Membrane Control of Ciliary Activity in the Protozoan Euplotes

1973 ◽  
Vol 58 (2) ◽  
pp. 437-462
Author(s):  
MILES EPSTEIN ◽  
ROGER ECKERT
Keyword(s):  
Membrane Potential ◽  
Normal Direction ◽  
Direct Access ◽  
Ciliary Activity ◽  
Cell Interior ◽  
Ciliary Beating ◽  
Ciliary Reversal ◽  
Intracellular Ca ◽  
Triton X ◽  
External Calcium

1. Membrane control of ciliary activity in the protozoan Euplotes was investigated by a combination of electrophysiological and cinematographic techniques. 2. The anal cirri, which are quiescent in the absence of stimulation, were selected for this study. 3. Membrane depolarization by means of injected current produced a reversal of the direction of beating (i.e. towards the cell anterior so as to make the ciliate swim backwards). Depolarization also increased the frequency of beating. Increasing depolarizations resulted in an increased number of reversed beats and increased frequency. 4. When the membrane potential was shifted beyond +70 mV, reversed beating did not occur until after the current pulse ended. 5. Depolarization did not evoke reversed beating when the external calcium (Ca) concentration was reduced to 10-6 M with EGTA. 6. Hyperpolarization caused the cirri to beat in a normal direction (i.e. towards the rear of the ciliate so as to cause the animal to swim forward). Increasing hyperpolarizations resulted in an increased number of forward beats and an increased frequency. 7. The cell was treated with the detergent Triton X-100 to permit Ca, Mg and ATP direct access through the extracted membrane to the cell interior. At Ca concentrations below 10-7 M, Mg-ATP-reactivated cilia of Triton-extracted cells beat normally. At Ca concentrations above approximately 10-7 M the reactivated beat resembled the reversed beat in the living cell. 8. The evidence suggests that membrane-regulated concentrations of intracellular Ca control the direction of ciliary beating. Thus, stimuli which produce an adequate Ca influx lead to ciliary reversal.

scholarly journals PKA induces Ca2+ release and enhances ciliary beat frequency in a Ca2+-dependent and -independent manner

1998 ◽  
Vol 275 (3) ◽  
pp. C790-C797 ◽  
Author(s):  
Alex Braiman ◽  
Orna Zagoory ◽  
Zvi Priel
Keyword(s):  
Beat Frequency ◽  
Ciliary Beat Frequency ◽  
Ciliary Activity ◽  
Experimental Conditions ◽  
Independent Manner ◽  
Ciliary Beating ◽  
Independent Mechanism ◽  
Intracellular Ca ◽  
Ciliary Beat

The intent of this work was to evaluate the role of cAMP in regulation of ciliary activity in frog mucociliary epithelium and to examine the possibility of cross talk between the cAMP- and Ca2+-dependent pathways in that regulation. Forskolin and dibutyryl cAMP induced strong transient intracellular Ca2+ concentration ([Ca2+]i) elevation and strong ciliary beat frequency enhancement with prolonged stabilization at an elevated plateau. The response was not affected by reduction of extracellular Ca2+concentration. The elevation in [Ca2+]iwas canceled by pretreatment with 1,2-bis(2-aminophenoxy)ethane- N, N, N′, N′-tetraacetic acid-AM, thapsigargin, and a phospholipase C inhibitor, U-73122. Under those experimental conditions, forskolin raised the beat frequency to a moderately elevated plateau, whereas the initial strong rise in frequency was completely abolished. All effects were canceled by H-89, a selective protein kinase A (PKA) inhibitor. The results suggest a dual role for PKA in ciliary regulation. PKA releases Ca2+ from intracellular stores, strongly activating ciliary beating, and, concurrently, produces moderate prolonged enhancement of the beat frequency by a Ca2+-independent mechanism.

Interactions of membrane potential and cations in regulation of ciliary activity in Paramecium

1976 ◽  
Vol 65 (2) ◽  
pp. 427-448
Author(s):  
H. Machemer
Keyword(s):  
Membrane Potential ◽  
High Frequency ◽  
Resting Potential ◽  
Ciliary Activity ◽  
Frequency Increase ◽  
External Concentration ◽  
Membrane Hyperpolarization ◽  
Ciliary Beating ◽  
Fixed Membrane ◽  
K Concentration

Ciliary activity in Paramecium was investigated in different external solutions using techniques of voltage clamp and high frequency cinematography. An increase in the external concentration of K, Ca or Mg ions decreased the resting potential. It had no effect on ciliary activity. When the membrane potential was fixed, an increase in external Ca or Mg and, to a lesser extent, an increase in K concentration, raised the frequency of normal beating or decreased the frequency of reversed beating of the cilia. Similar effects resulted from membrane hyperpolarization with constant ionic conditions. Increase in concentration of Ca, but not of Mg or K, enhanced hyperpolarization-induced augmentation of ciliary frequency. Increase in Ca concentration also specifically augmented the delayed increase in inward current during rapid hyperpolarizing clamp. The results support the view that [Ca]i regulates the frequency and direction of ciliary beating. It is suggested that the insensitivity of the ciliary motor system to elevations of the external concentrations of ions results from compensation of their effects on [Ca]i. Depolarization itself appears to increase [Ca]i while elevation of the external ion concentrations at a fixed membrane potential appears to decrease [Ca]i.

scholarly journals Electrophysiological Control of Reversed Ciliary Beating in Paramecium

1973 ◽  
Vol 61 (5) ◽  
pp. 572-587 ◽  
Author(s):  
Hans Machemer ◽  
Roger Eckert
Keyword(s):  
Membrane Potential ◽  
Electric Stimulation ◽  
Surface Membrane ◽  
Progressive Increase ◽  
Membrane Depolarization ◽  
Equilibrium Potential ◽  
Positive Potential ◽  
Ciliary Beating ◽  
Current Pulses ◽  
Ciliary Reversal

Quantitative relations between ciliary reversal and membrane responses were examined in electrically stimulated paramecia. Specimens bathed in 1 mM CaCl2, 1 mM KCl, and 1 mM Tris-HCl, pH 7.2, were filmed at 250 frames per second while depolarizing current pulses were injected. At current intensities producing only electrotonic shifts the cilia failed to respond. Stimuli which elicited a regenerative response were followed by a period of reversed ciliary beating. With increasing stimulus intensities the latency of ciliary reversal dropped from 30 to 4 ms or less, and the duration of reversal increased from 50 ms to 2.4 s or more; the corresponding regenerative responses increased in amplitude and rate of rise. With progressively larger intracellular positive pulses, electric stimulation became less effective, producing responses with a progressive increase in latency and decrease in duration of reversed beating of the cilia. When 100-ms pulses shifted the membrane potential to +70 mV or more, ciliary reversal was suppressed until the end of the pulse. "Off" responses then occurred with a latency of 2–4 ms independent of further increases in positive potential displacement. These results suggest that ciliary reversal is coupled to membrane depolarization by the influx of ions which produces the regenerative depolarization of the surface membrane. According to this view suppression of the ciliary response during stimulation occurs when the membrane potential approaches the equilibrium potential of the coupling ion, thereby retarding its influx. Previous data together with the present findings suggest that this ion is Ca2+.

Control of Ciliary Activities by Adenosinetriphosphate and Divalent Cations in Triton-Extracted Models of Paramecium Caudatum

1973 ◽  
Vol 58 (3) ◽  
pp. 657-676 ◽  
Author(s):  
YUTAKA NAITOH ◽  
HIROKI KANEKO
Keyword(s):  
Cell Membrane ◽  
Divalent Cations ◽  
Power Stroke ◽  
Ciliary Activity ◽  
Paramecium Caudatum ◽  
Cell Models ◽  
Effective Power ◽  
Ciliary Beating ◽  
Triton X ◽  
Beating Frequency

1. Cilia of Paramecium candatum extracted with Triton X-100 were reactivated in the presence of ATP and Mg2+. 2. The beating frequency of the reactivated cilia is a function of both the ATP and Mg2+ concentrations. 3. The reactivated cell models swam forward when the Ca2+ concentrations in the ATP-Mg2+ medium was kept below 10-7M. They swam backward when the Ca2+ concentration was above 10-6M. This was due to a reversed orientation of the effective power stroke of the reactivated cilia. 4. In the absence of Mg2+ the cilia failed to beat, even though ATP was present. If Ca2+ was then added the cilia assumed a new orientation, pointing toward the anterior without beating. 5. Ni2+ inhibited ciliary beating in the reactivated models, but has no influence on changes in the orientation of the cilia produced by ATP and Ca2+. This suggests that one ATP-activated system is responsible for beating, while another governs the direction of the effective stroke. 6. Mn2+ is half as effective as Mg2+ for inducing ciliary beating in the extracted models in the presence of ATP. 7. Salyrgan strongly inhibits Mg2+-ATP activated ciliary activity of the model. 8. Bioelectric control of ciliary activity by the cell membrane of live animals is discussed.

Role of the Cilia Membrane in Control of Microtubule Sliding

1980 ◽  
Vol 38 ◽  
pp. 612-615
Author(s):  
Edna S. Kaneshiro
Keyword(s):  
Control Mechanism ◽  
Beat Frequency ◽  
Surface Membrane ◽  
Ciliary Membrane ◽  
Ciliary Beating ◽  
Ciliary Reversal ◽  
Voltage Dependent ◽  
Reversal Mechanism ◽  
And Function

It is currently believed that ciliary beating results from microtubule sliding which is restricted in regions to cause bending. Cilia beat can be modified to bring about changes in beat frequency, cessation of beat and reversal in beat direction. In ciliated protozoans these modifications which determine swimming behavior have been shown to be related to intracellular (intraciliary) Ca2+ concentrations. The Ca2+ levels are in turn governed by the surface ciliary membrane which exhibits increased Ca2+ conductance (permeability) in response to depolarization. Mutants with altered behaviors have been isolated. Pawn mutants fail to exhibit reversal of the effective stroke of ciliary beat and therefore cannot swim backward. They lack the increased inward Ca2+ current in response to depolarizing stimuli. Both normal and pawn Paramecium made leaky to Ca2+ by Triton extrac¬tion of the surface membrane exhibit backward swimming only in reactivating solutions containing greater than IO-6 M Ca2+ Thus in pawns the ciliary reversal mechanism itself is left operational and only the control mechanism at the membrane is affected. The topographic location of voltage-dependent Ca2+ channels has been identified as a component of the ciliary mem¬brane since the inward Ca2+ conductance response is eliminated by deciliation and the return of the response occurs during cilia regeneration. Since the ciliary membrane has been impli¬cated in the control of Ca2+ levels in the cilium and therefore is the site of at least one kind of control of microtubule sliding, we have focused our attention on understanding the structure and function of the membrane.

Regulation of intracellular calcium in N1E-115 neuroblastoma cells: the role of Na+/Ca2+exchange

2002 ◽  
Vol 282 (5) ◽  
pp. C1000-C1008 ◽  
Author(s):  
Kara L. Kopper ◽  
Joseph S. Adorante
Keyword(s):  
Membrane Potential ◽  
Intracellular Calcium ◽  
Normal Cell ◽  
Buffer Capacity ◽  
Neuroblastoma Cells ◽  
Fold Increase ◽  
Scorpion Venom ◽  
Reverse Mode ◽  
Intracellular Ca

In fura 2-loaded N1E-115 cells, regulation of intracellular Ca2+ concentration ([Ca2+]i) following a Ca2+ load induced by 1 μM thapsigargin and 10 μM carbonylcyanide p-trifluoromethyoxyphenylhydrazone (FCCP) was Na+ dependent and inhibited by 5 mM Ni2+. In cells with normal intracellular Na+ concentration ([Na+]i), removal of bath Na+, which should result in reversal of Na+/Ca2+exchange, did not increase [Ca2+]i unless cell Ca2+ buffer capacity was reduced. When N1E-115 cells were Na+ loaded using 100 μM veratridine and 4 μg/ml scorpion venom, the rate of the reverse mode of the Na+/Ca2+ exchanger was apparently enhanced, since an ∼4- to 6-fold increase in [Ca2+]ioccurred despite normal cell Ca2+ buffering. In SBFI-loaded cells, we were able to demonstrate forward operation of the Na+/Ca2+ exchanger (net efflux of Ca2+) by observing increases (∼ 6 mM) in [Na+]i. These Ni2+ (5 mM)-inhibited increases in [Na+]i could only be observed when a continuous ionomycin-induced influx of Ca2+ occurred. The voltage-sensitive dye bis-(1,3-diethylthiobarbituric acid) trimethine oxonol was used to measure changes in membrane potential. Ionomycin (1 μM) depolarized N1E-115 cells (∼25 mV). This depolarization was Na+dependent and blocked by 5 mM Ni2+ and 250–500 μM benzamil. These data provide evidence for the presence of an electrogenic Na+/Ca2+ exchanger that is capable of regulating [Ca2+]i after release of Ca2+ from cell stores.

Differential effects of UTP, ATP, and adenosine on ciliary activity of human nasal epithelial cells

2001 ◽  
Vol 280 (6) ◽  
pp. C1485-C1497 ◽  
Author(s):  
Diane M. Morse ◽  
Jennifer L. Smullen ◽  
C. William Davis
Keyword(s):  
Epithelial Cells ◽  
Beat Frequency ◽  
Ciliary Beat Frequency ◽  
Ciliary Activity ◽  
Maximal Effect ◽  
Ciliary Beating ◽  
Mediated Effects ◽  
Human Nasal Epithelium ◽  
Human Nasal Epithelial Cells ◽  
Sustained Responses

The purinergic regulation of ciliary activity was studied using small, continuously superfused explants of human nasal epithelium. The P2Y2 purinoceptor (P2Y2-R) was identified as the major purinoceptor regulating ciliary beat frequency (CBF); UTP (EC50 = 4.7 μM), ATP, and adenosine-5′- O-(3-thiotriphosphate) elicited similar maximal responses, approximately twofold over baseline. ATP, however, elicited a post-peak sustained plateau in CBF (1.83 ± 0.1-fold), whereas the post-peak CBF response to UTP declined over 15 min to a low-level plateau (1.36 ± 0.16-fold). UDP also stimulated ciliary beating, probably via P2Y6-R, with a maximal effect approximately one-half that elicited by P2Y2-R stimulation. Not indicated were P2Y1-R-, P2Y4-R-, or P2Y11-R-mediated effects. A2B-receptor agonists elicited sustained responses in CBF approximately equal to those from UTP/ATP [5′-( N-ethylcarboxamido)adenosine, EC50 = 0.09 μM; adenosine, EC50 = 0.7 μM]. Surprisingly, ADP elicited a sustained stimulation in CBF. The ADP effect and the post-peak sustained portion of the ATP response in CBF were inhibited by the A2-R antagonist 8-( p-sulfophenyl)theophylline. Hence, ATP affects ciliary activity through P2Y2-R and, after an apparent ectohydrolysis to adenosine, through A2BAR.

scholarly journals Ciliomotor circuitry underlying whole-body coordination of ciliary activity in the Platynereis larva

2017 ◽  
Author(s):  
Csaba Verasztó ◽  
Nobuo Ueda ◽  
Luis A. Bezares-Calderón ◽  
Aurora Panzera ◽  
Elizabeth A. Williams ◽  
...  
Keyword(s):  
Beat Frequency ◽  
Ciliary Beat Frequency ◽  
Whole Body ◽  
Ciliary Activity ◽  
Ciliary Beating ◽  
Stop And Go ◽  
Multiciliated Cells ◽  
Platynereis Dumerilii ◽  
Catecholaminergic Cells ◽  
Ciliary Locomotion

AbstractCiliated surfaces harbouring synchronously beating cilia can generate fluid flow or drive locomotion. In ciliary swimmers, ciliary beating, arrests, and changes in beat frequency are often coordinated across extended or discontinuous surfaces. To understand how such coordination is achieved, we studied the ciliated larvae of Platynereis dumerilii, a marine annelid. Platynereis larvae have segmental multiciliated cells that regularly display spontaneous coordinated ciliary arrests. We used whole-body connectomics, activity imaging, transgenesis, and neuron ablation to characterize the ciliomotor circuitry. We identified cholinergic, serotonergic, and catecholaminergic ciliomotor neurons. The synchronous rhythmic activation of cholinergic cells drives the coordinated arrests of all cilia. The serotonergic cells are active when cilia are beating. Serotonin inhibits the cholinergic rhythm, and increases ciliary beat frequency. Based on their connectivity and alternating activity, the catecholaminergic cells may generate the rhythm. The ciliomotor circuitry thus constitutes a stop-and-go pacemaker system for the whole-body coordination of ciliary locomotion.

The resting membrane potential of Drosophila melanogaster larval muscle depends strongly on external calcium concentration

2010 ◽  
Vol 56 (3) ◽  
pp. 304-313 ◽  
Author(s):  
Jacob L. Krans ◽  
Karen D. Parfitt ◽  
Kristin D. Gawera ◽  
Patricia K. Rivlin ◽  
Ronald R. Hoy
Keyword(s):  
Drosophila Melanogaster ◽  
Membrane Potential ◽  
Calcium Concentration ◽  
Resting Membrane Potential ◽  
Larval Muscle ◽  
External Calcium

Voltage-dependence of ciliary activity in the ciliate Didinium nasutum

1995 ◽  
Vol 198 (12) ◽  
pp. 2537-2545 ◽  
Author(s):  
J Pernberg ◽  
H Machemer
Keyword(s):  
Membrane Potential ◽  
Voltage Dependence ◽  
Longitudinal Axis ◽  
Threshold Level ◽  
Swimming Behaviour ◽  
Ciliary Activity ◽  
Frequency Increase ◽  
Cyclic Activity ◽  
Beating Frequency ◽  
Ca2 Channels

In the gymnostome ciliate Didinium nasutum, swimming behaviour depends upon the cyclic activity of about 3000 cilia. The normal beating mode, resulting in forward swimming of the cell, is characterized by a posteriad effective beat (18 left of the longitudinal axis) at a frequency of approximately 15 Hz. Activation of depolarization-sensitive ciliary Ca2+ channels leads to an increase in intracellular Ca2+ concentration and a change in the beating mode. Following rapid reorientation, the effective stroke is anteriad (24 ° right of the longitudinal axis) and the beating frequency is about 26 Hz, resulting in fast backward swimming of the cell. In response to minor depolarizations, and hence small increases in cytoplasmic Ca2+ concentration, the cilia inactivate. Frequency increase and reversal in beat orientation share a single threshold level of membrane potential, since both changes of the beating mode occur simultaneously.

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