Optical approaches to ontogeny of electrical activity and related functional organization during early heart development

1991 ◽  
Vol 71 (1) ◽  
pp. 53-91 ◽  
Author(s):  
K. Kamino

Direct intracellular measurement of electrical events in the early embryonic heart is impossible because the cells are too small and frail to be impaled with microelectrodes; it is also not possible to apply conventional electrophysiological techniques to the early embryonic heart. For these reasons, complete understanding of the ontogeny of electrical activity and related physiological functions of the heart during early development has been hampered. Optical signals from voltage-sensitive dyes have provided a new powerful tool for monitoring changes in transmembrane voltage in a wide variety of living preparations. With this technique it is possible to make optical recordings from the cells that are inaccessible to microelectrodes. An additional advantage of the optical method for recording membrane potential activity is that electrical activity can be monitored simultaneously from many sites in a preparation. Thus, applying a multiple-site optical recording method with a 100- or 144-element photodiode array and voltage-sensitive dyes, we have been able to monitor, for the first time, spontaneous electrical activity in prefused cardiac primordia in the early chick embryos at the six- and the early seven-somite stages of development. We were able to determine that the time of initiation of the contraction is the middle period of the nine-somite stage. In the rat embryonic heart, the onset of spontaneous electrical activity and contraction occurs at the three-somite stage. In this review, a new view of the ontogenetic sequence of spontaneous electrical activity and related physiological functions such as ionic properties, pacemaker function, conduction, and characteristics of excitation-contraction coupling in the early embryonic heart are discussed.

Author(s):  
R H. Selinfreund ◽  
A. H. Cornell-Bell

Cellular electrophysiological properties are normally monitored by standard patch clamp techniques . The combination of membrane potential dyes with time-lapse laser confocal microscopy provides a more direct, least destructive rapid method for monitoring changes in neuronal electrical activity. Using membrane potential dyes we found that spontaneous action potential firing can be detected using time-lapse confocal microscopy. Initially, patch clamp recording techniques were used to verify spontaneous electrical activity in GH4\C1 pituitary cells. It was found that serum depleted cells had reduced spontaneous electrical activity. Brief exposure to the serum derived growth factor, IGF-1, reconstituted electrical activity. We have examined the possibility of developing a rapid fluorescent assay to measure neuronal activity using membrane potential dyes. This neuronal regeneration assay has been adapted to run on a confocal microscope. Quantitative fluorescence is then used to measure a compounds ability to regenerate neuronal firing.The membrane potential dye di-8-ANEPPS was selected for these experiments. Di-8- ANEPPS is internalized slowly, has a high signal to noise ratio (40:1), has a linear fluorescent response to change in voltage.


1959 ◽  
Vol 197 (4) ◽  
pp. 829-834 ◽  
Author(s):  
Dana C. Brooks

The spontaneous electrical activity of the ventromedial nucleus was studied in the cat under pentobarbital anesthesia and in the unanesthetized, unrestrained state. Under light pentobarbital anesthesia the activity of the nucleus is characterized by a predominant 9–15 cps, 50–100 µv component which is uniform from second to second. With small additional doses of anesthesia there is a selective depression of this activity; with recovery from light anesthesia this activity is gradually replaced by irregular, large, slow waves characteristic of sleep. When the unanesthetized animal is aroused 20–35 cps activity having an amplitude of 40 µv or more appears in the nucleus. While the pattern of activity during sleep resembles that seen elsewhere in the hypothalamus, the activity seen during barbiturate anesthesia and during arousal is confined to the nucleus and not seen in other parts of the diencephalon.


PLoS Genetics ◽  
2013 ◽  
Vol 9 (9) ◽  
pp. e1003793 ◽  
Author(s):  
Katharina Wystub ◽  
Johannes Besser ◽  
Angela Bachmann ◽  
Thomas Boettger ◽  
Thomas Braun

2005 ◽  
Vol 94 (6) ◽  
pp. 4131-4144 ◽  
Author(s):  
Ling Chen ◽  
Masahiro Sokabe

The effects of pregnenolone sulfate (PREGS), a putative neurosteroid, on the transmission of perforant path–granule cell synapses were investigated with an optical recording technique in rat hippocampal slices stained with voltage-sensitive dyes. Application of PREGS to the bath solution resulted in an acute augmentation of EPSP in a dose-dependent manner. The PREGS effect was dependent on the extracellular Ca2+ concentration ([Ca2+]o), but independent of NMDA receptor activation. PREGS caused a decrease in paired-pulse facilitation, which implies that PREGS positively modulates presynaptic neurotransmitter releases. Firmer support for this mechanism was that PREGS augmented the synaptically induced glial depolarization (SIGD) that reflects the activity of electrogenic glutamate transporters in glial cells during the uptake of released glutamate. The selective α7nAChR antagonist α-BGT or MLA prevented the SIGD increase by PREGS. Furthermore DMXB, a selective α7nAChR agonist, mimicked the PREGS effect on SIGD and antagonized the effect of PREGS. The presynaptic effect of PREGS was partially attenuated by the L-type Ca2+ channel (VGCC) blocker nifedipine. Based on these findings, we proposed a novel mechanism underlying the facilitated synaptic transmission by PREGS: this neurosteroid sensitizes presynaptic α7nAChR that is followed by an activation of L-type VGCC to increase the presynaptic glutamate release.


1959 ◽  
Vol 196 (2) ◽  
pp. 343-350 ◽  
Author(s):  
C. Y. Kao

The spontaneous electrical activity of uterine smooth muscle was rather variable when acute observations were made. Therefore, a series of chronic experiments was performed with implanted electrodes to monitor a group of myometrial cells under different physiological conditions for periods up to eight weeks. The results showed that consistent behavior of myometrial cells could be observed provided similar hormonal status was maintained. Action potentials were rare or absent in myometrium of oophorectomized animals but were caused to appear by estrogen. In the pregnant uterus, action potentials increased both in amplitude and frequency of discharge as parturition approached, reached a peak at that time, and then declined in the postpartum days. The results indicated that in estrogen treatment and in parturition activities of myometrial cells were more synchronous. There was suggestive evidence that there were central impulses initiating activity in the pregnant uterus, and that the responses of the myometrium were affected by the local conditions.


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