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Biomedical subjects

D Mercanti

Publications and source records attributed to D Mercanti.

At least 37 records · Page 2Linked to original sources
PubMed1995

Neurone decapping characterization by atomic force microscopy: a topological systematic analysis.

We tested a new approach to cell decapping on rat cerebellar neurones, and observed its effects on cell topography by atomic force microscopy (AFM). The results clearly demonstrate the effectiveness of our decapping approach, and also the ability of AFM to reveal fine details of the decapped cells. Specifically, varying the conditions and duration of the decapping process modifies the extent of the decapping. Such a method can be used to investigate the cytoplasm with surface sensitive techniques.

Animals↗
PubMed1995

Application of photoelectron spectromicroscopy to a systematic study of toxic and natural elements in neurons.

A systematic photoelectron spectromicroscopy study is presented of the spatial distribution of a toxic element, aluminium, iron or chromium, in neuron cultures, after exposure to a solution of the element. The study was performed by the X-ray secondary-emission microscopy (XSEM) version of photoelectron Spectromicroscopy. The distribution of the elements was investigated with two complementary approaches: digital subtraction imaging and individual X-ray absorption spectra from microscopic areas. The results coherently indicate different localization patterns for different elements, and, in particular, extreme localization of aluminium to a few rare cells identifiable as Purkinje neurons. In the case of iron-exposed specimens, the distribution analysis was extended to naturally present phosphorus, and used to estimate the XSEM sensitivity.

Journal Article↗
PubMed1995

Atomic Force microscopy of neuron networks.

We imaged uncoated neuron networks by an atomic force microscope in the repulsive regime of contact mode. Images of granule cells and their axons have been clearly revealed with details smaller than 20 nm. The good stability of the sample and the mechanical reproducibility of the microscope allowed the imaging of a neuron culture area of several square microns. By combining tens of images, we were able to reconstruct a highly defined neuronal network. Furthermore, the images were very reproducible over repeated scanning acquisition, demonstrating the mechanical and thermal stability of the instrument-sample system.

Animal Population Groups↗
PubMed1994

Aluminium in rat cerebellar primary cultures: glial cells and GABAergic neurones.

Experimental evidence of the preferential uptake of aluminium by GABAergic neurones and glial cells was provided by synchrotron spectromicroscopy studies. We observed rat cerebellar cultures enriched for GABAergic neurones or glial cells exposed to aluminium ions, detecting the presence and identifying the chemical status of aluminium on cell structures.

Absorptiometry, Photon↗
PubMed1994

Glutamate-induced protein phosphorylation in cerebellar granule cells: role of protein kinase C.

Protein phosphorylation in response to toxic doses of glutamate has been investigated in cerebellar granule cells. 32P-labelled cells have been stimulated with 100 microM glutamate for up to 20 min and analysed by one and two dimensional gel electrophoresis. A progressive incorporation of label is observed in two molecular species of about 80 and 43 kDa (PP80 and PP43) and acidic isoelectric point. Glutamate-stimulated phosphorylation is greatly reduced by antagonists of NMDA and non-NMDA glutamate receptors. The effect of glutamate is mimicked by phorbol esters and is markedly reduced by inhibitors of protein kinase C (PKC) such as staurosporine and calphostin C. PP80 has been identified by Western blot analysis as the PKC substrate MARCKS (myristoylated alanine-rich C kinase substrate), while antibody to GAP-43 (growth associated protein-43), the nervous tissue-specific substrate of PKC, failed to recognize PP43. Our results suggest that PKC is responsible for the early phosphorylative events induced by toxic doses of glutamate in cerebellar granule cells.

Animals↗
PubMed1993

Sodium, calcium and late potassium currents are reduced in cerebellar granule cells cultured in the presence of a protein complex conferring resistance to excitatory amino acids.

Whole-cell, patch-clamp recordings were used to study voltage-gated currents generated by cerebellar granule cells that were cultured in medium containing either 10% fetal calf serum (hereafter termed S + granules) or neurite outgrowth and adhesion complex (NOAC, hereafter called NOAC granules). NOAC is a protein complex found in rabbit serum that renders granules resistant to the excitotoxic action of excitatory amino acids. During depolarizing commands both S+ and NOAC granules generated Na+ and Ca2+ inward currents and an early and a late K+ outward currents. However, Na+ and Ca2+ inward currents and late outward K+ currents recorded in NOAC granules were smaller than those seen in S+ granules. Furthermore, although of similar amplitude, early K+ currents displayed different kinetics in the two types of neurons. Thus, these data demonstrate that the electrophysiological properties of cerebellar granules, and probably of other neuronal populations, depend upon serum components and raise the possibility that an analogous modulation might be operative in vivo, and play a role in development, synaptic plasticity or neuropathological processes.

4-Aminopyridine↗
PubMed1993

Recombinant human insulin-like growth factor I exerts a trophic action and confers glutamate sensitivity on glutamate-resistant cerebellar granule cells.

Cerebellar granule cells grown in the presence of a serum complex differentiate but are resistant to the lethal action of excitatory amino acids. When these cells are grown also in the presence of insulin-like growth factor I (IGF-I) they become fully susceptible to the toxic, lethal action of glutamate. The glutamate-sensitizing action of IGF-I is dependent on concentration (half-maximal effect at 2-4 ng/ml) and time (half-maximal effect at 2-4 days in vitro) and is paralleled by the appearance of functionally active, glutamate-activated, Ca2+ channels and of voltage-gated Na+ and late K+ channels. IGF-I-induced glutamate sensitivity is rapidly reversible (t1/2 = 30-60 min) after removal of this somatomedin. The action of IGF-I is not mimicked by IGF-II, nerve growth factor, basic or acidic fibroblast growth factor, platelet-derived growth factor, or tumor necrosis factor alpha. We postulate that the constitutive phenotype of cerebellar granule cells is glutamate-resistant and becomes responsive to excitatory amino acids under the action of epigenetic cues among which IGF-I may be one of those operative in vivo.

Animals↗
PubMed1993

Aluminium in rat cerebellar neural cultures.

A systematic microchemical analysis of unstained and uncoated neurone cultures was performed with synchrotron radiation photoemission spectromicroscopy after exposure to an aluminium solution. Clear evidence was found for localized aluminium uptake in a few cells. Their possible identification based on morphology is discussed.

Aluminum↗
PubMed1993

Differential involvement of protein kinase C in transmitter release and response to excitatory amino acids in cultured cerebellar neurons.

Cerebellar granule cells cultured in the presence of a differentiating factor isolated from rabbit serum exhibit, at variance with those cultured in fetal calf serum, an almost complete resistance to excitatory aminoacid (EAA)-induced cytotoxicity. We investigated the behaviour of protein kinase C (PKC), strongly implicated in EAA cytotoxicity, in the two types of culture. Phorbol esters, used to monitor the enzyme, enhanced the depolarization-evoked release of D-[3H]aspartate, but less effectively in factor-conditioned cells. EAAs increased phorbol esters binding in both cultures, but the effect was briefly lasting in factor-conditioned cells. The different behaviour of PKC is postulated to be causally related to different response to EAA of the cultures.

Amino Acids↗
PubMed1993

Nerve growth factor inhibits the expression of long-term potentiation in hippocampal slices.

Nerve growth factor and its receptor(s) are present in several parts of the hippocampus (CA1, CA3, dentate gyrus) but nothing is known about their function in this area which plays a fundamental role in learning and memory processes. NGF delivered exogenously to hippocampal slices causes a concentration-dependent, marked reduction in the expression (but not the induction) of long term potentiation (LTP) without altering basal synaptic transmission. The effect is already half maximal at 0.05-0.1 ng ml-1 NGF, is reversible after removal of this growth factor, and is also detectable with a modified version of NGF which has lost its neurite outgrowth promoting activity in PC12 cells. These findings point to a role for hippocampal NGF as a possible modulator of learning and memory processes. Such modulation would be mediated by high-affinity receptors functionally distinct from those promoting morphological differentiation of PC12 cells and other NGF target cells.

Animals↗
PubMed1993

Spinal cord transection in adult rats: effects of local infusion of nerve growth factor on the corticospinal tract axons.

The spinal cord of adult female rats was completely transected at the T8 level. Nerve growth factor (NGF) was administered at the lesion site via indwelling, implanted, osmotic minipumps. Purified NGF was supplied at doses of 100, 200, and 500 micrograms during a 30-day period. Control rats were treated with saline. At the end of the treatment, the proximal stump of corticospinal tract axons in the spinal cord was labeled with anterograde transported horseradish peroxidase (HRP) injected into the sensorimotor cortex. In control rats, the corticospinal tract axons ended abruptly, proximal to the zone of maximal damage. Sterile swellings developed at the axon tips, and no labeled axonal sprouts were apparent. On the contrary, in NGF-treated animals, the leading front of the corticospinal tract axons showed a trend of approaching the zone of maximal damage following abnormal paths through the dorsal-injured white matter. Axonal sprouts were seen more proximally, traveling toward the transection site in aberrantly located dorsal paths, completely outside the normal position of the corticospinal tract. NGF seems to partly restore the pattern of the regenerative behavior of the severed corticospinal tract axons after spinal cord transection in newborn rats, i.e., the induction of axonal sprouting in aberrantly located dorsal paths. An automated image analysis of the HRP reaction field close to the transection site demonstrated that the density of HRP-labeled axons in the corticospinal tract was significantly higher in the NGF-treated rats than in the control rats.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1993

Cerebellar neurones: differentiation and modulation of sensitivity to excitotoxic treatment.

The neurite outgrowth and adhesion complex (NOAC), isolated from rabbit sera has been dissociated in its major components by reverse-phase chromatography in HPLC by using a C18 column. SDS-PAGE analysis of the active fractions revealed the presence of three major bands of approximately 100, 70 and 50 kDa. Studies on the biological activity of NOAC were carried out on rat cerebellar granule cells. NOAC-cultured cells exhibit a marked resistance to excitotoxic stimuli carried by glutamate.

Animals↗
PubMed1992

Metal uptake in neurone cultures: a systematic study.

We present the first comparative study of the uptake of metal ions by neurons, performed for Zn, Cr, Co, Mo, Al, Ni, Mn and Cd. The study reveals substantial differences in the uptake of different metals, under similar exposure procedures. In particular, we found very large uptakes for aluminium and molybdenum. We also found significant effects of excitatory substances, in particular kainate, as stimulants of uptake of some of the metals.

Animals↗
PubMed1990

Effects of topically administered nerve growth factor on axonal regeneration in peripheral nerve autografts implanted in the spinal cord of rats.

The effect of exogenous nerve growth factor (NGF) on axonal regeneration into autologous peripheral nerve (PN) grafts implanted to the spinal cord (SC) of rats was assessed by retrograde labeling of the parent soma of the regenerating axons with horseradish peroxidase. NGF was delivered at the graft site over periods of 15 and 30 days by using indwelling osmotic minipumps. In control rats, the minipumps were filled with saline. At 15 days after grafting in the NGF-treated rats, the mean number of SC as well as dorsal root ganglion (DRG) neurons that regenerated their axons into the peripheral nerve grafts was increased 55.3 and 26.4 times, respectively, as compared to the control group values. At 30 days, SC and DRG neurons in the NGF-treated group were 10.9 and 3.1 times greater than in the control group. In the NGF-treated group, the regenerating SC neurons were located within a range of 7 to 13 mm from the graft site as compared to 1 to 7 mm in the control group. Finally, the analysis of the soma diameters of the regenerating neurons showed that NGF enhanced and maintained with time the regenerative response from small-sized DRG neurons. Therefore, NGF is thought to promote directly the regenerative potential of SC as well as DRG neurons and to exert an indirect glial cell-mediated effect at the SC-graft interface.

Animals↗
PubMed1987

Mitogenic effect of a human placental factor on astrocytes and glial precursors.

We have characterized and partially purified a new 'factor' present in human placenta which strongly stimulates the in vitro proliferation of two immunocytochemically characterized subtypes of astrocytes and of bipotential precursors of putative fibrous astrocytes and oligodendrocytes. This 'factor' has an apparent Mr of 60-80 kD and exhibits physicochemical and chromatographic properties characteristic of polypeptides. Our observations suggest that placenta-derived growth factors (PDMF) control the proliferation of glial cells and glial precursors during fetal development.

Animals↗