Dual determination of extracellular cholecystokinin and neurotensin fragments in rat forebrain: Microdialysis combined with a sequential multiple antigen radioimmunoassay

Maidment, Nigel T.; Siddall, Brenda; Rudolph, V.R.; Erdelyi, Elizabeth; Evans, Christopher J.

doi:10.1016/0306-4522(91)90105-w

Cited by 73 publications

(33 citation statements)

References 40 publications

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“…In agreement with previous observations (Maidment et al 1991), the majority of CCKLM collected in microdialysates under high K + conditions coeluted with the sulfated octapeptide CCK-8S. In addition, some CCKLM, corresponding to ~9% of total, coeluted with CCK-5, which is considered as the main metabolite of CCK (Rose et al 1988(Rose et al , 1996.…”

Section: Discussionsupporting

confidence: 88%

“…Previous studies showed that both met-enkephalin (ME) and CCK outflows can be measured in the nucleus accumbens using a microdialysis technique in awake, freely moving rats (Maidment et al 1989(Maidment et al , 1991Daugé et al 1996Daugé et al , 1999Bertrand et al 1997). Using this technique, we presently investigated whether interactions between endogenous opioids and CCK also exist physiologically in the nucleus accumbens.…”

Section: Introductionmentioning

confidence: 97%

See 1 more Smart Citation

Subcutaneous formalin enhances outflow of met-enkephalin- and cholecystokinin-like materials in the rat nucleus accumbens

Lapeyre

Mauborgne²,

Becker³

et al. 2001

Naunyn-Schmiedeberg's Archives of Pharmacology

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Using the microdialysis technique, the present study investigated the effects of a noxious stimulation on the extracellular levels of met-enkephalin and (sulfated octapeptide) cholecystokinin-like materials in the nucleus accumbens of freely moving rats. Injection of 50 microl of 5% formalin into the forepaw produced pain-related behaviours associated with an immediate and sustained (for approximately 2 h) increase (+27%) in the outflow of met-enkephalin-like material within the nucleus accumbens. This treatment also progressively enhanced the local outflow of cholecystokinin-like material that reached 200%-250% of the basal level at the end of the experiment, i.e. 4.5 h after formalin administration. Because naloxone (1.5 mg/kg i.p., 10 min prior to formalin injection) prevented the latter effect, it can be inferred that noxious stimulation-induced activation of cholecystokininergic neurotransmission in the nucleus accumbens probably resulted from the preceding activation of opioidergic systems. These data suggest that the nucleus accumbens may be another structure where interactions between opioids and cholecystokinin play a key role in the control of pain-processing mechanisms.

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Section: Discussionsupporting

confidence: 88%

Section: Introductionmentioning

confidence: 97%

Subcutaneous formalin enhances outflow of met-enkephalin- and cholecystokinin-like materials in the rat nucleus accumbens

Lapeyre

Mauborgne²,

Becker³

et al. 2001

Naunyn-Schmiedeberg's Archives of Pharmacology

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“…125 I] 96-well RIA in which antibody is prebound to polystyrene wells via protein A prior to addition of radioactive ligand and sample; individual wells are broken off at the end of the assay for placement into the gamma counter [8,9]. This assay has the merits of being both convenient and extremely sensitive; however, in our experience, not all RIAs are adaptable to this technique, and the assay times are much longer than those reported here.…”

Section: Discussionmentioning

confidence: 50%

Mini-RIA: adaptation of conventional 125I-labeled radioimmunoassay to a 96-tube format

Laurent

Lindberg

2002

Analytical Biochemistry

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Radioimmunoassay (RIA) employing iodinated ligands represents a popular measurement method for small molecules due to its excellent sensitivity and specificity. Yet performing RIAs of large numbers of tubes remains a tedious laboratory chore due to the need to individually handle tubes multiple times. We here present a method in which conventional 125 I-labeled RIA ([ 125 I] RIA) is adapted to a microtiter plate format, termed mini-RIA. Tubes are handled in batch for centrifugation or during the separation of antibody-bound ligand from free ligand. A simple draining device for batch decantation of free ligand from 96 minitubes is used. Optimal conditions for the mini-RIA were established using two workup methods-double-antibody immunoprecipitation and direct polyethylene glycol precipitation. Use of the mini-RIA method was found to result in a considerable savings in assay times; in addition, the sensitivity of the mini-RIA was improved over conventional RIA. The mini-RIA is particularly useful for assay of large numbers of samples derived from chromatographic methods, since aliquots can be transferred directly from the fraction collector into the minitubes using multiple channel pipettors. Because the method is flexible with regard to assay workup, we predict that most conventional [125 I] RIAs can be adapted to the mini-RIA format. Ó

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“…The earliest studies of neuropeptide release using microdialysis sampling employed radioimmunoassay (RIA), and this approach continues to be popular. RIA has been used for the determination of a number of different peptides, including neuropeptide Y [74] and substance P [75] , neurotensin [76] , and the opioid peptides [77] . Although immunoassays are highly sensitive, the assay itself is time consuming, and the antibodies often exhibit undesirable cross -reactivity, leading to erroneous results in some matrices.…”

Section: Non -Separation -Based Methods Of Analysismentioning

confidence: 99%

“…Using microbore and capillary columns in conjunction with electrochemical or fl uorescence detection makes it possible to detect femtomoles of catecholamines and amino acid neurotransmitters in brain dialysates [89 -91] . Off -line analysis of peptides using LC coupled to radioimmunoassays has also been employed to circumvent the cross -reactivity of the immunoassays [76] . Pharmacokinetic studies will also frequently employ off -line analysis, especially if multiple probes are used in a single animal [43,92] .…”

Section: Off -Line Analysis Using Liquid Chromatographymentioning

confidence: 99%

Analytical Considerations for Microdialysis Sampling

Nandi

Kuhnline

Lunte

2011

Applications of Microdialysis in Pharmaceutical Science

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Dual determination of extracellular cholecystokinin and neurotensin fragments in rat forebrain: Microdialysis combined with a sequential multiple antigen radioimmunoassay

Cited by 73 publications

References 40 publications

Subcutaneous formalin enhances outflow of met-enkephalin- and cholecystokinin-like materials in the rat nucleus accumbens

Subcutaneous formalin enhances outflow of met-enkephalin- and cholecystokinin-like materials in the rat nucleus accumbens

Mini-RIA: adaptation of conventional 125I-labeled radioimmunoassay to a 96-tube format

Analytical Considerations for Microdialysis Sampling

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