-1180, '88), demonstrate that cats have two groups of stapedius motoneurons which can be separated anatomically by the locations of their cell bodies or by the courses of their axons, and which, on the average, have different response properties.     

The locations of stapedius and tensor tympani motoneurons in the cat

The numbers and locations of motoneurons to the stapedius and tensor tympani muscles were determined by retrograde transport of horseradish peroxidase. Stapedius motoneurons lay outside the traditionally recognized facial nucleus, in several distinct locations: (1) in the interface between the facial nucleus and the superior olive; (2) in a thin, scattered lamina of somewhat smaller cells spread dorsal to the facial nucleus; and (3) in a cluster located ventromedial to the rostral third of the facial nucleus. Some cells also lay dorsal to the superior olive or scattered in the reticular formation, just medial to the descending loop of the facial nerve. Tensor tympani motoneurons also lay outside the traditionally recognized trigeminal motor nucleus, in an area just ventral to it. Both motoneuron pools were large, producing innervation ratios that establish stapedius and tensor tympani among the most finely innervated muscles yet studied. The degree of intermingling of large and small cells in these pools may explain, in part, why it has been easier to identify slow muscle fibers physiologically in tensor tympani than in stapedius.     

The central location of the motor neurons to the stapedius muscle in the cat

In two cats and 10 newborn kittens, retrograde transport of horseradish peroxidase was used to locate the motor neurons of the stapedius muscle. Following injection of tracer into the muscle, labeled neurons were observed in the interface between the facial nerve nucleus and the caudal end of the lateral superior olivary nucleus. Some marked neurons were also located dorsal and ventral to the facial nucleus. The number of labelled cells varied from a low of 34 to a high of 199.     

Musculotopic organization of the facial motor nucleus in Macaca fascicularis: a morphometric and retrograde tracing study with cholera toxin B-HRP

Morphometric and retrograde tracing methods were used to determine the location and number of motoneurons innervating individual facial muscles in Macaca fascicularis. Intramuscular injections of the cholera toxin B subunit-horseradish peroxidase conjugate produced discrete labeling patterns in the ipsilateral facial motor nucleus with good definition of somata and their processes. The facial nucleus extended rostrocaudally in the pons for about 2 mm, varying in shape and cross-sectional area along this axis. Motoneurons were clustered in subnuclei, but their boundaries were not sharp and they were not segregated by fiber bundles. The length, number, and area of subnuclei varied with rostrocaudal location. Retrograde labeling patterns revealed that individual muscles were innervated by longitudinal columns of motoneurons with each muscle region represented at all rostrocaudal levels of its column. The columns began at different rostrocaudal levels and varied in length. Columns for closely related muscles, such as the orbicularis oris and mentalis of the lower lip, tended to overlap, whereas columns for disparate muscles, such as the perioral and orbital, did not overlap. The dendritic processes of most motoneurons branched extensively among several different columns or subnuclei. Some dendrites extended outside of the nucleus into the surrounding tegmentum. Mean soma diameter (10.4-42.2 microns) was distributed unimodally, reflecting the absence of gamma motoneurons and lack of muscle spindles in the facial muscles. Large and small motoneurons were found in all regions of the nucleus, but the largest ones were located caudally and innervated muscles of the upper and lower lip. The perioral muscles also had more neurons, longer columns, and a lower cell density than the other muscle groups examined. These features may reflect the functions of the perioral muscles in facial expression and vocalization.     

Modification of the metabolism of cerebral serotonin (5-HT) induced in rats by the administration of 6-hydroxydopamine

The pathways of the acoustic reflexes activating the m. stapedius (St) and m. tensor tympani (TT) were studied with physiological and experimental neuroanatomical methods. The reflex activity was recorded in non-anesthetized rabbits with chronic lesions in the auditory pathways.The St reflex pathway was found to contain a direct pathway with 3–4 neurons: (1) the primary auditory afferent, (2) cells in the ventral cochlear nucleus (VCN) with their axons in the trapezoid body (TB) having a partially direct contact with the ipsilateral St motoneurons but mainly through (3) interneurons in the region of the ipsilateral and contralateral medial superior olive (MSO) leading to (4) the St motoneurons mainly medially in the facial motor nucleus. A parallel slower, pentobarbital sensitive pathway was found.The acoustic TT reflex pathway was also found to contain 4 neurons: (1) the primary auditory afferent, (2) neurons in the VCN passing the TB to the region of the ipsilateral and contralateral MSO, where (3) interneurons passed to (4) the motoneurons in the ventrolateral part of the trigeminal motor nucleus.The relation of the middle ear reflexes to the olivochochlear efferent system and the extrapyramidal system was discussed.     

Motoneurons of twitch and nontwitch extraocular muscle fibers in the abducens, trochlear, and oculomotor nuclei of monkeys

Eye muscle fibers can be divided into two categories: nontwitch, multiply innervated muscle fibers (MIFs), and twitch, singly innervated muscle fibers (SIFs). We investigated the location of motoneurons supplying SIFs and MIFs in the six extraocular muscles of monkeys. Injections of retrograde tracers into eye muscles were placed either centrally, within the central SIF endplate zone; in an intermediate zone, outside the SIF endplate zone, targeting MIF endplates along the length of muscle fiber; or distally, into the myotendinous junction containing palisade endings. Central injections labeled large motoneurons within the abducens, trochlear or oculomotor nucleus, and smaller motoneurons lying mainly around the periphery of the motor nuclei. Intermediate injections labeled some large motoneurons within the motor nuclei but also labeled many peripheral motoneurons. Distal injections labeled small and medium-large peripheral neurons strongly and almost exclusively. The peripheral neurons labeled from the lateral rectus muscle surround the medial half of the abducens nucleus: from superior oblique, they form a cap over the dorsal trochlear nucleus; from inferior oblique and superior rectus, they are scattered bilaterally around the midline, between the oculomotor nucleus; from both medial and inferior rectus, they lie mainly in the C-group, on the dorsomedial border of oculomotor nucleus. In the medial rectus distal injections, a "C-group extension" extended up to the Edinger-Westphal nucleus and labeled dendrites within the supraoculomotor area. We conclude that large motoneurons within the motor nuclei innervate twitch fibers, whereas smaller motoneurons around the periphery innervate nontwitch, MIF fibers. The peripheral subgroups also contain medium-large neurons which may be associated with the palisade endings of global MIFs. The role of MIFs in eye movements is unclear, but the concept of a final common pathway must now be reconsidered.     

Generation of motoneurons in the rabbit brainstem

Autoradiography of ³H-thymidine incorporation was used to determine the time of origin of motoneurons in the rabbit brainstem. With the exception of the facial nucleus, neurons of the branchial motor column originated earlier (days 9 and 10) than somatic motor column neurons (day 11). Labeling was obtained as early as embryonic day 8 for many motor nuclei and the mesencephalic trigeminal nucleus. The significance of temporal patterns of neurogenesis is discussed.     

Morphology and physiology of abducens motoneurons and internuclear neurons intracellularly injected with horseradish peroxidase in alert squirrel monkeys

Axons of abducens motoneurons and internuclear neurons were penetrated with HRP-filled glass microelectrodes in alert squirrel monkeys. The firing rate of these axons and spontaneous eye movements were recorded and the axons were then injected with HRP for subsequent visualization of the recorded cells. Soma-dendritic and axon and axonal terminal morphology were studied for possible correlation with firing frequency. The physiology of squirrel monkey abducens neurons is qualitatively similar to their counterparts in the rhesus monkey and the cat, being primarily correlated with the position and velocity of the eyes. The locations of moto- and internuclear neurons are similar in the squirrel monkey and cat as are the axonal projections and terminals. However, squirrel monkey abducens cells are smaller than their feline counterparts and have dendrites that are confined to the cellular borders of the abducens nucleus. The size of the soma and proximal dendrites of moto- and internuclear neurons are poorly correlated with either their threshold for recruitment or their tonic eye position sensitivity. However, cells with smaller dendritic trees tended to have higher saccadic eye velocity sensitivity than those with larger trees. Three types of internuclear neurons were distinguishable upon the basis of their axon collaterals. All cells terminated within the medial rectus subdivision of the oculomotor nucleus. One class of cells did not give rise to collaterals before projecting to the oculomotor nucleus and the other classes gave rise to collaterals that terminated in the intermediate and/or caudal interstitial nuclei of the median longitudinal fasciculus. Within the IIIrd nucleus internuclear terminations were usually confined to a single subgroup of medial rectus motoneurons.     

Intrinsic connections of layer III of striate cortex in squirrel monkey and bush baby: Correlations with patterns of cytochrome oxidase

This study used biocytin and horseradish peroxidase (HRP) to examine the intrinsic connections of the cytochrome oxidase (CO) rich blob and CO poor nonblob zones within layer III of striate cortex in two primate species, nocturnal prosimian bush babies (Galago crassicaudatus) and diurnal simian squirrel monkeys (Saimiri sciureus). Our main objective was to determine whether separate classes of lateral geniculate nucleus (LGN) cells projected to separate superficial layer zones or layers in either species. There were three significant findings. First, we confirm that layer III consists of three sublayers, IIIA, IIIB, and IIIC in both species. Layer IIIA receives input from layers IIIB, IIIC, and V, with little or no input from LGN recipient layers IV and VI. Layer IIIB receives its input from nearly every cortical layer. Layer IIIC, receives input principally from layers IVα [which receives its input from magnocellular (M) LGN cells] and from layers V and VI. Taken together with other findings on the extrinsic connections of these layers, our data suggest that IIIA and IIIC provide output to separate hierarchies of visual areas and IIIB acts as a set of interneurons. Second, we find that, as in macaque monkeys, cells in both IVβ and IVα of bush babies and squirrel monkeys projct to layer IIIB, converging within the blobs. These results suggest that information from all LGN cell classes [parvocellular (P), M, and the Koniocellular (K) or their equivalents] may be integrated within the blobs. Thus, blobs in all of these primates may perform a function that transcends visual niche differences. Third, our data show a species specific difference in the connections of the IIIB nonblobs; nonblobs receive indirect input via IVα from the LGN M pathway in bush babies but receive indirect input via IVβ from the LGN parvocellular (P) pathway in squirrel monkeys. These findings indicate that the role of nonblob zones within striate cortex differs from that of blob zones and takes into account visual niche differences. © 1993 Wiley-Liss, Inc.     

The organization of the extraocular motor nuclei in the atlantic stingray,Dasyatis sabina

Retrograde transport of HRP was used to determine the location and organization of the motor nuclei innervating the extrinsic eye muscles of the stingray, an elasmobranch fish. Oculomotor neurons are located both medial to and immediately ventrolateral to the MLF in the rostral midbrain. A ventral oculomotor nucleus was found among the IIIrd nerve rootlets close to the base of the midbrain. The dendrites of cells in the dorsal nucleus appear to be preferentially oriented in the transverse plane penetrating the MLF. Motoneuron pools innervating individual muscles are incompletely segregated in the dorsal group. However, the ventral nucleus innervates only the inferior oblique muscle. Dorsally, motoneurons innervating a single muscle are found on both sides of the MLF. In the caudal midbrain, the majority of trochlear motoneurons are located immediately ventrolateral to the MLF. Abducens motoneurons are scattered in the medulla from a ventrolateral position resembling the location of the nucleus in teleost fish to a dorsomedial position close to the MLF as in most other vertebrates. In contrast to other vertebrates, the medial rectus muscle is innervated by the contralateral oculomotor nucleus. Motoneurons innervating the other muscles have the same laterality as found in other vertebrates.     

Are cat lingual muscles dually innervated by the hypoglossal nerve and facial nerve?

The origins of the somatic motor nerve innervating the cat lingual muscles were studied using the horseradish peroxidase (HRP) method. Following HRP injection into the lingual muscles, labeled neurons were found not only in the hypoglossal nucleus but also in the facial nucleus, ipsilateral to the injection side. HRP-labeled neurons in the facial nucleus were principally observed in the ventromedial and ventrolateral divisions of the nucleus. This study suggests that cat lingual muscles are innervated by both hypoglossal motoneurons and some of the motoneurons in the ventromedial and ventrolateral divisions of the facial nucleus.     

Immunohistochemical distribution of substance P, serotonin, and methionine enkephalin in sexually dimorphic nuclei of the rat lumbar spinal cord

The purpose of the present study was to identify chemically some potential inputs to lumbar motoneurons of the rat in the spinal nucleus of the bulbocavernosus, ventral motor pool, dorsolateral nucleus, and retrodorsolateral nucleus. Substance P-like immunoreactivity and serotonin-like immunoreactivity were found in all four motor nuclei, with dense immunoreactive profiles surrounding motoneurons and their processes. Enkephalin-like immunoreactivity was restricted to the sexually dimorphic nuclei, the spinal nucleus of the bulbocavernosus, and the dorsolateral nucleus. Within the spinal nucleus of the bulbocavernosus, enkephalin-like immunoreactive profiles were apposed to the processes of motoneurons but not their somata. In contrast, enkephalin-like immunoreactivity surrounded motoneuron somata in the medial part but not the lateral part of the dorsolateral nucleus, in the location of motoneurons projecting to the ischiocavernosus muscle. Moreover, the density of serotonin-like immunoreactivity was also greater in the medial part of the dorsolateral nucleus. On the basis of the chemo-architecture and the connections of the dorsolateral nucleus, we suggest the division of this motor column into a medial part composed of ischiocavernosus motoneurons surrounded by enkephalin- and serotonin-like immunoreactivity and a lateral part that contains neurons that project to the sphincter urethrae muscle. Total spinal transection severely depleted both serotonin-like and substance P-like material in the lumbar ventral horn. No changes in the distribution of enkephalin-like immunoreactivity were observed following this lesion. It is therefore suggested that in the ventral horn, substance P- and serotonin-like material are derived from supraspinal tracts, whereas enkephalin-like material is derived from intrinsic nerve cell bodies of the spinal cord.     

Trigeminal mesencephalic neurons innervating functionally identified muscle spindles and involved in the monosynaptic stretch reflex of the lateral pterygoid muscle of the guinea pig

Location of the neurons in the trigeminal mesencephalic nucleus innervating stretch receptors of the lateral pterygoid muscle and the mode of their synaptic connection on the lateral pterygoid motoneurons of the guinea pig were studied physiologically as well as morphologically, in comparison with the trigeminal mesencephalic neurons innervating muscle spindles in the superficial masseter muscle, with the following results: stimulation of the caudal half of the trigeminal mesencephalic nucleus evoked monosynaptic excitatory postsynaptic potentials in the ipsilateral lateral pterygoid motoneurons. Stimulation of the lateral pterygoid nerve directly evoked spike potentials in the neurons located in the caudal half of the ipsilateral trigeminal mesencephalic nucleus, which responded with increased firing to stretch, and with silent period to twitch, of the ipsilateral lateral pterygoid muscle. Averaging of intracellular potentials of the lateral pterygoid motoneurons with extracellular spike potentials of these trigeminal mesencephalic neurons revealed excitatory postsynaptic potentials after a monosynaptic latency, but no inhibitory postsynaptic potentials. Injection of horseradish peroxidase into the lateral pterygoid muscle labeled 15-20 cells in the caudal half of the ipsilateral trigeminal mesencephalic nucleus, while 174-228 cells retrogradely labeled by horseradish peroxidase were found throughout the whole rostrocaudal extent of the ipsilateral trigeminal mesencephalic nucleus following injection of horseradish peroxidase into the masseter muscle. It was concluded that neurons in the caudal half of the trigeminal mesencephalic nucleus send their peripheral processes to stretch receptors, presumably muscle spindles, in the ipsilateral lateral pterygoid muscle and that their central processes have excitatory synapses on ipsilateral lateral pterygoid motoneurons, thus comprising the afferent limb of a monosynaptic stretch reflex arc of the lateral pterygoid muscle of the guinea pig.     

Localization of the calcium/calmodulin-dependent protein phosphatase,calcineurin, in the hindbrain and spinal cord of the rat

The calcium/calmodulin-dependent protein phosphatase calcineurin was localized at the light microscopic level in the rat hindbrain and spinal cord by using an antibody against the α-isoform of the catalytic subunit. Calcineurin was highly concentrated in axons, dendrites, and cell bodies of a subpopulation of α-motoneurons in hindbrain motor nuclei and the lateral motor column along the length of the spinal cord. These calcineurin-positive α-motoneurons appeared to be randomly distributed and represented approximately 25% of the total α-motoneuron pool in the motor trigeminal nucleus and the spinal cord lateral motor column. Within the facial nucleus, calcineurin-containing motoneurons were present in the medial and dorsal subdivision but not in the lateral and intermediate subdivision. In addition to the enrichment in motoneurons, calcineurin was enriched in cells of the superficial laminae of the spinal cord dorsal horn and its extension into the medulla, the caudal spinal trigeminal nucleus. Axonal staining in the white matter of the spinal cord was generally weak, except in the dorsolateral funiculus, where strongly calcineurin-positive axons formed a putative ascending tract that appeared to terminate uncrossed in the caudal lateral reticular nucleus of the medulla. This tract may originate from calcineurin-positive cells in the dorsolateral funiculus. We also compared the distribution of calcineurin with calcium/calmodulin-dependent kinase II in the spinal cord and found that the kinase is more widely expressed. Thus, calcineurin is highly restricted to a few locations in the hindbrain and spinal cord. Selective staining in facial subnuclei that innervate phasically active muscles suggests that calcineurin-positive motoneurons represent a subset of α-motoneurons innervating a metabolic subtype of muscle fibers, possibly fast-twitch fibers. © 1996 Wiley-Liss, Inc.     

Nucleus ambiguus motoneurons innervating the canine intrinsic laryngeal muscles by the fluorescent labeling technique

The localization of motoneurons innervating the canine intrinsic laryngeal muscles was investigated by the fluorescent labeling technique. Labeled cells were found in the ipsilateral nucleus ambiguus. The most rostral labeled neurons for the cricothyroid muscle, the posterior cricoarytenoid muscle, the thyroarytenoid muscle, and the lateral cricoarytenoid muscle were found at progressively more caudal levels, respectively, within the nucleus ambiguus. The rostral tip of the arytenoid muscle cell column was at the same level as the lateral cricoarytenoid muscle cell column. The cells labeled from the cricothyroid muscle occupied the ventral part of the nucleus at the rostral level of the nucleus. At the middle level of the nucleus, the cells from the posterior cricoarytenoid muscle occupied the ventral part of the nucleus and the cells from the thyroarytenoid muscle, the lateral cricoarytenoid muscle and the arytenoid muscle occupied the dorsal part of the nucleus. The existence of double-labeled cells which innervated both thyroarytenoid muscle and lateral cricoarytenoid muscle was detected.     

Cerebral cortical control of orbicularis oculi motoneurons

Cerebral cortical neural networks associated with eyelid movement play a critical role in facial animation, contribute to the regulation of blink frequency, and help prevent ocular injury. Eyelid closure depends, in part, on motoneurons that innervate the orbicularis oculi (OO) muscles. In this study, OO motoneuron cortical afferents were identified in rhesus monkeys with rabies virus, a retrograde transneuronal tracer. Virus was injected into the right OO muscle and immunohistochemically localized after 4-6 day transport intervals. Labeled motoneurons were limited to dorsal portions of the ipsilateral facial motor nucleus. After 4- and 4.5-day transport intervals, most labeled cortical neurons were localized to ventrolateral premotor (LPMCv), dorsolateral premotor (LPMCd), and motor (M1) cortices. Labeled neurons were more sparsely distributed in supplementary (M2), caudal (M4), and rostral (M3) cingulate motor cortices; the frontal eye fields (FEF); pre-supplementary motor cortex (pre-SMA); somatosensory cortices (areas 3a, 3b, and 1); and prefrontal cortex. At longer transport intervals (5-6 days), labeled neurons increased substantially in LPMCv, LPMCd, M2, M3, M4, pre-SMA, and FEF. Concentrations of labeled neurons also appeared in cortices along the lateral fissure and intraparietal sulcus. Overall, the densest collection of labeled neurons was localized to the caudal junction of LPMCd and LPMCv with M1. Rostral M3 was another focus of OO premotor neurons. Labeled neurons were distributed bilaterally in all motor cortical areas with a modest contralateral predominance for M2, LPMC, and M1. Thus, the cortical control of OO motor activity is distributed bilaterally among multiple motor areas.     

The development of the human motor trigeminal complex and accessory facial nucleus and their topographic relations with the facial and abducens nuclei

The development of the motor trigeminal complex and accessory facial nucleus was traced in 23 human fetuses (seven and one-half to 21 weeks, menstrual age; 20.7–p177.5 mm CR length) and a 75-day infant both by graphic reconstructions and cytologic methods. Positional relationships with the facial and abducens nuclei were determined. Differentiation begins at the junction of the posterior trigeminal and accessory facial nuclei (level of pontine flexure) and proceeds rostralward in the motor trigeminal complex and caudalward in the facial complex. The neurons that differentiate earliest complete their migration first and migrate the least from their original position. Thus the posterior trigeminal nucleus is most dorsally and the accessory facial cell groups are most medially located. Migration of the trigeminal complex is completed by eitht weeks and of the facial complex by ten to ten and one-half weeks. Correlations are made between the differentiation of the motor neurons of the trigeminal and facial nerves and: (1) the development of the muscles that they have been postulated to supply; (2) the known function of these muscles; (3) the development of reflex activity in these muscles; (4) the mammalian experimental and human pathologic evidence for their innervation. These correlations support the view that the posterior trigeminal nucleus supplies the anterior belly and the accessory facial nucleus the posterior belly of the digastric muscle and that the posterodorsal part of the motor trigeminal nucleus innervates the mylohyoid muscle. Furthermore, the sequence of differentiation of the trigeminal complex indicates that the ventral trigeminal muscles (floor of the mouth) are represented primarily dorsally and posteriorly in the trigeminal complex and the progressively more dorsal muscles increasingly farther ventrally and anteriorly. The three subdivisions of the motor trigeminal nucleus are probably related to the three major embryonic muscle masses that they supply early in development. Because the inconstant subgroups in the three subdivisions appear when the complixity of reflex activity increases, perhaps they represent functional units rather than single muscles. The position of the abducens nucleus varies widely and no consistent migration cephalad occurs. It may reach caudal motor trigeminal nuclear levels early in fetal life or remain at caudal levels of the facial nucleus postnatally. No accessory abducens nucleus was identified. The neurons in the location allocated to it in some mammals develop from the common cell column between the motor trigeminal nucleus and the facial nucleus and constitute the accessory facial nucleus. Possible explanations for the widely different number of cells in the posterior trigeminal and accessory facial nuclei are discussed.