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Radioimmunoassay for rat galanin: immunochemical and chromatographic characterization of immunoreactivity in tissue extracts
Linköping University, Department of Biomedicine and Surgery, Clinical Chemistry. Linköping University, Faculty of Health Sciences.
Linköping University, Department of Molecular and Clinical Medicine, Obstetrics and gynecology. Linköping University, Faculty of Health Sciences.
2000 (English)In: Scandinavian Journal of Clinical and Laboratory Investigation, ISSN 0036-5513, Vol. 60, no 5, 411-418 p.Article in journal (Refereed) Published
Abstract [en]

Galanin is a regulatory peptide with wide distribution in the central and peripheral nervous system and with numerous biological effects. Several radioimmunoassays based on antisera raised against porcine galanin have been used to measure immunoreactivity in rat tissues. However, considerable lack of parallelism has been observed between the porcine standard and rat tissue extracts, which may decrease the reliability of the quantitative data. The purpose of the present study was therefore to raise antibodies against rat galanin and establish a competitive radioimmunoassay for rat galanin. Two antisera, RatGal4 and RatGal5, were characterized in detail. The homogeneity of the immunoreactive material from several tissues was also investigated with column chromatography. At reverse-phase high-pressure liquid chromatography more than 95% of the immunoreactive material from rat CNS eluted as a single peak in the position of synthetic rat galanin, whereas almost half of the immunoreactive material from the intestine eluted in positions different from the synthetic peptide. Extracts of rat brains as well as jejunum diluted in parallel with the standard curve for both antisera. We conclude that measurements of rat galanin based on these antisera are therefore more reliable than those based on antisera raised against porcine galanin.

Place, publisher, year, edition, pages
2000. Vol. 60, no 5, 411-418 p.
National Category
Medical and Health Sciences
URN: urn:nbn:se:liu:diva-25424DOI: 10.1080/003655100750019323Local ID: 9869OAI: diva2:245753
Available from: 2009-10-07 Created: 2009-10-07 Last updated: 2012-09-06Bibliographically approved
In thesis
1. Effects of gonadal steroids on galanin and other neuropeptides in the rat brain
Open this publication in new window or tab >>Effects of gonadal steroids on galanin and other neuropeptides in the rat brain
2001 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

Many women experience mental or emotional changes when variations occur in the plasma concentrations of their ovarian hormones. These observations are supported by a large number of scientific reports concerning interactions between ovarian steroids and cerebral functions, e.g. mood, cognition and motor function. Activation of genes is a fundamental principle by which steroids exert effects in neurons, thereby regulating the expression of enzymes, receptor proteins, trophic factors and regulatory peptides. This thesis examines the hypothesis that sex steroid hormones can modify concentrations of peptides important for neuronal transmission in brain areas relevant for the above-mentioned modalities. A necessary starting point in this quest is to develop good quantitative data on neuropeptide concentrations in relation to sex steroid status. Accordingly, the general aim of the thesis is to investigate the influence of gender and sex steroid exposure on neuropeptide concentrations and identify one or several steroid-sensitive neuropeptides in extrahypothalamic brain regions of the rat.

Neuropeptide concentrations were measured with radioimmunoassay in extracts of extrahypothalarnic brain regions. A new radioimmunoassay, specific for rat galanin, was developed. Concentrations of neuropeptides were compared between sexes, across puberty and in ovariectomized animals treated with different doses and durations of estradiol, progesterone and norethisterone.

We found that differences in sex steroid exposure between gender, across puberty, and duting exogenous treatment correlate with differences in concentrations of neuropeptide immunoreactivities in several extrahypothalarnic regions of the rat brain. In particular galanin in rat hippocampus is sensitive to estrogen and can be increased in a dose-dependent manner with chronic estradiol treatment. It is likely that the increase mainly sterns from the ascending noradrenergic neuron system originating in locus cemleus. Chromatographic characterization shows that galanin, measured by the antisera Ga1Rat4 and Ga1Rat5, has a high degree of homogeneity in rat brain tissue. Norethisterone and progesterone have different effects on substance P and neuroldnin A in rat frontal cortex and striatum.

These findings could contribute to the understanding of how mood and cognition are affected by sex steroids in females and how estradiol interacts with neurons affected by Alzheimer's disease.

Place, publisher, year, edition, pages
Linköping: Linköpings universitet, 2001. 64 p.
Linköping University Medical Dissertations, ISSN 0345-0082 ; 673
National Category
Medical and Health Sciences
urn:nbn:se:liu:diva-25705 (URN)10081 (Local ID)91-7219-973-3 (ISBN)10081 (Archive number)10081 (OAI)
Public defence
2001-05-17, Elsa Brändströms sal, Hälsouniversitetet, Linköping, 09:00
Available from: 2009-10-08 Created: 2009-10-08 Last updated: 2012-09-06Bibliographically approved

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