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Biomedical subjects

Susanne Hilke

Publications and source records attributed to Susanne Hilke.

4 recordsLinked to original sources

A short estrogen-responsive N-terminal galanin homologue found in rat brain and gut with antiserum raised against rat galanin(1-16).

Galanin-like peptide (GALP) is currently the only known galanin(1-29) homologue. However, three different galanin receptors, of which GalR3 exhibits comparatively low affinity for galanin(1-29), and molecular heterogeneity of immunoreactive galanin are arguments for presence of other endogenous galanin homologues. Since antibodies recognize three-dimensional structures of 3-5 amino acids in a peptide, we raised antibodies in rabbits against galanin(1-16) conjugated to bovine serum albumin, looking for the presence of endogenous N-terminal galanin homologues in rat tissues. The antiserum selected had 7,830 times higher avidity for galanin(1-16) compared to galanin(1-29). A single immunoreactive component with a Stokes radius of about 8 amino acids was found. Immunohistochemistry strongly suggested that this immunoreactivity is localised in the same neurons as galanin(1-29). Furthermore, its concentration was increased in response to estrogen treatment in the same brain regions as galanin(1-29), although not as rapidly. The present results indicate the presence of a novel endogenous N-terminal galanin homologue.

Animals↗

Galanin in the hippocampal formation of female rats--effects of 17beta-estradiol.

17Beta-estradiol induced an increase in tissue concentrations of galanin in the hippocampal formation of ovariectomized rats. This increase was dose- and time dependent, and occurred already 60 min after steroid administration and was not blocked by Tamoxifen). There was also an increase in galanin in the pro-estrous phase in regularly cycling rats. The estrogen-induced rapid increase may at least in part be due to decreased release of galanin as demonstrated by in vivo microdialysis studies. Thus, sex steroid hormones may influence signalling molecules in brain areas of importance for cognitive functions.

Age Factors↗

Estrogen induces a rapid increase in galanin levels in female rat hippocampal formation--possibly a nongenomic/indirect effect.

Administration of 17beta-estradiol to ovariectomized rats increased the concentrations of galanin-like immunoreactivity (LI) in the hippocampal formation by 215% (P < 0.001) within 1 h. An increase of 125% (P < 0.05) was observed in the same brain region in the proestrous phase of a normal estrous cycle. Tamoxifen did not block the 17beta-estradiol-induced increase in the concentration of galanin-LI but resulted in a 62% decrease in the hypothalamus within 1 h. In vivo microdialysis in the dorsal hippocampal formation showed a decrease of extracellular galanin-LI (P < 0.001) 1-2 h after treatment with 17beta-estradiol, indicating a decreased release of galanin. For comparison, we studied the concentrations of neuropeptide Y, which were not influenced significantly in any of the regions studied. Taken together our results suggest that 17beta-estradiol inhibits galanin release, presumably from noradrenergic nerve terminals, and primarily via a nongenomic/indirect action, not necessarily involving the classical nuclear receptors ER-alpha or ER-beta. These rapid estrogen-induced changes in galanin release could influence transmitter signalling and plasticity in the hippocampal formation.

Analysis of Variance↗

Generic three-column parallel LC-MS/MS system for high-throughput in vitro screens.

A parallel three-column LC-MS/MS system for quantitative high-throughput in vitro screens is described. The robust novel system is composed of three LC pumps, an autosampler and a triple quadrupole mass spectrometer used in combination with three valves and three analytical LC columns in parallel configuration. Two of the three valves work in unison to select which column receives the injection, and the third valve selects which column is to be in line with the mass spectrometer. Improved sample throughput is achieved without sacrificing chromatographic separation quality or sensitivity. To demonstrate the applicability of the system, pools of five compounds (phenacetin, S-mephenytoin, bufuralol, midazolam and clomethiazole) were analyzed, together with an internal standard. The results show that the sample throughput can be increased significantly by reducing analysis time to 3 min per sample as compared to 8 min with a general gradient single-column system. Analysis of the five compounds shows an accuracy of 81-108% and a precision (given as relative standard deviation) of 1.5-14%. The system was further applied to samples from a metabolic stability assay in liver microsomes.

Chromatography, High Pressure Liquid↗