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In this study, age-related changes in the monoamine oxidases (MAO) were studied in the optic nerve (ON) of both young and aged male rats. The aim of the study was to assess the role of MAO in age-related changes in the rat ON and explain the mechanisms of neuroprotection mediated by MAO-B-specific inhibitors. Fifteen three month old and fifteen 26 month old Sprague-Dawley rats were used. The animals were killed by terminal anaesthesia. Staining of MAO, quantitative analysis of images, biochemical assays and statistical analysis of data were carried out. Samples of the ON were washed in water, fixed in Bowen fluid, dehydrated and embedded in Entellan. Histological sections were stained for MAO-enzymatic activities. The specificity of the reaction was evaluated by incubating control sections in a medium either without substrate or without dye. The quantitative analysis of images was carried out at the same magnification and the same lighting using a Zeiss photomicroscope. The histochemical findings were compared with the biochemical results. After enzymatic staining, MAO could be demonstrated in the ON fibres of both young and aged animals; however, MAO were increased in the nerve fibres of the elderly rats. These morphological findings were confirmed biochemically. The possibility that age-related changes in MAO levels may be attributed to impaired energy production mechanisms and/or represent the consequence of reduced energy needs is discussed., (© 2012 The Authors. International Journal of Experimental Pathology © 2012 International Journal of Experimental Pathology.)

Mitochondrial dysfunctions have been implicated in the pathophysiology of several age-related diseases including age-related macular degeneration (AMD), a progressive neurodegenerative disease affecting primarily the retinal pigment epithelium (RPE). The aims of our electron microscopic and morphometric studies were to reveal qualitative and quantitative alterations of mitochondria in human RPE from AMD and from age- and sex-matched controls. With increasing age a significant decrease in number and area of mitochondria, as well as loss of cristae and matrix density were found in both AMD and control specimens. These decreases were significantly greater in AMD than in normal aging. Alterations of mitochondria were accompanied by proliferation of peroxisomes and lipofuscin granules in both AMD and control specimens, although the difference between groups was significant only for peroxisomes. Unexpectedly, morphometric data showed that the RPE alterations seen in AMD may also develop in normal aging, 10-15 years after appearing in AMD patients. These findings suggest that (i) the severity of mitochondrial and peroxisomal alterations are different between AMD and normal aging, and (ii) the timing of damage to RPE may be critical for the development of AMD. We conclude that besides the well-documented age-related changes in mitochondrial DNA, alterations of mitochondrial membranes may also play a role in the pathogenesis of AMD. These membranes could be a new target for treatment of AMD and other age-related diseases.

Glycosaminoglycans play a central role in maintaining the normal outflow resistance in the human trabecular meshwork. We evaluated the possible morphological, histochemical and morphometrical age-related changes in glycosaminoglycans of the trabecular meshwork. Small human samples were drawn from 24 eyes after exitus from young and old humans. Samples were harvested from the anterior chamber of the eye, without any aesthetic damage for the face. They were divided into three fragments, each used for morphological, histochemical and ultrastructural staining. Quantitative analysis of images was performed to evaluate morphometrical data that were statistically analysed. Our findings demonstrate the following age-related changes: (1) deposition of fibrous granular material in the trabecular meshwork; (2) increased electron density of the structures; (3) strong decrease in the hyaluronic acid content, and (4) increase in sulphated proteoglycans. Glycosaminoglycans of human trabecular meshwork undergo age-related changes, as demonstrated by our morphological, histochemical and morphometrical results., (Copyright 2004 S. Karger AG, Basel)

Age-related changes of the catecholaminergic nerve fibers of the trachea, bronchial smooth muscle, lung capillaries and bronchus-associated lymphoid tissue (BALT) were studied in male Wistar rats aged 3 months (young), 12 months (adult) and 24 months (old/aged). Catecholamine histo- and immuno-fluorescence techniques were used, associated with image analysis and high pressure liquid chromatography with electrochemical detection of nor-epinephrine (nor-adrenaline). In young rats, blue-green fluorescent nerve fibers supply the trachea-bronchial smooth muscle and tracheal and bronchial glands. These structures are innervated by a delicate network of nerve fibers, being rich in varicosities. Pulmonary capillaries are sparsely innervated. The highest nor-epinephrine concentration was found in the trachea and bronchi, followed by BALT. The density and the pattern of noradrenergic nerve fibers of the trachea-bronchial tree or of the pulmonary vessels were similar in young and adult rats. In aged rats, a loss of noradrenergic nerve fibers, involving primarily the supply to the smooth muscle of the trachea-bronchial tree, was observed. Fluorescence microscopic techniques demonstrated a higher sensitivity than nor-epinephrine assay in detecting changes of the sympathetic nerve supply of the trachea-bronchial tree, pulmonary vessels and BALT. The possible significance of reduced noradrenergic nerve supply of the trachea-bronchial-pulmonary tree in aged rats is discussed., (Copyright 2004 Elsevier Ireland Ltd.)

Background: In a previous study, scanning electron microscopy (SEM) showed age-related changes in the rat retina. We carried out a study to evaluate age-related changes in the human retina., Methods: Samples of fresh retinal tissue obtained from younger (age 22 years or less) and older (age 66 years or more) donors were studied by means of traditional histologic methods and by SEM. Eight retinas were obtained from four donors whose corneas had been used for transplantation, and four retinas were obtained from four subjects whose eyes had been enucleated owing to injury. All morphologic results were subjected to quantitative analysis of images. The concentration of cytoplasmic (free) and structural (tissue-associated) protein in retinal tissue homogenates was determined by means of biochemical methods., Results: There was a decrease in all features studied with the exception of structural protein concentration. The mean retinal thickness (and standard error of the mean) was 426 (34.2) microm in the younger subjects and 261 (18.9) microm in the older subjects. The mean numbers of ganglion cells (and standard error of the mean) were 413.5/mm2 (32.3/mm2) and 256.2/mm2 (26.8/mm2) respectively, of capillaries 3.6/mm2 (1.4/mm2) and 1.8/mm2 (1.2/mm2) respectively, of synaptic bodies 122.4 (4.9) conventional units (CU)/area observed and 38.5 (1.6) CU/area observed respectively, of cellular processes 82.3 (3.1) CU/area observed and 13.1 (1.5) CU/ area observed respectively, and of intercellular connections 36.4 (2.5) CU/area observed and 14.3 (1.4) CU/area observed respectively. The mean concentration of total protein per milligram of fresh tissue (and standard error of the mean) was 92.1 (1.8) microg in the younger subjects and 78.7 (1.3) microg in the older subjects; the corresponding values for cytoplasmic protein were 27.6 (1.3) microg and 11.8 (0.8) microg, and for structural protein, 64.4 (1.6) microg and 86.9 (1.4) microg. All differences between the younger and older subjects were significant (p < 0.001) with the exception of mean concentration of cytoplasmic and of structural protein., Interpretation: The human retina undergoes specific changes with aging. SEM provides new morphometric information regarding age-related changes in photoreceptor cells, bipolar cells and ganglion cells that increases our understanding of this topic. Our results may be adopted as a model or as normal values when studying other changes that may occur in the human retina in pathological conditions.

The presence and distribution of acetylcholinesterase (AChE) and cholineacetyl transferase activities (Chat) were examined in the bronchus-associated lymphoid tissue (BALT) of juvenile, adult and old rats. Histoenzymatic and immunochemical methods were used in association with quantitative analysis of images and statistical analysis of the data. Our results showed that both AChE and Chat activities were primarily confined to the BALT lymphoid cells. Only a low level of activity was observed in the sub-pleural parenchyma of the lung and in the wall of the bronchus. Moreover, both AChE and Chat activities in the BALT are specifically located in the lymphoid cells. Histoenzymatic staining and corresponding values of quantitative analysis of images confirmed morphological and immunochemical results. Finally, the intensity of histoenzymatic staining for AChE and of immunochemical staining for Chat in BALT of rats strongly decreases with age. On the basis of our results we hypothesize that both AChE and Chat activities may play an important role in BALT and both these enzymes undergo specific age-related changes.

Age-related changes of the optic nerve were studied in 3-month-old (young), 12-month-old (adult) and 24-month-old (aged) male Sprague-Dawley rats. Cross sections of the intracranial portion of the optic nerves of animals of different age groups were stained with haematoxylin-eosin and examined under a light microscope at low and high magnification. Other sections were stained with crystal violet for demonstration of glial cells. A third group of sections were stained immunohistochemically to detect glial fibrillary acidic protein (GFAP) which is a marker for localizing and characterizing astrocytes. All morphological results were subjected to the quantitative analysis of images and to statistical analysis to identify significant morphometrical data. Tissue protein concentrations were determined on homogenized fragments of optic nerve. Our results demonstrate the following age-related changes: (1) increase of the optic nerve sheaths (meningeal membranes); (2) increased number of astrocytes; (3) increase of areal density of GFAP immunoreactivity; (4) increased diameter and area of the optic nerve; (5) decreased number of nerve fibres; (6) decreased-size of nerve fibres and (7) decrease of the nerve fibres/meningeal membrane ratio from 3:1 to 1:1. Moreover, the protein amount does not change with age. The rat optic nerve, therefore, appears sensitive to ageing processes and can be considered as a useful model for the studies on neuronal ageing.

Age-related changes of the monoaminoxidases, evaluated by enzymatic staining, quantitative analysis of images, biochemical assay and statistical analysis of data were studied in cerebellar cortex of young (3-month-old) and aged (26-month-old) male Sprague-Dawley rats. The enzymatic staining shows the presence of monoamino-oxidases within the molecular and granular layers as well as within the Purkinje neurons of the cerebellum of young and aged animals. In molecular layer, and in Purkinje neurons the levels of monoaminooxidases were strongly increased in old rats. The granular layer showed, on the contrary, an age-dependent loss of enzymatic staining. These morphological findings were confirmed by biochemical results. The possibility that age-related changes in monoaminooxidase levels may be due to impaired energy production mechanisms and/or represent the consequence of reduced energetic needs is discussed.

Background: Recent morphologic research has demonstrated the presence of nerve fibres of different diameters in the human optic nerve. The purpose of this study was to investigate age-related changes in fibres of the human optic nerve., Methods: We studied the left optic nerve of 50 male cadaveric donors, 16 aged 18 to 22 years (mean 20 [standard deviation 1.2] years) and 34 aged 68 to 76 years (mean 72 [standard deviation 1.6] years). The samples were carefully harvested during autopsy from the intracranial portion of the optic nerve. Each nerve was cut into four 4-mm segments. After morphologic, histochemical and immunohistochemical staining, the optic nerve fibres were counted and measured. Each segment was evaluated under light microscopy for microanatomic details, glial cells and glial fibrillary acidic protein (GFAP) staining. The protein content was determined under biochemical analysis. We performed morphometric analysis by examining the optic nerve images quantitatively., Results: Compared with the younger group, in the older group there was an increase in mean diameter of the optic nerve (p < 0.001), due to an increase in the optic nerve:meningeal membrane ratio. There was also an increase in mean optic nerve area (p < 0.001) and in mean number of astrocytes and the related GFAP-immunoreactive area (p < 0.001). The mean number of nerve fibres of large diameter (greater than 4 pm) was decreased (p < 0.001). There was no difference in mean protein content of the fibres between the two groups., Interpretation: The human optic nerve is sensitive to the aging process and may be considered as a model for studies on neuronal aging.

1. The distribution of dopamine D2 receptors in rat heart and coronary vessels and the possible age-related changes in D2 receptor density were studied. The pharmacological characteristics and the anatomical location of dopamine D2-like receptor sites in rat heart and coronary vessels were investigated using combined binding techniques and light microscopy autoradiography. 2. Samples of heart and coronary vessels were harvested from young and old rats. On frozen slices, dopaminergic D2 receptors were labelled by means of a selective D2 ligand, namely [3H]-spiroperidol (spiperone). Inhibition studies were performed using unlabelled agonists and/or labelled and unlabelled antagonists to define pharmacological specificity of the binding. Physiological experiments were performed to demonstrate the selective antagonism between D2 receptors and many dopaminergic drugs. 3. [3H]-Spiroperidol was bound to sections of rat heart and coronary artery (in a manner consistent with the labelling of dopamine D2-like receptors) with an equilibrium dissociation constant of approximately 2.4 +/- 0.7 nmol/L and a maximum capacity of binding sites of 65.8 +/- 4.5 fmol/mg protein. Experiments performed on sections of coronary veins did not allow the evaluation of specific binding. Autoradiography, observed with light microscopy, showed the development of specific silver grains within the whole wall of rat heart and coronary artery. The greater sensitivity to displacement by amisulpride, bromocriptine, domperidone, haloperidol, raclopride and L-sulpiride than to displacement by N-propyl-norapomorphine, quinpirole and clozapine suggests that the binding sites observed in these experiments are likely to belong to the dopamine D2 receptor subtype. 4. Comparing results in young and old rats, we observed numerous significant age-related changes, including a decrease in D2 receptors localized in rat heart and coronary artery wall. These D2 receptors show a specific location, in close relationship with dopaminergic nerve fibres. They decrease with age and their role remains unknown.

Age-related changes of the optic nerve fibres were studied in 3-month-old (young), 12-month-old (adult) and 24-month-old (aged) male Sprague-Dawley rats. The optic nerve was harvested with particular care from the intracranial portion. Cross sections from the optic nerve of animals of different age groups were stained with toluidine blue and examined under a light microscope at low and high magnification. Other sections were stained for the demonstration of glial cells using the method described by Holzer. A third group of sections were stained by the immunohistochemical method to detect glial fibrillary acidic protein, which is a marker for localising and characterising astrocytes. All these morphological results were subjected to the quantitative analysis of images and to statistical analysis of the values to identify significant morphometric data. Biochemical dosages of proteins were also performed on homogenised fragments of the optic nerve. Our results demonstrate that the following age-related changes can be observed: (1) an increase in meningeal membranes, (2) an increased number of astrocytes, (3) an increase in areal density of glial fibrillary acidic protein immunoreactivities, (4) an increased thickness of the entire optic nerve and an increased area of the nerve, (5) a decreased number of nerve fibres and (6) a decrease in the nerve fibre/meningeal membrane ratio from 3:1 to 1:1. Moreover, the amount of protein does not change with age. The rat optic nerve, therefore, appears sensitive to aging processes., (Copyright 2001 S. Karger AG, Basel)

Purpose: To describe the changes in rat retina occurring with ageing by means of histological methods, scanning electron microscopic observations and morphometrical data; and to study by means of biochemical methods the amount of protein content in retinal tissues., Methods: Samples of fresh retinal tissue obtained from young, adult, and aged rats were studied by means of traditional histological methods and by scanning electron microscopy. Particular attention was paid to morphometrical data and to the changes which occur with ageing. With the aid of a quantitative analysis of images, a large amount of morphometrical data was collected. Moreover, the amount of protein content in retinal tissues has been determined., Results: Retinal thickness significantly decreases with age. The ganglion cells seem to be more vulnerable to age-related loss than other retinal cells. The number of retinal capillaries is diminished with age. The intercellular connections between photoreceptors, the number of cellular processes, and the number of synaptic bodies of the bipolar cells also decrease significantly with age. These results were all confirmed by scanning electron microscopy observations and morphometrical findings. Biochemical dosage of proteins demonstrates that retinal tissues decrease with age., Conclusions: All morphological, morphometrical, ultrastructural and biochemical data are concordant in demonstrating that the retinal tissues of rats undergo specific changes with age. Our findings are in agreement with those described by previous authors and underline that the rat retina can be considered an optimal model for studies on neuronal maturation and/or neuronal ageing. Since our data have confirmed that many changes occur in rat retina with ageing, we can hypothesize that rat retina is particularly sensitive to developmental changes and to senile decay.

Acetylcholinesterase (AChE) localization in the human thymus has been studied by biochemical and morphological methods during development and aging. The occurrence, the amount and the distribution of acetylcholinesterase and the changes with age were examined in 24 human thymuses. The whole human thymus was removed during autopsies in males of the following age-groups: prenatal of six months, new-born, infant, young, adult and elderly. The thymuses were weighed, measured and dissected: the microanatomical details were stained with Eosin-orange, nervous structures were identified by means of Bodian's method. Protein content was determined with biochemical methods. Histoenzymatical and biochemical demonstration of acetylcholinesterase was performed. The morphological results obtained were submitted to quantitative image analysis. Our results show that the thymic microenvironment changes with age; moreover, an increase of acetylcholinesterase-positive structures can be observed with age. Biochemical results are in agreement with morphological results and both are confirmed by the outcome of quantitative analysis of images. Acetylcholinesterase activity in human thymus may play a key role in thymic functions.

The occurrence and distribution of GABA-transaminase (GABA-t) activity were examined in the thymus of juvenile, adult and aged rats, using enzyme-histochemical and biochemical methods. Quantitative image analysis showed that specific GABA-t reactivity was localized in the wall of the arteries and, to a lesser extent, to the veins. Only a low activity could be observed in association with the subcapsular and medullary part of the parenchyma of the thymus. Many structures resembling nerve fibers also showed low positive reactivity. Biochemical results gave the following decreasing order of GABA-t activity: arteries, veins, whole thymus and parenchyma. Histoenzymatic staining and related values of quantitative analysis of images, are in agreement with the biochemical results; moreover, they demonstrated that the intensity of histoenzymatic staining for GABA-t in thymus of rats strongly decreases with age. GABA-t in thymic tissue is concentrated in blood vessels and particularly in the arteries. Therefore, our findings do not support the earlier assumptions that GABA-t is exclusively concentrated in cerebral vessels. Moreover, the decrease of GABA-t activity during age in thymic tissues may be related to the decrease of thymic microvessels as consequence of the thymic involution. On the contrary, the thymic macrovessels show an elevated staining in all ages with an apparent increase due to the thymic involution.

The expression of gp75LNGFR (low-affinity receptor of the nerve growth factor (NGF) family of neurotrophins) immunoreactivity was studied in the cerebellar cortex of male Wistar rats of 3 months (young), 12 months (adult) and 24 months of age using immunohistochemical techniques and a monoclonal antibody against rat-gp75LNGFR. The percentage of Purkinje neurons displaying gp75LNGFR immunoreactivity (IR), and the intensity of gp75LNGFR IR in the cytoplasm of Purkinje and granule neurons, and in the neuropil of molecular layer of the cerebellar cortex were determined by quantitative image analysis and microdensitometry. The number of Purkinje neurons displaying gp75LNGFR IR and the intensity of gp75LNGFRIR in the cytoplasm of Purkinje neurons was significantly higher in rats of 12 months of age in comparison with 24- and 3-month-old rats. No significant changes were observed in the intensity of gp75LNGFRIR in the granule neuron layer of young, adult and old rats. In the molecular layer, the highest gp75LNGFRIR was found in young animals. It was reduced as a function of age. The present results show that changes in gp75LNGFRIR observed as a function of age affect to a different extent the three layers of the rat cerebellar cortex.

The rat cerebellar cortex represents an interesting animal model for the analysis of age-dependent changes in brain microanatomy and function. Moreover, the cerebellar cortex contains detectable amounts of nerve growth factor (NGF) and express NGF receptors, which are sensitive to aging. Previous studies of our group have shown that treatment with choline alfoscerate (alpha-glyceryl-phosphorylcholine) countered the loss of nerve cells and fibers occurring with age in the cerebellar cortex. The present study was designed to assess whether treatment for 6 months with a daily dose of 100 mg/kg of choline alfoscerate has any effect on the expression of NGF receptor immunoreactivity in male Wistar rats of 24 months of age. Twelve-month-old rats were used as an adult reference group. NGF receptor immunoreactivity which was developed in the 3 layers of the cerebellar cortex in adult rats was decreased in the neuropil of the molecular layer and in the cytoplasm of Purkinje neurons of rats of 24 months. The number of NGF receptor immunoreactive Purkinje neurons was also lower in the oldest age group, whereas the NGF receptor immunoreactivity in the cytoplasm of granule neurons was unchanged. Treatment with choline alfoscerate increased NGF receptor immunoreactivity in the molecular layer and in the cytoplasm of Purkinje neurons as well as the number of immunoreactive Purkinje neurons but was without effect on NGF receptor immunoreactivity in the granule neurons. These results suggest that choline alfoscerate treatment may increase the expression of NGF receptors in the rat cerebellar cortex.

The influence of aging on glutathione levels and distribution in the heart was studied in male Sprague-Dawley rats of 3 (young), 12 (adult) and 24 (old) months of age using biochemical and histofluorescence techniques, respectively. Biochemical assays of reduced glutathione (GSH) in the right and left ventricles and in the septum showed a significant decrease in GSH levels in adult in comparison with young animals. No further changes were noticeable between adult and old rats. GSH histofluorescence revealed a rather homogeneous distribution of the product of histochemical reaction within both right and left atria in 3-month-old rats. In 12-month-old rats a reduction of GSH histofluorescence in comparison with younger animals was noticeable. The loss is more consistent in the epicardial portion of the right atrium and in the endocardial region of the left atrium. In the atria of 24-month-old rats GSH reactivity was homogeneously distributed throughout the atrial wall and was significantly lower than in young or adult rats. In 3-month-old rats GSH histofluorescence was slightly lower in the epicardial than in the endocardial portions of both ventricles. In adult rats a significant decrease of GSH histofluorescence was noticeable in comparison with 3-month-old rats. The loss is particularly pronounced within the endocardial region of the left ventricle. In 24-month-old rats GSH histofluorescence showed no significant differences between adult rats. However, GSH was more homogeneously distributed throughout the ventricular wall than in adult animals. The significance of these data is discussed in relation to the role that GSH plays in protecting the myocytes against free radical damage.

The influence of ageing on the noradrenergic innervation of the ovary was studied in female Wistar rats using high pressure liquid chromatography (HPLC) with electrochemical detection and catecholamine histofluorescence techniques. Old age was accompanied by a significant decrease in ovarian noradrenaline levels. In young animals (3-month-old) noradrenergic nerve fibres were distributed primarily to blood vessels and in lesser amounts to the interstitial glands. In aged animals (24-month-old) perivascular noradrenergic fibres were reduced by more than 40%; interstitial gland nerve fibres were reduced by approximately 20%. The possibility that impaired noradrenergic ovarian innervation occurring in old age may be in some way related with age-dependent failure in reproductive activity is discussed.

The dopamine (DA) D-1 and D-2 receptors coupled to 3',5'-cyclic adenosine monophosphate (cAMP) generation were studied in membrane particles of the frontal cortex in young (3-month-old), adult (12-month-old) and aged (24-month-old) male Sprague-Dawley rats. Activation of D-1 receptors with DA, apomorphine or fenoldopam enhanced accumulation of cAMP in the frontal cortex of young rats. The stimulatory effect elicited by DA on cAMP generation declined by about 20% in adult rats. No further decline in cAMP accumulation was noticeable in aged animals. The response to dopaminergic agonists was blocked by the D-1 receptor antagonist SCH 29390 in the three age groups examined. The presence of D-2 receptors, negatively coupled to cAMP generation, was demonstrated by incubating frontal cortex membrane particles with SCH 23390 and then with DA. This inhibitory response, was also elicited with D-2 receptor agonists quinpirole or bromocriptine in the absence of SCH 23390 in which these compounds produced a decrease in cAMP. The decrease in cAMP caused following D-2 receptor stimulation was shown to be enhanced with age. No difference was observed between the three age groups of animals in the activation of cAMP production by forskolin. The present data suggest a selective decrease in the coupling between the D-1 receptor and cAMP generation in the frontal cortex of adult and aged rats and of an age-dependent increase in the coupling between the D-2 receptor and cAMP inhibition. The functional consequences of these biochemical changes may have important implications in the aging of the rat frontal cortex.

Vasoactive intestinal polypeptide (VIP) levels and distribution were studied in the penis of young-adult (3-month-old) and old (30-month-old) Wistar rats by radioimmunoassay and immunofluorescence. No significant changes in tissue VIP concentrations or distribution occurred in the penis of old rats compared to young-adult rats. The present data indicate that the age-related impairment of male sexual function is not dependent on modifications of the VIP-ergic innervation of penile tissue.

The effect of ageing on the density and pattern of adrenergic innervation of rat uterine tubes was studied using combined glyoxylic acid induced fluorescence and quantitative image analysis techniques. With age a reduction in the number of nerve fibers of the circular muscle layer of the isthmic portion was observed. No significant differences in the density of innervation were found at the level of blood vessels associated with uterine tubes.

Substance P levels in the lung of young (3-month-old), adult (12-month-old), senescent (20-month-old) and aged (28-month-old) male Wistar rats were determined by radioimmunoassay. Levels of substance P were constant until 28 months of age when these become significantly higher (approximately more than 2 times, p less than 0.001) than the values found in younger rats. Whether the increase of substance P content in the lung of old rats may reflect changes in the density of substance P containing nerve fibres or in the content of the peptide in single nerve fibres is discussed.

The effects of senescence and of long-term hydergine treatment on the density and the pattern of muscarinic cholinergic receptors in rat hippocampus were studied using combined radioreceptor binding and autoradiography with [3H]-quinuclidinyl-benzilate ([3H]-QNB) as a ligand. [3H]-QNB accumulated in the hippocampus of 3 month old rats, primarily in the stratum oriens of the CA1 and CA2 fields, followed in descending order by the molecular layer of granule cells of the dentate gyrus and the CA3 field. The density and pattern of [3H]-QNB binding sites in the hippocampus of 16 month old rats were the same. In contrast, the density of [3H]-QNB binding sites in the hippocampus of 22 months old rats was significantly reduced, mainly in the granule cells of the dentate gyrus, next in the CA3 and in the CA1-CA2 fields. Hydergine treatment (0.6 mg/kg and 1 mg/kg, p.o.), started at 16 months of age and continued for 6 months, caused a dose-related restoration of the number of [3H]-QNB binding sites, primarily in the CA1 and CA2 fields and in lesser amounts of the dentate gyrus. The possibility that the therapeutic effects of hydergine administration on memory function in the elderly might be related to an effect on cholinergic muscarinic receptors of the ageing hippocampus is discussed.

The effect of ageing on the density and pattern of noradrenergic nerves in the perivascular nerve plexus supplying the vasa nervorum of the rat sciatic nerve was studied using combined catecholamine histofluorescence and quantitative image analysis techniques. The density of noradrenergic fibres around arteries and arterioles of the rat sciatic nerve vasa nervorum increased in the old animals. In contrast, no changes in perivascular nerve fibres supplying the veins and venules were found in the vasa nervorum of old rats. The increase in old age of noradrenergic innervation of arteries and arterioles of the vasa nervorum may be related to the pathogenesis of some peripheral nerve diseases.