Noëlle Bons

Senile plaques and neurofibrillary changes in the brain of an aged lemurian primate, Microcebus murinus

In some aged Microcebus brains (8- to 11-year-old animals) dramatic atrophy is found, particularly of the cortex, the hippocampus, the basal ganglia, the brainstem and the cerebellum, associated with a conspicuous increase in the size of the cerebral ventricles. These morphological changes are accompanied by certain histological profiles indicative of pathology. In the cortex, these histological changes consist of 1) a large number of senile plaques composed of degenerated neurites sometimes surrounding an amyloid plaque, 2) amyloid deposits in the vascular walls and 3) dense bundles of argyrophilic filaments in numerous pyramidal neurons. All these lesions resemble changes associated with Alzheimers disease in man. The degenerative changes observed in the Microcebus brain are accompanied by behavioral changes. At the moment these preliminary studies, carried out on the smallest of all primates, do not prove that the degeneration is of the Alzheimer type, but do indicate that Microcebus murinus may well be a good model for the study of cerebral aging, providing a comparison with cerebral ageing in humans. The size, life span and cost of the animal provide further advantages when compared with other nonhuman primates.

Identification of amyloid beta protein in the brain of the small, short-lived Lemurian primate Microcebus murinus

The deposition of amyloid beta (A beta) protein in the brain has been demonstrated immunocytochemically in the small Lemurian primate Microcebus murinus. Both meningocerebral vascular deposits and cortical parenchymal deposits occur. All eight aged (> 8 years old) Microcebus examined showed vascular amyloid deposits, whereas only four exhibited parenchymal plaques. The vascular amyloid infiltrated the tunica media of the leptomeningeal and cortical arteries and arterioles and was also found in capillaries. A beta was observed to be deposited in three general forms in the cortical neuropil: round or elliptical plaques that were thioflavin-negative but sometimes showed a central concentration of A beta immunoreactivity; round plaques with a densely immunoreactive core that was thioflavin-positive; extensive ribbon-like infiltrations enclosing multiple cortical blood vessels. These observations, taken together with previous descriptions of age-related neurodegenerative changes in Microcebus, indicate that this species undergoes a beta-amyloid-associated neuropathology highly similar to that seen in Alzheimers disease. We conclude that this lemurian primate of small size and relatively short life expectancy, provides a compelling animal model of some principal features of Alzheimers disease.

A stereotaxic atlas of the grey lesser mouse lemur brain (Microcebus murinus)

In response to the growing interest in the prosimian Microcebus murinus for studies on cerebral aging, the stereotaxic atlas of its brain was carried out in view of further anatomical, biochemical, electrophysiological, and behavioral investigations as well as for therapeutic experiments. This primate, which could be a valuable model for neuroscientific studies in various domains, presents numerous physiological advantages (e.g., size, cost, and ability to breed) compared to rodents, which can be used as nonprimate models, and simians. The atlas, valid for adult microcebes of every age and both sexes, consists of 54 frontal plates and 28 sagittal plates. For the establishment of stereotaxic coordinates and for drawings and photographs, 10 adult specimens of Microcebus murinus were used. The brains were frozen, cut into sections of 50 microm thickness, every fourth section being stained with Nissl. First, sections were projected and the outlines of the different structures, nuclei, and fibers were drawn. Then, the accuracy of the analysis was improved by detailed observation directly by microscope and also by computer analysis. Finally, the photographs of the sections were scanned and processed using the software Photoshop and Illustrator. For testing coordinates, several verifications were made. Experiments on lesions and injections of different substances were carried out in specific regions of the brain and brains implanted with needles were fixed in formol and embedded in paraffin wax.

Tissue Localization of Overproduced Esterases in the Mosquito Culex pipiens (Diptera: Culicidae)

Abstract We have investigated the tissue distribution of overproduced esterases A (A1 and A2) and B (B1 and B2) in strains of Culex pipiens L. by immunocytochemistry. S-LAB mosquitoes, lacking overproduced esterases, were used as reference. Tissues showing a strong specific reaction (fluorescence) were observed with anti-esterase A1 antiserum in S54 (with A1) and BOUAKE (with A2) strains, and with anti-esterase B1 antiserum in TEM-R and EDIT (with B1) and BOUAKE (with B2) strains. Overproduction of esterases A and B was tissue-specific. The most constant pattern for the two types of esterases was their overproduction in the alimentary canal and Malpighian tubes, although fluorescence varied in intensity depending on strains and developmental stages. There was no difference in the tissue distribution of esterases A1 and A2. In contrast, esterases B pattern was highly variable among strains. Differences between TEM-R and EDIT were explained by the different overall overproduction and number of copies of the amplified gene (10-fold higher in TEM-R). The most striking difference in esterase B1 and B2 tissue localization concerned the nervous system where neurons were intensely fluorescent in TEM-R and EDIT (B1), but not in BOUAKE (B2). All esterase B positive tissues in TEM-R contained large quantities of esterase B1 mRNA (in situ hybridization), indicating that at least part of the protein revealed by immunochemistry was produced in the tissues where it was observed. Our results are discussed in terms of the protection that the different esterases can confer during exposition to organophosphorous insecticides.

Topographical distribution of CRF immunoreactivity in the pigeon brain

The distribution of corticotropin-releasing factor (CRF) immunoreactivity was demonstrated by immunocytochemistry in intact and colchicine-treated pigeons. Colchicine injections were administered at different times related to the circadian activity of the CRF-adrenocorticotropin (ACTH)-corticosterone axis. Three CRF antisera were used, two directed against synthetic rat CRF and one directed against synthetic ovine CRF. No fundamental differences appeared in the pigeon brain with respect to the specific CRF antiserum used. The most effective colchicine injection times corresponded to hypersecretion in the corticotropic axis. CRF-immunopositive neurons were scattered throughout the pigeon brain. In addition to the paraventricular hypothalamic system, which is involved in adenohypophysial ACTH regulation, several other hypothalamic and extrahypothalamic areas showed CRF neurons. The distribution suggests that CRF may also act as a modulator and a neurotransmitter. Two hypothalamic paraventricular nucleus-median eminence CRF pathways are described here. Moreover, CRF-immunopositive reactions were observed in specific areas of cerebral ventricle walls, suggesting that CRF may be released into the cerebral fluid.

Prion immunoreactivity in brain, tonsil, gastrointestinal epithelial cells, and blood and lymph vessels in lemurian zoo primates with spongiform encephalopathy

We report on two animals of a non-human primate species Eulemur fulvus mayottensis, housed in the local zoo and fed over a number of years with a food containing cattle meat, that developed serious neurological symptoms associated with prion immunoreactivity in brain and various viscera. Microscopy of the brains showed neuronal vacuolation with patchy/perivacuolar immunolabelling with an abnormal isoform of prion protein (IR-PrP), an important characteristic of spongiform encephalopathy. For the first time, we report the presence in the same severely ill animals of IR-PrP in the gastrointestinal tract, detected by immunocytochemistry with mono- and polyclonal antibodies directed against various parts of the PrP. Strong PrP labelling was observed in the epithelial cells lining the pharyngeal and gastrointestinal lumen. The tonsils and the walls of the lymph and blood vessels below the intestinal epithelium were also labelled. There were no such immunoreactions in healthy lemurians killed as controls, i.e. a younger congener of the same species housed under the same conditions, and others belonging to the smaller species Microcebus murinus, reared in the laboratory and never fed on commercial food products containing cattle meat. These results demonstrate a strong PrP accumulation in the brain, the gastrointestinal tract and underlying lymphoreticular structures in these primates living in a zoological park and suffering from a spongiform encephalopathy.

Choline acetyltransferase and somatostatin levels in aged Microcebus murinus brain.

beta-Amyloid protein (beta-AP) deposits, analoguous to those found in Alzheimers disease (AD) are observed in the brain of aging Microcebus murinus. Because choline acetyltransferase (ChAT) activity and somatostatin (SRIH) content are consistently decreased in AD, we tested whether such changes could be observed in middle aged to aged Microcebus cerebral cortex and whether they were accompanied by beta-AP deposits. A positive correlation was observed between age and ChAT activity. By HPLC, SRIH immunoreactivity eluted as four peaks, two of which being identical with SRIH-28 and SRIH-14 while the other two likely represented precursor forms. Cortical SRIH content was not significantly affected by age. ChAT activity and SRIH content were not significantly correlated. Amyloid angiopathy was observed in every brain examined and the presence of cortical lesions analoguous to senile plaques observed in the oldest case only which did not demonstrate important alterations in ChAT and somatostatin levels.

M1 Muscarinic Agonists as a Therapeutic Strategy in Alzheimer’s Disease

M1 muscarinic receptors (M 1 mAChR) have an important role in cognitive processing relevant to Alzheimer’s disease brains (AD)1–5. M1 mAChR are relatively unchanged in AD1–5 and therefore may serve as a target for an anti-dementia drug treatment. However, some of the tested muscarinic agonists were not highly M1 selective, had major clinical limitations and showed disappointing clinical results in AD 4,6,7. Thus the proof of clinical concept could not be shown.