Yolanda León

Cell signalling by inositol phosphoglycans from different species

The discovery of glycosyl-phosphatidylinositol (GPI) molecules and their products has given new insight into the field of signal transduction. In the last decade a novel mechanism of protein attachment to membranes has emerged, which involves a covalent linkage of the protein to the glycan moiety of a GPI. The discovery that GPI-anchored proteins are ubiquitous throughout the eukaryotes was followed by the observation that uncomplexed GPI molecules are implicated in signal transduction for a diversity of hormones and growth factors. The hydrolysis of free-GPI generates a novel second messenger: the inositol phosphoglycan (IPG). The aim of this article is to review the role of IPG and IPG-like molecules in signal transduction and to discuss future research directions.

Insulin-like growth factor 1 is required for survival of transit-amplifying neuroblasts and differentiation of otic neurons

Neurons that connect mechanosensory hair cell receptors to the central nervous system derive from the otic vesicle from where otic neuroblasts delaminate and form the cochleovestibular ganglion (CVG). Local signals interact to promote this process, which is autonomous and intrinsic to the otic vesicle. We have studied the expression and activity of insulin-like growth factor-1 (IGF-1) during the formation of the chick CVG, focusing attention on its role in neurogenesis. IGF-1 and its receptor (IGFR) were detected at the mRNA and protein levels in the otic epithelium and the CVG. The function of IGF-1 was explored in explants of otic vesicle by assessing the formation of the CVG in the presence of anti-IGF-1 antibodies or the receptor competitive antagonist JB1. Interference with IGF-1 activity inhibited CVG formation in growth factor-free media, revealing that endogenous IGF-1 activity is essential for ganglion generation. Analysis of cell proliferation cell death, and expression of the early neuronal antigens Tuj-1, Islet-1/2, and G4 indicated that IGF-1 was required for survival, proliferation, and differentiation of an actively expanding population of otic neuroblasts. IGF-1 blockade, however, did not affect NeuroD within the otic epithelium. Experiments carried out on isolated CVG showed that exogenous IGF-1 induced cell proliferation, neurite outgrowth, and G4 expression. These effects of IGF-1 were blocked by JB1. These findings suggest that IGF-1 is essential for neurogenesis by allowing the expansion of a transit-amplifying neuroblast population and its differentiation into postmitotic neurons. IGF-1 is one of the signals underlying autonomous development of the otic vesicle.

Glycosyl Inositol Derivatives Related to Inositolphosphoglycan Mediators: Synthesis, Structure, and Biological Activity

17 paginas, 12 figuras, 4 esquemas, 7 tablas.-- Supporting information for this article is available on the WWW under http://www.wiley-vch.de/home/chemistry/ or from the author.

Cell death in the nervous system: Lessons from insulin and insulin-like growth factors

Programmed cell death is an essential process for proper neural development. Cell death, with its similar regulatory and executory mechanisms, also contributes to the origin or progression of many or even all neurodegenerative diseases. An understanding of the mechanisms that regulate cell death during neural development may provide new targets and tools to prevent neurodegeneration. Many studies that have focused mainly on insulin-like growth factor-I (IGF-I), have shown that insulin-related growth factors are widely expressed in the developing and adult nervous system, and positively modulate a number of processes during neural development, as well as in adult neuronal and glial physiology. These factors also show neuroprotective effects following neural damage. Although some specific actions have been demonstrated to be anti-apoptotic, we propose that a broad neuroprotective role is the foundation for many of the observed functions of the insulin-related growth factors, whose therapeutical potential for nervous system disorders may be greater than currently accepted.

Synthesis and investigation of the possible insulin-like activity of 1d-4-O- and 1d-6-O-(2-amino-2-deoxy-α-d-glucopyranosyl) myo-inositol 1-phosphate and 1d-6-O-(2-amino-2-deoxy-α-d-glucopyranosyl) myo-inositol 1,2-(cyclic phosphate)

The synthesis of the glycosyl-myo-inositol 1-phosphates 1 and 2 and of the glycosyl-myo-inositol 1,2-(cyclic phosphate) 3, starting from previously synthesized intermediates, is reported. Compound 3 was found to display proliferative effects on the early developing inner ear of chick embryo.

Programmed cell death in the development of the vertebrate inner ear.

Programmed cell death is known to be an essential process for accurate ontogeny during the normal development of the inner ear. The inner ear is a complex sensory organ responsible for equilibrium and sound detection in vertebrates. In all vertebrates, the inner ear develops from a single ectodermic patch on the surface of the embryo’s head, which undergoes a series of morphological changes to give rise to the complex structure of the adult inner ear. Enlargement and morphogenesis of the inner ear primordium is likely to depend on cellular division, growth, migration, differentiation and apoptosis. Here we describe the regions of programmed cell death that contribute to the final morphological aspect of the adult inner ear. The few studies that focus on the molecules that control this process during inner ear development indicate that the molecules and intracellular signaling pathways activated during the apoptotic response in the inner ear are similar to the previously described for the nervous system. In this review, we will describe some of the growth factors and key pathways that regulate pro- and anti-apoptotic signals and how they cross talk to determine the apoptotic or survival fate of cells in the development of the inner ear.

Programmed cell death in the developing inner ear is balanced by nerve growth factor and insulin-like growth factor I

Nerve growth factor induces cell death in organotypic cultures of otic vesicle explants. This cell death has a restricted pattern that reproduces the in vivo pattern of apoptosis occurring during inner ear development. In this study, we show that binding of nerve growth factor to its low affinity p75 neurotrophin receptor is essential to achieve the apoptotic response. Blockage of binding to p75 receptor neutralized nerve-growth-factor-induced cell death, as measured by immunoassays detecting the presence of cytosolic oligonucleosomes and by TUNEL assay to visualize DNA fragmentation. Nerve growth factor also induced a number of cell-death-related intracellular events including ceramide generation, caspase activation and poly-(ADP ribose) polymerase cleavage. Again, p75 receptor blockade completely abolished all of these effects. Concerning the intracellular pathway, ceramide increase depended on initiator caspases, whereas its actions depended on both initiator and effector caspases, as shown by using site-specific caspase inhibitors. Conversely, insulin-like growth factor I, which promotes cell growth and survival in the inner ear, abolished apoptosis induced by nerve growth factor. Insulin-like growth factor cytoprotective actions were accomplished, at least in part, by decreasing endogenous ceramide levels and activating Akt. Taken together, these results strongly suggest that regulation of nerve-growth-factor-induced apoptosis in the otocysts occurs via p75 receptor binding and is strictly controlled by the interaction with survival signalling pathways.

Induction of cell growth by insulin and insulin-like growth factor-I is associated with Jun expression in the otic vesicle.

The present report investigates the cellular mechanisms involved in the regulation of cell proliferation by insulin and insulin‐like growth factor‐I (IGF‐I) in the developing inner ear. The results show that insulin and IGF‐I stimulate cell proliferation in the otic vesicle. This effect is associated with the induction of the expression of the nuclear proto‐oncogene c‐jun. The temporal profile of Jun expression coincided with the proliferative period of growth of the otic vesicle. IGF‐I promoted the hydrolysis of a membrane glycosyl‐phosphatidylinositol, which was characterised as the endogenous precursor for inositol phosphoglycan (IPG). Both purified IPG and a synthetic analogue, 6‐O‐(2‐amino‐2‐deoxy‐α‐D‐glucopyranosyl)‐D‐myo‐inositol‐1,2‐cyclic phosphate (C3), were able to mimic the effects of IGF‐I on Jun expression. Anti‐IPG antibodies blocked the effects of IGF‐I, which were rescued by the addition of IPG or its analogue. These results suggest that the sequence involving the hydrolysis of membrane glycolipids and the expression of c‐jun and c‐fos proto‐oncogenes is part of the mechanism that activates cell division in response to insulin and IGF‐I during early organogenesis of the avian inner ear. The implications of these observations for otic development and regeneration are briefly discussed. J. Comp. Neurol. 398:323–332, 1997.

Strict regulation of c-Raf kinase levels is required for early organogenesis of the vertebrate inner ear

Regulation of organogenesis involves a dynamic balance of the mechanisms regulating cell division, differentiation and death. Here we have investigated the pattern of expression of c-Raf kinase in the inner ear during early developmental stages and the consequences of manipulating c-Raf levels by misexpression of c-raf viral vectors in organotypic cultures of otic vesicle explants. We found that otic vesicles expressed c-Raf and its level remained constant during embryonic days 2 and 3 (E2-E3). c-Raf activity was increased in response to insulin like growth factor-I (IGF-I) and the activation by IGF-I of the c-Raf kinase pathway was a requirement to turn on cell proliferation in the otic vesicle. Overexpression of c-raf in E2.5 explants increased the proliferative response to low serum and IGF-I and blocked differentiation induced by retinoic acid. The increase in c-Raf levels also prevented nerve growth factor (NGF)-dependent induction of programmed cell death. Consistent with these results, the expression of a dominant negative c-Raf mutant potentiated retinoic acid action and decreased the rate of cell proliferation. We conclude that a strict control of c-Raf levels is essential for the co-ordination of the biological processes that operate simultaneously during early inner ear development.

Anti-apoptotic actions of insulin-like growth factors: lessons from development and implications in neoplastic cell transformation.

Insulin-like growth factor-I (IGF-I) is widely expressed during development, and is actively involved in the regulation of cell growth, proliferation, and differentiation. Underlying these activities is the capacity of IGF-I to promote survival in a variety of cell types, including those of the nervous system. However, in adult tissues deregulation of the IGF system can cause undesired cell survival and therefore excessive cell proliferation. Here, we review the contribution of IGF-I in developmental processes with a focus on the development of the inner ear, as well as pathological implications resulting from IGF-I deregulation during cancer.