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A Rakowska

Publications and source records attributed to A Rakowska.

7 recordsLinked to original sources
PubMed1998

ATP-Induced shape change of nuclear pores visualized with the atomic force microscope.

Bidirectional transport of molecules between nucleus and cytoplasm through the nuclear pore complexes (NPCs) spanning the nuclear envelope plays a fundamental role in cell function and metabolism. Nuclear import of macromolecules is a two-step process involving initial recognition of targeting signals, docking to the pore and energy-driven translocation. ATP depletion inhibits the translocation step. The mechanism of translocation itself and the conformational changes of the NPC components that occur during macromolecular transport, are still unclear. The present study investigates the effect of ATP on nuclear pore conformation in isolated nuclear envelopes from Xenopus laevis oocytes using the atomic force microscope. All experiments were conducted in a saline solution mimicking the cytosol using unfixed nuclear envelopes. ATP (1 mM) was added during the scanning procedure and the resultant conformational changes of the NPCs were directly monitored. Images of the same nuclear pores recorded before and during ATP exposure revealed dramatic conformational changes of NPCs subsequent to the addition of ATP. The height of the pores protruding from the cytoplasmic surface of the nuclear envelope visibly increased while the diameter of the pore opening decreased. The observed changes occurred within minutes and were transient. The slow-hydrolyzing ATP analogue, ATP-gamma-S, in equimolar concentrations did not exert any effects. The ATP-induced shape change could represent a nuclear pore "contraction."

Adenosine Triphosphate↗
PubMed1998

[Erythropoietin in the pathogenesis of anemia in patients with multiple myeloma].

UNLABELLED: Anemia is a common symptom in multiple myeloma (MM) patients but the pathogenesis of it is still unknown. The aim of the study was to explain the causes of anemia in MM patients. Peripheral blood count, bone marrow aspirate, iron and ferritin level, serum erythropoietin (EPO) level, T cell subsets and in vitro CFU-E count were analyzed in the group od 31 MM patients. Erythropoietin and iron deficiency in the study group were not observed. EPO serum level was not significantly different in patients with multiple myeloma and in comparison to patients with sideroblastic anemia with solid tumors. Absolute CD8 T lymphocyte count was not significantly increased in the study group. CFU-E colonies count in vitro was not decreased in these patients. CONCLUSIONS: In the study group of the MM patients anemia probably does not depend on EPO production. Diminished proliferative response of erythropoietic cells on normal serum level of EPO and abnormal iron utilisation probably occur in these patients. Replacement of normal erythropoiesis by tumor plasma cells is probably not decisive.

Adult↗
PubMed1997

The atomic force microscope detects ATP-sensitive protein clusters in the plasma membrane of transformed MDCK cells.

Plasma membrane proteins are supposed to form clusters that allow 'functional cross-talk' between individual molecules within nanometre distance. However, such hypothetical protein clusters have not yet been shown directly in native plasma membranes. Therefore, we developed a technique to get access to the inner face of the plasma membrane of cultured transformed kidney (MDCK) cells. The authors applied atomic force microscopy (AFM) to visualize clusters of native proteins protruding from the cytoplasmic membrane surface. We used the K+ channel blocker iberiotoxin (IBTX), a positively charged toxin molecule, that binds with high affinity to plasma membrane potassium channels and to atomically flat mica. Thus, apical plasma membranes could be 'glued' with IBTX to the mica surface with the cytosolic side of the membrane accessible to the scanning AFM tip. The topography of these native inside-out membrane patches was imaged with AFM in electrolyte solution mimicking the cytosol. The plasma membrane could be clearly identified as a lipid bilayer with the characteristic height of 4.9 +/- 0.02 nm. Multiple proteins protruded from the lipid bilayer into the cytosolic space with molecule heights between 1 and 20 nm. Large protrusions were most likely protein clusters. Addition of the proteolytic enzyme pronase to the bath solution led to the disappearance of the proteins within minutes. The metabolic substrate ATP induced a shape-change of the protein clusters and smaller subunits became visible. ADP or the non-hydrolysable ATP analogue, ATP-gamma-S, could not exert similar effects. It is concluded that plasma membrane proteins (and/or membrane associated proteins) form 'functional clusters' in their native environment. The 'physiological' arrangement of the protein molecules within a cluster requires ATP.

Adenosine Triphosphate↗
PubMed1997

Using atomic force microscopy to investigate patch-clamped nuclear membrane.

Nuclear patch clamp is an emerging research field that aims to disclose the electrical phenomena underlying macromolecular transport across the nuclear envelope (NE), its properties as an ion barrier and its function as an intracellular calcium store. The authors combined the patch clamp technique with atomic force microscopy (AFM) to investigate the structure-function relationship of NE. In principle, patch clamp currents, recorded from the NE can indicate the activity of the nuclear pore complexes (NPCs) and/or of ion channels in the two biomembranes that compose the NE. However, the role of the NPCs is still nuclear because the observed NE current in patch clamp experiments is lower than expected from the known density of the NPCs. Therefore, AFM was applied to link patch clamp currents to structure. The membrane patch was excised from the nuclear envelope and, after electrical evaluation, transferred from the patch pipette to a substrate. We could identify the native nuclear membrane patches with AFM at a lateral and a vertical resolution of 3 nm and 0.1 nm, respectively. It was shown that complete NE together with NPCs can be excised from the nucleus after their functional identification in patch clamp experiments. However, we also show that membranes of the endoplasmic reticulum can contaminate the tip of the patch pipette during nuclear patch clamp experiments. This possibility must be considered carefully in nuclear patch clamp experiments.

Animals↗
PubMed1995

Ca2+ antagonists reveal a novel inhibitory effect of caerulein on guinea pig ileum.

The peptide caerulein (CK) is a potent muscle stimulant of guinea pig ileum and its effect is Ca2+-dependent. Ca2+ antagonist drugs administered to the organ perfusion solution diminished the twitches of the electrically stimulated ileum. Subsequent administration of caerulein reinforced this inhibition of contraction and acetylcholine (ACh) release by the ileum. This effect was antagonized by cholecystokinin antagonists (i.e., proglumide and loxiglumide) and by dibutyryl-cyclic-guanosine monophosphate, which overcame the stimulant effect of the peptide. These findings indicated that in the guinea pig ileum, caerulein exerts two opposite effects, both of which are mediated by the same receptor: a stimulation effect, with an increase in acetylcholine release when the organ is incubated in normal saline, and a relaxation effect, with a reduction in acetylcholine release when the ileum is bathed with calcium antagonist drugs.

Acetylcholine↗
PubMed1991

Changes in motor unit action potentials during the fatigue test.

The studies were performed on medial gastrocnemius motor units of the rat. Changes in amplitude, duration and latency of motor unit action potentials were monitored in the course of fatigue test in the three types of motor unites: FF (fast twitch, fatigable), FR (fast twitch, resistant), and S (slow twitch, resistant) motor unit. In a majority of the motor units the amplitude of action potential decreased. The most pronounced decrease took place in FF units, less pronounced one in FR units, while in S type motor units the decrease was insignificant. However, in a part of investigated motor units the amplitude of their action potential transiently increased at the beginning of the fatigue test and, then decreased or was maintained at the same level till the end of the fatigue test. In a few motor units, particularly the slow ones, no changes in action potential amplitude were observed. In the studied material, a decrease in action potential amplitude was on the average less pronounced than a decrease in tetanus tension in the course of the fatigue test. In a majority of studied motor units the duration of action potential was becoming prolonged, which was most pronounced in FF type motor units and least pronounced in S type ones. In few slow motor units the duration showed no alterations. Changes in action potential duration were much more marked than changes in their amplitude. The changes in action potential in fast motor units were less pronounced than changes in their tetanus tension, while in the slow motor units they resembled each other in intensity. Comparison of motor unit action potential changes with changes in their tetanic tension in the course of fatigue test demonstrated no clear-cut relationship between the two phenomena.

Action Potentials↗