J.A. Aguirre

Transgenic expression and activation of PGC-1α protect dopaminergic neurons in the MPTP mouse model of Parkinson’s disease

Mitochondrial dysfunction and oxidative stress occur in Parkinson’s disease (PD), but little is known about the molecular mechanisms controlling these events. Peroxisome proliferator-activated receptor-gamma coactivator-1α (PGC-1α) is a transcriptional coactivator that is a master regulator of oxidative stress and mitochondrial metabolism. We show here that transgenic mice overexpressing PGC-1α in dopaminergic neurons are resistant against cell degeneration induced by the neurotoxin MPTP. The increase in neuronal viability was accompanied by elevated levels of mitochondrial antioxidants SOD2 and Trx2 in the substantia nigra of transgenic mice. PGC-1α overexpression also protected against MPTP-induced striatal loss of dopamine, and mitochondria from PGC-1α transgenic mice showed an increased respiratory control ratio compared with wild-type animals. To modulate PGC-1α, we employed the small molecular compound, resveratrol (RSV) that protected dopaminergic neurons against the MPTP-induced cell degeneration almost to the same extent as after PGC-1α overexpression. As studied in vitro, RSV activated PGC-1α in dopaminergic SN4741 cells via the deacetylase SIRT1, and enhanced PGC-1α gene transcription with increases in SOD2 and Trx2. Taken together, the results reveal an important function of PGC-1α in dopaminergic neurons to combat oxidative stress and increase neuronal viability. RSV and other compounds acting via SIRT1/PGC-1α may prove useful as neuroprotective agents in PD and possibly in other neurological disorders.

Protection but maintained dysfunction of nigral dopaminergic nerve cell bodies and striatal dopaminergic terminals in MPTP-lesioned mice after acute treatment with the mGluR5 antagonist MPEP

The mGluR5 antagonist MPEP was used to study the role of mGluR5 in MPTP-induced injury of the nigrostriatal DA neurons. The findings indicate that acute blockade of mGluR5 may result in neuroprotective actions against MPTP neurotoxicity on nigral DA cell bodies and striatal DA terminals using stereological analysis of TH immunoreactivity and microdensitometry. Biochemical analysis showed no restoration of DA levels and metabolism indicating a maintained reduction of DA transmission.

On the role of P2X(7) receptors in dopamine nerve cell degeneration in a rat model of Parkinson's disease: studies with the P2X(7) receptor antagonist A-438079.

The role of the ATP-gated receptor, P2X7, has been evaluated in the unilateral 6-OHDA rat model of Parkinson’s disease using the P2X7 competitive antagonist A-438079. Nigral P2X7 immunoreactivity was mainly located in microglia but also in astroglia. A-438079 partially but significantly prevented the 6-OHDA-induced depletion of striatal DA stores. However, this was not associated with a reduction of DA cell loss. Blockade of P2X7 receptors may represent a novel protective strategy for striatal DA terminals in Parkinson’s disease and warrants further future investigation.

c-Fos expression in supramammillary and medial mammillary nuclei following spatial reference and working memory tasks.

To investigate brain substrates of spatial memory, neuronal expression of c-Fos protein was studied. Two groups of rats were trained in two spatial memory tasks in the Morris water maze, where the rats have to apply a reference memory rule or a working memory rule. In addition to the experimental groups, two control groups were used to study c-fos activation not specific to the memory processes studied. After immunohistochemical procedures, the number of c-Fos positive neuronal nuclei was quantified in the mammillary body (MB) region (medial mammillary nucleus [MMn] and supramammillary nucleus [SuM]). The results have shown that some MMn neurons expressed c-Fos nuclear immunoreactivity related to spatial working memory but not to spatial reference memory. The increased number of c-Fos immunoreactive neuronal nuclei in the SuM was related to spatial training but not to either working or reference memory demands of the tasks.

Distribution of neuropeptide Y-like immunoreactive cell bodies and fibers in the brain stem of the cat

By using intratissue injections of colchicine and an indirect immunoperoxidase technique, we studied the distribution of cell bodies and fibers containing neuropeptide Y-like immunoreactivity in the brain stem of the cat. The densest clusters of immunoreactive perikarya were observed in the following nuclei: anteroventral cochlear, lateral reticular (internal and external divisions), dorsal tegmental, inferior colliculus and dorsal nucleus of the lateral lemniscus. By contrast, the nuclei abducens, the nucleus of the trapezoid body, preolivary, interpeduncularis, infratrigeminal, gigantocellular tegmental field, coeruleus and dorsal motor nucleus of the vagus had the lowest density. Finally, a moderate density of neuropeptide Y-like immunoreactive cell bodies was found in the nuclei: lateral tegmental field, laminar spinal trigeminal, praepositus hypoglossi, superior colliculus, lateral vestibular and motor trigeminal. In addition, a mapping of the neuropeptide Y-like immunoreactive fibers was carried out. Thus, the densest network of immunoreactive fibers was observed in the laminar spinal trigeminal nucleus. The nuclei periaqueductal gray, inferior central, praepositus hypoglossi, postpyramidal raphe, dorsal raphe, incertus and medial vestibular contained a moderate density of immunoreactive fibers, whereas the nuclei interpeduncularis, inferior colliculus, superior central, gracile, retrorubral, Kölliker-Fuse, dorsal tegmental, ambiguus and alaminar spinal trigeminal had the lowest density of neuropeptide Y-like immunoreactive fibers. The anatomical location of neuropeptide Y-like immunoreactivity suggests that the peptide could play an important role in several physiological functions, e.g., those involved in cardiovascular, auditory, motor, visual, nociceptive and somatosensory mechanisms.

Supramammillary and adjacent nuclei lesions impair spatial working memory and induce anxiolitic-like behavior.

The present study assesses the involvement of the supramammillary and adjacent nuclei in spatial memory and anxiety-like behaviors. Rats with electrolytic lesions in the supramammillary nucleus were pre- and post-operatively trained in two spatial memory tasks and two anxiety tasks. Spatial memory tasks were performed in an open field with seven different goal positions containing the reward. Anxiety-like behaviors were tested in the elevated T-maze. In the spatial reference memory task, neither lesioned nor sham-lesioned groups were impaired. In the working memory task, lesioned animals were permanently impaired in their ability to solve the delayed-matching-to-position task. This working memory deficit is not related to increased proactive interference. It could be related to impairment of the rats ability to reorganize spatial stimuli. Consequently, rats were not able to achieve an optimal performance level to solve spatial tasks with continuous changes in the place location. In the elevated T-maze, lesioned rats reduced passive avoidance response but no changes in the escape response were observed. These results suggest a clear involvement of the supramammillary nucleus in working memory and behavioral inhibition but not in either spatial reference memory or in escape responses.

Effects of medial prefrontal cortex lesions on anxiety-like behaviour in restrained and non-restrained rats.

The medial prefrontal cortex has been associated with fear, anxiety and stress regulation, and has recently been suggested to play a crucial role in the development of behavioural changes in response to stress. In this study, we evaluated medial prefrontal cortex (mPFC) involvement in both anxiety-like behaviour and increased anxiety-like responses induced by uncontrollable restraint. Rats with mPFC electrolytic lesions (n=7) and sham-lesioned (n=8) were tested in the elevated T-maze (ETM). Restrained rats with mPFC lesions (n=8) and sham-lesioned rats (n=6) were tested in the elevated T-maze 24h after restraint. Both two-trial passive avoidance and one-trial escape behaviours were assessed. The results revealed that mPFC lesions impair passive avoidance, but not escape behaviour. In addition, decreased anxiety-like behaviour in both passive avoidance and escape behaviours were observed in restrained rats with mPFC lesions. Our results suggest that mPFC is important in mediating both anxiety-like behaviour expression and long-term anxiogenic-like effects induced by acute restraint.

Anatomical distribution of beta-endorphin (1-27) in the cat brainstem: an immunocytochemical study

 Using an indirect immunoperoxidase technique, we studied the location of β-endorphin (1–27) fibres and cell bodies in the cat brainstem. The highest density of immunoreactive fibres was found in the lateral and medial parabrachial nuclei and in the locus coeruleus; a moderate density was observed in the periaqueductal gray and the central reticular nucleus, and a low density was observed in the interpeduncular nucleus, the nucleus incertus, the raphe pallidus nucleus, the paralemniscal reticular nucleus, the laterodorsal tegmental nucleus, the pericentral division of the dorsal tegmental nucleus and the lateral reticular nucleus. Immunoreactive neurons were observed in the superior central nucleus, the pericentral division of the dorsal tegmental nucleus, the interpeduncular nucleus, the nucleus incertus and the dorsal raphe nucleus. Our results point to a more widespread distribution of β-endorphin (1–27)-immunoreactive perikarya in the cat brainstem in comparison with previous studies carried out in the same region of other mammals. The distribution of β-endorphin (1–27)-immunoreactive fibres and perikarya is compared with the location of other neuropeptides in the cat brainstem. Moreover, our findings reveal that β-endorphin (1–27)-immunoreactive structures are widely distributed in the cat brainstem, suggesting that the peptide might be involved in several physiological functions.

Immunohistochemical mapping of enkephalins, NPY, CGRP, and GRP in the cat amygdala.

This immunohistochemical study shows a wide distribution of neuropeptides in the cat amygdala. Neuropeptide Y is present along the whole amygdaloid complex, and fibers and cell bodies containing neuropeptide Y are observed in all the nuclei studied. Leucine-enkephalin-, gastrin-releasing peptide/bombesin-, and calcitonin gene-related peptide-immunoreactive fibers and perikarya are observed only in discrete nuclei of the amygdaloid complex, whereas only fibers -but no cell bodies- containing methionine-enkephalin-Arg6-Gly7-Leu8 have been observed. No immunoreactivity has been found for gamma-melanocyte-stimulating hormone, dynorphin A (1-17), or galanin. These data are compared with those reported in the amygdala of other mammals.

Neurokinin A-like immunoreactivity in the cat brainstem

We have studied the distribution of neurokinin A-like immunoreactive cell bodies and fibers in the cat brainstem. The densest clusters of perikarya containing the peptide were observed in the periaqueductal gray, inferior colliculus, postpyramidal nucleus of the raphe, medial nucleus of the solitary tract and in the lateral reticular nucleus. By contrast, the interpeduncular nucleus, magnocellular part of the red nucleus, central tegmental field, cuneiform nucleus, dorsal tegmental nucleus, nucleus sagulum and the medial and inferior vestibular nuclei had the lowest density, whereas a moderate density of immunoreactive cell bodies was found in the superior colliculus, medial division of the dorsal nucleus of the raphe, nucleus incertus, locus coeruleus and in the Kölliker-Fuse area. The highest density of immunoreactive fibers was observed in the substantia nigra, periaqueductal gray, marginal nucleus of the brachium conjunctivum, medial vestibular nucleus, medial nucleus of the solitary tract, laminar spinal trigeminal nucleus, inferior colliculus, medial division of the dorsal nucleus of the raphe, locus coeruleus, dorsal tegmental nucleus and in the spinal trigeminal tract. A moderate density of immunoreactive fibers was found in the dorsal motor nucleus of the vagus and in the postpyramidal nucleus of the raphe and a low density in the cuneiform nucleus, Kölliker-Fuse area, nucleus sagulum, inferior and superior central nuclei, lateral reticular nucleus and in the lateral and magnocellular tegmental fields.