A new procedure for haptenizing adenosine leading to a more specific radioimmunoassay method

Newby, Andrew C.; Sala, G

doi:10.1042/bj2080603

Cited by 14 publications

(8 citation statements)

References 24 publications

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“…Two to three portions of 50-100 mg each were separately processed and assayed for adenosine bv RIA as described in the text. This enzyme converts adenosine to inosine whose affinity to the antibody is 30000 times lower than that of adenosine (Newby & Sala 1982). The en¬ zyme is specific for adenosine.…”

Section: Resultsmentioning

confidence: 99%

Assay of adenosine in human adipose tissue

Ranta

Kiviluoto

Newby³

et al. 1985

Acta Endocrinologica

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A method is described to assay the adenosine content of human adipose tissue. Tissue pieces of 10\p=n-\20 mg were frozen within 1 sec of excision by a conchotome. The pieces were pooled and ground in liquid nitrogen, divided into samples weighing 50\p=n-\100 mg each and placed in perchloric acid. Neutralized defatted extracts were treated by Dowex AG-1 to remove nucleotides and lyophilized. The adenosine content was measured by RIA using antiserum raised against laevulinic acid ( 02' , 3' \ x=r eq-\ adenosine-acetal)-albumin conjugate. The adenosine content in human abdominal sc adipose tissue was 0.56 \ m=+-\ 0.08 nmol/g of tissue (mean \ m=+-\SEM). This shows that adenosine is present in human adipose tissue at concentrations that have been shown to have regulatory effects on cAMP accumulation and lipolysis.

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Section: Resultsmentioning

confidence: 99%

Assay of adenosine in human adipose tissue

Ranta

Kiviluoto

Newby³

et al. 1985

Acta Endocrinologica

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“…Staining was most intense in perikarya and select fiber regions. The antisera employed in the present study have previously been shown by radioimmunoassay to be both sensitive and selective for adenosine (26). Several control experiments, as previously described for immunocytochemical staining of the rat brain (1), were conducted to ensure that the immunoreactivity observed was specific for adenosine itself (data not shown).…”

Section: Resultsmentioning

confidence: 99%

“…Paraffin tissue sections (8 Am) were stained using a modification of the avidin-biotinperoxidase complex (ABC) technique (11,24,25). The production and characterization of specific antisera to adenosine using the adenosine analogue laevulinic acid (O2'3'_ adenosine acetal) has been described previously (26). Briefly, tissue was deparaffinized, rehydrated, and immunocytochemically stained as follows: (i) incubation with 0.2% Triton X-100 in 0.…”

Section: Methodsmentioning

confidence: 99%

Endogenous adenosine and adenosine receptors localized to ganglion cells of the retina.

Braas¹,

Zarbin²,

Snyder³

1987

Proc. Natl. Acad. Sci. U.S.A.

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Using specific sensitive antisera against adenosine, we have immunocytochemically localized endogenous adenosine to specific layers of rat, guinea pig, monkey, and human retina. Highest adenosine immunoreactivity was observed in ganglion cells and their processes in the optic nerve fiber layer. Substantial staining was also found throughout the inner plexiform layer and in select cells in the inner nuclear layer. Adenosine Al receptors, labeled with the agonists L-[3H]phenylisopropyladenosine and 1251-labeled hydroxyphenylisopropyladenosine, were autoradiographically localized. The highest levels of binding sites occurred in the nerve fiber, ganglion cell, and inner plexiform layers of the retina in all the species examined. The distribution of adenosine Al receptor sites closely parallels that of retinal neurons and fibers containing immunoreactive adenosine. These results suggest a role for endogenous adenosine as a coneurotransmitter in ganglion cells and their fibers in the optic nerve.The purine nucleoside adenosine influences numerous physiological processes, including effects on neuronal communication within the central nervous system. These observations suggest that adenosine may serve a role as a neurotransmitter or neuromodulator. Adenosine and its analogues inhibit the release of other neurotransmitters (1-7) and depress both spontaneous and stimulated neuronal depolarization (8). The actions of adenosine are mediated by two receptor subtypes: high affinity Al receptors inhibit adenylate cyclase activity, whereas low affinity A2 receptors stimulate the enzyme activity (9,10). Immunocytochemical localizations of endogenous adenosine in neuronal perikarya and fibers in discrete regions of the brain (11) support a neurotransmitter or neuromodulator role.

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“…The production and characterization of this antisera has been previously described (Newby & Sala, 1982). The production and characterization of this antisera has been previously described (Newby & Sala, 1982).…”

Section: Antiseramentioning

confidence: 99%

Comparison of adenosine uptake and endogenous adenosine-containing cells in mammalian retina

1989

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Autoradiographic techniques were used to label [ 3 H]-adenosine and [ 3 H]-cyclohexyladenosine accumulating cells in rabbit, mouse, and ground squirrel retinas. Immunohistochemical methods revealed the distribution of cells that stained for endogenous adenosine. Comparisons of these two markers revealed for all three species that the distribution of specific subpopulations of retinal cells that store or accumulate the purine nucleoside, adenosine, is similar. For all three species, cells localized in the ganglion cell layer accumulated adenosine and exhibited adenosine-like immunoreactivity (ALIR). A smaller proportion of cells localized in the inner nuclear layer were labeled for ALIR, while a larger proportion of cells in this layer accumulated adenosine. Subtle differences between species are presented. However, the general similarities of the distribution of these two putative purinergic markers supports the evidence that a discrete adenosinergic neurotransmitter/modulatory system is present in the retina.

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A new procedure for haptenizing adenosine leading to a more specific radioimmunoassay method

Cited by 14 publications

References 24 publications

Assay of adenosine in human adipose tissue

Assay of adenosine in human adipose tissue

Endogenous adenosine and adenosine receptors localized to ganglion cells of the retina.

Comparison of adenosine uptake and endogenous adenosine-containing cells in mammalian retina

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