[The academic benefit and the system for the promotion of the researcher. Stimuli for excellence].
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Biomedical subjects
F Valverde
Publications and source records attributed to F Valverde.
Occurrence of a differential expression of the glyceraldehyde-3-phosphate dehydrogenase gene in muscle and liver from euthermic and induced hibernating jerboa (Jaculus orientalis).
A cDNA clone which contains the near-complete open reading frame (ORF) encoding glyceraldehyde-3-phosphate dehydrogenase (GAPDH, EC 1.2.1.12) was obtained by screening a muscle cDNA library of jerboa (Jaculus orientalis), a true hibernating rodent, with a PCR-amplified 0.5-kb genomic DNA probe from an internal region of the gene. The 1.1-kb cDNA clone consists of a 927-bp ORF which codifies for 309 aa, about 93% of the original GapC gene encoding the 36-kDa protein, and a 3'-noncoding region of 167 bp. The full-length aa sequence of GAPDH was achieved by sequencing the N-terminal region of the purified protein completing the missing part in the cDNA clone. Both nt and aa sequences exhibit a high degree of homology to other mammalian GAPDHs. The expression of the GapC gene was studied in skeletal muscle and liver of euthermic and hibernating jerboas both on the mRNA level by Northern blot hybridization using the cDNA clone as a probe and on the protein level by Western blot immunodetection using an antibody raised against muscle GAPDH. A clear decrease (about threefold) in the amount of GapC mRNA, a single 1.2-kb transcript, was observed in muscle of hibernating jerboa when compared with the same tissue from the euthermic animal. This mRNA level decrease directly correlates with a reduction in both protein amount and specific activity in crude protein extracts. In contrast, both GAPDH protein and GapC mRNA levels remained unchanged in liver from euthermic and hibernating jerboas although the enzymatic activity was also about threefold lower in the hibernating tissue. These result, together with previous data obtained from protein studies [Soukri et al. (1995) Biochim. Biophys. Acta 1243, 161-168 and (1996) 1292, 177-187] indicate that jerboa GAPDH is regulated by different mechanisms during hibernation in these tissues, that is, at transcriptional level in muscle and at posttranslational level in liver. The reduced GAPDH activity should result in both cases in a decrease of the glycolytic flux that would eventually contribute to the dramatic metabolic depression of this dormant state.
Amino Acid Sequence↗
Dynamics of cell migration from the lateral ganglionic eminence in the rat.
From previous developmental studies, it has been proposed that the neurons of the ventrolateral cortex, including the primary olfactory cortex, differentiate from progenitor cells in the lateral ganglionic eminence. The objective of the present study was to test this hypothesis. The cells first generated in the forebrain of the rat migrate to the surface of the telencephalic vesicle by embryonic day (E) 12. Using [3H]thymidine, we found that most of these cells contributed to the formation of the deep layer III of the primary olfactory cortex. To study the migratory routes of these cells, we made localized injections of the carbocyanine fluorescent tracers Dil and DiA into various parts of the lateral ganglionic eminence in living embryos at E12-E14 and subsequently maintained the embryos in a culture device for 17-48 hr. After fixation, most migrating cells were located at the surface of the telencephalic vesicle, whereas others were seen coursing tangentially into the preplate. Injections made at E13 and in fixed tissue at E15 showed that migrating cells follow radial glial fibers extending from the ventricular zone of the lateral ganglionic eminence to the ventrolateral surface of the telencephalic vesicle. The spatial distribution of radial glial fibers was studied in Golgi preparations, and these observations provided further evidence of the existence of long glial fibers extending from the ventricular zone of the lateral ganglionic eminence to the ventrolateral cortex. We conclude that cells of the primary olfactory cortex derive from the lateral ganglionic eminence and that some early generated cells migrating from the lateral ganglionic eminence transgress the cortico-striatal boundary entering the preplate of the neocortical primordium.
Animals↗
Presence of LHRH (luteinizing hormone-releasing hormone) fibers in the optic nerve, optic chiasm and optic tract of the adult rat.
In mammals LHRH (luteinizing hormone-releasing hormone) is synthesized and released by a set of neurons that have their embryonic origin in the olfactory placode. We have observed that, besides their classical location, LHRH fibers can also be seen in the optic nerve and optic chiasm. Some LHRH fibers could also be traced in the optic tract. The possible course of these projections, and their functional significance are discussed.
Age Factors↗
Evidence for a posttranslational covalent modification of liver glyceraldehyde-3-phosphate dehydrogenase in hibernating jerboa (Jaculus orientalis).
The specific activity of D-glyceraldehyde-3-phosphate (G3P) dehydrogenase (phosphorylating) (GPDH, EC 1.2.1.12) found in liver of induced hibernating jerboa (Jaculus orientalis) was 2-3-fold lower than in the euthermic animal. However, the comparative analysis of the soluble protein fraction of these tissues by SDS-PAGE and Western blotting showed no significant changes in the intensity of the 36 kDa protein band of the GPDH subunit. After using the same purification procedure, the GPDH from liver hibernating jerboa exhibited lower values for both apparent optimal temperature and specific activity than the enzyme from the euthermic animal. Similar non-linear Arrhenius plots were obtained, but the Ea values calculated for the GPDH from hibernating tissue were higher. Although in both purified enzyme preparations four isoelectric GPDH isoforms were resolved by chromatofocusing, those of hibernating liver exhibited more acidic pI values (pI 7.3-6.1) than the hepatic isoforms of euthermic animals (pI 8.7-8.1). However, all liver GPDH isoforms exhibited similar native and subunit molecular masses and cross-reacted with an antibody raised against muscle GPDH. The comparison of the kinetic parameters of both purified preparations and the main isoforms isolated from euthermic and hibernating tissues showed the decreased catalytic efficiency of hibernating enzyme being exclusively due to a lower Vmax for both substrates G3P and NAD+. Phosphodiesterase treatment of cell-free extracts increased GPDH activity in the case of hibernating liver only. The pI of the main isoform purified from this tissue, about 6.9, changed after this treatment to an alkaline value (pI 8.44) similar to those of the euthermic GPDH isoforms. Differential ultraviolet absorption spectra of these isoforms indicated that a substance absorbing at 260 nm, that was released by the phosphodiesterase digestion, was present in the enzyme of hibernating tissue. Incubation of purified GPDH with the NO-releasing agent sodium nitroprussite produced under conditions that promote mono-ADP-ribosylation a dramatic decrease of activity (up to 60%) of both euthermic and phosphodiesterase-treated hibernating preparations but only a marginal inhibition of the hibernating enzyme. These data suggest that the liver GPDH of hibernating jerboa exhibits a posttranslational covalent modification, being probably a mono-ADP-ribosylation. The resulting inhibition of enzyme activity could contribute to the wide depression of the glycolytic metabolic flow associated with mammalian hibernation.
Alkaline Phosphatase↗
Early onset of the rat olfactory bulb projections.
Using the fluorescent carbocyanine tracer DiI, we examined in detail the early development of the projections emanating from the rat olfactory bulb. The study commenced at embryonic day 13 when the first fibres can be detected and ended at embryonic day 20, when all major fibre systems have been established. The first axons arising from the prospective olfactory bulb area are seen at embryonic day 13. Labelled fibres are provided with elaborate axonal growth cones advancing through the ventrolateral part of the telencephalic vesicle. At embryonic day 14, while the main fibre tract has not developed much further, some isolated fibres are located quite distally from the prospective olfactory bulb. These early fibres apparently course within a narrow cell-free space that extends caudally along the ventrolateral part of the telencephalic vesicle. At embryonic day 15, a number of labelled fibres form a compact bundle, corresponding to the lateral olfactory tract, that ultimately reaches the prospective primary olfactory cortex. The fibres do not stop growing, but continue to extend caudally at embryonic day 17. The results of this study provide new information on the development of axonal tracts in the olfactory system. We show that the olfactory tract projection develops earlier than the morphological appearance of the olfactory bulbs. This suggests that the early development of olfactory projections might not depend on the arrival of the olfactory epithelium axons and thus, could be governed by factors intrinsic to the neurons and/or cues present in the target environment.
Animals↗
Early olfactory fiber projections and cell migration into the rat telencephalon.
The formation and development of primary olfactory axons was studied in the rat embryo using acetylcholinesterase histochemistry, immunocytochemistry for neuron-specific beta-tubulin (TuJ1) and growth associated protein 43 (GAP43), and a fluorescent tracer DiI. Olfactory axons extend from the olfactory receptor neurons localized in the olfactory epithelium. These fibers grow to reach and enter the olfactory bulbs, where they form the first relay and integrative synaptic station in the olfactory system: the olfactory glomerulus. In this communication we address the development of primary olfactory fibers: first from the olfactory placode and later from the olfactory epithelium. Olfactory fibers enter the olfactory bulbs apparently in a disordered manner but soon arrange themselves in hook shaped aggregates of fibers, with many boutons (immature synaptic terminals), to form the glomeruli. We detected this kind of structure for the first time at embryonic day 16. The olfactory receptor cells are usually anchored in the basal lamina of the olfactory epithelium but some of them, after reaching their targets, lose their epithelial attachment, leave the olfactory epithelium and migrate to and enter the olfactory bulbs. The traffic of cells between the olfactory epithelium and the brain lasts late into embryonic development. We describe four types of migratory mechanism used by different populations of cells to reach their targets in the telencephalic vesicle and propose the existence of migrating cells that enter the telencephalon. These data were corroborated by injections into the olfactory epithelium a of murine retrovirus carrying the Escherichia coli lac-Z gene.
Acetylcholinesterase↗
Characterization of muscle glyceraldehyde-3-phosphate dehydrogenase isoforms from euthermic and induced hibernating Jaculus orientalis.
The specific activity of D-glyceraldehyde-3-phosphate (G3P) dehydrogenase (phosphorylating) (GPDH, EC 1.2.1.12) found in skeletal muscle of induced hibernating jerboa (Jaculus orientalis) was 3-4-fold lower than in the euthermic animal. The comparative analysis of the soluble protein fraction of these tissues by SDS-PAGE and Western blotting showed a significant decrease in the intensity of a protein band of about 36 kDa, the GPDH subunit, in hibernating jerboa. After using the same purification procedure, the GPDH from muscle of hibernating jerboa exhibited lower values for both apparent optimal temperature and specific activity than the enzyme from the euthermic animal. Non-linear Arrhenius plots were obtained in both cases, but the Ea values calculated for the GPDH from hibernating tissue were higher. Although in both purified enzyme preparations three isoelectric GPDH isoforms, exhibiting pI values in the range 8.2-7.5, were resolved by chromatofocusing, clear differences were observed in these preparations concerning the relative contribution to the total enzymatic activity of the two main isoforms, named GPDH I (pI values, 8.1-8.2) and GPDH II (pI values, 7.8-7.9). Thus, whereas GPDH I was the major isoform purified from euthermic muscle, accounting for more than 90% of the total activity, the amount of activity due to GPDH II reached up to 65% in preparations of hibernating jerboa. All isoforms exhibited similar native and subunit molecular masses and cross-reacted with an anti-GPDH antibody raised against the GPDH I. However, the two muscle GPDH isoforms prevailing under hibernating conditions exhibited a decreased catalytic efficiency when compared with the corresponding major isoforms purified from euthermic animals, as indicated by their different specific activities and kinetic parameters, i.e. relatively high Km and low Vmax values. Since the glycolytic flow has been found to be widely reduced in skeletal muscle of induced hibernating jerboa, the changes in the GPDH isoforms described in the present study could provide a molecular basis to explain some of the metabolic changes associated with mammalian hibernation.
Animals↗
Olfactory bulb ensheathing glia: a unique cell type with axonal growth-promoting properties.
The olfactory bulb (OB) is a structure of the central nervous system (CNS) in which axonal growth occurs throughout the lifetime of the organism. A major difference between the OB and the remaining CNS is the presence of ensheathing glia in the first two layers of the OB. Ensheathing glia display properties that might be involved in the process of regeneration and they appear to be responsible for the permissibility of the adult OB to axonal growth. In fact, transplants of ensheathing glia can be used as promoters of axonal regeneration within the adult CNS. The axonal growth-promoting properties of ensheathing glia make the study of this cell type interesting for understanding the mechanisms underlying axonal regeneration. Several groups have studied OB ensheathing cells extensively in an attempt to classify them within any of the known glial groups. However, this cell type does not exhibit the phenotypic features of any glial population described thus far. In this article we review the characteristics that differentiate ensheathing glia from other peripheral and central glial populations as well as the properties that involve them in axonal regeneration. The evidence suggests that ensheathing glia are unique, have their own identity, and do not belong to any previously described glial type.
Animals↗
The telencephalic vesicles are innervated by olfactory placode-derived cells: a possible mechanism to induce neocortical development.
During early embryonic development, the olfactory placode is the source of different cell types migrating toward the telencephalic vesicle. Among these cell types are the ensheathing cells, the luteinizing hormone-releasing hormone-producing cells and the olfactory marker protein-immunoreactive cells. We have identified a novel group of olfactory placode-derived migratory cells using an antibody against beta-tubulin to label neurons and acetylcholinesterase histochemistry to label posmitotic cells. In this paper we describe the morphology, migration and fate of this novel group of cells. The first neurons detected in the rostral prosencephalon with acetylcholinesterase and anti-beta-tubulin antibody are localized in the olfactory placodes at embryonic day 11 in the rate. At embryonic day 12, anti-beta-tubulin antibody-positive cells were observed in the mesenchymal tissue between the olfactory pit and the rostral pole of the telencephalic vesicle. Anti-beta-tubulin antibody-positive cells were seen running superficially over the pial (dorsal) side of the telencephalic vesicle at embryonic day 13. The majority of these cells have a bipolar profile with short leading and trailing processes, suggesting that they are migratory elements. However, some of these cells showed elaborate processes extending for quite long distances, overlying the pial surface of the telencephalic vesicle. A mass of cells extending over the telencephalic vesicle from the developing olfactory epithelium were observed at embryonic day 13 using acetylcholinesterase histochemistry. Some of these acetylcholinesterase-positive cells were identified as neurons with the specific neuronal marker anti-beta-tubulin antibody. On embryonic day 12, neurons from the olfactory epithelium send axonal fibers toward the telencephalic vesicles. Most of these fibers spread over the anteroventral pole of the vesicles but others entered deep into the telencephalon, reaching the germinal ventricular zone. We also show that fibers run rostrocaudally over the surface of the telencephalic vesicles. We suggest that these cells and fibers, apparently originating in the olfactory placode and migrating through non-conventional routes, might play a significant role in the earliest stages of telencephalic vesicle development.
Acetylcholinesterase↗
Persistence of early-generated neurons in the rodent subplate: assessment of cell death in neocortex during the early postnatal period.
In the rat, the deepest neocortical layer forms a conspicuous cell band known as layer Vlb. Cells in layer Vlb are among the first to differentiate, and it has been regarded as an homolog to the subplate of primates and carnivores. Cell death has been considered a universal feature of subplate cells. In order to assess the validity of this assertion, we examined the sequence of generation and the extent of cell death in layer Vlb. This was achieved using injections of 3H-thymidine and two methods for the direct visualization of apoptotic figures. Single injections of 3H-thymidine were performed between E12 and E15 (E0 is the day of insemination), and brains were examined at different postnatal ages between P1 and P63. The number of heavily labeled cells were counted in layer Vlb in six standard, equally spaced coronal sections in each brain. Single injections at E12 labels about 3% of the entire population of layer Vlb cells, 17% at E13, 30% at E14, and < 1% at E15. Our results indicate that the absolute number of heavily labeled cells in layer Vlb remains constant. The analysis of variance (one-way ANOVA) showed that the difference among the group means was not significant from P1 to P63 after injections at either E12, E13, or E14. In order to confirm these results, we evaluated the distribution of pyknotic (apoptotic) cell bodies in the neocortex. Apoptotic cells were visualized in Nissl preparations and by histochemical staining using an in situ apoptosis detection kit. The analysis was performed in rats from E18 to P15. Both methods gave comparable results. We found that the amount of cell death in layer Vlb is neither particularly prominent nor significantly different from that which occurs in the remaining neocortical layers, apart from layer II and in the white matter of the corpus callosum. We conclude that neuronal death does not play any significant role in the rodent subplate.
Aging↗
Development and early postnatal maturation of the primary olfactory cortex.
Tritiated thymidine autoradiography was used to study the origin and distribution of neurons in the primary olfactory cortex of the rat. The principal interest was devoted to animals injected at embryonic day 12 (E12) and sacrificed at different pre- and postnatal ages. The first generated neurons appearing at E12 were studied from E15 to P63. Animals sacrificed at E15 show a group of heavily labeled cells occupying a large area of the ventro lateral region of the telencephalic vesicle. At E16 this group differentiates into the principal cells of the accessory olfactory bulb and cells of the prospective primary olfactory cortex (POC). At E18-E20 the ventral tip of the cortical plate apparently divides this group into a superficial part corresponding to layer I and a deep part, corresponding to cells located in the adult in layer III. Labeled cells in layer I were found flanking the lateral olfactory tract (TOL), but rarely in the adult suggesting that they disappear or transform postnatally. Golgi observations were carried out from E15 to postnatal day 8. The morphology of different cells were studied. Layer I contains polymorphic cells resembling Cajal-Retzius cells. Among other cell types, layer II includes kinds of pyramidal cells lacking basal dendrites known as semilunar cells and intrinsic neurons. Layer III contains pyramidal cells having more than one apical dendrite ascending to the surface.
Animals↗
Successful treatment of antimony-resistant cutaneous leishmaniasis with liposomal amphotericin B.
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Adult↗
Congenital dyserythropoietic anemia type III with unbalanced globin chain synthesis.
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Anemia, Macrocytic↗
Characterization of neuronal cell varieties migrating from the olfactory epithelium during prenatal development in the rat. Immunocytochemical study using antibodies against olfactory marker protein (OMP) and luteinizing hormone-releasing hormone (LH-RH).
The development of neurons located outside the olfactory epithelium was studied by using antisera against olfactory marker protein (OMP) and luteinizing hormone-releasing hormone (LH-RH) in the rat. The study was restricted to the localization of these cells in the nasal cavity and in the region of the olfactory bulb during development. We describe groups of cells that stain positively for OMP located principally on the ventro-lateral aspect of the olfactory bulbs. A comparison is made with the LH-RH-immunoreactive system of cells which predominate on the medial aspect following the known trajectory of the nervus terminalis. OMP-immunoreactive cells appeared along the course of the olfactory fibers when they were first detected at embryonic day 16. These cells became restricted to a small group above the cribriform plate, ventral to the olfactory bulbs that seemed to disappear shortly after birth. It is concluded that these cells, which like the LH-RH cells have most probably migrated from the olfactory placode, represent a group of intervening neurons between the olfactory receptor cells and the olfactory bulb, serving as hints for olfactory axons to reach their targets during prenatal development.
Animals↗
[The first case of thalassemia intermedia in Spain due to the interaction of 3 alpha genes with beta-thalassemia minor].
We have identified the case of a 9-months-old girl with heterozygotic thalassemia and triplication of alpha genes of globin (alpha alpha alpha 3.7). Molecular defect of thalassemia was a mutation without sense of 39 codon. Patient's phenotype was an intermediate thalassemia with moderate splenomegaly and marked unbalance on the globin chains. This is the first case of intermediate thalassemia, through this mechanism, described in Spain.
Female↗
Transient pattern of exuberant projections of olfactory axons during development in the rat.
The purpose of our study was twofold: (1) to trace the development of the olfactory axons from early embryonic stages until the mature pattern of connectivity and (2) to determine whether a transient penetration of them exists beyond the olfactory glomeruli. Two techniques were employed: DiI applied in the olfactory epithelium after aldehyde fixation, and olfactory marker protein (OMP) immunostaining. At E13 and E14 olfactory axons were observed spreading over the telencephalic vesicle and entering deeply into the prospective olfactory bulb, extending near the ventricular zone. Growth cones were seen at the end of these axons. At E15, the bundles of olfactory axons form a network, in which axons, growth cones and cells were seen. Some of these axons entered the olfactory bulb. Using OMP immunostaining olfactory axons were observed along the external plexiform layer, the mitral cell layer and in the granular layer from E19 to P6. At P9 some OMP immunoreactive axons were observed in the external plexiform layer. No OMP immunostained axons could be observed outside the glomeruli at P10. Our conclusions are that a transient immature pattern of early invasion over the telencephalic vesicle and of the olfactory bulb by olfactory axons occurs in the olfactory system. By the second postnatal week the glomerular layer reaches its mature configuration, and no olfactory fibers are seen outside the glomerular layer.
Animals↗
Development of the main efferent cells of the olfactory bulb and of the bulbar component of the anterior commissure.
The development of the efferent cells of the main olfactory bulb and the development of the bulbar part of the anterior commissure were studied in the rat from E16 to P7. DiI was used in fixed tissues as a neuronal tracer. From E16 onwards cells located in the olfactory bulb anlage were stained in a Golgi-like appearance. The morphological changes of these cells were: from E16 to P4, re-orientation from a tangential position to a radial position, elongation of the principal dendrite and spreading out of the secondary dendrites. From P4 onwards, there was a lack of migrating mitral cells in the inner part of the bulb. At E16 some fibers of the anterior commissure reached the midline, the number of fibers increased slowly until P0/P1. At P2 there was an explosive increase in the number of fibers crossing the midline and reaching the contralateral bulb. The development in two stages is hypothesized.
Animals↗