Catherine Buisseret-Delmas

Serotonergic projections from the nodose ganglia to the nucleus tractus solitarius: an immunohistochemical and double labeling study in the rat

Possible projections of serotonin (5-HT)-immunoreactive neurons in the nodose ganglia (NG) to the nucleus tractus solitarius (NTS) were investigated in the rat using a double labeling method combining retrograde transport and 5-HT immunohistochemistry. After injection of a complex of colloidal gold-apo-horseradish peroxidase into the medio-caudal and commissural parts of the NTS, most of the 5-HT-immunoreactive neurons were found to be labelled by the gold complex. The present study provides direct evidence for the existence, in the rat, of a serotonergic NG-NTS system. This system may be involved in the regulation of blood pressure and vigilance states.

Trigeminal Projections to Hypoglossal and Facial Motor Nuclei in the Rat

This study was undertaken to identify the trigeminal nuclear regions connected to the hypoglossal (XII) and facial (VII) motor nuclei in rats. Anterogradely transported tracers (biotinylated dextran amine, biocytin) were injected into the various subdivisions of the sensory trigeminal complex, and labeled fibers and terminals were searched for in the XII and VII. In a second series of experiments, injections of retrogradely transported tracers (biotinylated dextran amine, gold‐horseradish peroxidase complex, fluoro‐red, fluoro‐green) were made into the XII and the VII, and labeled cells were searched for in the principal sensory trigeminal nucleus, and in the pars oralis, interpolaris, and caudalis of the spinal trigeminal nucleus. Trigeminohypoglossal projections were distributed throughout the ventral and dorsal region of the XII. Neurons projecting to the XII were found in all subdivisions of the sensory trigeminal complex with the greatest concentration in the dorsal part of each spinal subnucleus and exclusively in the dorsal part of the principal nucleus. Trigeminofacial projections reached all subdivisions of the VII, with a gradual decreasing density from lateral to medial cell groups. They mainly originated from the ventral part of the principal nucleus. In the spinal nucleus, most of the neurons projecting to the VII were in the dorsal part of the nucleus, but some were also found in its central and ventral parts. By using retrograde double labeling after injections of different tracers in the XII and VII on the same side, we examined whether neurons in the trigeminal complex project to both motor nuclei. These experiments demonstrate that in the spinal trigeminal nucleus, neurons located in the pars caudalis and pars interpolaris project by axon collaterals to XII and VII. J. Comp. Neurol. 415:91–104, 1999.

Trigemino‐reticulo‐facial and trigemino‐reticulo‐hypoglossal pathways in the rat

This study was undertaken to identify premotor neurons in the pontomedullary reticular formation serving as relay neurons between the sensory trigeminal complex and the motor nuclei of the VIIth and XIIth nerves. Trigeminoreticular projections were first investigated after injections of anterogradely transported tracers (biotinylated dextran amine, biocytin) into single subdivisions of the sensory trigeminal complex. The results show that the trigeminoreticular projections were abundant from the pars interpolaris (5i) and caudalis (5c) and moderate from pars oralis (5o) of the spinal trigeminal nucleus. Injections into the 5i and 5c produce dense anterograde labeling (1) in the dorsal medullary reticular field; (2) in the parvocellular reticular field, medially adjacent to the 5i; and (3) more rostral in the region dorsal and lateral to the superior olivary nucleus. Some labeled terminals were also found in the intermediate reticular field, whereas only light anterograde labeling was observed in the gigantocellular and oral pontine reticular formation. The 5o sends fibers and terminals throughout the whole reticular formation, with no clear preferential projections within a particular field. Only light projections originated from the principal nucleus (5P). In a second series of experiments, we examined whether premotor neurons in the reticular formation are afferented by trigeminal fibers. Double labeling was performed by injection of an anterograde tracer in the 5i and 5c and retrograde tracer (gold–horseradish peroxidase complex) into the VII or the XII motor nucleus on the same side. Retrogradely labeled neurons in contact with anterogradely labeled boutons were found throughout the reticular formation with predominance in the parvocellular and intermediate reticular fields. These experiments demonstrate the existence of trigeminal disynaptic influences, via reticular neurons of the pontomedullary reticular formation, in the control of orofacial motor behaviors. J. Comp. Neurol. 429:80–93, 2001.

Primary trigeminal afferents to the vestibular nuclei in the rat: existence of a collateral projection to the vestibulo-cerebellum.

Projections from the mesencephalic trigeminal nucleus to the vestibular nuclei were analyzed using retrograde and anterograde tracing methods. The results show that neurons in the caudal part of the trigeminal mesencephalic nucleus project mainly to the medial, inferior and lateral vestibular nuclei and moderately to the peripheral part of the superior vestibular nucleus. Using the double-labeling technique we demonstrate that individual neurons of the mesencephalic nucleus send collaterals to the vestibular nuclei and the vestibulo-cerebellum. These results suggest that these anatomical connections are involved in mechanisms of eye-head coordination.

Trigeminovestibular and trigeminospinal pathways in rats: Retrograde tracing compared with glutamic acid decarboxylase and glutamate immunohistochemistry

This study identified neurons in the sensory trigeminal complex with connections to the medial (MVN), inferior (IVN), lateral (LVN), and superior (SVN) vestibular nuclei or the spinal cord. Trigeminovestibular and trigeminospinal neurons were localized by injection of retrograde tracers. Immunohistochemical processing revealed γ‐aminobutyric acid (GABA)‐ and glutamate‐containing neurons in these two populations. Trigeminovestibular neurons projecting to the MVN and the IVN were in the caudal principal nucleus (5P), pars oralis (5o), interpolaris (5i), and caudalis (5c) and scattered throughout the rostral 5P. Projections were bilateral to the IVN, with an ipsilateral dominance to the MVN, except from the rostral 5P, which was contralateral. Neurons projecting to the LVN were numerous in the ventral caudal 5P and the 5o and less abundant in the rostral 5P, 5i, and 5c. Our results suggested that only 5P and 5o project to the dorsal LVN. Neurons projecting to the SVN were in the dorsal 5P, 5o, and 5i but not in 5c. Trigeminospinal neurons were mainly in the ventral 5o and 5i and in the lateral 5c, rarely or never in 5P. Among trigeminovestibular neurons, most of the somas were immunoreactive for glutamate, but some reacted for GABA. Among trigeminospinal neurons, the number of somas immunoreactive for each of the two amino acids was similar. Trigeminal terminals were observed in contact with vestibulospinal neurons in the IVN and LVN, giving evidence of a trigeminovestibulospinal pathway. Therefore, inhibitory and excitatory facial inputs may contribute through trigeminospinal or trigeminovestibulospinal pathways to the control of head/neck movements. J. Comp. Neurol. 496:759–772, 2006.

Trigemino-solitarii-facial pathway in rats

This study was undertaken to identify premotor neurons in the nucleus tractus solitarii (NTS) serving as relay neurons between the sensory trigeminal complex (STC) and the facial motor nucleus in rats. Trigemino‐solitarii connections were first investigated following injections of anterograde and/or retrograde (biotinylated dextran amine, biocytin, or gold‐HRP) tracers in STC or NTS. Trigemino‐solitarii neurons were abundant in the ventral and dorsal parts of the STC and of moderate density in its intermediate part. They project throughout the entire rostrocaudal extent of the NTS with a strong lateral preponderance. Solitarii‐trigeminal neurons were observed mostly in the rostral and rostrolateral NTS. They mainly project to the ventral and dorsal parts of the spinal trigeminal nucleus but not to the principal nucleus. Additional neurons located in the middle NTS were found to project exclusively to the spinal trigeminal nucleus pars caudalis. No solitarii‐trigeminal cells were observed in the caudal NTS. In addition, evidence was obtained of NTS retrogradely labeled neurons contacted by anterogradely labeled trigeminal terminals. Second, solitarii‐facial projections were analyzed following injections of anterograde and retrograde tracers into the NTS and the facial nucleus, respectively. NTS neurons, except those of the rostrolateral part, reached the dorsal aspect of the facial nucleus. Finally, simultaneous injections of anterograde tracer in the STC and retrograde tracer in the facial nucleus gave retrogradely labeled neurons in the NTS receiving contacts from anterogradely labeled trigeminal boutons. Thus, the present data demonstrate for the first time the existence of a trigemino‐solitarii‐facial pathway. This could account for the involvement of the NTS in the control of orofacial motor behaviors. J. Comp. Neurol. 487:176–189, 2005.

Reticular premotor neurons projecting to both facial and hypoglossal nuclei receive trigeminal afferents in rats

The distribution of premotor neurons projecting to motor nuclei of both the VIIth (VII) and XIIth (XII) nerves was examined in the pontomedullary reticular formation (RF) of the rat by using retrograde double labeling. After injection of two different tracers in the VII and the XII, most of the double labeled neurons were found caudally in the dorsal RF whereas rostrally they were located in the ventral RF. In some experiments, additional injections of an anterograde tracer were made in the sensory trigeminal nuclei. Anterogradely labeled trigeminal boutons were found in contact with retrogradely double labeled neurons throughout the pontomedullary RF. These neurons were mainly encountered ventral to the trigeminal motor nucleus and dorsal to the VII. Functionally, this region is known to be involved in eye protection mechanisms.

Projections from the Superior Colliculus to the Trigeminal System and Facial Nucleus in the Rat

To determine the influence of the superior colliculus (SC) in orienting behaviors, we examined SC projections to the sensory trigeminal complex, the juxtatrigeminal region, and the facial motor nucleus in rats. Anterograde tracer experiments in the SC demonstrated predominantly contralateral colliculotrigeminal projections. Microinjections in the deep layers of the lateral portion showed labeled nerve fibers and terminals in the ventromedial parts of the caudal principal nucleus and of the rostral oral subnucleus and in the medial part of the interpolar subnucleus. Some terminals were also observed in the juxtatrigeminal region and in the dorsolateral part of the facial motor nucleus contralaterally, overlying the orbicularis oculi motoneuronal region. Verification by retrograde tracer injections into the trigeminal target regions showed labeled SC neurons mostly in lateral portions of layers 4–7. When the juxtatrigeminal region was involved, a remarkable increase of labeled neurons was observed, having a patch‐like arrangement with a decreasing gradient from lateral to medial SC portions. Retrograde tracer injections in the dorsolateral VII nucleus showed bilateral labeled neurons mainly in the deep lateral SC portion. Retrograde BDA microinjections into the same trigeminal or juxtatrigeminal regions, followed by gold‐HRP into the dorsolateral VII nucleus, demonstrated a significant number of SC neurons in deep layers 6–7 projecting to both structures by axon collaterals. These neurons are mediolaterally grouped in patches along the rostrocaudal SC extent; a subset of them are immunoreactive for glutamic acid decarboxylase (GAD). They could be involved in the coordination of facial movements. Simultaneous anterograde and retrograde tracer injections into the lateral SC portion and the VII nucleus respectively localized trigeminofacial neurons receiving collicular input in the trigeminal principal nucleus and pars oralis. Therefore the SC should play a crucial role in regulating motor programs of both eye and eyelid movements. J. Comp. Neurol. 478:233–247, 2004.

Organization of trigeminocollicular connections and their relations to the sensory innervation of the eyelids in the rat

Relationships between the trigeminal component of blinking and the superior colliculus (SC) were studied in rats. To localize primary afferent eyelid projections in the sensory trigeminal complex, neuronal tracing experiments were performed as well as analysis of c‐Fos protein expression after supraorbital (SO) nerve stimulation. Labelled nerve fibers were found to enter ventrally within the ipsilateral sensory trigeminal complex. Labelled boutons were observed at the junction of the principal nucleus (5P) and the pars oralis (5o) and in the pars caudalis (5c). The c‐Fos immunoreactivity was observed in neurons located in the ipsilateral ventral parts of 5P, 5o, and the pars interpolaris (5i) and bilaterally in 5c. Injections in 5P, 5o, 5i, and 5c resulted in anterogradely labelled fibers, with a contralateral preponderance, within the intermediate and deeper SC layers. Injections in 5P or 5o showed anterogradely labelled nerve fibers, profusely terminating in small patches in the medial and central portions of SC layer 4. Subsequently, dense labelling was found in the lateral portion of SC layers 4–7, without patch‐like organization. Injections in SC showed retrogradely labelled neurons predominantly within the contralateral part of the sensory trigeminal complex (28% in 5P, 20% in 5o, 50% in 5i, and 2% in 5c). Colocalization of the retrograde tracer after SC injections and c‐Fos immunoreactivity in neurons demonstrated that some 5P, 5o, and 5i neurons receive SO nerve inputs and project to SC. This implies that intermediate and deeper SC layers receive sensory information from the eyelids and may be directly involved in the regulation of eye–eyelid coordination. J. Comp. Neurol. 448:373–387, 2002.

Localization of Trigeminal, Spinal, and Reticular Neurons Involved in the Rat Blink Reflex

Electrical stimulation of the supraorbital nerve (SO) induces eyelid closure by activation of orbicularis oculi muscle motoneurons located in the facial motor nucleus (VII). Neurons involved in brainstem central pathways implicated in rat blink reflex were localized by analyzing c‐Fos protein expression after SO stimulation in conjunction with tracing experiments. A retrograde tracer (gold‐horseradish peroxidase [HRP]) was injected into the VII. The distribution patterns of activated c‐Fos‐immunoreactive neurons and of neurons exhibiting both c‐Fos immunoreactivity and gold‐HRP labeling were determined in the sensory trigeminal complex (STC), the cervical spinal cord (C1), and the pontomedullary reticular formation. Within the STC, c‐Fos immunoreactivity labeled neurons in the ipsilateral ventral part of the principal nucleus, the pars oralis and interpolaris, and bilaterally in the pars caudalis. Colocalization of gold‐HRP and c‐Fos immunoreactivity was observed in neurons of ventral pars caudalis layers I–IV and ventral pars interpolaris. In C1, SO stimulation revealed c‐Fos neurons in laminae I–V. After additional injections in VII, the double‐labeled c‐Fos/gold‐HRP neurons were concentrated in laminae IV and V. Although c‐Fos neurons were found throughout the pontomedullary reticular formation, most appeared rostrally around the motor trigeminal nucleus and in the ventral parvocellular reticular nucleus medial to the fiber bundles of the seventh nerve. Caudally, c‐Fos neurons were in the lateral portion of the dorsal medullary reticular field. In addition, these reticular areas contained double‐labeled neurons in electrically stimulated rats that had received gold‐HRP injections in the VII. The presence of double‐labeled neurons in the STC, C1, and the reticular formation implies that these neurons receive sensory information from eyelids and project to the VII. These double‐labeled neurons could then be involved in di‐ or trisynaptic pathways contributing to the blink reflex. J. Comp. Neurol. 467:173–184, 2003.