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001-es BibID:BIBFORM004938
Első szerző:Balkay László (biofizikus)
Cím:Flow cytometric determination of intracellular free potassium concentration / Balkay, L., Marian, T., Emri, M., Krasznai, Z., Tron, L.
Dátum:1997
ISSN:0196-4763
Megjegyzések:A flow cytometric assay has been developed for determination of intracellular free potassium concentration ([K+]i). Investigated cells, loaded with the fluorescent pH indicator 2',7bis-(2-carboxyethyl)-5(and-6)-carboxyfluorescein (BCECF), are incubated in the presence of nigericin, and the intracellular pH is measured. The ionophore maintains the same ratio value [H+]/[K+] on both sides of the cytoplasmic membrane, so [K+]i can be evaluated from the measured intracellular pH (pHi) and the known parameters of the buffer. Application of the method revealed that the intracellular potassium concentration is significantly higher in lymphocytes than in proliferating cells of HUT-78, U266, and JY cell lines. A surprisingly low (60 mM) [K+]i concentration was observed with sperm cells of common carp. This method allows measurements on individual cells, provides data of excellent statistics, and still does not require large amounts of material. These features offer remarkable advantages over other techniques used for intracellular K+ measurements, such as steady-state fluorescence, atom absorption photometry, or energy-dispersive x-ray analysis.
Tárgyszavak:Orvostudományok Elméleti orvostudományok idegen nyelvű folyóiratközlemény külföldi lapban
analysis
Animal
Biophysics
Buffers
Cell Line
Cells
diagnostic use
Dyes
Flow Cytometry
Fluorescein
Fluoresceins
Fluorescence
Fluorescent Dyes
Human
Hungary
Hydrogen-Ion Concentration
Ionophores
Lymphocytes
metabolism
Mice
Nigericin
Potassium
Rats
Megjelenés:Cytometry. - 28 : 1 (1997), p. 42-49. -
További szerzők:Márián Teréz (1950-) (radiobiológus) Emri Miklós (1962-) (fizikus) Krasznai Zoltán (1950-) (biofizikus) Trón Lajos (1941-) (biofizikus)
Internet cím:elektronikus változat
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2.

001-es BibID:BIBFORM004948
Első szerző:Márián Teréz (radiobiológus)
Cím:Role of extracellular and intracellular pH in carp sperm motility and modifications by hyperosmosis of regulation of the Na+/H+ exchanger / Marian, T., Krasznai, Z., Balkay, L., Emri, M., Tron, L.
Dátum:1997
ISSN:0196-4763
Megjegyzések:Recently developed flow cytometric methods (Tron et al.: Mol Immunol 27:1307-1311, 1990) to measure the intracellular pH (pHi) and intracellular potassium concentration in mammalian cells by using the fluorescent pH-indicator dye 2',7bis-(2-carboxyethyl)-5 (and-6)-carboxyfluorescein (BCECF) were adopted for measuring these parameters in carp sperm. The intracellular potassium concentration of the carp sperm was 62.4 +/- 5.3 mM. This is very similar to the potassium concentration of the seminal plasma (87 +/- 16 mM), and it suggests a depolarized state of the sperm cell in the semen. An average pHi value of 7.06 +/- 0.11 was obtained by measuring sperm samples taken from ten animals. Changes in the ionic composition of the environment did not alter pHi. Sperm motility was initiated by transferring the cells to an environment of 110 mOsm osmolality. This hypoosmotic shock induced fast changes in the membrane structure that could be reversed by restoring physiologic osmolality. Activation was accompanied by a fast alkalinization of the sperm cells. This pH change was amiloride sensitive, suggesting the involvement of the Na+/H+ exchanger in the activation process. Alkalinization of acid-loaded sperm cells depended on the osmolality of the environment. Equilibrium pHi of these cells in hyperosmotic buffers was substantially lower relative to cells in an isoosmotic environment. Effects of the extracellular and intracellular pH on carp sperm motility were also examined. Extracellular pH below 5.5 abolished sperm motility completely. Alkaline extracellular pH did not alter the duration of sperm motility even at extreme values (pHe = 9.6). Duration of the flagellar motion did not depend on the pHi between values of 6.5 and 8.5; however, it was significantly reduced both below and above this range. No motility was observed below pHi = 6.0 or above pHi = 9.5 with a 10 min incubation time at these pH values prior to activation. Effects of the extracellular and intracellular pH on sperm motility were partially reversible.
Tárgyszavak:Orvostudományok Elméleti orvostudományok idegen nyelvű folyóiratközlemény külföldi lapban
Amiloride
Animal
Buffers
Carps
Cell Membrane Permeability
Cells
chemistry
cytology
Dyes
Extracellular Space
Flow Cytometry
Fluorescein
Fluoresceins
Fluorescent Dyes
Hungary
Hydrogen-Ion Concentration
Intracellular Fluid
Light
Male
methods
Motion
Osmolar Concentration
physiology
Potassium
Scattering,Radiation
Semen
Sodium-Hydrogen Antiporter
Sperm Motility
Spermatozoa
Support,Non-U.S.Gov't
egyetemen (Magyarországon) készült közlemény
Megjelenés:Cytometry. - 27 : 4 (1997), p. 374-382. -
További szerzők:Krasznai Zoltán (1950-) (biofizikus) Balkay László (1963-) (biofizikus) Emri Miklós (1962-) (fizikus) Trón Lajos (1941-) (biofizikus)
Internet cím:elektronikus változat
Borító:
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