Expression of certain proteins in the subfornical organ and cerebrospinal fluid of spontaneously hypertensive rats.

The objective of this study was to analyze the proteins in the cerebrospinal fluid of spontaneously hypertensive rats and to study their possible role in the relationship between hydrocephalus, arterial hypertension and variations in the subfornical organ. Brains and cerebrospinal fluid from control Wistar-Kyoto rats and spontaneously hypertensive rats sacrificed with chloral hydrate were used. Cerebrospinal fluid and extract of subfornical organ were processed by protein electrophoresis. Antisera against protein bands of 141, 117 and 48 kDa and Concanavalin A were used for immunohistochemical and western blot study of the subfornical organ, adjacent circumventricular structures and cerebrospinal fluid. Ventricular dilation in the spontaneously hypertensive rats and the presence of quite a lot of protein bands in the cerebrospinal fluid of the hypertensive rats, which were either not observed or scarcely present in the cerebrospinal fluid of the Wistar-Kyoto rats, were confirmed. The subfornical organ, third ventricle ependyma and choroideus plexus showed immunoreactive material for antibodies against 141kDa, 117 and 48 kDa proteins band (anti-B1, anti-B2 and anti-B3). The larger amount of the immunoreactive material was found in the subfornical organ of the spontaneously hypertensive rat. Our results and the alterations observed by other authors in the subfornical organ in hydrocephalic and hypertensive rats support the possibility that this circumventricular organ, some proteins of the cerebrospinal fluid and ventricular dilation could be connected with the physiopathology of this type of hypertension.

Alterations of the cerebrospinal fluid proteins and subcommissural organ secretion in the arterial hypertension and ventricular dilatation. A study in SHR rats.

The aim of this work was to analyze the proteins in the cerebrospinal fluid (CSF) of spontaneously hypertensive rats, to study their possible role in the relationship between hydrocephalus, arterial hypertension and alterations in the subcommissural organ. Brains from control Wistar-Kyoto rats (WKY) and spontaneously hypertensive rats (SHR) sacrificed with chloral hydrate were used. Antiserums against some cerebrospinal fluid protein bands and Reissner's fiber (RF) were used for immunohistochemical study of the SCO. Ventricular dilation was observed in the lateral and third ventricle of the SHR. Third ventricle ependyma showed immunoreactive material (IRM) for antibody against 141 kDa protein band anti-B1 and 117 protein band anti-B2 and the SCO of the SHR showed a decrease of the IRM when compared with WKY rats. An alteration in the expression of anti-RF was found to compare the SCO of the WKY and SHR groups. Our results demonstrate that hydrocephalus and hypertension are interconnected in this kind of rat which produce alterations in SCO secretions and some proteins of the CSF.

Postnatal development of the secretory activity of the goat subcommissural organ. A Reissner's fibre and angiotensin II immunohistochemical study.

Subcommissural organ (SCO) secretory activity of the goat (variations of Capra hircus, that live in arid conditions) was examined during the postnatal development, using specific antibodies against the Reissner's fibre (AFRU) and angiotensin II (AAGII). The SCO was strongly stained with the anti-glycoproteins that form the Reissner's fibre and lightly marked with the anti-angiotensin II. The AFRU-immunoreactivity (ir) was found in the ependymal and hypendymal cells and in the ventricular and peripheral secretory routes of the goat SCO. The amount AFRU increases at 6 months and decreases at adult age. In contrast, the anti-angiotensin II-ir was mainly found in the adult age, not being practically observed at one postnatal month. The AAGII-ir was mainly found in ependymal cells in which AFRU-ir was downregulated. In addition, we detected the presence of double immunostained for AFRU and AAGII in ependymocytes of the pre-commissural and subcommissural parts. In conclusion the present results may suggest a functional interrelation between AAGII and the secretory activity of the SCO of this kind of goat.

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.

Ontogenic development of the human subcommissural organ

The structure of the human subcommissuralorgan during its ontogenic development in 24human embryos and foetuses ranging from 6 to40 weeks of gestation (WG), and three adulthuman brains from 27-, 65- and 70-year old subjectswas investigated using both qualitative andquantitative methods. Concurrently, the appearanceof the subcommissural organ, pineal glandand mesocoelic recess was determined by studyingtheir structure, length and volume. Thehuman SCO appears at the beginning of 8th WG,which confirms previous results; the completematuration of the SCO occurs at the 15th WG andthe following three parts can be distinguished:the precommissural part, located in the rostralzone of the posterior commissure (PC) andextending to the pineal recess; the subcommissuralpart, located under the PC, and the retrocommissuralpart, located in the caudal zone ofthe PC, in the mesocoelic recess and at thebeginning of the Sylvian aqueduct. The reductionin size of the SCO begins after the 17th WGand this decrease in size begins in the precommissural,continues in the subcommissural, andfinishes in the retrocommissural part. The regressionand atrophies of the SCO begin after birth,and the SCO disappears completely after the ageof 30. The mesocoelic recess starts to form at thebeginning of the 10th WG, and is completely formedby the 14th WG and this is where the retrocommissuralpart of the SCO is located. In the 40th WG the regression of the mesocoelic recessbegins and this takes place at the same time asthe regression of the SCO. A parallel developmentbetween the SCO and the pineal wasfound. Thus, we observed the first appearance ofthe pineal recess in the 7-8th WG; during the 10thWG a compact mass of cells appeared in the rostralpart of pineal recess and by the 15th WG thepineal gland (PG) had acquired an almost definitiveaspect

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Connection between the arcuate nucleus and the posterior lobe of the hypophysis. An angiotensin II immunohistochemical study in rats

The aim of this study was to perform an immunohistochemical study of the, angiotensinergic pathway from the arcuate nucleus (AN) to the posterior lobe of the hypophysis (PLH) of 10-week-old matched normotensive Wistar Kyoto rats (WKY), using our own policlonal antibody raised in mice against Angiotensin II (mouseantiangiotensin II, MAAII). Cells and fibers in the rostroventral and dorsocaudal parts of the AN, the internal zone of the median eminence and PLH showed immunoreactive material for antiangiotensin II. Angiontensin II fibers originating in the anteroventral part of the AN, crossing median eminence (ME) and infundibular stem and arriving at the PLH were also observed (AU)

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Somatotipo de referencia de luchador canario / Canarian wrestler reference somatotype

La mayoría de las modalidades de lucha clasifican a los practicantes agrupándolos por peso. En Lucha Canaria los mejores luchadores son denominados "puntales" (A, B y C), sin considerar sus pesos. Existen pocos trabajos morfológicos sobre luchadores de la modalidad canaria, lo que supone una falta de información científica que podría limitar su rendimiento. No existe un Somatotipo de Referencia (SR) para éstos luchadores, de ahí que el principal objetivo de este trabajo sea establecer el citado somatotipo. Se han medido doscientos diecinueve (219) luchadores siguiendo las directrices de la International Society for the Advancement of Kinanthropometry (ISAK) y el Grupo Español de Cincantropometría (GREC). Con ellos se hicieron cuatro grupos (A, B, C y NC) atendiendo a la clasificación establecida por su rendimiento en competición. El grupo A lo constituyeron 5 luchadores puntales A, que es la categoría más alta en este deporte. El grupo B lo fonnaron 9 luchadores puntales B, el grupo C 28 luchadores puntales C y, finalmente, los luchadores No Clasificados (grupo NC) que fueron 177 individuos tomados al azar. Estos últimos fueron además divididos en 5 subgrupos atendiendo a su peso: 60K, 70K, 8OK, 90K y mas de 1OOK.El SR se calculó utilizando el Método Antropométrico de Heath- Carter. Nuestro estudio puso de manifiesto que los luchadores más viejos eran los puntales B, y los no clasificados los más jóvenes. Los puntales A eran los más altos y los más pesados. Todos ellos fueron endo-mesomorfos, con la excepción de¡ subgrupo 60K de no clasificados (ecto-mesomorfo). Globalmente, el componente mesomorfo fue dominante, y la endomorfj'a mayor que la ectomorfia. Podemos concluir que, sin considerar el éxito en competición, la fonna actual de clasificación otorga el rango de punta¡ a aquellos luchadores con un somatotipo de (5.9-8.9-0. l). En ambos casos, puntales y no clasificados, la distribución de los somatotipos según el componente dominante reveló no ser muy similares, como se desprende del SDD y de¡ SAD. Sin embargo, se requieren nuevos estudios para detemiinar el SR de los no clasificados (AU)

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.

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.

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.

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.

Sources of GABAergic input to the inferior colliculus of the rat.

We have studied the GABAergic projections to the inferior colliculus (IC) of the rat by combining the retrograde transport of horseradish peroxidase (HRP) and immunohistochemistry for gamma-amino butyric acid (GABA). Medium-sized (0.06-0.14 microliter) HRP injections were made in the ventral part of the central nucleus (CNIC), in the dorsal part of the CNIC, in the dorsal cortex (DCIC), and in the external cortex (ECIC) of the IC. Single HRP-labeled and double (HRP-GABA)-labeled neurons were systematically counted in all brainstem auditory nuclei. Our results revealed that the IC receives GABAergic afferent connections from ipsi- and contralateral brainstem auditory nuclei. Most of the contralateral GABAergic input originates in the IC and the dorsal nucleus of the lateral lemniscus (DNLL). The dorsal region of the IC (DCIC and dorsal part of the CNIC) receives connections mostly from its homonimous contralateral region, and the ventral region from the contralateral DNLL. The commissural GABAergic projections originate in a morphologically heterogeneous neuronal population that includes small to medium-sized round and fusiform neurons as well as large and giant neurons. Quantitatively, the ipsilateral ventral nucleus of the lateral lemniscus is the most important source of GABAergic input to the CNIC. In the superior olivary complex, a smaller number of neurons, which lie mainly in the periolivary nuclei, display double labeling. In the contralateral cochlear nuclei, only a few of the retrogradely labeled neurons were GABA immunoreactive. These findings give us more information about the role of GABA in the auditory system, indicating that inhibitory inputs from different ipsi- and contralateral, mono- and binaural auditory brainstem centers converge in the IC.

Alcohol intake effects on the dorsal vagal complex of the rat: a cellular morphometric study.

We have analyzed the morphometric effects on the dorsal vagal complex (DVC) of the rat of alcohol exposure and/or hypoproteic diet intake during 8 weeks. In the area postrema (AP), alcohol treatment (combined with normal isoproteic or hypoproteic diet) caused a significant decrease in karyometric parameters. In the dorsal motor nucleus of the vagus (DMV) and nucleus tractus solitari (NTS), the alcohol isoproteic intake (AI) produced an increase in neuron size (expressed by an increase in the neuronal nuclear area and the cell/neuropil coefficient). The hypoproteic diets produced a reduction in the global volume of each structure of the DVC which was accompanied by a decrease in global brain volume. These results indicate that after 8 weeks of treatment, alcohol is the main cause of the morphometric alteration found in the DVC, while variations in the amount of protein intake appear to produce global effects on the whole brain.

Effects of chronic alcohol exposure on the morphometric development of medial preoptic area neurons of the male mouse.

We have performed a karyometric study of the medial preoptic area of male mice from mothers that ingested chronically a solution of 20% of ethanol added to the drinking water. Pups then were exposed prenatally to alcohol. After parturition, pups were also exposed to alcohol, first through their mother's milk and after weaning by direct ingestion of the same solution of 20% of alcohol until the day of sacrifice. Animals were sacrificed at the 25th, 35th, 45th, 55th and 100th day and the results compared with those obtained in another group of control animals, sacrificed at the same ages. Chronic alcohol exposure reduces the studied nuclear sizes (perimeter, area and maximum diameter) in adult animals of 100 days of life, but does not produce significative changes in nuclear sizes of younger animals. However, nuclear shape, another of the nuclear parameters analysed, did show significative alterations in relation with the puberal age. These morphometric effects could be due to the reduction of plasmatic testosterone levels produced by alcohol and/or to a direct toxic effect of the alcohol on central nervous system neurons.

Postnatal development of NADPH-diaphorase activity in the superior colliculus and the ventral lateral geniculate nucleus of the rat.

We have studied the postnatal development of dihydronicotinamide adenine dinucleotide phosphate diaphorase (NADPH-d) activity in the superior colliculus (SC) and the ventral lateral geniculate nucleus (LGv) of the rat. We describe two different developmental patterns of NADPH-diaphorase activity. The first pattern, observed in the deep layers of the SC, shows a transient activity during the first week which progressively decreases during the following two weeks. The second pattern is observed in the superficial layers of the SC and in the LGv. They become positive during the first week, their NADPH-d activity increases progressively during the second and third weeks, reaching the adult pattern at the fourth week. On the whole, the developmental chronology of the laminar distribution of NADPH-d in the SC displays an inside-out pattern. Our results suggest that NADPH-d activity may play different roles at different stages of the developing nervous system.

The effects of chronic administration of captopril on the mouse subfornical organ and area postrema.

We have studied by morphometric procedures the chronic effect of captopril on the subfornical organ (SFO) and area postrema (AP) of the adult mouse. Oral administration of captopril does not produce any change in the size of individual nuclei of the ependymocytes and neurons in both centers. However, there are other quantitative effects of captopril on the global volume of the SFO and on the neuropil and vascular elements of both the SFO and AP which present a significant increase. It is suggested that this increase is due to metabolic processes at the level of both circumventricular organs.

Alcohol effects on the morphometric development of the subfornical organ and area postrema of the albino mouse.

We have studied the development of the nuclear sizes of ependymocytes and neurons of two circumventricular organs of the male alcoholic mouse: the Subfornical Organ (SFO) and the Area Postrema (AP), comparing the results with a control group. The global volume of both centers was also studied. The results show that the SFO, a structure related to the control of fluid balance, responds to alcoholism with an increase of the global volume. This increase could be related to the variations of salt-water balance and/or blood pressure in chronic alcoholism. However, the size of cell nuclei in the SFO is not affected. In contrast, the AP responds to chronic alcoholism like other nervous centres, with a decrease of the nuclear size of its cells. The global volume of AP does not change.

Development of the median eminence in the male mouse. Karyometric effect of neonatal and prepuberal castration.

We have studied the karyometric development of the ependymocytes of the median eminence and ependyma adjacent of the arcuate nucleus in a control group of male albino mice from the 5th to the 160th postnatal day, and in other two experimental groups of animals that were castrated at two different days: the first and the 20th day of life. We have found differences in the spontaneous development of both ependymocytes showing the median eminence ependyma a more closely relationship to changes of gonadal hormone levels around puberty. In both zones, the response of the ependymocytes to neonatal castration was clearly more significative than that obtained after prepuberal castration, with lower values in the castrated animals than in the control mice. We suggest that this could be related to nervous and hormonal mechanisms.

Effects of postnatal administration of ethanol on the M. gastrocnemius of the albino mouse.

A combined morphometric-histochemical (mATPase) study of the effects of ethanol during postnatal development on the m. gastrocnemius has been performed in the albino mouse. The experimental group received ethanol in the drinking water until sacrifice at the age of 40 days. Based on the fiber composition, three different areas are distinguished in the m. gastrocnemius of the mouse. The typical location of these areas does not change after ethanol administration. However, postnatal administration of ethanol produces a selective atrophy and a decrease of the number of type IIb (fast glycolytic) fibers. Concurrently, the number of type IIa (fast oxidative-glycolytic) fibers increases.