PubMed HealthSearch

Biomedical subjects

H Marrero

Publications and source records attributed to H Marrero.

13 recordsLinked to original sources

Nerve impulses increase glial intercellular permeability.

Coordinating the activity of neurons and their satellite glial cells requires mechanisms by which glial cells detect neuronal activity and change their properties as a result. This study monitors the intercellular diffusion of the fluorescent dye Lucifer Yellow (LY), following its injection into glial cells of the frog optic nerve, and demonstrates that nerve impulses increase the permeability of interglial gap junctions. Consequently, the spatial buffer capacity of the neuroglial cell syncytium for potassium, other ions, and small molecules will be enhanced; this may facilitate glial function in maintaining homeostasis of the neuronal microenvironment.

Animals

Facilitation of sodium currents in frog neuroglia by nerve impulses: dependence on external calcium.

Facilitation of voltage-gated sodium currents in glial membranes by nerve impulses has been studied by using both the whole cell and loose-patch clamp techniques in the isolated intact optic nerve of the frog. During facilitation there is a shift in the voltage dependence of glial Na+ channels such that a given depolarization produces a larger inward Na+ current. Decreasing external calcium from 4 times normal to 0.2 times normal produced a similar shift in the current-voltage relation. Increasing the external calcium concentration to 4-5 times normal blocks facilitation. In reduced calcium, 0.1-0.2 times normal, the peak of facilitation was unaffected, but its decay was slowed. The addition of 1 mM nickel and 2 mM cobalt or 2 mM cadmium, to prevent depletion of extracellular calcium that might result from voltage-dependent entry of calcium into the axons, did not block the facilitation. The results suggest that, even though facilitation is blocked by high extracellular calcium, a decrease in extracellular calcium produced by axon impulses is not the cause of the facilitation.

Animals

Changes in ultrastructure and voltage-dependent currents at the glia limitans of the frog optic nerve following retinal ablation.

The surface of the frog optic nerve consists of astrocytic processes separated by narrow extracellular clefts underlying a pial sheath of loose connective tissue. Macroscopic voltage dependent currents can be recorded from this surface using the loose patch-clamp technique. In this study the changes in ultrastructure and voltage dependent Na currents have been studied for up to 1 year following removal of the retina. During the first 1-4 weeks, many of the myelinated and unmyelinated axons of the retinal ganglion cells degenerate, and the debris is phagocytosed by macrophages and glial cells. However, some morphologically intact axons remain even 12 weeks after surgery. Finally, after 16 weeks all the axons have disappeared, leaving a nerve consisting only of glial cells, some of which contain phagosomes. At 40-52 weeks after enucleation, the nerve persists, at 20-40% of the normal diameter, consisting mostly of normal looking astrocytes. The amplitude of the voltage dependent Na currents recorded from nerves during the first 1-4 weeks after enucleation, with the pial sheath intact, decreases by about 50%. After 8 weeks, the Na current recorded from the surface is about 30% of control. At 16-52 weeks after removal of the retina, when there are no intact axons, the Na current is reduced by 90%. If, however, the pial sheath is stripped away, the Na currents recorded from the glial surface are 40-50% of control during this same 16- to 52-week period, suggesting that in the all-glia nerve, the currents are shunted by the relatively thicker pial sheath.(ABSTRACT TRUNCATED AT 250 WORDS)

Action Potentials

Cardiovascular autonomic function in anthracycline-treated breast cancer patients.

Preclinical studies suggest that in addition to the well-known direct damage to the myocardium, anthracycline antineoplastic drugs exert toxic effects on the cardiovascular autonomic system as well. To investigate whether this phenomenon occurs in the clinic, we carried out noninvasive, widely used tests of cardiovascular autonomic physiology in 55 women with stage II or III breast cancer. In all, 31 were being treated with anthracycline-containing chemotherapy regimens, and 24 who were receiving CMF (cyclophosphamide, Methotrexate, and fluorouracil) served as controls. Of 279 tests conducted in anthracycline (A)-treated patients, 123 were abnormal, vs 54 of 216 tests carried out in 24 controls (44% vs 25%; P less than 0.005). Abnormal variations in heart rate on standing and in diastolic blood pressure during handgrip was found in 25 (81%) and 17 patients receiving A, vs 9 (37%; P less than 0.005) and 5 (21%; P less than 0.0001), respectively, in controls. The incidence of abnormal tests was significantly higher in A-treated patients greater than 60 years of age (41%) vs 67%; P less than 0.05). Radionuclide ventriculography was carried out in 19 patients who showed abnormal tests of cardiovascular autonomic function after greater than or equal to 6 courses of a-containing chemotherapy; only 1 of them had abnormal cardiac contractility (global hypokinesia), suggesting that abnormal tests of cardiovascular autonomic function may occur in the absence of a detectable deterioration in left ventricular ejection fraction. A large number of factors may alter cardiovascular autonomic function in cancer patients, including age, radiation therapy to the chest, and multidrug treatment. Even after correcting for the most obvious of these, chemotherapy with anthracyclines is associated with a significantly higher percentage of abnormal tests for cardiovascular autonomic function. Although indirect and semi-quantitative, our results are compatible with the idea of A-induced cardiac autonomic dysfunction.

Antibiotics, Antineoplastic

Facilitation of voltage-gated ion channels in frog neuroglia by nerve impulses.

The functions of glial cells in the nervous system are not well defined, with the exception of myelin production by oligodendrocytes, uptake of amino-acid synaptic transmitters, and a contribution to extracellular potassium homeostasis. Neuroglia have receptors for neurotransmitters which may be involved in neuron-glia interactions. Recent studies have demonstrated voltage-gated ion channels in glial membranes. In a study of the optic nerve of the frog, small areas of the surface were examined with the loose patch-clamp method, and voltage-gated Na+ and K+ channels, presumably located in the membranes of the astrocytes forming the glia limitans, were identified. We now report that nerve impulses in the axons of the frog optic nerve transiently alter the properties of the voltage-dependent membrane channels of the surface glial cells (astrocytes), a demonstration of a new form of neuron-glia interaction.

Action Potentials

Voltage-gated currents recorded from the surface of the frog optic nerve.

Macroscopic voltage-dependent currents were recorded from the surface of the intact optic nerve of Rana pipiens using the loose patch clamp technique. Depolarizing steps of more than 40 mV produced sodium-dependent TTX sensitive inward currents and a 4-AP and sodium sensitive fast outward current in addition to a slower outward current. Since the surface of the nerve is a glia limitans, it appears that the membranes of these astrocytes contain both voltage-sensitive sodium and potassium channels.

Animals

Conformational changes in bacteriorhodopsin studied by infrared attenuated total reflection.

We report on a new method based on Fourier transform infrared (FTIR)-difference spectroscopy for studying the conformational changes occurring during the photocycle of bacteriorhodopsin. Previous studies have been made by measuring the absorbance of an infrared (IR) beam transmitted through a thin hydrated purple membrane film. In contrast, the present study utilizes the technique of attenuated total reflection (ATR). Purple membrane is fixed on the surface of a germanium internal reflection crystal and immersed in a buffer whose pH and ionic composition can be varied. Measurements of the amide I and II absorbance with light polarized parallel and at 45 degrees to the crystal surface reveals that the membrane is highly oriented. An ATR-FTIR-difference spectrum of the light to dark (bR570 to bR548) transition is similar but not identical to the transmittance FTIR-difference spectrum. This disagreement between the two methods is shown to be due in the ATR case to the absorption of transition moments oriented predominantly out of the membrane plane. Raising the pH of La3+ substituted purple membrane films from 6.8 to 8.0 slows the M-decay rate sufficiently so that a bR570 to M412 difference spectrum can be obtained with steady state illumination at room temperature. A comparison of this difference spectrum with that obtained at -23 degrees C using the transmittance method reveals several changes that cannot be attributed to out-of-plane transition moments. An increase in the intensity of peaks in the amide I and II regions agrees with recent time-resolved kinetic FTIR-difference measurements and indicates that a localized protein conformational change involving the peptide backbone of bR occurs which is not evident at the lower temperature.

Bacteriorhodopsins

Fourier transform infrared difference spectra of intermediates in rhodopsin bleaching.

The membrane protein rhodopsin is the primary light receptor in vision. Fourier transform infrared difference spectroscopy is sensitive to conformational changes in both the protein and the retinylidene chromophore of rhodopsin. By blocking rhodopsin bleaching at specific intermediates, it is possible to elucidate some of the primary molecular events of vision.

Animals

Infrared evidence that the Schiff base of bacteriorhodopsin is protonated: bR570 and K intermediates.

It is possible, by using Fourier-transform infrared (FTIR) difference spectroscopy, to detect the conformational changes occurring in both the protein and the chromophore of bacteriorhodopsin during the photocycle. In contrast to Raman spectroscopy, a laser is unnecessary and hence the problem of a perturbing probe beam is eliminated. Furthermore, the relatively high signal-to-noise ratio obtainable with FTIR enables measurements to be made in minutes over a large spectral range. In the study reported in this paper, we used this method to examine the state of protonation of the retinylidene Schiff base in light-adapted bR570 and in K, the first intermediate in the photocycle. Resonance Raman spectroscopy provides evidence that bR570 is protonated, but these results have been questioned on the basis of theoretical and experimental grounds. FTIR difference spectral changes in the bR570-to-K transition clearly indicate that bR570 contains a protonated Schiff base. In contrast, the K intermediate displays a Schiff base that is altered but still is associated to some degree with a proton. Because the low-temperature FTIR difference spectrum of bR570 and K is similar to the recently reported low-temperature resonance Raman spectra of bR570 and K [Braiman, M. & Mathies, R. (1982) Proc. Natl. Acad. Sci. USA 79, 403-407], we can assign most, but not all, vibrational changes in the bR570-to-K transition to the chromophore. These results are consistent with a simple model of the first step in the photocycle which involves a movement of the Schiff base proton away from a counterion.

Bacteriorhodopsins