Octanoic acid-induced coma and reticular formation energy metabolism

The medium chain fatty acid octanoic acid was injected i.p. into 20-22 g Swiss-Albino mice at a dose of 15 mumol/g. This dose produced a reproducible response consisting of a 3-4 min period of drowsiness, followed by coma. These mice as well as suitable controls were sacrificed by rapid submersion in liquid N2, or by microwave irradiation in a 7.3 kW microwave oven. Tissue from the reticular formation and the inferior colliculus was prepared for microanalysis of the energy metabolites glucose, glycogen, ATP and phosphocreatine. Results from this study showed a selective effect on energy metabolism in cells of the reticular formation. Both glucose and glycogen were elevated in the coma and precoma state. In addition, ATP and phosphocreatine were decreased in the reticular formation during coma. These results show a selective effect of octanoic acid on energy metabolism in the reticular formation both in the precoma stage, and during overt coma. The selective vulnerability of the reticular formation to metabolic insult may act in a beneficial manner to the animal by inducing coma. This lowers the overall demand for energy, thereby placing the animal in a milieu in which there is an increased chance for correction of the perturbation.     

Neuronal activity in the inferior colliculus and bordering structures during vocalization in the squirrel monkey

In four squirrel monkeys (Saimiri sciureus), the inferior colliculus, together with the neighboring superior colliculus, reticular formation, cuneiform nucleus and parabrachial area, were explored with microelectrodes, looking for neurons that might be involved in the discrimination between self-produced and external sounds. Vocalization was elicited by kainic acid injections into the periaqueductal gray of the midbrain. Acoustic tests were carried out with ascending and descending narrow-band noise sweeps spanning virtually the whole hearing range of the squirrel monkey. Altogether 577 neurons were analyzed. Neurons that both were audiosensitive and fired in advance of self-produced vocalization were found almost exclusively in the pericentral nuclei of the inferior colliculus and the adjacent reticular formation. Only the latter, however, contained, in addition, neurons that fired during external acoustic stimulation, but remained quiet during self-produced vocalization. These findings suggest that the reticular formation bordering the inferior colliculus is involved in the discrimination between self-produced and foreign vocalization on the basis of a vocalmotor feedforward mechanism.     

HRP study of the organization of auditory afferents ascending to central nucleus of inferior colliculus in cat

The ascending auditory projections to central nucleus of inferior colliculus its ventrolateral and dorsomedial subdivisions (IC_VI, and IC_DM) have been studied in cat using both pressure and electrophoretic injections of horseradish peroxidase (HRP). The results indicate that the predominant ascending projections to inferior colliculus orginate in (1) contralateral cochlear nucleus, (2) contralateral and ipsilateral lateral superior olive, (3) ipsilateral medial superior olive, (4) ipsilateral ventral nucleus of the lateral lemniscus, (5) ipsilateral and contralateral dorsal nucleus of the lateral lemniscus, and (6) contralateral inferior colliculus. In addition, ipsilateral cochlear nucleus, ipsilateral and contralateral intermediate nucleus of the lateral lemniscus, ipsilateral, and to a lesser extent contralateral, periolivary nuclei project to inferior colliculus. Of these nuclei, the lateral superior olive projects exclusively to IC_VL and ipsilateral cochlear nucleus and contralateral inferior colliculus project mostly, if not exclusively, to IC_DM. Many of these projections demonstrate a cochleotopic organization and frequently a nucleotopic organization as well. A cochleotopic organization of the projections is apparent for cochlear nucleus and superior olivary complex. A nucleotopic organization suggests that the heaviest terminations of contralateral inferior colliculus are medial and dorsal in inferior colliculus, of medial superior olive are dorsal and lateral, of superior olivary complex are rostral, of cochlear nucleus are caudal, and of ventral nucleus of the lateral leminiscus are caudal.     

Descending nerve tracts in the spinal cord of the rat. I. Fibers from the midbrain

Unilateral stereotaxic lesions were made in the superior colliculus, the inferior colliculus, the red nucleus or the reticular formation of the midbrain of rats. The descending fibers from these sites were studied using the method of Nauta ('57). One crossed and two uncrossed tracts descend from the superior colliculus. These tracts terminate mainly in the reticular formation of the medulla oblongata. Only a small number of tecto-spinal fibers were found. These were confined to the upper contralateral cervical segments of the cord. The tecto-spinal fibers terminate on cells in the ventral gray matter of the cord, corresponding in position to those in Rexed's lamina VIII in the cat. One uncrossed descending tract passes from the inferior colliculus and terminates in the reticular formation of the medulla oblongata. No fibers were contributed to the tecto-spinal tract by the inferior colliculus. Rubro-spinal fibers decussate completely in the ventral tegmental decussation and descend in the lateral funiculus of the cord into lumbar segments and on interneurons which correspond in position to those in Rexed's laminae V and VI in the cat. From the reticular formation of the midbrain, ipsilateral fibers course in the ventral funiculus of the cord, descending to the ninth or tenth thoracic segments. These fibers terminate on cells corresponding in position to those in Rexed's lamina VIII in the cat. Considerable preterminal degeneration, mainly ipsilateral, was found in the brain stem of the animals with lesions in the midbrain reticular formation. The findings are discussed in relation to what has been reported previously in the rat, and in other animals.     

Ascending tracts of the lateral columns of the rat spinal cord: a study using the silver impregnation and horseradish peroxidase techniques.

The location of projection areas and cells of origin of the ascending fiber tracts of the spinal cord lateral columns were examined in the rat. Projection areas were localized after unilateral microtransection of lateral column fibers at C2 or T10, using silver impregnation of preterminal and fiber degeneration. Cells of origin were localized by unilateral microtransection and subsequent application of horseradish peroxidase (HRP). Two groups of fibers projected to the dorsal medulla. One group projected to nucleus intercalatus, commissuralis, and the dorsal column nuclei. The second group projected via the inferior cerebellar peduncle to the vestibular complex, with additional fibers continuing dorsally to the cerebellum. The most extensive system of ascending fibers projected to the reticular formation. Most spinoreticular fibers coursed through the ventral hindbrain and projected to the lateral reticular nucleus, ventral reticular nucleus, nucleus gigantocellularis, and nucleus subceruleus. Spinotectal and spinocentral gray fibers coursed through the ventral portion of the medulla and then dorsally through the pons. Spinocentral gray fibers projected to the caudal portion of the central gray matter, ipsilaterally. Spinotectal fibers projected to the intercollicular nucleus and adjacent portions of the superior colliculus, bilaterally. Two projections to the thalamus were observed after anterolateral column transection. Preterminal degeneration was observed in the ventrobasal complex ipsilaterally, and bilaterally in the intralaminar nuclei. In conjunction with previous results the present HRP data suggest that the cells of origin of spinothalamic tract fibers were situated in laminae IV, V, and VI. The location of spinal cord cells of origin of additional ascending tracts is discussed.     

Electron microscopy of classically stained astrocytes

The ascending connections of the lateral lemniscus were studied in the cat and squirrel monkey (Saimiri sciureus). In both species, the central nucleus of the inferior colliculus receives a massive projection from the lateral lemniscus. Only a few lemniscal fibers were found to terminate in the external nucleus of the inferior colliculus. The commissure of the lateral lemniscus originates from the dorsal nucleus of the lateral lemniscus and projects to the contralateral dorsal nucleus and the contralateral central nucleus of the inferior colliculus. No lemniscal fibers were seen ascending in the inferior brachium or terminating in the principal division of the medial geniculate body. A bundle of fibers was observed, however, which passed medial to the inferior brachium and terminated in the magnocellular or internal division of the medial geniculate. The bundle degenerated after lesions confined to the lateral lemniscus and is probably identical with the central acoustic tract of earlier workers. Evidence is presented that the fibers of the bundle are of spinal origin. Since the lemniscal fibers which ascend to the thalamus appear to be non-auditory, it is suggested that the inferior colliculus is an obligatory relay station in the classical auditory system and that the inferior brachium is the only ascending pathway of the system to project upon the thalamus.     

Afferent and efferent connections of the laterodorsal tegmental nucleus in the rat.

The connections of the laterodorsal tegmental nucleus (LDTg) have been investigated using anterograde and retrograde lectin tracers with immunocytochemical detection. Inputs to LDTg were found from frontal cortex, diagonal band, preoptic areas, lateral hypothalamus, lateral mamillary nucleus, lateral habenula; the interpeduncular nucleus, ventral tegmental area, substantia nigra and retrorubral fields; the medial terminal nucleus, interstitial nucleus, supraoculomotor central grey, medial pretectum, nucleus of the posterior commissure, paramedian pontine reticular formation, paraabducens and paratrochlear region; the parabrachial nuclei and nucleus of the tractus solitarius. Terminal labelling from PHA-L injections of LDTg was found in infralimbic, cingulate and hippocampal cortex, lateral septum, septofimbrial and triangular nuclei, horizontal limb of diagonal band and preoptic areas; in the anterior, mediodorsal, reuniens, centrolateral, parafascicular, paraventricular and laterodorsal thalamic nuclei, rostral reticular thalamic nucleus, and zona incerta; the lateral habenula and the lateral hypothalamus. A number of brainstem structures apparently associated with visual functions were also innervated, mainly the superior colliculus, medial pretectum, medial terminal nucleus, paramedian pontine reticular formation, inferior olive, supraoculomotor, paraabducens and supragenual regions, prepositus hypoglossi and nucleus of the posterior commissure. Also innervated were substantia nigra compacta, ventral tegmental area, interfascicular nucleus, interpeduncular nucleus, dorsal and medial raphe, pedunculopontine tegmental region, parabrachial nuclei, and nucleus of the tractus solitarius. These findings suggest the LDTg to be a highly differentiated part of the ascending "reticular activating" system, concerned not only with specific cortical and thalamic regions, especially those associated with the limbic system, but also with the basal ganglia, and visual (particularly oculomotor) mechanisms. Additional links with the habenula-interpeduncular system are discussed in this context.     

Mesencephalic and diencephalic afferents to the superior colliculus and periaqueductal gray substance demonstrated by retrograde axonal transport of horseradish peroxidase in the cat

The mesencephalic and diencephalic afferent connections to the superior colliculus and the central gray substance in the cat were examined by means of the retrograde transport of horseradish peroxidase (HRP). After deep collicular injections numerous labeled cells were consistently found in the parabigeminal nucleus, the mesencephalic reticular formation, substantia nigra pars reticulata, the nucleus of posterior commissure, the pretectal area, zona incerta, and the ventral nucleus of the lateral geniculate body. A smaller number of cells was found in the inferior colluculus, the nucleus of the lateral lemniscus, the central gray substance, nucleus reticularis thalami, the anterior hypothalamic area, and, in some cases, in the contralateral superior colliculus, Forel's field, and the ventromedial hypothalamic nucleus. Only the parabigeminal nucleus and the pretectal area showed labeled cells following injections in the superficial layers of the superior colliculus. In the cats submitted to injections in the central gray substance, labeled cells were consistently found in the contralateral superior colliculus, the mesencephalic reticular formation, substantia nigra parts reticulata, zona incerta and various hypothalamic areas, especially the ventromedial nucleus. In some cases, HRP-positive cells were seen in the nucleus of posterior commissure, the pretectal area, Forel's field, and nucleus reticularis thalami. A large injection in the mediodorsal part of the caudal mesencephalic reticular formation, which included the superior colliculus and the central gray substance, resulted in numerous labeled cells in nucleus reticularis thalami. The findings are discussed with respect to the suggested functional division of the superior colliculus into deep and superficial layers. Furthermore, the possible implications of labeled cells in zona incerta and the reticular thalamic nucleus are briefly discussed.     

Projections from cortex to tectum in the tree shrew, Tupaia glis.

Sensory neocortex of the tree shrew was divided into three main areas: the visual field, the auditory field, and the somatic field which includes motor cortex. Cortical cells which project to the tectum were identified by injecting HRP into superficial or deep layers of the superior colliculus and into various parts of the inferior colliculus. The main result is that these descending projections are well organized according to their origin in the three main sensory fields of the cortex. (1) Auditory field: labeled cells are found only in the core or auditory koniocortex, after injections of HRP in the pericentral area of the inferior colliculus; labeled cells are found in auditory belt areas after injections in posterior parts of the intermediate and deep layers of the superior colliculus, adjacent to the inferior colliculus. (2) Somatic field: labeled cells are also found in the somatic field after injections in the intermediate and deep layers of the superior colliculus, so that auditory and somatic fields probably overlap to some extent. The results do not exclude the possibility that somatic koniocortex has an exclusive target in the intermediate or deep layers of the superior colliculus. (3) Visual field: labeled cells are found only in the core or striate cortex after injections in the superficial layers of the superior colliculus. Labeled cells are found in the visual belt after injections in the rostral parts of the intermediate layers of the superior colliculus. When these results are related to ascending sensory pathways a picture emerges of a series of circuits or loops which interconnect parallel sensory pathways. These loops eventually reach the deep layers of the superior colliculus which of course have indirect access to motor neurons.     

Brainstem Monitoring in the Neurocritical Care Unit: A Rationale for Real-Time,Automated Neurophysiological Monitoring

Patients with severe traumatic brain injury or large intracranial space-occupying lesions (spontaneous cerebral hemorrhage, infarction, or tumor) commonly present to the neurocritical care unit with an altered mental status. Many experience progressive stupor and coma from mass effects and transtentorial brain herniation compromising the ascending arousal (reticular activating) system. Yet, little progress has been made in the practicality of bedside, noninvasive, real-time, automated, neurophysiological brainstem, or cerebral hemispheric monitoring. In this critical review, we discuss the ascending arousal system, brain herniation, and shortcomings of our current management including the neurological exam, intracranial pressure monitoring, and neuroimaging. We present a rationale for the development of nurse-friendly—continuous, automated, and alarmed—evoked potential monitoring, based upon the clinical and experimental literature, advances in the prognostication of cerebral anoxia, and intraoperative neurophysiological monitoring.

     

Superior paraolivary nucleus in the pigmented guinea pig: separate classes of neurons project to the inferior colliculus and the cochlear nucleus.

The superior paraolivary nucleus is a large component of the superior olivary complex in rodents and a major source of input to the inferior colliculi and the cochlear nuclei. In the present study, retrograde transport of the fluorescent tracers Fluoro-Gold, Fluoro-Ruby (tetramethyl rhodamine conjugated to dextran), fluorescein-coated microspheres, and Fast Blue were used to reveal the morphology and collateral projection patterns of cells in the superior paraolivary nucleus. The ascending projections to the inferior colliculus from the superior paraolivary nucleus arise mainly from round, multipolar cells, including large cells that project exclusively to the inferior colliculi and not to the cochlear nuclei. Projections to the ipsilateral and contralateral inferior colliculi arise from cells with similar morphology and, in fact, many of the cells that project contralaterally project ipsilaterally as well. Projections to the ipsilateral and contralateral cochlear nuclei arise primarily from cells that do not have collicular projections. On average, the somas of these cells are significantly smaller and more elongated than those that project to the inferior colliculi. Overlap between these ascending and descending systems is restricted to a small percentage of cells that send collateral projections to both the ipsilateral cochlear nucleus and the ipsilateral inferior colliculus. These cells are small and moderately elongated. Thus the ascending and descending projections examined here arise largely from different cells that belong to different morphological classes.     

Connectional basis for frequency representation in the nuclei of the lateral lemniscus of the bat Eptesicus fuscus

To study the role of the lateral lemniscus as a link in the ascending auditory pathway, injections of neuronal tracers were placed in the anteroventral cochlear nucleus (AVCN) and in the inferior colliculus of the bat Eptesicus fuscus. To correlate the anatomical results with tonotopic organization, the characteristic frequency of cells at each injection site was determined electrophysiologically. Pathways from AVCN diverge to 3 major targets in the lateral lemniscus, the intermediate nucleus and 2 divisions of the ventral nucleus (VNLL). Projections from these 3 nuclei then converge at the inferior colliculus. One cell group is particularly notable for its cytoarchitectural appearance. It is referred to here as the columnar area of VNLL because its cells are organized as a tightly packed matrix of columns and rows. The connections of the columnar area are organized in sheets that are precisely related to the tonotopic organization of both AVCN and the inferior colliculus. Sheets of cells in the dorsal part of the columnar area receive projections from low-frequency parts of AVCN and project to low-frequency parts of the inferior colliculus. These sheets of connections occupy successively more ventral locations as the tonotopic focus of the injection site increases in frequency. The entire range of frequencies audible to the bat is systematically represented along the dorsal-ventral dimension of the columnar area. Because each column is only 20-30 cells in height, frequency representation must be compressed in this dimension. Within the columnar area there is an overrepresentation of frequencies between 25 and 50 kHz, which corresponds roughly to the range of the FM echo-location call in Eptesicus. The connections of the other nuclei of the lateral lemniscus are not as precisely related to the tonotopic organization of the system as are those of the columnar area.     

Sources of projections to subdivisions of the inferior colliculus in the rat

Brainstem and forebrain projections to major subdivisions of the rat inferior colliculus were studied by using retrograde and anterograde transport of horseradish peroxidase. Retrograde label from injection into the external cortex of the inferior colliculus appears bilaterally in cells of the inferior colliculus, as well as in other brainstem auditory groups including the ipsilateral dorsal nucleus of the lateral lemniscus and contralateral dorsal cochlear nucleus. The external cortex is the only collicular subdivision where an injection labels cells in the contralateral cuneate nucleus, gracile nucleus, and spinal trigeminal nucleus. Other projecting cells to the external cortex are found in the lateral nucleus of substantia nigra, the parabrachial region, the deep superior colliculus, the midbrain central gray, the periventricular nucleus, and area 39 of auditory cortex. Injection of the dorsal cortex of inferior colliculus heavily labels pyramidal cells of areas 41, 20, and 36 of the ipsilateral neocortex. Anterograde label from a large injection of auditory cortex is densely distributed in the dorsal cortex, lesser so in the external cortex, and only slightly in the central nucleus. Labelled cells appear in the central nucleus, dorsal cortex, and external cortex, primarily ipsilaterally, following dorsal cortex injection. Relatively few cells from other brainstem auditory groups show projections to the dorsal cortex. Injection of the central nucleus of the inferior colliculus results in robust labelling of nuclei of the ascending auditory pathway including the anteroventral, posteroventral, and dorsal cochlear nuclei (mainly contralaterally), and bilaterally the lateral superior olive, lateral nucleus of the trapezoid body, dorsal nucleus of the lateral lemniscus, and the central nucleus, dorsal cortex, and external cortex of the colliculus. The medial superior olive, superior paraolivary nucleus, and ventral nucleus of the trapezoid body essentially show ipsilateral projections to the central nucleus. The differential distribution of afferents to the inferior colliculus provides a substrate for functional parcellation of collicular subdivisions.     

Functional activation in the auditory system of the rat produced by arousing reticular stimulation: a 2-deoxyglucose study

The 2-deoxyglucose (2-DG) autoradiographic method was used to map the activity in the auditory pathway during behaviorally arousing electrical stimulation of the mesencephalic reticular formation (RET). Uptake of 2-DG during RET stimulation was compared to the effect of a frequency-modulated tone (4-5 kHz, 60 dB SPL) and to controls without stimulation. The major finding was a specific pattern of increased metabolic activation throughout the auditory pathway evoked during RET stimulation. The observed increases in 2-DG uptake were always greater in RET-stimulated rats as compared to sound-stimulated or control rats. The dorsal cochlear nucleus (DCN) showed the largest incorporation of 2-DG among the auditory nuclei of the brainstem in RET-stimulated rats. In the central nucleus of the inferior colliculus a layered pattern made of 3 discrete bands of high 2-DG uptake was visible in RET-stimulated rats. The medial geniculate (MG) and the auditory cortex (AC) also showed highly significant increases in 2-DG uptake induced by RET stimulation. The method provided correlations between classical morphological schemes of parcellation on nuclei and functionally defined areas of increased 2-DG uptake. Our observations represent the first anatomical demonstration of the activating effects of RET stimulation in a sensory system, and they support the concepts of arousing reticular mechanisms for sensory control.     

Efferent connections of the subthalamic region in the rat. II. The zona incerta

The efferent connections of the zona incerta (ZI) were studied experimentally in the rat by the aid of the autoradiographic tracer technique.Small microelectrophoretic injections of tritiated proline and leucine practically confined to the ZI were found to label a widespread, predominantly ipsilateral system of descending and ascending fibers distributed to reticular structures of the brain stem (mesencephalic reticular formation, nucleus tegmenti pedunculopontinus pars compacta, parabrachial area, nuclei reticularis pontis oralis, pontis caudalis, gigantocellularis and medullae oblongatae, pars ventralis), precerebellar nuclei (nucleus reticularis tegmenti pontis, pontine nuclei and inferior olivary complex), the middle and deep layers of the superior colliculus, the pretectum (anterior, posterior and medial pretectal nuclei), perioculomotor nuclei (interstitial nucleus of Cajal, nucleus of Darkschewitsch and nuclei of the posterior commissure), the parvocellular portion of the red nucleus, the central gray substance, the nucleus tegmenti dorsalis lateralis, the ventral horn of the cervical spinal cord, non-specific thalamic nuclei (parafascicular, centralis medius, paracentralis centralis lateralis and ventromedial thalamic nuclei, nucleus reuniens), basal ganglia (entopeduncular nucleus and globus pallidus), hypothalamic structures (posterior hypothalamic nucleus, dorsal and lateral hypothalamic areas), and a subpallidal district of the substantia innominata. Isotope injections centered in Forel's field H₁ resulted in the labeling of a similar set of projections. Some of the possible functional correlates of these connections are briefly discussed.     

Efferent tectal pathways of the opossum (Didelphis virginiana)

Efferent tectal pathways have been determined for the opossum, Didelphis virginiana, by employing the Nauta-Gygax technique ('54) on animals with tectal lesions of varying sizes. The superior colliculus projected tectothalamic fascicles to the suprageniculate nucleus, the central nucleus of the medial geniculate body, the lateral posterior thalamus, the pretectal nucleus, the ventral lateral geniculate nucleus, the fields of Forel and zona incerta, the parafascicular complex, the paracentral thalamic nucleus and in some cases to restricted areas of the anterior thalamus. Degenerating fibers from superior collicular lesions showed profuse distribution to the deeper layers of the superior colliculus on both sides and to the midbrain tegmentum, but only minimally to the red nucleus and substantia nigra. Fibers of tectal origin did not distribute to the motor nuclei of the oculomotor or trochlear nerves. At pontine levels, efferent fascicles from the superior colliculus were present as an ipsilateral tectopontine and tectobulbar tract and as a crossed predorsal bundle. The tectopontine tract ended mostly within the lateral and ventral basal pontine nuclei, whereas the ipsilateral tectobulbar tract distributed to certain specific areas of the reticular formation throughout the pons and medulla, minimally to the most medial portion of the motor nucleus of the facial nerve and to the nucleus of the inferior olive. The predorsal tract contributed fascicles to certain nuclei of the pontine raphe, extensively to the medial reticular formation of the pons, to the central and ventral motor tegmental nuclei of the reticular formation within the pons and medulla, to the paraabducens region, minimally to cells within restricted portions of the motor nucleus of the facial nerve, to certail specific regions of the caudal medulla and to the cervical cord as far caudally as the fourth segment. The tectospinal fascicles were few but some ended related to the spinal accessory nucleus and the ventral medial nucleus of the ventral horn. Lesions of the inferior colliculus resulted in degenerating fibers which distributed rostrally to the rostral nucleus of the lateral lemniscus and parabrachial region, to the suprageniculate nucleus, the parabigeminal nucleus and to the central nucleus of the medial geniculate body. The inferior colliculus also contributed fibers to the ipsilateral tectopontine and tectobulbar tracts. The latter bundle was traced as far caudally as the medulla and may arise from cells of the superior colliculus which are situated dorsal to the nucleus of the inferior colliculus.     

Alterations in cytochrome c oxidase activity and energy metabolites in response to kainic acid-induced status epilepticus

The effects of kainic acid (KA)-induced limbic seizures have been investigated on cytochrome c oxidase (COx) activity, COx subunit IV mRNA abundance, ATP and phosphocreatine (PCr) levels in amygdala, hippocampus and frontal cortex of rat brain. Rats were killed either 1 h, three days or seven days after the onset of status epilepticus (SE) by CO₂ and decapitation for the assay of COx activity and by head-focused microwave for the determination of ATP and PCr. Within 1 h COx activity and COx subunit IV mRNA increased in all brain areas tested between 120% and 130% of control activity, followed by a significant reduction from control, in amygdala and hippocampus on day three and seven, respectively. In amygdala, ATP and PCr levels were reduced to 44% and 49% of control 1 h after seizures. No significant recovery was seen on day three or seven. Pretreatment of rats with the spin trapping agent N-tert-butyl-α-phenylnitrone (PBN, 200 mg kg⁻¹, i.p.) 30 min before KA administration had no effect on SE, but protected COx activity and attenuated changes in energy metabolites. Pretreatment for three days with the endogenous antioxidant vitamin E (Vit-E, 100 mg/kg, i.p.) had an even greater protective effect than PBN. Both pretreatment regimens attenuated KA-induced neurodegenerative changes, as assessed by histology and prevention of the decrease of COx subunit IV mRNA and COx activity in hippocampus and amygdala, otherwise seen following KA-treatment alone. These findings suggest a close relationship between SE-induced neuronal injury and deficits in energy metabolism due to mitochondrial dysfunction.     

Neuroanatomical and neuropharmacological study of opioid pathways in the mesencephalic tectum: effect of mu(1)- and kappa-opioid receptor blockade on escape behavior induced by electrical stimulation of the inferior colliculus

Deep layers of the superior colliculus (DLSC), the dorsal and ventral periaqueductal gray matter (PAG), and inferior colliculus (IC) are midbrain structures involved in the generation of defensive behavior. beta-Endorphin and Leu-enkephalin are some neurotransmitters that may modulate such behavior in mammals. Light microscopy immunocytochemistry with streptavidin method was used for the localization of the putative cells of defensive behavior with antibodies for endogenous opioids in rat brainstem. Midbrain structures showed positive neurons to beta-endorphin and Leu-enkephalin in similar distributions in the experimental animals, but we also noted the presence of varicose fibers positive to endogenous opioids in the PAG. Neuroanatomical techniques showed varicose fibers from the central nucleus of the inferior colliculus to ventral aspects of the PAG, at more caudal levels. Naloxonazine and nor-binaltorphimine, competitive antagonists that block mu(1)- and kappa-opioid receptors, were then used in the present work to investigate the involvement of opioid peptide neural system in the control of the fear-induced reactions evoked by electrical stimulation of the neural substrates of the inferior colliculus. The fear-like responses were measured by electrical stimulation of the central nucleus of the inferior colliculus, eliciting the escape behavior, which is characterized by vigorous running and jumping. Central administration of opioid antagonists (2.5 microg/0.2 microl and 5.0 microg/0.2 microl) was performed in non-anesthetized animals (Rattus norvegicus), and the behavioral manifestations of fear were registered after 10 min, 2 h, and 24 h of the pretreatment. Naloxonazine caused an increase of the defensive threshold, as compared to control, suggesting an antiaversive effect of the antagonism on mu(1)-opioid receptor. This finding was corroborated with central administration of nor-binaltorphimine, which also induced a decrease of the fear-like responses evoked by electrical stimulation of the inferior colliculus, since the threshold of the escape behavior was increased 2 and 24 h after the blockade of kappa-opioid receptor. These results indicate that endogenous opioids may be involved in the modulation of fear in the central nucleus of the inferior colliculus. Although the acute treatment (after 10 min) of both naloxonazine and nor-binaltorphimine causes nonspecific effect on opioid receptors, we must consider the involvement of mu(1)- and kappa-opioid receptors in the antiaversive influence of the opioidergic interneurons in the dorsal mesencephalon, at caudal level, after chronic (2-24 h) treatment of these opioid antagonists. The neuroanatomical study of the connections between the central nucleus of the inferior colliculus and the periaqueductal gray matter showed neuronal fibers with varicosities and with terminal bottons, both in the pericentral nucleus of the inferior colliculus and in ventral and dorsal parts of caudal aspects of the periaqueductal gray matter.     

Relationship of opioid receptors with GABAergic neurons in the rat inferior colliculus

The inferior colliculus is a critical structure for processing auditory information and receives ascending and descending synaptic auditory projections. In addition to GABAergic and glutamatergic innervations, other neurotransmitter systems are also reported in the inferior colliculus, including opioid peptides. In the present study, the relative distribution of each type of opioid receptor, mu (MOR), delta (DOR) and kappa (KOR) within GABAergic neurons in the inferior colliculus was examined. GABA immunoreactivity was expressed by small, medium and large neurons and distributed in the central nucleus and the pericentral nucleus of the inferior colliculus. Immunostaining for MOR, DOR and KOR receptors was found in both disc-shaped cells and stellate cells. Punctiform beta-endorphin immunolabelling was observed in the proximity of GABA-positive neurons. Co-localization of GABA and MOR receptors was observed in neurons and nerve terminals in the central nucleus, dorsal cortex and external cortex of the inferior colliculus. Quantification of the co-localization patterns determined that a higher proportion of GABA neurons was associated with MOR receptors compared with KOR or DOR receptors.