Acetylcholine synthesis and possible functions during sea urchin development (original) (raw)
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Journal of Experimental Zoology, 1983
Acetylcholinesterase (AChE) activity has been studied in Artemia salina embryos and larvae by quantitative, histochemical, and electrophoretic methods. An AChE activity is present in dehydrated encysted gastrulae and remains low in rehydrated developing embryos, showing at hatching an increase that becomes faster during the nauplius stages (twice the gastrula activity at the nauplius I1 stage), and continues linearly in the first metanauplius stages. The histochemical staining, faint and diffuse in dehydrated gastrulae, in more advanced embryos becomes localized in cell membranes and stronger in differentiating tissues; most AChE activity is localized in the nervous and muscular cells, and a fainter one in the differentiating metanauplius segments. Dehydrated cysts show one electrophoretic AChE band, that is no longer detectable in rehydrated embryos and larvae, while two other bands appear, that migrate more slowly; in advanced larval stages, the more cathodic band is predominant. In dehydrated cysts, isoelectric focusing separates three AChE bands with isoelectric points (IP) at pH 5.0, 5.7, and 5.9; in hydrated embryos and larvae, the band at pH 5.0 is absent and another is seen at pH 5.5. Eserine, DFP, and BW284c51 inhibit the enzyme activity strongly and cause paralysis of larvae; also tubocurarine blocks the motility. These results are discussed in relation to the differentiation of the neuromuscular system and to the development of motility. As a hypothesis, in early embryonic stages, before any neuromuscular differentiation is evident, a cholinergic-like system might regulate non-neural cell activities and interactions, associated with embryogenetic events, This work is dedicated to our master, Professor Antonio Min-Address reprint requests to Dr. M Raineri, Istituto di Anato ganti.
Caryologia, 2006
The distribution of acetylcholinesterase (AChE) activity has been studied in ovary, unfertilized egg, embryos and larva of Ciona intestinalis and Ascidia malaca (Ascidiacea, Urochordata) by biochemical and histochemical methods. The biochemical results have shown that the AChE activity is detected in ovary, and in the egg before fertilization; it rises from 8-cell stage up to gastrula, neurula, and above all at swimming larva. Cholinesterase activity has been histochemically localized and the results of this study agree with the biochemical data. The distribution of the enzyme activity in ovary and in different stages of development suggests also a non-classical role on maturation of female gametes, on cell adhesiveness and interaction, on differentiation, on secretion and apoptosis process.
Developmental Brain Research, 2003
The ontogeny of cholinergic neurons has been studied in the brain of the urodele amphibian Pleurodeles waltl by means of choline acetyltransferase (ChAT) immunohistochemistry. Embryonic and larval stages were studied. The earliest ChAT immunoreactive (ChATi) cells were the primary motoneurons in the upper spinal cord, at embryonic stage 29. Slightly later, before hatching, the cranial nerve motor nuclei were immunopositive as well as non-motor populations in the developing inferior reticular nucleus, the nucleus of the solitary tract, the dorsal column nucleus and the retina. At initial larval stages, ChATi cells were located for the first time in the suprachiasmatic nucleus and the isthmic tegmentum. In addition, moderate immunoreactivity appeared in the Mauthner cells. During the period of active larval life the cholinergic systems maturated progressively and new ChATi cell groups were found in the caudal telencephalon and the habenula. Also extensive fiber labeling occurred at active larval stages. No transient ChAT expression was observed and even the Mauthner cells maintained their immunoreactivity after metamorphosis. A general caudorostral spatio-temporal sequence of appearance of cholinergic structures was found in the brain. Comparison of the results observed in the urodele with previous data available only in amniotes shows numerous similarities and suggests a conservative developmental pattern of cholinergic systems in vertebrates.
Comparative biochemistry and physiology. C, Comparative pharmacology and toxicology, 1989
1. The changes in the consistence of the spine-test articular capsule, or ligament, of the primary spines of Eucidaris tribuloides induced by acetylcholine (ACh) have been studied. Two complementary techniques were used: (a) "forced-vibration", which detects variations in the stiffness of the ligament along a single diametral plane; and (b) "forced-rotation" which records the spatial distribution of those changes. 2. ACh (1 microM to 1 mM) caused a rapid increase in the resistive force opposed by the ligament to passive stretching. Similar effects were elicited by several monoquaternary, N-substituted derivatives of trimethylammonium. 3. The opposite effect, i.e. softening, was induced by decamethonium, dimethylphenylpiperazine, and 2-ketoamyltrimethylammonium. 4. The involvement in these effects of ACh-binding groups with pharmacological properties similar to those of the "anionic sites" of nicotinic ACh receptors is suggested.
Identification of a sperm receptor on the surface of the eggs of the sea urchin Arbacia punctulata
The Journal of cell biology, 1977
The possibility that the surface of the egg of the sea urchin Arbacia punctulata contains a species-specific receptor for sperm has been investigated. The extent of fertilization of eggs of A. punctulata, which is proportional to the number of sperm, is unaffected by the presence of either eggs or membranes prepared from eggs of Strongylocentrotus purpuratus. In marked contrast, membranes prepared from eggs of A. punctulata quantitatively inhibit fertilization of A. punctulata eggs by A. punctulata sperm. Several lines of evidence indicate that this inhibition is due to the presence of a membrane-associated glycoprotein that binds to the sperm, thus preventing them from interacting with receptor on the surface of the eggs. First, eggs treated with trypsin are incapable of being fertilized, although they can be activated with the Ca2+ ionophore A23187. Moreover, membranes prepared from eggs pretreated with trypsin do not inhibit fertilization of eggs. Second, receptor isolated in sol...
Cell Calcium, 2002
The existence of a response to acetylcholine (ACh) and cholinomimetic drugs in sea urchin eggs and zygotes was investigated in two sea urchin species: Paracentrotus lividus and Lytechinus pictus. The calcium sensitive fluorescent probe, Fura-2 dextran, was employed to investigate the regulation of cytosolic free calcium concentration ([Ca 2+ ] i ) by cholinomimetic drugs in unfertilised and fertilised eggs of both the sea urchin species. Exposure to cholinomimetic agonists/antagonists, either extracellularly or intracellularly, had no effect either on resting [Ca 2+ ] i levels in the unfertilised sea urchin egg, or on the transient [Ca 2+ ] i increase at fertilisation. However, following fertilisation, extracellular application of ACh receptors agonists, such as ACh and carbachol, predominantly muscarinic agonist, but not nicotine, induced a significant increase in [Ca 2+ ] i , which was partially inhibited by atropine. As a consequence of exposure after fertilisation to the agonists of ACh receptors, chromatin structure was transiently affected. The hypothesis is proposed that muscarinic receptors may be involved in the (presumably Ca 2+ -dependent) modulation of the nuclear status during the first cell cycles.
Brain Research, 1986
In the hatchetfish, the Mauthner cell (M-cell) is thought to be cholinergic based on electrophysiological studies using cholinergic agents and on the localization of acetylcholinesterase (ACHE) and a-bungarotoxin to M-cell-giant fiber synapses. Immunocytochemical studies have shown that mammalian and non-mammalian cholinergic neurons stain positive for choline acetyltransferase (CHAT), the enzyme responsible for synthesizing acetylcholine. We processed tissue from the goldfish (Carassius auratus) for the immunohistochemical detection of ChAT using the monoclonal antibody AB8 and the peroxidase-antiperoxidase procedure. ChAT immunoreactivity was found in selected areas of the goldfish brain including the cranial nerve nuclei and the ventral horn motoneurons of the spinal cord. Interestingly, the M-cell soma which stains positive for AChE was ChAT negative. This immunohistochemical evidence does not support cholinergic functioning of the Mauthner cell.
Chemico-Biological Interactions, 2005
In zebrafish, the onset of acetylcholinesterase (AChE) expression was detected by RT-PCR at 4 hpf (hours postfertilization). The aryl acylamidase (AAA) associated with AChE, a serotonin sensitive activity with unknown physiological function, was significantly higher than the esterase activity on zebrafish embryos homogenates at 4-12 h development (test-t = 3.523; d.f. = 4). Remarkably, the ratio of AAA/AChE activity decreased 210-fold from 4 to 144 h development, indicating a distinct embryonic role of AAA during early embryogenesis. The AAA activity was sensitive to eserine and serotonin, ensuring its association with AChE. This is the first report of AAA activity on fish, establishing zebrafish as a model to study AAA on development.