Variations of the grid and place cells in the entorhinal cortex and dentate gyrus of 6 individuals aged 56 to 87 years.

INTRODUCTION: The relationship between the entorhinal cortex (EC) and the hippocampus has been studied by different authors, who have highlighted the importance of grid cells, place cells, and the trisynaptic circuit in the processes that they regulate: the persistence of spatial, explicit, and recent memory and their possible impairment with ageing. OBJECTIVE: We aimed to determine whether older age causes changes in the size and number of grid cells contained in layer III of the EC and in the granular layer of the dentate gyrus (DG) of the hippocampus. METHODS: We conducted post-mortem studies of the brains of 6 individuals aged 56-87 years. The brain sections containing the DG and the adjacent EC were stained according to the Klüver-Barrera method, then the ImageJ software was used to measure the individual neuronal area, the total neuronal area, and the number of neurons contained in rectangular areas in layer III of the EC and layer II of the DG. Statistical analysis was subsequently performed. RESULTS: We observed an age-related reduction in the cell population of the external pyramidal layer of the EC, and in the number of neurons in the granular layer of the DG. CONCLUSION: Our results indicate that ageing causes a decrease in the size and density of grid cells of the EC and place cells of the DG.

Changes in the choroid plexuses and brain barriers associated with high blood pressure and ageing.

INTRODUCTION: The choroid plexuses, blood vessels, and brain barriers are closely related both in terms of morphology and function. Hypertension causes changes in cerebral blood flow and in small vessels and capillaries of the brain. This review studies the effects of high blood pressure (HBP) on the choroid plexuses and brain barriers. DEVELOPMENT: The choroid plexuses (ChP) are structures located in the cerebral ventricles, and are highly conserved both phylogenetically and ontogenetically. The ChPs develop during embryogenesis, forming a functional barrier during the first weeks of gestation. They are composed of highly vascularised epithelial tissue covered by microvilli, and their main function is cerebrospinal fluid (CSF) production. The central nervous system (CNS) is protected by the blood-brain barrier (BBB) and the blood-CSF barrier (BCSFB). While the BBB is formed by endothelial cells of the microvasculature of the CNS, the BCSFB is formed by epithelial cells of the choroid plexuses. Chronic hypertension induces vascular remodelling. This prevents hyperperfusion at HBPs, but increases the risk of ischaemia at low blood pressures. In normotensive individuals, in contrast, cerebral circulation is self-regulated, blood flow remains constant, and the integrity of the BBB is preserved. CONCLUSIONS: HBP induces changes in the choroid plexuses that affect the stroma, blood vessels, and CSF production. HBP also exacerbates age-related ChP dysfunction and causes alterations in the brain barriers, which are more marked in the BCSFB than in the BBB. Brain barrier damage may be determined by quantifying blood S-100ß and TTRm levels.

Variations of the grid and place cells in the entorhinal cortex and dentate gyrus of 6 individuals aged 56 to 87 years. / Variaciones de las células de cuadrícula y de posicionamiento de la corteza entorrinal y del giro dentado de 6 humanos de 56 a 87 años.

INTRODUCTION: The relationship between the entorhinal cortex and the hippocampus has been studied by different authors, who have highlighted the importance of grid cells, place cells, and the trisynaptic circuit in the processes that they regulate: the persistence of spatial, explicit, and recent memory and their possible impairment with ageing. OBJECTIVE: We aimed to determine whether older age causes changes in the size and number of grid cells contained in layer III of the entorhinal cortex and in the granular layer of the dentate gyrus of the hippocampus. METHODS: We conducted post-mortem studies of the brains of 6 individuals aged 56-87 years. The brain sections containing the dentate gyrus and the adjacent entorhinal cortex were stained according to the Klüver-Barrera method, then the Image J software was used to measure the individual neuronal area, the total neuronal area, and the number of neurons contained in rectangular areas in layer III of the entorhinal cortex and layer II of the dentate gyrus. Statistical analysis was subsequently performed. RESULTS: We observed an age-related reduction in the cell population of the external pyramidal layer of the entorhinal cortex, and in the number of neurons in the granular layer of the dentate gyrus. CONCLUSION: Our results indicate that ageing causes a decrease in the size and density of grid cells of the entorhinal cortex and place cells of the dentate gyrus.

Changes in the choroid plexuses and brain barriers associated with high blood pressure and ageing. / Variaciones de los plexos coroideos y las barreras cerebrales en la hipertensión arterial y el envejecimiento.

INTRODUCTION: The choroid plexuses, blood vessels, and brain barriers are closely related both in terms of morphology and function. Hypertension causes changes in cerebral blood flow and in small vessels and capillaries of the brain. This review studies the effects of high blood pressure (HBP) on the choroid plexuses and brain barriers. DEVELOPMENT: The choroid plexuses (ChP) are structures located in the cerebral ventricles, and are highly conserved both phylogenetically and ontogenetically. The ChPs develop during embryogenesis, forming a functional barrier during the first weeks of gestation. They are composed of highly vascularised epithelial tissue covered by microvilli, and their main function is cerebrospinal fluid (CSF) production. The central nervous system (CNS) is protected by the blood-brain barrier (BBB) and the blood-CSF barrier (BCSFB). While the BBB is formed by endothelial cells of the microvasculature of the CNS, the BCSFB is formed by epithelial cells of the choroid plexuses. Chronic hypertension induces vascular remodelling. This prevents hyperperfusion at HBPs, but increases the risk of ischaemia at low blood pressures. In normotensive individuals, in contrast, cerebral circulation is self-regulated, blood flow remains constant, and the integrity of the BBB is preserved. CONCLUSIONS: HBP induces changes in the choroid plexuses that affect the stroma, blood vessels, and CSF production. HBP also exacerbates age-related ChP dysfunction and causes alterations in the brain barriers, which are more marked in the BCSFB than in the BBB. Brain barrier damage may be determined by quantifying blood S-100ß and TTRm levels.

Effect of the arterial hypertension and captopril treatment on the angiotensin II content in the subfornical organ. A study in SHR rats.

We studied the effects of spontaneous high blood pressure and the captopril treatment on the subfornical organ (SFO) of rats. The brains of control Wistar-Kyoto rats (WKY), WKY rats treated with captopril (WKY-T), spontaneously hypertensive rats (SHR) and SHR rats treated with captopril (SHR-T) were processed immunohistochemically using anti-angiotensin II as primary antibody. Immunorective material (IRM) for angiotensin II was observed in a group of neurons and some cells of the ependymal layer of the SFO in WKY rats. The angiotensin II immunoreactive (AGII-ir) in the SHR rats was decreased, showing positive reaction only in a few neurons, while captopril treatment induced an increase in immunoreactive material in hypertensive rats, but contrarily, the expression of AGII-ir in the WKY-T group was scarce. The variations of the angiotensin II observed in the SFO could be owing to an interaction between the hypertension and its captopril treatment.

Effect of hypertension on the angiotensin II fibres arriving at the posterior lobe of the hypophysis of the rat. An immunohistochemical study.

We studied immunohistochemically the posterior lobe of the hypophysis (PL) of 15-week-old spontaneously hypertensive rats (SHR) and of matched normotensive Wistar Kyoto rats (WKY), by using our own polyclonal antibody raised in mice against Angiotensin II (mouse-antiangiotensin II, MAAII). The blood pressure, water intake and volume of the PL were also recorded. The SHR rats were hypertensive, drank more water and showed a clear hypertrophy of their hypophysial PL. Also the PL of the SHR animals showed an increase in the immunoreactivity to the anti-angiotensin II antibody in the fibres arriving at the PL, with respect to the PL of WKY rats. This increase is compatible with the hyperactivity of the brain RAS, depletion of vasopressin content in the PL and increase in plasmatic levels of vasopressin described in SHR rats with respect to normotensive animals, as angiotensin II could locally stimulate vasopressin release to plasma from the neurohypophysis.

Effects of alcohol and aging on the cingular (area 24) and frontal (area 6) cortical areas of the mouse.

We have studied the morphometric changes of the neurons of the cingular area 24 and frontal area 6 of the mouse, produced by age and/or chronic alcohol intake. The parameters analyzed were nuclear area of these cortical neurons and cellular density (cell/neuropil coefficient). We detected a decrease in the number of neurons with age in practically all layers of the control animals. In the animals that chronically ingested the alcoholic solution, we also detected a decrease in the number of neurons with age, but only in layer V of the frontal cortex and in layer VI of the cingular area 24. The comparison between the control and the alcoholic group showed that alcohol intake caused an increase in the nuclear area of the neurons in layer II-III of the frontal cortex at 180 days, while in the cingular cortex the increase in nuclear area of its neurons was significative at 180 days in layer II-III and at 35 and 180 days in layers V and VI. We think that these changes are the expression of the neuronal plasticity in both cortical areas in response to the alcohol exposure.

The effects of chronic administration of captopril on the mouse median eminence.

The effects of Captopril (an angiotensin-converting enzyme inhibitor) on the median eminence (ME) of the male albino mouse have been examined using morphometric and immunohistochemical procedures. We measured the nuclear area of the ependymocytes of the ME and of the glial cells of the reticular external zone of the ME. We also determined the cell/neuropil coefficient (CNC), which expresses the relation between cellular area and neuropil of the ME, and the global volume of the ME in each animal. For the immunohistochemical study we used rabbit antiarginine-vasopressin, and compared the results in the different groups of mice. We detected an increased in the immunoreactive material (arginine-vasopressin, A-V) and an increase in the global volume of the organ and also an increase of the neuropil of the ME after the longest exposure to the drug. These alterations could be related to the inhibition of the brain angiotensin II by captopril and the accumulation of vasopressin in the fibrous tract that runs from the paraventricular nucleus (PVN) to the neurohypophysis.

Effect of hypertension and captopril treatment on the vasopressin in the rat median eminence and posterior lobe of the hypophysis. An immunohistochemical study.

The present study analyses the effects of hypertension and/or its oral treatment with captopril (angiotensine-converting enzyme inhibitor) on the rat median eminence (ME) and the posterior lobe of the hypophysis (PL). After an immunohistochemical reaction using an antibody against arginine-vasopressin, we compared by densitometry the amount of vasopressin immunoreactive material (vasopressin-ir) of these centers in 4 groups of animals: control Wistar Kyoto rats (WKY), spontaneously hypertensive rats (SHR), WKY rats treated with captopril (WKY-T) and SHR rats also treated with the same drug (SHR-T). Captopril was administrated at a dosage of 0.1 mg/ml in the drinking water from the 8th to the 15th weeks. We have found that the rats showing the lowest level of vasopressin-ir, in both ME and PL, were those from the SHR group, the concentration increasing after oral captopril treatment (SHR-T), although without reaching the values of WKY rats. Then, ACE inhibition by captopril influences vasopressin content in brain areas where the hormone is concentrated before being released, which supports the hypothesis that suggests a central modulatory effect of ACE inhibitors, contributing to their therapeutic action on hypertension.

Postnatal development of the Ammon's horn (CA1 and CA3 fields). A karyometric and topographic study.

We have performed a karyometric study of the pyramidal neurons of CA1 and CA3 fields of the Ammon's horn, in male mice aged from the 5th to the 190th postnatal day. Nuclear sizes were measured with the aid of a Magiscan Analysis System, used in an interactive form, in both superficial and deep layers of the stratum pyramidal in those fields. The measurements were made at three different topographic levels: rostral; intermediate; and caudal, to detect any possible difference related to the topography of the neuron in the same field. We have found that both CA1 and CA3 fields are correlated in the postnatal development of their nuclear pyramidal sizes and that all topographic levels of the hippocampus reach their highest karyometric sizes at the 10th-15th postnatal day. Caudal levels show higher karyometric values than the other levels and some differences between neurons of the superficial and deep layers of both fields are also described here and analysed in relation to the different ontogenetic gradients of these cells.

Cortical projections of the nucleus centralis superior and the adjacent reticular tegmentum in the mouse.

After horseradish peroxidase-injections into different cortical areas of the mouse, labeled neurons were observed in the n. centralis superior and in the bilateral adjacent reticular tegmentum, especially in the zone dorsal to the medial lemniscus and the n. reticularis tegmenti pontis. Labeled cells in the n. centralis superior were nearly always concurrent with labeled paramedian cells, the topographical distribution of which was independent of the site of injection.

Quantitative development of the subcommissural organ in hypothyroid mice.

The postnatal development of the global volume of the subcommissural organ (SCO) and of the karyometric changes of the ependymocytes in the SCO and the adjacent ventricle is studied in male albino mice aged from 25 to 160 days, and in a hypothyroid group treated with propylthiouracil with and without interruption of the treatment at 35 days. Hypothyroidism produces a decrease of the global volume of the SCO and of the nuclear size of the ependymocytes in the SCO and the adjacent ventricle.

Direct projections from the reticular formation of the medulla oblongata to the anterior cingulate cortex in the mouse and the rat.

After injections of horseradish peroxidase into the anterior cingulate cortex of the mouse and the rat, labeled neurons were observed in different nuclei of the reticular formation of the rostral medulla oblongata: ventral part of the nucleus gigantocellularis, nucleus paragigantocellularis and nucleus lateralis reticularis. Other labeled neurons lay scattered outside the nuclear boundaries, often close to the ventral-most border of the medulla. The projection to the cingulate cortex is sparse, with rarely more than 3 cells per case, but constant. Injections into other cortical areas (visual, motor and somatosensory cortex) did not produce labeling in the medullary reticular formation.

Effects of hypothyroidism on the karyometric development of the paraventricular and ventromedial nuclei of the hypothalamus in the mouse.

We studied the karyometric development of 4 subdivisions of the paraventricular nucleus (rostral, caudal, medial and lateral) and 3 subdivisions of the ventromedial nucleus of the hypothalamus (ventral, central and dorsal) in a hypothyroid group of male albino mice treated with propylthiouracil with or without interruption of the treatment at the 35th postnatal day. Hypothyroidism produces an increase of the nuclear-size values of the paraventricular and ventromedial nuclei, and the continued treatment increases this effect. The caudal and the rostral subdivisions of the paraventricular nucleus show more significant changes of their nuclear sizes compared to the medial and lateral subdivisions. The ventromedial nucleus responds similarly to hypothyroidism in all its subdivisions.

Alterations of the subcommissural organ in the hydrocephalic human fetal brain.

We have studied the subcommissural organ of two hydrocephalic brains, of 20 and 21 gestational weeks and of two normal brains, aged 19 and 23 gestational weeks. Both hydrocephalic cases presented a size reduction of the subcommissural organ compared to the normal cases; only in one case, there were also alterations of the morphological components of the subcommissural organ, suggesting different pathogenic relationships between hydrocephalus and dysplasia of the subcommissural organ.

Karyometric changes in the subcommissural organ of male mice after gonadectomy.

A karyometric study of the ependymal cells of the subcommissural organ (SCO) was made in male albino mice castrated at 20 days and examined at 25, 35, 45, 55 and 85 days of life, and in control groups of the respective ages. The values of the nuclear areas and perimeters of SCO cells as well as those of the global volume of the SCO are significantly lower in the experimental groups, and the nuclear membrane shows less invaginations. No changes are observed after gonadectomy in the subjacent thalamic ependyma.

Brainstem projections to the medial preoptic region containing the luteinizing hormone-releasing hormone perikarya in the rat. An immunohistochemical and retrograde transport study.

The afferent projections to the anterior medial preoptic area (MPA) from the brainstem have been studied, in female Wistar rats, by retrograde tracing with horseradish peroxidase (HRP). The HRP was injected by iontophoresis into the preoptic region containing the luteinizing hormone-releasing hormone (LHRH) perikarya. The brain sections including the MPA were reacted with diaminobenzidine (DAB) to reveal the injection site; the LHRH cells were then immunohistochemically identified using DAB with ammonium nickel sulphate. When the injection site incorporated the LHRH cells, the brainstem sections were reacted with the DAB nickel solution to detect lysosomal HRP and then immunohistochemically processed to locate the adrenaline-synthesizing cells using DAB alone. The results confirm the brainstem projections to the MPA from the central grey matter, ventral tegmental area, subcoeruleus area, the dorsal raphe nucleus, the lateral parabrachial nucleus, the raphe pontis nucleus, the raphe obscurus nucleus, the region of the paragigantocellular nucleus and the nucleus of the solitary tract. Given the considerable evidence implicating the ascending adrenergic systems in the regulation of LHRH, we focused our attention on the afferents from the locus coeruleus, area postrema and the adrenaline-synthesizing cell groups (C1-3). The only cells which were retrogradely labelled and immunopositive for phenylethanolamine N-methyltransferase were found in C3.

Changes in the secretory activity of the subcommissural organ of spontaneously hypertensive rats.

The subcommissural organ (SCO) is a glandular circumventricular organ secreting glycoproteins into the cerebrospinal fluid. The SCO of 15-week-old spontaneously hypertensive rats (SHR) and of matched normotensive Wistar-Kyoto rats (WKY) was studied immunocytochemically by using an antibody against the glycoproteins secreted by the SCO. The blood pressure, water intake and volume of brain ventricles of SHR and WKY rats were also recorded. The SHR were hypertensive, drank more water and did not display dilatation of the brain ventricles. The SCO of the SHR rats showed a drastic decrease of the immunoreactive material stored in the rough endoplasmic reticulum whereas the amount of immunoreactive apical secretory granules did not vary with respect to the SCO of WKY rats. These changes are compatible with an increased secretory activity of the SCO of the SHR rats. It is suggested that the changes in the SCO of SHR rats, and their hypertensive state, are interrelated phenomena.

Morphometric changes in alcoholic mice of neurons of areas 6 and 17 and ependyma of the subcommissural organ.

We have performed a karyometric study of the postnatal development of the subcommissural organ (SOC), the subjacent thalamic ependyma, and the pyramidal neurons of layer V of the motor and visual cortical areas, in alcoholic male albino mice, aged 25-100 days. Ethyl alcohol was added to the drinking water at a concentration of 20%, from birth until the day of sacrifice. Our results show that alcoholism affects the SOC ependyma and the motor cortex, and, apparently, does not affect the ventricular thalamic ependyma and the visual cortex. We suggest a neuroendocrine mechanism to explain the SOC answer, and a functional deprivation to explain the changes in the motor cortex.

Effect of ethanol on liver cell-development in the male albino mouse.

The aim of this study is to determine the influence of ethanol on the development of periportal and pericentral hepatocytes and their nuclei, comparing it with the development of these cells and nuclei in a control, age-matched population. In male albino mice fed with 20% ethanol added to drinking water and sacrificed at day 25, 35, 85 and 180 of life haematoxylin-eosin stained liver specimens were studied with the aid of a LEITZ ASM semiautomatic autoanalyzer. The nuclear area of at least 25 hepatocytes with evident nucleoli was measured at each of 3 periportal and 3 pericentral fields. The area of the cells of the same fields was estimated by dividing the area of the fields by the number of hepatocyte nuclei present in them. Both cellular and nuclear areas of periportal and pericentral hepatocytes of the 25-day-old animals were smaller than those of age-matched controls. By contrast, the experimental group showed a striking increase both in cellular and nuclear size in the pericentral hepatocytes, but not in the periportal ones. Thus, ethanol seems to inhibit the initial growth of cells and nuclei; but, after maturation, it causes a marked increase not only in cellular size of pericentral hepatocytes, especially in 180-day-old animals, but also in nuclear size, already evident in the 35-day-old mice.