Nitrite and Nitrate Levels in Individual Molluscan Neurons: Single‐Cell Capillary Electrophoresis Analysis

Cruz, Lou Ann; Moroz, Leonid L.; Gillette, Rhanor; Sweedler, Jonathan V.

doi:10.1046/j.1471-4159.1997.69010110.x

Cited by 70 publications

(52 citation statements)

References 4 publications

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“…been described previously (Cruz et al, 1997;Floyd et al, 1998;Moroz et al, 1999). Briefly, isolated individual tentacles, the entire manubrium, and small areas of umbrella of equal size were dissected, immediately transferred to 1-2 ml of 400 mM glucose solution in a 35-mm Petri dish, and observed under an inverted microscope (Zeiss, Axiovert 100, Thornwood, NY).…”

Section: Nitrite and Nitrate Assaymentioning

confidence: 99%

“…Detection was carried out on a capillary at 10.2 cm from the outlet by using a UV detector (Thermo Separations Group, Model SC10) set at 214 nm. Peak identity and calibration were accomplished by comparing the migration times with those of standards as has been described in detail elsewhere (Cruz et al, 1997). Five animals were used for this assay.…”

Section: Nitrite and Nitrate Assaymentioning

confidence: 99%

“…Nitrites and nitrates, both breakdown products of NO oxidation (Stamler et al, 1992;Lewis and Deen, 1994;Hogg et al, 1995), can be accumulated in cells and tissues with a high level of NOS activity. Recently, this assay has been successful in identifying the presence of NOS in the mammalian central nervous system (Salter et al, 1996) and in individual molluscan neurons (Cruz et al, 1997). In Aglantha, a high level of nitrite was detected in the tentacles (which had the highest levels of NADPH-d activity) but not in the gonads or other predicted NOS-negative areas.…”

Section: Nos In Aglanthamentioning

confidence: 99%

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Nitric oxide regulates swimming in the jellyfish Aglantha digitale

Moroz

Meech²,

Sweedler

et al. 2004

J of Comparative Neurology

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The cnidarian nervous system is considered by many to represent neuronal organization in its earliest and simplest form. Here we demonstrate, for the first time in the Cnidaria, the neuronal localization of nitric oxide synthase (NOS) in the hydromedusa Aglantha digitale (Trachylina). Expression of specific, fixative-resistant NADPH-diaphorase (NADPH-d) activity, characteristic of NOS, was observed in neurites running in the outer nerve ring at the base of the animal and in putative sensory cells in the ectoderm covering its tentacles. At both sites, diphenyleneiodonium (10(-4) M) abolished staining. Capillary electrophoresis confirmed that the NO breakdown products NO2- and NO3- were present at high levels in the tentacles, but were not detectable in NADPH-d-negative areas. The NADPH-d-reactive neurons in the tentacles send processes to regions adjacent to the inner nerve ring where swimming pacemaker cells are located. Free-moving animals and semi-intact preparations were used to test whether NO is involved in regulating the swimming program. NO (30-50 nM) and its precursor L-arginine (1 mM) stimulated swimming, and the effect was mimicked by 8-Br-cGMP (50-100 microM). The NO scavenger PTIO (10-100 microM) and a competitive inhibitor of NOS, L-nitroarginine methyl ester (L-NAME, 200 microM), significantly decreased the swimming frequency in free-moving animals, while its less-active stereoisomer D-nitroarginine methyl ester (D-NAME, 200 microM) had no such effect. 1H-[1,2,4]oxadiazolo[4,3-a]quinoxaline-1-one (ODQ, 5-20 microM), a selective inhibitor of soluble guanylyl cyclase, suppressed spontaneous swimming and prevented NO-induced activation of the swimming program. We suggest that an NO/cGMP signaling pathway modulates the rhythmic swimming associated with feeding in Aglantha, possibly by means of putative nitrergic sensory neurons in its tentacles.

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Section: Nitrite and Nitrate Assaymentioning

confidence: 99%

Section: Nitrite and Nitrate Assaymentioning

confidence: 99%

Section: Nos In Aglanthamentioning

confidence: 99%

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Nitric oxide regulates swimming in the jellyfish Aglantha digitale

Moroz

Meech²,

Sweedler

et al. 2004

J of Comparative Neurology

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“…Some gastropod species that have large, easily accessible and identifiable NO-ergic neurons provide experimental advantages for such studies. The first work on determining nitrite and nitrate levels in individual neuronal cells has been published by Sweedler's group [387]. Individual NO-positive buccal neurons from the sea slug Pleurobranchaea californica were electrophoretically separated, and NO 2 − and NO 3 − were measured by direct UV absorbance detection.…”

Section: Single-neuron and Neuronal Tissue Hpce Of No Metabolitesmentioning

confidence: 99%

Bioanalytical profile of the l-arginine/nitric oxide pathway and its evaluation by capillary electrophoresis

Boudko

2007

Journal of Chromatography B

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This review briefly summarizes recent progress in fundamental understanding and analytical profiling of the L-arginine/nitric oxide (NO) pathway. It focuses on key analytical references of NO actions and on the experimental acquisition of these references in vivo, with capillary electrophoresis (CE) and high-performance capillary electrophoresis (HPCE) comprising one of the most flexible and technologically promising analytical platform for comprehensive high-resolution profiling of NO-related metabolites. Second aim of this review is to express demands and bridge efforts of experimental biologists, medical professionals and chemical analysis-oriented scientists who strive to understand evolution and physiological roles of NO and to develop analytical methods for use in biology and medicine.

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“…Yet, often, biological samples can be limited in volume such as those collected via push-pull perfusion [16] or single cells [24,25]. Therefore, exploring the use of small volume samples with the HT method is important.…”

Section: Reservoir Tubementioning

confidence: 99%

Hadamard transform CE-UV detection for biological samples

et al. 2005

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A Hadamard transform-capillary electrophoresis-UV (HT-CE-UV) detection technique is described for the analysis of biological samples. Pseudorandom injections of sample and buffer according to a simplex matrix obtained from the corresponding Hadamard matrix is performed with conventional capillaries. Alternating injections are achieved with a novel capillary "T" connector created by drilling conventional capillary dimensions through a 1-cm diameter polycarbonate disc. This connector design coupled with a switching system allows for rapid, electrokinetic injections of solution into alternating sample and buffer capillary arms for UV detection. The standard mixtures of nitric oxide (NO) metabolites, nitrite and nitrate, dissolved in physiological saline solution are injected into the separation capillary according to an 83-element injection sequence to obtain a signal-to-noise ratio (S/N) enhancement of ca. 4.5 over a single injection. Nitrite, being the less concentrated metabolite in NO detection and thereby more difficult to detect, was calibrated with the HT-CE-UV method and a limit of detection (LOD) of 0.56 microM was obtained. Rat blood plasma was analyzed with this detection system and demonstrated to be comparable with NO metabolite concentrations of previously published results. This HT-CE-UV method is described where a unique reservoir tube design that contains 8-microL standard nitrite sample volumes is placed over the end of the capillary arm to explore low volume limits for biological samples.

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Nitrite and Nitrate Levels in Individual Molluscan Neurons: Single‐Cell Capillary Electrophoresis Analysis

Abstract: Cell and tissue concentrations of NO 2-and

Cited by 70 publications

References 4 publications

Nitric oxide regulates swimming in the jellyfish Aglantha digitale

Nitric oxide regulates swimming in the jellyfish Aglantha digitale

Bioanalytical profile of the l-arginine/nitric oxide pathway and its evaluation by capillary electrophoresis

Hadamard transform CE-UV detection for biological samples

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