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Biomedical subjects

A Caceres

Publications and source records attributed to A Caceres.

At least 19 recordsLinked to original sources

Evidence for the involvement of Tiam1 in axon formation.

In cultured neurons, axon formation is preceded by the appearance in one of the multiple neurites of a large growth cone containing a labile actin network and abundant dynamic microtubules. The invasion-inducing T-lymphoma and metastasis 1 (Tiam1) protein that functions as a guanosine nucleotide exchange factor for Rac1 localizes to this neurite and its growth cone, where it associates with microtubules. Neurons overexpressing Tiam1 extend several axon-like neurites, whereas suppression of Tiam1 prevents axon formation, with most of the cells failing to undergo changes in growth cone size and in cytoskeletal organization typical of prospective axons. Cytochalasin D reverts this effect leading to multiple axon formation and penetration of microtubules within neuritic tips devoid of actin filaments. Taken together, these results suggest that by regulating growth cone actin organization and allowing microtubule invasion within selected growth cones, Tiam1 promotes axon formation and hence participates in neuronal polarization.

Actin Cytoskeleton↗

Furanocoumarins from the aerial parts of Dorstenia contrajerva.

A new glycosylated furanocoumarin, alpha-L-rhamnopyranosyl-(1-->6)-beta-D-glucopyranosyl-bergaptol (1), has been isolated from Dorstenia contrajerva together with three known furanocoumarins, catechin and epicatechin. Their structures were established using high field 2D NMR techniques.

Coumarins↗

Tau protein function in axonal formation.

Tau protein is a predominantly neuronal microtubule-associated protein that is enriched in axons and is capable of promoting microtubule assembly and stabilization. In the present article we review some of the key experiments directed to obtain insights about tau protein function in developing neurons. Aspects related to whether or not tau has essential, unique, or complementary functions during axonal formation are discussed.

Animals↗

Studies on the constituents of Gliricidia sepium (Leguminosae) leaves and roots: isolation and structure elucidation of new triterpenoid saponins and aromatic compounds.

Our research program on the Central American fooder plant Gliricidia sepium led to the discovery of two new triterpene saponins (1 and 2) and known aromatic compounds. The new compounds possess 3beta,21beta, 24-trihydroxy-22-oxoolean-12-ene as an aglycon. The oligosaccharide moiety linked to C-3 of the aglycon contained two pyranoses (glucuronic acid and xylose); in addition the glucose residue of both 1 and 2 is also linked to C-21. Structure elucidation of these new compounds through the extensive use of 1D and 2D NMR techniques have provided detailed information about the sapogenin and the saccharide chains, inclusive of sugar sequence and the position of glycosylation.

Central America↗

New 12a-hydroxyrotenoids from gliricidia sepium bark

The investigation of a methanolic extract of Gliricidia sepium bark afforded, in addition to vestitol and 2'-O-methylvestitol, three new 12a-hydroxyrotenoids, gliricidol (1), 2-methoxygliricidol (2), and gliricidin (3). The structures of 1-3 were elucidated by analysis of their spectroscopic data. Compounds 1-3 exhibited activity against Artemia salina larvae.

Journal Article↗

Iridoids from Lippia graveolens.

An investigation of the leaves of Lippia graveolens from Guatemala provided 10 iridoid and secoiridoid glucosides as well as their ester derivatives. Minor constituents were loganin, secologanin, secoxyloganin, dimethylsecologanoside, loganic acid, 8-epi-loganic acid and caryoptoside. Major constituents were the novel iridoids caryoptosidic acid and lippioside I and II consisting of caryoptosidic acid esterified at the C-6 position of glucose with p-coumaroyl or caffeoyl residues, respectively. Their structures were mainly elucidated by NMR spectroscopy.

Journal Article↗

Kinesin-mediated organelle translocation revealed by specific cellular manipulations.

The distribution of membrane-bound organelles was studied in cultured hippocampal neurons after antisense oligonucleotide suppression of the kinesin-heavy chain (KHC). We observed reduced 3,3'-dihexyloxacarbocyanine iodide (DiOC6(3)) fluorescent staining in neurites and growth cones. In astrocytes, KHC suppression results in the disappearance of the DiOC6(3)-positive reticular network from the cell periphery, and a parallel accumulation of label within the cell center. On the other hand, mitochondria microtubules and microfilaments display a distribution that closely resembles that observed in control cells. KHC suppression of neurons and astrocytes completely inhibited the Brefeldin A-induced spreading and tubulation of the Golgi-associated structure enriched in mannose-6-phosphate receptors. In addition, KHC suppression prevents the low pH-induced anterograde redistribution of late endocytic structures. Taken collectively, these observations suggest that in living neurons, kinesin mediates the anterograde transport of tubulovesicular structures originated in the central vacuolar system (e.g., the endoplasmic reticulum) and that the regulation of kinesin-membrane interactions may be of key importance for determining the intracellular distribution of selected organelles.

Actin Cytoskeleton↗

Molecular karyotype variation in Leishmania (Viannia) peruviana: indication of geographical populations in Peru distributed along a north-south cline.

Forty-one Leishmania peruviana isolates were selected along a north-south transect which crossed areas endemic for uta in three different biogeographical regions in the Peruvian Andes. The isolates were analysed by molecular karyotyping and hybridization with three chromosome-derived DNA probes. All the isolates could be distinguished from L. braziliensis by their pLb-134 hybridization patterns. However, the patterns with the other probes (pLb-168 and -22) could be used to cluster the Peruvian isolates in discrete groups (karyodemes) which varied in their level of similarity with L. braziliensis. The geographical distribution of these karyodemes supports the hypothesis that eco-graphical isolation has contributed to the heterogeneity of L. peruviana.

Animals↗

Intraneuronal compartments of the amyloid precursor protein.

The amyloid precursor protein (APP) is the parent molecule from which beta-amyloid protein is cleaved and deposits as amyloid fibrils in the senile plaques of Alzheimer's disease. Its primary structure resembles a receptor; however, no ligand has been identified. In growing hippocampal neurons APP is localized to growth cones. APP immunoreactivity was highly enriched in the axons of mature cultured neurons, where it appears as a specialization of the axonal membrane. Its anterograde translocation occurs via a kinesin-based motor. Following cytosolic acidification, APP colocalizes with late endosomes that get redistributed from the neuronal cell body to the processes. APP colocalizes in cultured hippocampal neurons to clathrin-immunoreactive clusters of vesicular-like structures. The finding lends additional credence to the possibility that APP could function as a receptor.

Amyloid beta-Protein Precursor↗

Expression of the class III beta-tubulin isotype in developing neurons in culture.

The expression of the class III beta-tubulin isotype was studied in cultured brain neurons by means of a monoclonal antibody (TuJ1). The results obtained indicate that during early axonal outgrowth most of the class III beta-tubulin is not incorporated into microtubules, a phenomenon which is also observed under conditions which alter the rate and extent of the neurite outgrowth response. On the other hand, a dramatic increase in its incorporation into microtubules is observed after the neurons have differentiated their neurites as axons and dendrites. In addition, the appearance of colchicine-resistant microtubules containing this isotype, a phenomenon which occurs late in neurite development, is highly coincident with the appearance of stable microtubules containing high molecular weight microtubule-associated proteins (MAPs). This pattern is different from that of the accumulation and incorporation of other beta-tubulin isotypes into microtubules. Taken collectively, our results indicate that differences exist in the in vivo utilization of tubulin isotypes in developing brain neurons and suggest that the class III beta-tubulin isotype is not a primary factor involved in the regulation of microtubule assembly during early neurite outgrowth, but that it may be important for maintaining further neurite elongation and/or determining some unique binding property of MAPs to specific microtubule subsets.

Animals↗

Suppression of MAP2 in cultured cerebellar macroneurons inhibits minor neurite formation.

We show here that antisense MAP2 oligonucleotides inhibit neurite outgrowth in cultured cerebellar macroneurons. Unlike control neurons, which first extend a lamellipodial veil followed by a consolidation phase during which the cells extend minor neurites, MAP2-suppressed cells persist with lamellipodia and later become rounded. The induction of microtubules containing tyrosinated tubulin, which parallels neurite outgrowth in control neurons, was blocked under antisense conditions. The small but significant increase in acetylated microtubules was not affected. In contrast, the suppression of tau, which selectively blocks axonal elongation, completely prevented the increase of acetylated microtubules, but did not modify the induction of labile microtubules. These results suggest that MAP2 and tau have different functions: the initial establishment of neurites depends upon MAP2, whereas further neurite elongation depends upon tau and microtubule stabilization.

Analysis of Variance↗

Tyrosinated and detyrosinated microtubules in axonal processes of cerebellar macroneurons grown in culture.

We have used the monoclonal antibody YL 1/2 (Tyr) specific for tyrosinated tubulin, and a polyclonal antibody (Glu) specific for detyrosinated tubulin to visualize the distribution of microtubules and microtubule assembly sites during axonal outgrowth. Cerebellar macroneurons growing in culture initially extend several short and thin neurites which have the potential to differentiate either as axons or dendrites (Ferreira and Caceres: Developmental Brain Research 49:205-213, 1989). At the onset of axonal outgrowth the Tyr antibody labels the minor neurites, the axon, and its growth cone, while the Glu antibody only shows immunoreactivity in the axonal shaft. After nocodazole treatment, the Tyr staining disappears, whereas that produced by the Glu antibody remains practically unchanged. When nocodazole was removed, tyrosinated microtubules reappeared first at the tip of the axon, in a more distal region than that occupied by detyrosinated microtubules; another focal site of tyrosinated tubulin incorporation was detected in the cell body. Incorporation of tyrosinated tubulin into growing axons was also studied after taxol treatment. After long incubation periods in the presence of taxol, the Tyr staining disappeared from the axon but remained in the cell body; however, immunoreactivity in this site was negative when the cells were preincubated in the presence of protein synthesis inhibitors. Release from taxol results in the reappearance of Tyr immunoreactivity at the distal end of the axon. Taken collectively, the present results indicate 1) that in cerebellar macroneurons axonal differentiation is accompanied by a temporal and spatial differentiation of microtubules and 2) that there is an active site of tyrosinated tubulin assembly at the tip of axonal processes, and they suggest that the highly tyrosinated domain in this region is a consequence of rapid microtubule turnover and tubulin tyrosine ligase activity.

Alkaloids↗

Plants used in Guatemala for the treatment of respiratory diseases. 1. Screening of 68 plants against gram-positive bacteria.

Respiratory ailments are important causes of morbidity and mortality in developing countries. Ethnobotanical surveys and literature reviews conducted in Guatemala during 1986-88 showed that 234 plants from 75 families, most of them of American origin, have been used for the treatment of respiratory ailments. Three Gram-positive bacteria causing respiratory infections (Staphylococcus aureus, Streptococcus pneumoniae and Streptococcus pyogenes) were used to screen 68 of the most commonly used plants for activity. Twenty-eight of these (41.2%) inhibited the growth of one or more of the bacteria tested. Staphylococcus aureus was inhibited by 18 of the plant extracts, while 7 extracts were effective against Streptococcus pyogenes. Plants of American origin which exhibited antibacterial activity were: Gnaphalium viscosum, Lippia alba, Lippia dulcis, Physalis philadelphica, Satureja brownei, Solanum nigrescens and Tagetes lucida. These preliminary in vitro results provide scientific basis for the use of these plants against bacterial respiratory infections.

Anti-Bacterial Agents↗

Plants used in Guatemala for the treatment of dermatophytic infections. 1. Screening for antimycotic activity of 44 plant extracts.

Skin infections are common diseases in developing countries, of which dermatophytoses are of particular concern in the tropics, especially in infants. Through ethnobotanical surveys and literature review 100 plants were detected as being used in Guatemala for the treatment of dermatophytoses. Of these, 44 plants were screened for in vitro activity against the most common dermatophytes (Epidermophyton floccosum, Microsporum canis, Microsporum gypseum, Trichophyton mentagrophytes and Trichophyton rubrum). Results showed that aqueous extracts from 22 of the plants tested inhibit one or more of the dermatophytes. The most commonly inhibited dermatophytes were E. floccosum (43.2%), T. rubrum (36.0%), and T. mentagrophytes (31.8%); the less inhibited were M. canis (22.7%) and M. gypseum (24.0%). Plants of American origin which exhibited anti-dermatophyte activity were: Byrsonima crassifolia, Cassia grandis, Cassia occidentalis, Diphysa carthagenensis, Gliricidia sepium, Piscidia piscipula, Sambucus mexicana, Smilax regelii, Solanum americanum and Solanum nigrescens. Fungicidal and fungistatic activities as well as the minimal inhibitory concentration were demonstrated. These results provide a scientific basis for the use of these plants for the treatment of dermatophyte infections in man.

Antifungal Agents↗

Tau protein and the establishment of an axonal morphology.

Dissociated neuronal cultures from several regions of the nervous system elaborate two populations of neurites which have features of axons and dendrites. The microtubule-associated protein tau appears to segregate to the axon in some of these culture systems, however it does not do so until after the development of morphological polarity. Despite this observation, tau very likely has some role in the development of polarity because in cultured cerebellar macroneurons taken from the rat embryonic day 15 primordial cerebellum, the inhibition of tau expression by antisense techniques resulted in the failure of a single minor neurite to elongate and form an axon-like neurite. Tau antisense given continuously for up to 72 h kept neurons locked in a stage with minor neurites only; however when released from the effects of the antisense they fully recovered. The administration of tau antisense after the development of polarity resulted in the loss of the axon-like neurite, while dendrite-like neurites continued to grow. Together these results suggest that dendritic differentiation in cerebellar macroneurons requires the prior elaboration of an axon-like structure.

Animals↗

Estrogen-enhanced neurite growth: evidence for a selective induction of Tau and stable microtubules.

Estrogen stimulates the neurite outgrowth response of medial basal hypothalamic neurons maintained in culture. We show here that one effect of estrogen is to promote an increase in tau, but not in tubulin, microtubule-associated protein 1a (MAP-1a), or MAP-2 protein levels. This response precedes and accompanies an increase in stable microtubules and in neurite length. Taken collectively, our data suggest that estrogen-enhanced neurite growth is mediated by a selective induction of microtubule-stabilizing factors, namely, the tau proteins.

Animals↗

The effect of tau antisense oligonucleotides on neurite formation of cultured cerebellar macroneurons.

Tau, a microtubule-associated protein (MAP) enriched in axons, may have a role in the generation and maintenance of an axonal morphology. Neurons from embryonic day 15 rat cerebellum in culture elaborate two morphologically distinct neurite populations--one with nontapering, elongated axonlike neurites and the other with tapered dendritelike neurites that branch frequently and are selectively stained with antibodies to MAP2. Tau antisense oligonucleotides were utilized in two ways: (1) continuous application of antisense every 24 hr for variable periods of time or (2) application of antisense that was delayed until neurite differentiation was underway. In both cases, 24 hr after the administration of the antisense, tau protein was not detected immunocytochemically. When the antisense was given continuously directly after plating, the neurites persisted as simple minor outgrowths. When antisense was added 72 hr after plating, axonlike neurites were lost, while the remaining neurites continued to grow and increase in complexity. We concluded that the initial establishment of an elongated axonlike neurite is a prerequisite for further neurite maturation; however, once the axon is established, the remaining neurites are able to grow independently and assume a tapered dendritelike appearance.

Animals↗