Miriam González

Distribution patterns of estrogen receptor α and β in the human cortex and hippocampus during development and adulthood

The expression of estrogen receptors (ERs) in the developing and adult human brain has not been clearly established, although estrogens are crucial for neuronal differentiation, synapse formation, and cognitive functions. By using immunohistochemistry, we have studied the distribution of ERα and ERβ in human cerebral cortex and hippocampus from early prenatal stages to adult life. ERα was detected in the cortex at 9 gestational weeks (GW), with a high expression in proliferating zones and the cortical plate. The staining intensity decreased gradually during prenatal development but increased again from birth to adulthood. In contrast, ERβ was first detected at 15 GW in proliferating zones, and at 16/17 GW, numerous ERβ immunopositive cells were also observed in the cortical plate. ERβ expression persisted in the adult cortex, being widely distributed throughout cortical layers II–VI. In addition, from around 15 GW to adulthood, ERα and ERβ were expressed in human hippocampus mainly in pyramidal cells of Ammons horn and in the dentate gyrus. Western blotting and immunohistochemistry in the adult cerebral cortex and hippocampus revealed lower protein expression of ERα compared with ERβ. Double immunostaining showed that during fetal life both ERs are expressed in neurons as well as in radial glia, although only ERα is expressed in the Cajal‐Retzius neurons of the marginal zone. These observations demonstrate that the expression of ERα and ERβ displays different spatial‐temporal patterns during human cortical and hippocampal development and suggest that both ERs may play distinct roles in several processes related to prenatal brain development. J. Comp. Neurol. J. Comp. Neurol. 503:790–802, 2007.

VDAC and ERα interaction in caveolae from human cortex is altered in Alzheimer's disease

Voltage-dependent anion channel (VDAC) is a mitochondrial porin also found in the neuronal membrane (pl-VDAC), where its function may be related to redox homeostasis and apoptosis. Murine models have evidenced pl-VDAC into caveolae in a complex with estrogen receptor alpha (mERalpha), which participates in neuroprotection against amyloid beta (Abeta), and whose integration into this hydrophobic domain remains unclear. Here, we have demonstrated in caveolae of human cortex and hippocampus the presence of pl-VDAC and mERalpha, in a complex with scaffolding caveolin-1 which likely provides mERalpha stability at the plasma membrane. In Alzheimers disease (AD) brains, VDAC was accumulated in caveolae, and it was observed in dystrophic neurites of senile plaques, whereas ERalpha was expressed in astrocytes surrounding the plaques. Together with previous data in murine neurons demonstrating the participation of pl-VDAC in Abeta-induced neurotoxicity, these data suggest that the channel may be involved in membrane dysfunctioning observed in AD neuropathology.

Voltage-dependent anion channel (VDAC) participates in amyloid beta-induced toxicity and interacts with plasma membrane estrogen receptor α in septal and hippocampal neurons

Voltage-dependent anion channel (VDAC) is a porin known by its role in metabolite transport across mitochondria and participation in apoptotic processes. Although traditionally accepted to be located within mitochondrial outer membrane, some data has also reported its presence at the plasma membrane level where it seems to participate in regulation of normal redox homeostasis and apoptosis. Here, exposure of septal SN56 and hippocampal HT22 cells to specific anti-VDAC antibodies prior to amyloid beta (Aβ) peptide was observed to prevent neurotoxicity. In these cell lines, we identified a VDAC form associated with the plasma membrane that seems to be particularly abundant in caveolae. The two membrane-related isoforms of estrogen receptor α (mERα) (80 and 67 kDa), known in SN56 cells to participate in estrogen-induced neuroprotection against Aβ injury, were also observed to be present in caveolae. Interestingly, we demonstrated for the first time that both VDAC and mERα interact at the plasma membrane of these neurons as well as in microsomal fractions of the corresponding murine septal and hippocampal tissues. These proteins were also shown to associate with caveolin-1, thereby corroborating their presence in caveolar microdomains. Taken together, these results suggest that VDAC-mERα association at the plasma membrane level may participate in the modulation of Aβ-induced cell death.

Alternative estrogen receptors homologous to classical receptor α in murine neural tissues

Abstract Although it is widely accepted the existence of putative estrogen receptors (ERs) localized at extranuclear domains in the brain, their molecular identity is still unclear. We have previously demonstrated in a murine septal cell line the existence of a membrane-related ER (mER) that participates in estrogen-mediated neuroprotection. To investigate the molecular structure of mER, we have used a battery of antibodies raised against different domains of the classical ERα to immunoblot with plasma membrane fractions from septal SN56 and hippocampal HT22 cell lines, and microsomal fractions of mouse septal and hippocampal tissues. The results confirmed that mER is the homologue of its intracellular counterpart ERα, suggesting the possibility that both nuclear and extranuclear receptors may share a common origin.

Modulation of Aβ-induced neurotoxicity by estrogen receptor alpha and other associated proteins in lipid rafts

Some evidences have demonstrated the participation of estrogen receptors (ERs) in rapid, non-genomic actions of estrogen to promote neuroprotection against different toxic agents. However, there is still very little information about the structural nature of these receptors and the manner these proteins may be integrated into the plasma membrane. One of the plausible possibilities is that they may be localized in lipid rafts microstructures where they would be associated with other, still unknown, molecules which may modulate their physiological activities related to cell survival. In this work, we have identified in caveolar fractions of murine septal and hippocampal neurons a membrane-related ER shown to physically interact with, both, a voltage-dependent anion channel and scaffold protein caveolin-1.

Effects of nitric oxide donors sodium nitroprusside and 3-morpholino-sydnonimine on prolactin secretion in conscious rats

In the present study, we have examined the possible involvement of the central nitric oxide (NO) pathway in the control of prolactin secretion in vivo. The effects of intracerebroventricular (i.c.v.) injections of L-arginine (L-Arg), a precursor of NO, N omega-nitro-L-arginine methyl ester (L-NAME), an inhibitor of NO synthase (NOS), and of sodium nitroprusside (SNP) and 3-morpholino-sydnonimine (SIN-1), NO donors, on basal prolactin levels were studied in conscious male rats. Microinjections of L-Arg (100 and 500 mu g) or L-NAME (100 and 500 mu g) did not modify plasma prolactin levels, however i.c.v. injections of both SNP (1, 5, 10 and 20 mu g) and SIN-1 (1, 10 and 100 mu g) induced dose-dependent increases in these levels although SNP was much more potent than SIN-1. These results suggest a role of NO in the control of prolactin secretion.

Nitric oxide inhibits tyrosine hydroxylase of rat median eminence

Tyrosine hydroxylase (TH) is the rate-limiting enzyme in the biosynthesis of catecholamines. In a previous report we found that intracerebroventricular administration of nitric oxide (NO) generator sodium nitroprusside (SNP) to conscious male rats inhibited dose-dependently the TH activity of the median eminence (ME). In the present study we have tested the in vitro effects of SNP on TH activity, its possible mediator and action mechanism. Exposure of the ME TH to SNP (50, 100 and 500 microM) caused concentration-dependent inhibition of its enzyme activity. Addition of; reduced hemoglobin Hb (10 microM), a NO scavenger, superoxide dismutase SOD (1000 units/ml), a superoxide scavenger enzyme, or uric acid UA (300 microM), a peroxynitrite scavenger, did not affect the enzyme activity by themselves, but prevented the inhibitory effect of SNP 500 microM. However, the presence of methylene blue MB (100 microM), a guanylyl cyclase inhibitor, did not alter either basal enzyme activity or the inhibitory action of SNP 500 microM. These results suggest that this action of SNP on TH of the ME would be mediated by peroxynitrite generated by the reaction of NO with superoxide.

Sodium nitroprusside inhibits the tyrosine hydroxylase activity of the median eminence in the rat

Nitric oxide (NO) has been involved in the modulation of various neuroendocrine responses. This work is a study of dose-response and time-course of the effect of intracerebroventricular (i.c.v.) administration of (NO) generator sodium nitroprusside (SNP), on tyrosine hydroxylase (TH) activity of the median eminence (ME) and serum prolactin levels, performed on conscious male rats. SNP (1, 5 and 10 microg) inhibited the TH activity of the ME, 15 min following injection in a dose-dependent way, although the effect was only significant with the highest dose, and also increased in a dose-dependent manner the serum prolactin levels. Both actions were transient but vanished at different times following injection of 10 microg of SNP. These results suggest that NO, released from SNP, inhibits the tuberoinfundibular dopaminergic neurons of the basal hypothalamus to stimulate prolactin secretion.

A Facial Aging Simulation Method Using flaccidity deformation criteria

Due to the fact that the aging human face encompasses skull bones, facial muscles, and tissues, we render it using the effects of flaccidity through the observation of family groups categorized by sex, race and age. Considering that patterns of aging are consistent, facial ptosis becomes manifest toward the end of the fourth decade. In order to simulate facial aging according to these patterns, we used surfaces with control points so that it was possible to represent the effect of aging through flaccidity. The main use of these surfaces is to simulate flaccidity and aging consequently

Origin of Adenohypophysial Lobes and Cells from Rathke’s Pouch in Chicken (Gallus gallus) and Japanese Quail (Coturniz coturniz japonica). Expression of Calcium‐Binding Proteins

A histological and immunochemical study of adenohypophysis development in two bird species: chicken (Gallus gallus) and Japanese quail (Coturnix coturnix japonica) was carried out, focussing firstly morphologically on the origin of its different lobes, then secondly on the differentiation of hormone‐producing cells from the adenohypophysial anlage and their involvement in the differentiation of three calcium‐binding proteins. The results of the morphological development study show how the origin of the adenohypophysis in chicken is totally ectodermic, whilst in Japanese quail the endoderm, in the form of Sessel’s pouch, participates in forming the rostral zone of the anterior lobe. After studying the organogenesis and spatio/temporal differentiation of the hormone‐producing cells proceeding from the adenohypophysial anlage, a regionalization model is proposed for the origin of the different lobes and cell types as well as time sequence, fundamentally the origin of cell regionalization in the adult adenohypophysis. In this process, at least in the two bird species studied, the results obtained from expressing the calcium‐binding proteins, calbindin D 28K, calretinin and parvalbumin show a characteristic distribution pattern for each, suggesting distinct functions.