Isolation of Sensory Neurons of <em>Aplysia californica</em> for Patch Clamp Recordings of Glutamatergic Currents

The marine gastropod mollusk Aplysia californica has a venerable  history as a model of nervous system function, with particular significance in  studies ...

University of Miami

The marine toxin, okadaic acid (OA) is produced by dinoflagellates of the genera Prorocentrum and Dinophysis and is the causative agent of the syndrome known as diarrheic shellfish poisoning (DSP). In addition, OA acts as both a tumor promoter, attributed to OA-induced inhibition of protein phosphatases as well as an inducer of apoptosis. To better understand the potentially divergent toxicological profile of OA, the concentration dependent cytotoxicity and alterations in gene expression on the human liver tumor cell line HepG2 upon OA exposure were determined using RNA microarrays, DNA fragmentation, and cell proliferation assays as well as determinations of cell detachment and cell death in different concentrations of OA. mRNA expression was quantified for approximately 15,000 genes. Cell attachment and proliferation were both negatively correlated with OA concentration. Detached cells displayed necrotic DNA signatures but apoptosis also was broadly observed. Data suggest that OA has a concentration dependent effect on cell cycle, which might explain the divergent effects that at low concentration OA stimulates genes involved in the cell cycle and at high concentrations it stimulates apoptosis.

GENE EXPRESSION PROFILING OF HUMAN LIVER CARCINOMA (HepG2) CELLS EXPOSED TO THE MARINE TOXIN OKADAIC ACID.

Les toxines marines, l&#39;acide okada&iuml;que (AO) est produite par les dinoflagell&eacute;s des genres Prorocentrum et Dinophysis et est l&#39;agent causal du syndrome appel&eacute; diarrh&eacute;iques shellfish poisoning (DSP). En outre, OA agit comme un promoteur tumeur, attribu&eacute; &agrave; l&#39;inhibition induite par l&#39;OA des prot&eacute;ines phosphatases comme un inducteur de l&#39;apoptose. Pour mieux comprendre le profil toxicologique potentiellement divergent de OA, la cytotoxicit&eacute; d&eacute;pendante de concentration et les modifications dans l&#39;expression des g&egrave;nes sur la lign&eacute;e de cellules de tumeur h&eacute;patique humaine HepG2 lors de l&#39;exposition de l&#39;arthrose sont d&eacute;termin&eacute;es au moyen des RNA microarrays, fragmentation de l&#39;ADN et les analyses de prolif&eacute;ration de cellules ainsi que d&eacute;cisions de d&eacute;tachement cellulaire et de la mort cellulaire dans des concentrations diff&eacute;rentes d&#39;OA. expression de l&#39;ARNm a &eacute;t&eacute; quantifi&eacute;e pour environ 15 000 g&egrave;nes. Fixation des cellules et la prolif&eacute;ration ont &eacute;t&eacute; tous deux n&eacute;gativement corr&eacute;l&eacute;es avec la concentration de l&#39;OA. Les cellules individuelles affichent n&eacute;crotiques signatures ADN mais l&#39;apoptose a &eacute;t&eacute; observ&eacute;e aussi largement. R&eacute;sultats sugg&egrave;rent qu&#39;OA a un effet d&eacute;pendant de la concentration sur le cycle cellulaire, ce qui pourrait expliquer les effets divergents qui, &agrave; faible concentration OA, stimule les g&egrave;nes impliqu&eacute;s dans le cycle cellulaire et &agrave; des concentrations &eacute;lev&eacute;es, il stimule l&#39;apoptose.

Profil d&#39;EXPRESSION g&eacute;nique de cellules de carcinome du foie humain (HepG2) expos&eacute;s &agrave; l&#39;acide OKADA&Iuml;QUE de toxines marines.

La tossina marina, acido okadaico (OA) &egrave; prodotta da dinoflagellati del genere Prorocentrum e Dinophysis ed &egrave; l&#39;agente eziologico della sindrome nota come diarroiche shellfish poisoning (DSP). Inoltre, OA agisce come sia un promotore tumorale, attribuito a OA-indotta di inibizione della fosfatasi della proteina cos&igrave; come un induttore di apoptosi. Per capire meglio il profilo tossicologico potenzialmente divergente di OA, la concentrazione dipendente citotossicit&agrave; e alterazioni nell&#39;espressione genica sulla linea cellulare di tumore epatico umano HepG2 all&#39;esposizione OA sono stati determinati utilizzando RNA microarray, frammentazione del DNA e dosaggi di proliferazione delle cellule nonch&eacute; le determinazioni di distacco delle cellule e delle cellule morte in diverse concentrazioni di OA. espressione del mRNA &egrave; stato quantificato per circa 15.000 geni. Accessorio cellulare e proliferazione sono stati negativamente correlate con concentrazione di OA. Cellule Detached visualizzato necrotiche firme DNA ma apoptosis inoltre ampiamente &egrave; stato osservato. I dati suggeriscono che OA ha un effetto dipendente dalla concentrazione sul ciclo cellulare, che potrebbe spiegare gli effetti divergenti che a bassa concentrazione OA stimola i geni coinvolti nel ciclo cellulare e ad alte concentrazioni stimola l&#39;apoptosi.

Profilo di espressione genica delle cellule di CARCINOMA EPATICO umano (HepG2) esposte all&#39;acido OKADAICO tossina marina.

As, Cr, and Cu represent one potential combination of multiple metals/metalloids exposures since these three elements are simultaneously leached from chromated copper arsenate (CCA)-treated wood, a common product used for building construction, at levels that can be potentially harmful. This study investigated the neurotoxicity of As associated with CCA-treated wood when accompanied by Cr and Cu. The toxicity was evaluated on the basis of a cytotoxicity model using human neuroblastoma cell line SK-N-SH. The cells were cultured with CCA-treated wood leachates or with solutions containing arsenate [As(V)], divalent copper [Cu(II)], trivalent chromium [Cr(III)] alone or in different combinations of the three elements. The toxicity was evaluated using variations in cell replication compared to controls after 96 h exposure. Among the three elements present in wood leachates, As played the primary role in the observed toxic effects, which were exerted through multiple pathways, including the generation of oxidative stress. DOM affected the absorption of metals/metalloids into the test cells, which however did not obviously appear to impact toxicity. As toxicity was enhanced by Cu(II) and inhibited by Cr(III) at concentrations below U.S. EPA's allowable maximum contaminant levels in drinking waters. Thus assessing As toxicity in real environments is not sufficient if based solely on the result from As.

Arsenic toxicity in the human nerve cell line SK-N-SH in the presence of chromium and copper.

Comme, Cr et Cu repr&eacute;sentent une combinaison potentielle de plusieurs expositions de m&eacute;taux/m&eacute;tallo&iuml;des, &eacute;tant donn&eacute; que ces trois &eacute;l&eacute;ments sont lessiv&eacute;s simultan&eacute;ment de l&#39;ars&eacute;niate de cuivre chromat&eacute; (ACC)-trait&eacute; de bois, un produit courant utilis&eacute; pour la construction de b&acirc;timents, &agrave; des niveaux qui peut &ecirc;tre potentiellement dangereux. Cette &eacute;tude examine la neurotoxicit&eacute; du comme associ&eacute; de bois trait&eacute; au CCA lorsqu&#39;ils sont accompagn&eacute;s par le Cr et Cu. La toxicit&eacute; a &eacute;t&eacute; &eacute;valu&eacute;e sur la base d&#39;un mod&egrave;le de cytotoxicit&eacute; &agrave; l&#39;aide de neuroblastome humain lign&eacute;e de cellules SK-N-SH. Les cellules sont cultiv&eacute;es avec les lixiviats de bois trait&eacute; au CCA solutions contenant de l&#39;ars&eacute;niate [As (v)], cuivre divalent [Cu (II)], du chrome trivalent [CR] seul ou dans diff&eacute;rentes combinaisons de trois &eacute;l&eacute;ments. La toxicit&eacute; a &eacute;t&eacute; &eacute;valu&eacute;e &agrave; l&#39;aide des variations dans la r&eacute;plication cellulaire par rapport aux contr&ocirc;les apr&egrave;s exposition de 96 heures. Parmi les trois &eacute;l&eacute;ments pr&eacute;sents dans les lixiviats de bois, a jou&eacute; le r&ocirc;le principal dans les effets toxiques observ&eacute;s, qui ont &eacute;t&eacute; exerc&eacute;es par des voies multiples, notamment la g&eacute;n&eacute;ration du stress oxydatif. DOM affect&eacute; l&#39;absorption des m&eacute;taux/m&eacute;tallo&iuml;des dans les h&eacute;maties-tests, qui toutefois ne pas &eacute;videmment figuraient &agrave; la toxicit&eacute; de l&#39;impact. Comme la toxicit&eacute; a &eacute;t&eacute; am&eacute;lior&eacute;e de Cu (II) et inhib&eacute;e par le CR (III) &agrave; des concentrations inf&eacute;rieures de l&#39;EPA maximal admissible de contaminants dans les eaux potables. &Eacute;valuation ainsi que la toxicit&eacute; dans des environnements r&eacute;els ne suffit pas si fond&eacute;es uniquement sur le r&eacute;sultat de comme.

Toxicit&eacute; de l&#39;arsenic dans la nerf lign&eacute;e cellulaire humaine SK-N-SH en pr&eacute;sence de chrome et de cuivre.

Come Cr e Cu rappresentano un potenziale combinazione di esposizioni multiple di metalli/metalloidi poich&eacute; questi tre elementi sono contemporaneamente lisciviati da arsenato di rame cromatura (CCA)-trattati di legno, un prodotto comune utilizzato per la costruzione, a livelli che pu&ograve; essere potenzialmente dannoso. Questo studio ha studiato la neurotossicit&agrave; del come associato con legno trattato CCA quando accompagnata da Cr e Cu. La tossicit&agrave; &egrave; stata valutata sulla base di un modello di citotossicit&agrave; mediante neuroblastoma umano cella linea SK-N-SH. Le cellule sono state coltivate con percolati legno trattato CCA o con soluzioni contenenti arseniato [As(V)], rame bivalente [cu], cromo trivalente [CR] da sola o in diverse combinazioni di tre elementi. La tossicit&agrave; &egrave; stata valutata utilizzando le variazioni nella replica delle cellule rispetto ai controlli dopo l&#39;esposizione 96 h. Tra i tre elementi presenti nel legno percolati, come ha svolto il ruolo principale in effetti tossici osservati, che erano esercitati attraverso percorsi multipli, compreso la generazione dello stress ossidativo. DOM influenzato l&#39;assorbimento di metalli/metalloidi nelle cellule del test, che tuttavia non ha fatto ovviamente compaiono alla tossicit&agrave; di impatto. Come la tossicit&agrave; &egrave; stata rafforzata da cu e inibita da CR (III) a concentrazioni inferiori US EPA contaminante massimo consentito nelle acque potabili. Quindi valutare come tossicit&agrave; negli ambienti reali non &egrave; sufficiente se basato esclusivamente sul risultato da As.

Justin B. Greer

Division of Marine Biology and Fisheries,
Rosenstiel School of Marine and Atmospheric Sciences,
Division of Marine Biology and Fisheries, Rosenstiel School of Marine and Atmospheric Sciences

University of Miami

Isolation of Sensory Neurons of Aplysia californica for Patch Clamp Recordings of Glutamatergic Currents

Justin B. Greer

.css-o250i3{font-size:18px;font-family:Open Sans,sans-serif;line-height:21.6px;}Division of Marine Biology and Fisheries,Rosenstiel School of Marine and Atmospheric Sciences,Division of Marine Biology and Fisheries, Rosenstiel School of Marine and Atmospheric Sciences

University of Miami

Isolation of Sensory Neurons of Aplysia californica for Patch Clamp Recordings of Glutamatergic Currents

Division of Marine Biology and Fisheries,
Rosenstiel School of Marine and Atmospheric Sciences,
Division of Marine Biology and Fisheries, Rosenstiel School of Marine and Atmospheric Sciences