Identification of sex hormone-binding globulin in the human hypothalamus

Gonadal steroids are known to influence hypothalamic functions through both genomic and non-genomic pathways. Sex hormone-binding globulin ( SHBG) may act by a non-genomic mechanism independent of classical steroid receptors. Here we describe the immunocytochemical mapping of SHBG-containing neurons and nerve fibers in the human hypothalamus and infundibulum. Mass spectrometry and Western blot analysis were also used to characterize the biochemical characteristics of SHBG in the hypothalamus and cerebrospinal fluid (CSF) of humans. SHBG-immunoreactive neurons were observed in the supraoptic nucleus, the suprachiasmatic nucleus, the bed nucleus of the stria terminalis, paraventricular nucleus, arcuate nucleus, the perifornical region and the medial preoptic area in human brains. There were SHBG-immunoreactive axons in the median eminence and the infundibulum. A partial colocalization with oxytocin could be observed in the posterior pituitary lobe in consecutive semithin sections. We also found strong immunoreactivity for SHBG in epithelial cells of the choroid plexus and in a portion of the ependymal cells lining the third ventricle. Mass spectrometry showed that affinity-purified SHBG from the hypothalamus and choroid plexus is structurally similar to the SHBG identified in the CSF. The multiple localizations of SHBG suggest neurohypophyseal and neuroendocrine functions. The biochemical data suggest that CSF SHBG is of brain rather than blood origin. Copyright (c) 2005 S. Karger AG, Base

Neuronal and glial regeneration after focal cerebral ischemia in rat, an immunohistochemical and electron microscopical study

Objectives: Unilateral middle cerebral artery occlusion (MCAO) is an established rat model for stoke studies. It induces focal cerebral ischemia, prior to necrotic and apoptotic loss of tissue in a circumscribed cortical area, paralleled by temporary motor impairment.Methods: Here we examined tissue samples from the peri-infarct zone of rats that had survived unilateral MCAO for up to 90 min. With immunohistochemistry we stained sections for proliferation markers Ki 67 and PCNA and for intermediate filament protein nestin. Electron microscopy was employed to assess ultrastructural changes.Results: All MCAO animals developed pronounced lesions in the motor cortex. Numerous cells in the immediate peri-infarct area and scattered cells which seem to have migrated into the infarcted lesion stained positively for Ki 67 and PCNA. Electron microscopy revealed that cells in the lesion site proliferate along the blood vessels. Most of these cells had the ultrastructural features of fibrillary astrocytes while some of the cells were clearly neurons. Endothelia were in part fenestrated. Some of the surrounding cells showed immunostaining for PCNA, indicating proliferation. Oligodendroglia and myelination could not be seen in the lesion site. Single neuronal contacts exhibited the ultrastructural features of synapses. Reformation of cortical layers could not be observed.Conclusions: We concluded that in spite of extensive proliferation; neuronal and glial regeneration occurs after MCAO only to a small extent. Revascularization seems to be an important initial step. The observed functional recovery of experimental animals may be due to neuronal plasticity in young rats rather than structural regeneration.Keywords: Middle cerebral artery occlusion, Rat stroke model, Neuronal regeneration, Revascularization in brain, Electron microscopy, Proliferation marker

Changes of sex hormone-binding globulin/SHBG expression in the hypothalamo-hypophyseal system of rats during pregnancy, parturition and lactation

Sex hormone-binding globulin (SHBG) is expressed in hypothalamic magnocellular neurons. High co-localization rates of SHBG with oxytocin have been observed in the hypothalamus, indicating that SHBG plays a role in pregnancy, parturition and lactation. Further studies have shown that hypothalamic SHBG expression is malleable to changing steroid conditions. In this study, we have examined SHBG levels in the supraoptic and paraventricular hypothalamic nuclei and in the posterior pituitary lobe of late pregnant, parturient and early lactating rats by in situ hybridization, immunocytochemistry, and ELISA. Immunocytochemical and biochemical analysis showed that the SHBG levels increased during late pregnancy in hypothalamic nuclei. During parturition, SHBG levels fell in the magnocellular nuclei but increased in the posterior pituitary lobe. SHBG levels increase again during lactation. At day six of lactation, there was no significant difference in SHBG levels compared to normal cycling female rats, which served as control in this study. In situ hybridization showed increased SHBG mRNA signal during late pregnancy. The highest SHBG expression was observed during parturition. Our data indicate that hypothalamic SHBG expression changes during pregnancy, parturition and lactation, parallel to ovarian steroid and co-localized OT levels. This may in part be linked to known steroid actions on synthesis and secretion of magnocellular hypothalamic peptide hormones, important for the control of parturition and lactation.United States Department of Health & Human Services National Institutes of Health (NIH) - USA NIH National Institute of Mental Health (NIMH)-R21 MH06981

Distribution of 1,25-Dihydroxyvitamin D3 Receptor Immunoreactivity in the Rat Brain and Spinal Cord

A complete mapping study on the 1,25-dihydroxyvitamin D3 receptor immunoreactivity within the rat central nervous system was performed with a monoclonal and a polyclonal antibody. Specific immunostaining was observed within both nuclear and cytoplasmic compartments of a variety of cells in the cerebellum, mesopontine area, diencephalon, cortex, spinal cord, and limbic system. Both monoclonal and polyclonal antibodies provided similar staining patterns. The monoclonal antibody stained distinct domains within the nuclei of all and the cytoplasm of specific neuronal cell types, like motor neurons, Purkinje cells, and pyramidal cells of the cortex more clearly than the polyclonal antibody. The expression of vitamin D3 receptor in the rat central nervous system was confirmed by in situ hybridisation. The widespread distribution of vitamin D3 receptor in distinct portions of the sensory, motor, and limbic brain systems suggests multiple functional properties of 1,25-dihydroxyvitamin D3 in the central nervous system

Distribution of 1,25-Dihydroxyvitamin D3 Receptor Immunoreactivity in the Rat Olfactory System

1. The rat olfactory system contains numerous target sites for 1,25-dihydroxyvitamin D3, as determined by receptor protein (VDR) immunocytochemistry and in situ hybridization. 2. Nuclear and cytoplasmic VDR immunoreactivity as well as the corresponding hybridization signal was observed in neurons in the olfactory epithelium, the olfactory bulb, and throughout the limbic system in locations also known to be glucocorticoid targets. 3. The widespread distribution of VDR indicates the distinct functional importance of 1,25-dihydroxyvitamin D3 for olfactory perception

Changing caveolin-1 and oxytocin receptor distribution in the ageing human prostate

Several observations suggest that caveolin-1 has an important role in control of cell proliferation and cancerogenesis. For instance, oxytocin provokes a proliferative response in the prostate tissue when the oxytocin receptor is localized mainly in caveolin-1-enriched domains and an anti-proliferative effect when the same receptor is not localized in caveolae. Moreover, oxytocin concentrations are elevated in prostate tissue of patients with benign prostatic hyperplasia (BPH). In this study the expression pattern of the molecules caveolin-1, oxytocin receptor, androgen receptor and p21 (cell cycle arrest indicator) was investigated in the prostate tissue of BPH patients and of young controls.We found that both caveolin-1 and oxytocin receptor expression is drastically increased with age in both smooth muscle and epithelium of the prostate. We also found a significantly increased co-localization of the oxytocin receptor with caveolin-1 in both the muscle and the epithelium, especially in BPH patients. Androgen receptor and p21 staining was found throughout the prostate but did not change significantly with age or in BPH patients. We conclude that oxytocin may have a proliferative effect on the prostate tissue through the caveolae-associated receptors and thus contribute to BPH. This process seems to be androgen receptor independent