Molecular Nature of β-Nerve Growth Factor (NGF)-Like Immunoreactive Substance(s) in Mouse Plasma

Shinoda, Ichizo; Takeuchi, Rie; Kurobe, Masayuki; Furukawa, Shoei; Hayashi, Kyozo

doi:10.3164/jcbn.5.135

Cited by 2 publications

(3 citation statements)

References 16 publications

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“…This assay provided a reliable determination of NGF levels in serum and it became clear that in rat and mouse serum there is less than 5 ng of NGF per ml, in agreement with results obtained from bioassays of NGF activity. Up until now the only clear case in which the presence of NGF has been detected in serum is in mouse where most of it is probably derived from the extreme amounts of NGF found in the submandibular gland (Aloe et al, 1986;Shinoda et al, 1988). Shinoda et al (1988), using the sensitive two-site EIA found serum levels of NGF in the male mouse to be 0.07 ng/ml and for the female mouse 0.01 ng/ml.…”

Section: Discussionmentioning

confidence: 99%

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Two‐site enzyme immunoassay for βNGF applied to human patient sera

Lorigados

Söderström

Ebendal

1992

J of Neuroscience Research

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Nerve growth factor (NGF) supports sympathetic and sensory neurons in the peripheral nervous system and serves functions in the development and maintenance of cholinergic neurons in the basal forebrain. NGF distribution can be studied with the use of a sensitive two-site enzyme immunoassay (EIA). The monoclonal antibody 27/21 to mouse NGF was recently shown to effectively block the activity of both recombinant human NGF and native mouse NGF, and a two-site EIA using monoclonal antibody 27/21 was optimized. We have now applied this assay to examine NGF levels in normal human serum and serum from Parkinson, Alzheimer, and Huntington patients. To further test the specificity of conjugate binding, dilutions of the human sera were preincubated with an excess of monoclonal NGF antibody 27/21 in solution. With this strategy it was possible to completely block the signal obtained using the two-site EIA. Furthermore, we show that recombinant BDNF and NT-3 do not cross-react with monoclonal antibody 27/21 under our conditions. We found low levels of specific NGF immunoreactivity in normal human sera (0.4 +/- 0.1 ng/ml). Significantly lower levels of NGF were found in sera from patients with Parkinson's and Huntington's disease whereas sera from Alzheimer patients showed only slight reductions in the NGF level. Two patients who had received intracerebral NGF infusions (one with Parkinson's and other with Alzheimer's disease) showed significantly elevated serum levels of NGF during the period of infusion. Due to an inhibitory activity in human serum, it was impossible to demonstrate the low levels of NGF activity in the human serum samples using explanted embryonic sympathetic ganglia, even after concentration by pressure dialysis. Thus, the serum levels are below the limit to evoke a response in NGF-sensitive neurons and thus to expect any physiological effect. Nevertheless, the levels measured may be used as indicators in clinical conditions such as Parkinson's and Huntington's disease.

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Section: Discussionmentioning

confidence: 99%

“…It was also demonstrated that high molecular weight species that bind 1251-labelled mouse PNGF exist in human serum. Moreover, it has been suggested that NGF may circulate in plasma complexed through disulfide linkage with macromolecular components (Shinoda et al, 1988).…”

Section: Discussionmentioning

confidence: 99%

Two‐site enzyme immunoassay for βNGF applied to human patient sera

Lorigados

Söderström

Ebendal

1992

J of Neuroscience Research

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“…As a2M is abundant in serum (1.5-4.2 mg in human sera) and NGF is in low concentration in the body, it is likely that NGF released into the blood stream is completely absorbed by a,M. Only in mice NGF is detected in serum or plasma by radioimmunoassay (Aloe et al, 1986;Lakshmanan, 1986) or EIA (Shinoda et al, 1988). In this case, the circulating factor is considered to be derived from the submaxillary gland, which gland in mice contains an extremely large amount of NGF.…”

Section: Introductionmentioning

confidence: 99%

Developmental changes in nerve growth factor level in rat serum

Murase

Takeuchi

Iwata

et al. 1992

J of Neuroscience Research

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In serum, nerve growth factor (NGF) forms a complex with alpha 2-macroglobulin (alpha 2M), which formation inhibits the immunoreactivity between NGF and its antibodies. For measuring the serum level of NGF, it is thus necessary to liberate NGF from the NGF-alpha 2M complex and prevent reformation of such complex. The pretreatment of rat serum with 1 M guanidine hydrochloride for a few hours and operation of the enzyme immunoassay (EIA) in the presence of guanidine hydrochloride provided a reliable means for determination of the NGF level in serum. By this procedure we followed the serum NGF level in rats developmentally. It increased from prenatal day 2 to postnatal day 5 and decreased slightly at postnatal week 3, thereafter remaining constant throughout adulthood. In pregnant rats, the NGF level in serum increased threefold to fivefold before birth and then decreased rapidly. These data suggest that serum NGF level may reflect the demand for this molecule during establishment of the peripheral nervous system.

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Molecular Nature of β-Nerve Growth Factor (NGF)-Like Immunoreactive Substance(s) in Mouse Plasma

Abstract: A highly sensitive two-site enzyme immunoassay system macromolecule(s) through disulfide linkage(s) in mouse plasma.

Cited by 2 publications

References 16 publications

Two‐site enzyme immunoassay for βNGF applied to human patient sera

Two‐site enzyme immunoassay for βNGF applied to human patient sera

Developmental changes in nerve growth factor level in rat serum

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