scholarly journals ATP signaling in the integrative neural center of Aplysia californica

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
János Györi ◽  
Andrea B. Kohn ◽  
Daria Y. Romanova ◽  
Leonid L. Moroz

AbstractATP and its ionotropic P2X receptors are components of the most ancient signaling system. However, little is known about the distribution and function of purinergic transmission in invertebrates. Here, we cloned, expressed, and pharmacologically characterized the P2X receptors in the sea slug Aplysia californica—a prominent neuroscience model. AcP2X receptors were successfully expressed in Xenopus oocytes and displayed activation by ATP with two-phased kinetics and Na+-dependence. Pharmacologically, they were different from other P2X receptors. The ATP analog, Bz-ATP, was a less effective agonist than ATP, and PPADS was a more potent inhibitor of the AcP2X receptors than the suramin. AcP2X were uniquely expressed within the cerebral F-cluster, the multifunctional integrative neurosecretory center. AcP2X receptors were also detected in the chemosensory structures and the early cleavage stages. Therefore, in molluscs, rapid ATP-dependent signaling can be implicated both in development and diverse homeostatic functions. Furthermore, this study illuminates novel cellular and systemic features of P2X-type ligand-gated ion channels for deciphering the evolution of neurotransmitters.

2021 ◽  
Author(s):  
János Györi ◽  
Andrea Kohn ◽  
Daria Romanova ◽  
Leonid Moroz

Abstract ATP and its ionotropic P2X receptors are components of the most ancient signaling systems. However, little is known about the distribution and function of purinergic transmission in invertebrates. Here, we cloned, expressed, and pharmacologically characterized P2X receptors in the sea slug Aplysia californica – the prominent neuroscience model. acP2X receptors were successfully expressed in Xenopus oocytes and were displayed activation by ATP with two-phased kinetics and Na+-dependence. Pharmacologically, they were quite different from other P2X receptors. The ATP analog, Bz-ATP, was a less effective agonist than ATP, and PPADS was a more potent inhibitor of the acP2X receptors than the suramin. acP2X were uniquely expressed within the cerebral F-cluster, which contains multiple secretory peptides (e.g., insulins, interleukins, and potential toxins), ecdysone-type receptors, and a district subset of ion channels. We view F-cluster as the multifunctional integrative center, remarkably different from other neurosecretory cells. acP2X receptors were also found in the chemosensory structures and the early cleavage stages. Therefore, in molluscs, rapid ATP-dependent signaling can be implicated both in development and diverse homeostatic functions. Furthermore, this study illuminates novel cellular and systemic features of P2X-type ligand-gated ion channels for deciphering evolution of neurotransmitters.


2020 ◽  
Author(s):  
János Györi ◽  
Andrea B. Kohn ◽  
Leonid L. Moroz

AbstractATP and its ionotropic P2X receptors are components of one of the most ancient signaling systems. However, little is known about the distribution and function of purinergic transmission in invertebrates. Here, we cloned, expressed, and pharmacologically characterized P2X receptors in the sea slug Aplysia californica – a prominent model in cellular and system neuroscience. We showed that ATP and P2X receptors are essential signaling components within the unique bioenergetic center located in the CNS of Aplysia, also known as the cerebral F-cluster of insulin-containing neurons. Functional P2X receptors were successfully expressed in Xenopus oocytes to characterize their ATP-dependence (EC50=306μM), two-phased kinetics, ion selectivity (Na+-dependence), sensitivity to the ATP analog Bz-ATP (~20% compare to ATP) and antagonists (with PPADS as a more potent inhibitor compared to suramin). Next, using RNA-seq, we characterized the expression of P2X receptors across more than a dozen Aplysia peripheral tissues and developmental stages. We showed that P2X receptors are predominantly expressed in chemosensory structures and during early cleavage stages. The localization and pharmacology of P2X receptors in Aplysia highlight the evolutionary conservation of bioenergetic sensors and chemosensory purinergic transmission across animals. This study also provides a foundation to decipher homeostatic mechanisms in development and neuroendocrine systems.Graphical AbstractWe show that ATP and its ligand-gated P2X receptors are essential signaling components within both the chemosensory systems and the unique bioenergetic center, present in the CNS of the sea slug Aplysia californica – a prominent model in neuroscience. Expression and pharmacology of P2X receptors in Aplysia confirms the preservation of evolutionary conserved bioenergetic sensors across animals and provide new tools to decipher homeostatic mechanisms in neuro-endocrine systems in general.


2020 ◽  
Author(s):  
János Györi ◽  
Andrea B. Kohn ◽  
Daria Y. Romanova ◽  
Leonid L. Moroz

AbstractATP and its ionotropic P2X receptors are components of one of the most ancient signaling systems. However, little is known about the distribution and function of purinergic transmission in invertebrates. Here, we cloned, expressed, and pharmacologically characterized P2X receptors in the sea slug Aplysia californica – the prominent model in cellular and system neuroscience. These functional P2X receptors were successfully expressed in Xenopus oocytes and displayed activation by ATP (EC50=306 μM) with two-phased kinetics as well as Na+-dependence. The ATP analog, Bz-ATP, was a less effective agonist (~20%) than ATP, and PPADS was a more potent inhibitor of the P2X receptors than the suramin. We showed that P2X receptors are uniquely expressed within Aplysia’s cerebral bioenergetic center (also known as F-cluster). Using RNA-seq, we found that the F-cluster contains more than a dozen unique secretory peptides, including three insulins, interleukins, and potential toxins, as well as ecdysone-type receptors and a district subset of ion channels. This structure is one of the most prominent integrative centers in the entire CNS and remarkably different from the morphologically similar neurosecretory center (bag cluster) involved in egg-laying behavior. Using RNA-seq, we also characterized the expression of P2X receptors across more than a dozen Aplysia peripheral tissues and developmental stages. We showed that P2X receptors are predominantly expressed in chemosensory structures and during early cleavage stages. The localization and pharmacology of P2X receptors in Aplysia highlight the evolutionary conservation of bioenergetic sensors and chemosensory purinergic transmission across animals. This study also provides a foundation to decipher homeostatic mechanisms in development and neuroendocrine systems.Graphical AbstractWe show that ATP and its ligand-gated P2X receptors are essential signaling components within both the chemosensory systems and the unique integrative neurosecretory center, present in the CNS of the sea slug Aplysia – a prominent model in neuroscience. Expression and pharmacology of P2X receptors in Aplysia confirms the preservation of evolutionary conserved bioenergetic sensors across animals and provide new tools to decipher homeostatic mechanisms in neuro-endocrine systems in general.


Development ◽  
1992 ◽  
Vol 115 (4) ◽  
pp. 1175-1182 ◽  
Author(s):  
T. Akiyama ◽  
M. Okada

Mitochondria of early Drosophila embryos were observed with a transmission electron microscope and a fluorescent microscope after vital staining with rhodamine 123, which accumulates only in active mitochondria. Rhodamine 123 accumulated particularly in the posterior pole region in early cleavage embryos, whereas the spatial distribution of mitochondria in an embryo was uniform throughout cleavage stages. In late cleavage stages, the dye showed very weak and uniform accumulation in all regions of periplasm. Polar plasm, sequestered in pole cells, restored the ability to accumulate the dye. Therefore, it is concluded that the respiratory activity of mitochondria is higher in the polar plasm than in the other regions of periplasm in early embryos, and this changes during development. The temporal changes in rhodamine 123-staining of polar plasm were not affected by u.v. irradiation at the posterior of early cleavage embryos at a sufficient dosage to prevent pole cell formation. This suggests that the inhibition of pole cell formation by u.v. irradiation is not due to the inactivation of the respiratory activities of mitochondria. In addition, we found that the anterior of Bicaudal-D mutant embryos at cleavage stage was stained with rhodamine 123 with the same intensity as the posterior of wild-type embryos. No pole cells form in the anterior of Bic-D embryos, where no restoration of mitochondrial activity occurs in the blastoderm stage. The posterior group mutations that we tested (staufen, oskar, tudor, nanos) and the terminal mutation (torso) did not alter staining pattern of the posterior with rhodamine 123.


Author(s):  
Meysam T. Chorsi ◽  
Pouya Tavousi ◽  
Caitlyn Mundrane ◽  
Vitaliy Gorbatyuk ◽  
Kazem Kazerounian ◽  
...  

Abstract Natural nanomechanisms such as capillaries, neurotransmitters, and ion channels play a vital role in the living systems. But the design principles developed by nature through evolution are not well understood and, hence, not applicable to engineered nanomachines. Thus, the design of nanoscale mechanisms with prescribed functions remains a challenge. Here, we present a systematic approach based on established kinematics techniques to designing, analyzing, and controlling manufacturable nanomachines with prescribed mobility and function built from a finite but extendable number of available "molecular primitives." Our framework allows the systematic exploration of the design space of irreducibly simple nanomachines, built with prescribed motion specification by combining available nanocomponents into systems having constrained, and consequently controllable motions. We show that the proposed framework has allowed us to discover and verify a molecule in the form of a seven link, seven revolute (7R) close loop spatial linkage with mobility (degree of freedom) of one. Furthermore, our experiments exhibit the type and range of motion predicted by our simulations. Enhancing such a structure into functional nanomechanisms by exploiting and controlling their motions individually or as part of an ensemble could galvanize development of the multitude of engineering, scientific, medical, and consumer applications that can benefit from engineered nanomachines.


1971 ◽  
Vol 26 (8) ◽  
pp. 816-821 ◽  
Author(s):  
Larry E. Bockstahler

Incorporation of uridine in cleavage stage eggs of the sea urchin Paracentrotus lividus was investigated. It was shown by ion exchange and thin layer chromatography that most of the uridine taken up during the 16-cell stage was converted into UTP with some incorporation into UDP and UMP. Conversion of uridine to these phosphorylated nucleosides occurred throughout early cleavage stages. A very small amount of uridine taken up by cleavage stage eggs is incorporated into RNA heterogeneous in size. This RNA was examined by polyacrylamide gel electrophoresis.


2017 ◽  
Vol 5 (12) ◽  
pp. e13290 ◽  
Author(s):  
Anne Hahn ◽  
Johannes Faulhaber ◽  
Lalita Srisawang ◽  
Andreas Stortz ◽  
Johanna J Salomon ◽  
...  

Development ◽  
1959 ◽  
Vol 7 (2) ◽  
pp. 173-192
Author(s):  
A. A. Neyfakh

It is generally accepted at present that during cleavage in echinoderms, amphibians, and fishes, the nuclei do not have specific functions in regulating development, their role being at this time restricted to participation in the processes of cleavage (Schechtman & Nishihara, 1955). Eggs devoid of nuclei sometimes begin cleavage which may proceed up to the stage of the late blastula. Extirpation or inactivation of nuclei may be achieved through the separation of the nuclear region of the egg by means of centrifugation (Harvey, 1940); through extirpation of the female nucleus followed by fertilization with sperm inactivated by a heavy dose of radiation (Briggs, Green, & King, 1951); through spontaneous degeneration of the male nucleus during artificial androgenesis (Stauffer, 1945); and by means of other techniques. Exposure of early cleavage stages in amphibians (Mangold & Peters, 1956; Sanides, 1956) and fishes (Neyfakh, 1956a) to heavy doses of ionizing radiation also leads to arrest of development at the late blastula stage.


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