Pyritinol facilitates the recovery of cortical cholinergic deficits caused by nucleus basalis lesions

The effect of a nootropic, Pyritinol, on the recovery of cortical cholinergic deficits induced by injury of the nucleus basalis has been tested on two groups of unilateral quisqualic acid nbM-lesioned rats. The first group had a 30 nmol lesion producing a cortical cholinergic impairment at 21 days, with a spontaneous recovery at 45 days. The second group had a 50 nmol lesion that produced a deeper cholinergic deficit, which did not recover at 45 days. Pyritinol enhanced the recovery in the 30 nmol group of animals on the 21st day after surgery. The recovery was measured as an increase in the activities of acetylcholinesterase (AChE), cholineacetyltransferase (ChAT) and the high affinity choline uptake system, and the histochemical densities of the cortical AChE network and the M2 receptor. Histochemical analysis of the nbM enabled cortical recovery to be related to the number of surviving neurons and also to their hypertrophy and AChE-ChAT hyperactivity. Pyritinol enhanced recovery in 30 nmol lesioned animals but in the other group, with a lower number of surviving neurons and a lower ability of the cells to become hypertrophic, the drug was unable to promote cortical recovery.Peer Reviewe

Morphometric study on the development of magnocellular neurons of the supraoptic nucleus utilising immunohistochemical methods

Vasopressin (VP)- and oxytocin (OXY)-producing neurons, components of the rat supraoptic nucleus, have been located with immunohistochemical methods, with the purpose of studying their morphofunctional characteristics during different phases of life (embryonic, juvenile, adult and senile). To carry out this study, an IBAS I (Kontron) computerised image analyser has been utilised. The hormone VP is first detected in the neuronal cytoplasm of 21 day old rat embryos and the hormone OXY appears in the neuronal cytoplasm later, in the newborn phase. The neuronal area with a positive reaction for the two neurohormones has been evaluated and it has been found that the quantity of reaction substance is proportional to the age. In the adult period, VP neurons possess a reaction area (198 μm2) greater than that of OXY neurons (153 μm2). In the SON, there are two neuronal shapes, fusiform and round; these shapes coexist in both hormonal types of neurons. Until Day 15 of postnatal development, the SON neurons are intermixed in the interior of the nucleus but in this period a neuronal redistribution is initiated. In the adult phase, OXY neurons are situated preferentially in the anterior, posterior and dorsal parts of VP neurons in the ventral and posterior parts, with both neurons being present in the intermediate part of the SON.Peer Reviewe

Morphometric and neurosecretory changes in supraoptic neurons after D-amphetamine treatment

Several morphological and immunochemical characteristics of the neurosecretory neurons of the supraoptic nucleus (SON) have been studied of rats treated for 1 month with D-amphetamine sulfate (AMP) (8 mg/kg weight, daily). An increase of SON volume (11%) has been observed as a consequence of the growth of the dorsoventral axis. Neurosecretory neurons increased their nucleolar area (11.4%), their nuclear area (8.3%), and their cytoplasmatic area (18.3%). Vasopressin immunoreaction did not show any differences between treated and control animals, but oxytocin immunostaining displayed an important increase (23.7%) in the neuronal cytoplasm of the treated rats. The SON hypertrophy of the AMP-treated rats corresponded to the hypertrophy/hyperfunction of its oxytocinergic neurons, and could be considered as a new mechanism of the action of the AMP. The results are discussed in relation to the plastic features of the SON and its central (neuronal) and peripheral (hormonal) function.Peer Reviewe

Cytophotometric study on acetylcholinesterase activity in mammalian hypothalamus.

Peer Reviewe

Presence of Calcitonin Gene-Related Peptide in Intraepithelial Nerve Fibers and Motor End-Plates of the Cat Esophagus: A Light and Electron Microscopic Study

The morphology and distribution of the motor end-plates in the striated muscle and the terminal nerve fibers in the epithelium of the wall of the esophagus, which contain calcitonin gene-related peptide, were studied by light and electron microscopic immunocytochemistry. Varicose immunoreactive nerve fibers arising from the subepithelial plexus were seen to penetrate into the epithelium where they ended in terminal boutons. These nerve fibers lost their Schwann cells just at the point of penetration into the epithelium. Characteristically, the epithelial cells of the spinous layer showed prominent tonofilaments in the part of the cytoplasm in contact with the immunoreactive nerve varicosities, but membrane specializations between these structures were not observed. In the striated muscle of the esophageal wall there were small, elliptical, immunoreactive motor end-plates, which contained a small number of axonal clear vesicles and mitochondria. They were associated with relatively short and rarely branched junctional folds, reduced postjunctional surfaces and few organelles in the underlying sarcoplasm, features characteristic of the neuromuscular junctions of slow-fatiguing red muscle fibers. The two types of immunoreactive nerve endings, epithelial and muscular, presumably participate in afferent and efferent limbs respectively of the neural control of esophageal motility. The relationship between immunoreactive nerve terminals and epithelial cells in the spinous layer exhibiting prominent tonofilaments allowed us to speculate about the existence of two different patterns of reception to sensory stimuli. The intraepithelial fibers that end in the middle layer of the epithelium could be related to mechanoreceptor reflexes, while those that end in the upper layer may be related to thermoreceptor reflexes or facilitate information about the chemical and other characteristics of foods

Chemical heterogeneity in adult rat cerebellar Purkinje cells as revealed by zebrin I and low-affinity nerve growth factor receptor immunocytochemical expression following injury

Cerebellar Purkinje cells in rat express low-affinity nerve growth factor receptor during development, but rarely in normal adult animals. However, after either mechanical injury or colchicine treatment during adulthood, these cells re-express low-affinity nerve growth factor receptor-immunoreactive protein. Two Purkinje cell subpopulations were defined in normal adult cerebellum by the presence or the absence of zebrin I antigen. Nevertheless, it remains an open question as to whether low-affinity nerve growth factor receptor-immunoreactive protein can be expressed by a]l damaged Purkinje cells, independent of their location and their staining with antibodies against intrinsic molecular markers that reveal Purkinje cell heterogeneity, such as zebrin I. In this study, a serial-section immunocytochemical mapping of the expression zebrin I and low-affinity nerve growth factor receptor, using specific monoclonal antibodies, was carried out in colchicine-treated rats. After mechanical damage of the cerebellar cortex, co-localization of these antigens at the cellular level was also analysed in thin adjacent sections, and by using a combined immunocytochemical staining method in individual sections. The findings revealed the existence of three sub-sets of Purkinje cells: (1) two complementary groups distinctly immunoreactive to one antibody, but not to the other and (2) a third group that contained double-labelled cells. In contrast, co-expression of both antigens was never observed following mechanical lesions. The seemingly independent response to mechanical injury of Purkinje cells located in different zebrin-defined compartments, indicates that particular subpopulations of Purkinje cells may respond differentially to traumatic injury.Peer Reviewe

Subcellular localization of the inositol 1,4,5-trisphosphate receptor, P400, in the vestibular complex and dorsal cochlear nucleus of the rat

The subcellular localization of the inositol 1,4,5-trisphosphate receptor protein P400, was studied in the vestibular complex, an area to which Purkinje cells project, as well as in neurons of the dorsal cochlear nucleus and in ectopic Purkinje cells of adult rat brain. The receptor was demonstrated by electron microscopical immunocytochemistry using the avidin-biotin peroxidase complex procedure, with the monoclonal antibody 4C11 raised against mouse cerebellar inositol 1,4,5-trisphosphate receptor protein. Immunoreactivity was found in preterminal fibres and terminal boutons in the nuclei of the vestibular complex, generally associated with the subsurface systems and stacks or fragments of smooth endoplasmic reticulum. Ectopic Purkinje cells and cartwheel cells of the dorsal cochlear nucleus also displayed immunoreactivity, but this was much less intense in the latter. The results of the present study suggest that this receptor protein, involved in the release of Ca2+, is located in sites that enable it to influence the synthesis, transport and release of neurotransmitters.Peer Reviewe

Presence of Calcitonin Gene-Related Peptide in Intraepithelial Nerve Fibers and Motor End-Plates of the Cat Esophagus: A Light and Electron Microscopic Study

The morphology and distribution of the motor end-plates in the striated muscle and the terminal nerve fibers in the epithelium of the wall of the esophagus, which contain calcitonin gene-related peptide, were studied by light and electron microscopic immunocytochemistry. Varicose immunoreactive nerve fibers arising from the subepithelial plexus were seen to penetrate into the epithelium where they ended in terminal boutons. These nerve fibers lost their Schwann cells just at the point of penetration into the epithelium. Characteristically, the epithelial cells of the spinous layer showed prominent tonofilaments in the part of the cytoplasm in contact with the immunoreactive nerve varicosities, but membrane specializations between these structures were not observed. In the striated muscle of the esophageal wall there were small, elliptical, immunoreactive motor end-plates, which contained a small number of axonal clear vesicles and mitochondria. They were associated with relatively short and rarely branched junctional folds, reduced postjunctional surfaces and few organelles in the underlying sarcoplasm, features characteristic of the neuromuscular junctions of slow-fatiguing red muscle fibers. The two types of immunoreactive nerve endings, epithelial and muscular, presumably participate in afferent and efferent limbs respectively of the neural control of esophageal motility. The relationship between immunoreactive nerve terminals and epithelial cells in the spinous layer exhibiting prominent tonofilaments allowed us to speculate about the existence of two different patterns of reception to sensory stimuli. The intraepithelial fibers that end in the middle layer of the epithelium could be related to mechanoreceptor reflexes, while those that end in the upper layer may be related to thermoreceptor reflexes or facilitate information about the chemical and other characteristics of foods

Distribution of catecholaminergic afferent fibres in the rat globus pallidus and their relations with cholinergic neurons

The topographical distribution of catecholaminergic nerve fibres and their anatomical relationship to cholinergic elements in the rat globus pallidus were studied. Peroxidase-antiperoxidase and two-colour immunoperoxidase staining procedures were used to demonstrate tyrosine hydroxylase (TH), dopamine β-hydroxylase (DBH), phenylethanolamine N-methyltransferase (PNMT) and choline acetyltransferase (ChAT) immunoreactivities, combined with acetylcholinesterase (AChE) pharmacohistochemistry. TH immunoreactive nerve fibres were seen to enter the globus pallidus from the medial forebrain bundle. The greatest density of such fibres was found in the ventral region of the globus pallidus, which was also characterized by the greatest density of ChAT immunoreactive neurons. TH immunoreactive nerve fibres showed varicose arborizations and sparse boutons, which were occasionally seen in close opposition to cholinergic structures. In all regions of the globus pallidus, there were also larger, smooth TH immunoreactive nerve fibres of passage to the caudate putamen. A smaller number of DBH immunoreactive nerve fibres and terminal arborizations were found in the substantia innominata, internal capsule and in the globus pallidus bordering these structures. A few PNMT immunoreactive nerve fibres in the substantia innominata and internal capsule did not enter the globus pallidus. Electron microscopy revealed TH immunoreactive synaptic profiles in the ventromedial area of the globus pallidus corresponding to the nucleus basalis magnocellularis of Meynert (nBM). These made mainly symmetrical and only a few asymmetrical synaptic contacts with dendrites containing AChE reaction product. The results indicate that cholinergic structures in the nBM are innervated by dopaminergic fibres and terminals, with only a very small input from noradrenergic fibres.Peer Reviewe