Upregulation of [3H]methyllycaconitine binding sites following continuous infusion of nicotine, without changes of alpha7 or alpha6 subunit mRNA: an autoradiography and in situ hybridization study in rat brain.

It is well established that exposure of experimental animals to nicotine results in upregulation of the alpha4beta2-subtype of neuronal nicotinic acetylcholine receptors (nAChRs). The aim of this study was to determine the effect of nicotine on the levels of alpha7-nAChRs in rat brain, for which only partial information is available. Rats were infused with nicotine (3 mg/kg/day) or saline for 2 weeks and their brains processed for receptor autoradiography with [3H]methyllycaconitine (MLA), a radioligand with nanomolar affinity for alpha7-nAChRs.

Chronic nicotine treatment changes the axonal distribution of 68 kDa neurofilaments in the rat ventral tegmental area.

Region-specific decreases of neurofilament proteins (NF) were described in the ventral tegmental area (VTA) of rats treated chronically with morphine, cocaine or alcohol. In a previous study, we demonstrated that NF levels were also changed in the VTA after chronic treatment with nicotine. The aim of this study was to clarify the submicroscopic basis of decreased immunoreactivity for NF-68, NF-160 and NF-200, as determined by using NR4, BF10 and RT97 antibodies, respectively.

Allosteric modulation of [3H]-CGP39653 binding through the glycine site of the NMDA receptor: further studies in rat and human brain.

Binding of D,L-(E)-2-amino-4-[(3)H]-propyl-5-phosphono-3-pentenoic acid ([(3)H]-CGP39653), a selective antagonist at the glutamate site of the NMDA receptor, is modulated by glycine in rat brain tissue. We have further investigated this phenomenon in rodent and human brain by means of receptor binding and quantitative autoradiography techniques. In rat cerebral cortical membranes the glycine antagonist 3-[2-(Phenylaminocarbonyl)ethenyl]-4,6-dichloro-indole-2-carboxylic acid sodium salt (GV150526A) did not change basal [(3)H]-CGP39653 binding, but competitively reversed the high affinity component of [(3)H]-CGP39653 binding inhibition by glycine, with a pK(B) value of 8.

Distribution of the messenger RNA for the small conductance calcium-activated potassium channel SK3 in the adult rat brain and correlation with immunoreactivity.

Small conductance calcium-activated potassium channels are voltage independent potassium channels which modulate the firing patterns of neurons by activating the slow component of the afterhyperpolarization. The genes encoding a family of small conductance calcium-activated potassium channels have been cloned and up to now three known members have been described and named small conductance calcium-activated potassium channel type 1, small conductance calcium-activated potassium channel type 2 and small conductance calcium-activated potassium channel type 3; the distribution of their messenger RNA in the rat CNS has already been performed but only in a limited detail. The present study represents the first detailed analysis of small conductance calcium-activated potassium channel type 3 mRNA distribution in the adult rat brain and resulted in a strong to moderate expression of signal in medial habenular nucleus, substantia nigra compact part, suprachiasmatic nucleus, ventral tegmental area, lateral septum, dorsal raphe and locus coeruleus.

Chronic nicotine treatment decreases neurofilament immunoreactivity in the rat ventral tegmental area.

Region-specific decreases of neurofilament proteins have been described in the ventral tegmental area of rats chronically treated with either morphine or cocaine. The aim of the present study was to assess if the levels of neurofilament proteins are changed in the ventral tegmental area by chronic treatment with nicotine. Immunoreactivity for NF-68, NF-160 and NF-200 was determined using NR4, BF10 and RT97 antibodies, respectively.

Receptor binding characteristics of the novel NMDA receptor glycine site antagonist [3H]GV150526A in rat cerebral cortical membranes.

Binding of the glycine site antagonist 3-[2-(Phenylamino-carbonyl)ethenyl]-4,6-dichloro-indole-2-carboxylic acid sodium salt ([3H]GV150526A) was characterised in rat cerebral cortical membranes. Saturation experiments indicated the existence of a high affinity binding site, with a pK(d) value of 9.08 (K(d)=0.

NMDA NR1 subunit mRNA and glutamate NMDA-sensitive binding are differentially affected in the striatum and pre-frontal cortex of Parkinson's disease patients.

Changes in the levels of mRNA for the NR1 subunit of the glutamate NMDA receptor and in NMDA-sensitive glutamate binding were investigated in consecutive sections of the prefrontal cortex and striatum of control and Parkinson's disease (PD) post-mortem brain using in-situ hybridisation and receptor autoradiography. Both markers of NMDA receptors were found to be relatively unaffected when measured by microdensitometry in the prefrontal cortex of control and PD brains. At a cellular level, a subpopulation of small and medium neurons in the superficial layers of the prefrontal cortex of the PD group showed a decreased expression of NMDA NR1 mRNA, with the maximal decrease in cortical layer IV.

On the relationship of neuropeptide Y Y1 receptor-immunoreactive neuronal structures to the neuropeptide Y-immunoreactive nerve terminal networks. A double immunolabelling analysis in the rat brain.

Neuropeptide Y is the most abundant peptide in the mammalian central nervous system and exhibits a variety of potent neurobiological functions. In the present study, double immunolabelling histochemistry was performed, using previously characterized antibodies against neuropeptide Y and the neuropeptide Y Y1 receptor subtype, to clarify the cellular distribution of Y1 receptors in the rat brain in relation to the neuropeptide Y-immunoreactive systems. Based on fluorescence and confocal laser microscopy analysis, morphological evidence is presented that the perikaryal and dendritic Y1 receptor-like immunoreactivity demonstrated in discrete regions of the tel-, diencephalon and of the lower brain stem, shown to be cytoplasmic and membrane associated, in many brain regions is not co-distributed with the neuropeptide Y-immunoreactive terminal network.

N-terminal splice variants of the NMDAR1 glutamate receptor subunit: differential expression in human and monkey brain.

The distribution of the mRNA coding for the two N-terminal splice variants of the N-methyl-D-aspartate receptor 1 (NMDAR1) subunit of the glutamate NMDA receptor was studied on whole-hemisphere human and macaca fascicularis brain sections by in-situ hybridisation. Synthetic oligonucleotides directed against NMDAR1a and NMDAR1b variants showed a specific distribution that was similar in human and monkey brain, with the NMDAR1a isoform present in the majority of the NMDA receptors, and the NMDAR1b variant present at high levels only in the cortex and dentate gyrus of the hippocampus. The distribution of the mRNAs for the NMDAR1pan and NMDAR1a subunit reported in this study support previous findings in rodent brain, while the restricted distribution of the NMDAR1b variant found in human and monkey suggests some important differences in the composition of the NMDA receptor in rodents and primates.

[3H]MK-801 binding and the mRNA for the NMDAR1 subunit of the NMDA receptor are differentially distributed in human and rat forebrain.

The distributions of [3H]MK-801 binding and the NMDA NR1 subunit mRNA were studied using receptor autoradiography and in-situ hybridization in rat and human brain whole-hemisphere coronal sections. Receptor protein detected by radioligand autoradiography and the mRNA for the key subunit of the receptor presented similar distributions in the forebrain, with a few areas showing an imbalance between the levels of mRNA and receptor protein. Human frontal cortex showed a relative abundance of NMDAR1 mRNA as compared to [3H]MK-801 binding.

NPY Y1 receptor like immunoreactivity exists in a subpopulation of beta-endorphin immunoreactive nerve cells in the arcuate nucleus: a double immunolabelling analysis in the rat.

Double immunolabelling immunohistochemistry in the arcuate nucleus of the rat demonstrates that neuropeptide Y (NPY) Y1 receptor like immunoreactivity is strongly present in a subpopulation of beta-endorphin immunoreactive nerve cell bodies, while the small NPY immunoreactive nerve cell bodies located medially lack NPY Y1 receptor like immunoreactivity. The NPY Y1 like immunoreactive nerve cell bodies lie in an arcuate area rich in NPY immunoreactive nerve terminals forming an uniform plexus. It is postulated that NPY Y1 receptors in beta-endorphin neurons may mediate some actions of NPY on motivational processes and pain control as well as on hypophyseal hormone secretion, involving at the least in part a regulation of the tubero-infundibular DA neurons.

Localization of neuropeptide Y Y1 receptor-like immunoreactivity in catecholaminergic neurons of the rat medulla oblongata.

Neuropeptide Y receptors in the medulla oblongata participate in central cardiovascular control. The neuropeptide Y1 receptor subtype gene and amino acid sequence have been identified by molecular cloning studies. In this study, a C-terminal peptide representing amino acids 355-382 of the neuropeptide Y1 receptor was synthesized and cross-linked to thyroglobulin to produce an antibody against a partial sequence of the neuropeptide Y1 receptor, used to localize neuropeptide Y1 receptor-like immunoreactivity in the catecholaminergic neurons of the medulla oblongata.

Increased vasopressor actions of intraventricular neuropeptide Y-(13-36) in spontaneously hypertensive versus normotensive Wistar-Kyoto rats. Possible relationship to increases in Y2 receptor binding in the nucleus tractus solitarius.

The C-terminal NPY fragment (13-36)[NPY-(13-36)], a Y2 receptor agonist, elicits vasopressor responses upon central administration. The cardiovascular responses of NPY-(13-36) together with the distribution of NPY receptor subtypes within the nucleus tractus solitarius (nTS) have therefore been studied in spontaneously hypertensive rats (SHR). NPY-(13-36) was injected intracerebro-ventricularly in different doses (7.

Expression of the mouse and rat mas proto-oncogene in the brain and peripheral tissues.

We isolated the mas proto-oncogene from a mouse genomic library. Sequence analysis showed that it contains an open reading frame without intervening sequences. The amino acid sequence deduced confirms the seven-transmembrane-domain structure and exhibits 97% and 91% amino acid homology with the rat and the human Mas, respectively.

On the plasticity of the cerebellar renin-angiotensin system: localization of components and effects of mechanical perturbation.

This study focuses on the renin-angiotensin system (RAS) in the cerebellar cortex and changes within this system after mechanically induced cerebellar injury. Using radioactive and non-radioactive in situ hybridization and immunocytochemistry angiotensinogen mRNA, angiotensinogen, angiotensin II and, for the first time, N-terminal angiotensin fragment (1-7) immunoreactivities, respectively, were demonstrated in the rat cerebellum. Angiotensinogen mRNA and angiotensinogen immunoreactivity (IR) were both present in glial cell populations of all layers, especially in the Purkinje and granular cell layers and within the cerebellar nuclei.

Cellular expression of angiotensin type-1 receptor mRNA in the kidney.

Angiotensin II has multiple renal effects that are important in the regulation of renal hemodynamics and electrolyte secretion, and binding sites for angiotensin II have been demonstrated in different cells of the kidney. In the present study the cellular localization of mRNA for the angiotensin type 1 (AT1) subtype of the angiotensin II receptor was studied in adult rat kidney using a cRNA probe and in situ hybridization. Strong labeling was demonstrated in tubule cells of the inner and outer stripe of the outer medulla.

The renin-angiotensin system in the brain: an update 1993.

The renin-angiotensin system is considered to be one of the most important hormonal systems in the regulation of blood pressure and body fluid homeostasis. Ever since this system has been demonstrated to be present also in the brain, vast efforts have been made in investigating its central impact and function. The last few years, and especially the development of non-peptidic angiotensin II receptor subtype specific antagonists and the subsequent pharmacological characterization of these subtypes, brought this field of research a large step forward.

Prenatal development of glucocorticoid receptor gene expression and immunoreactivity in the rat brain and pituitary gland: a combined in situ hybridization and immunocytochemical analysis.

By means of in situ hybridization and immunocytochemical techniques it has been possible to follow the prenatal development of glucocorticoid receptor (GR) messenger RNA (mRNA) expression and GR immunoreactivity (IR) in the rat brain from embryonic day (E) 15 to 22. A 700-base-pair GR cDNA fragment was used for RNA probe generation. In the immunocytochemical analysis a mouse monoclonal antibody (IgG2a) against the rat liver GR was used in combination with the indirect fluorescence technique or the avidin-biotin immunoperoxidase method.

Regional expression of angiotensinogen mRNA in the brain of one-week-old, adult and old male rats.

The purpose of this study was to investigate possible regional differences in the distribution of angiotensinogen-mRNA in the postnatal versus the aging animal using in situ hybridization and computer-assisted microdensitometry. An essentially identical regional distribution pattern of angiotensinogen-mRNA in the brains of postnatal, adult and old rats was demonstrated. Substantial differences in angiotensinogen expression were observed in brain areas of postnatal versus adult and old animals.

Glucocorticoid regulation of angiotensinogen gene expression in discrete areas of the male rat brain. An in situ hybridization study.

The regulation of angiotensinogen gene expression by glucocorticoids has been described in several studies. Kalinyak and Perlman reported on a 60% increase of angiotensinogen expression in the rat brain after dexamethasone treatment with a single, high-dose injection. The purpose of the present study was to investigate whether a general upregulation of angiotensinogen expression or a region-specific upregulation underlies these findings.

Cellular localization of angiotensin type 1 receptor and angiotensinogen mRNAs in the subfornical organ of the rat brain.

The cellular localization of angiotensin type 1 receptor (AT 1) and angiotensinogen mRNA expression in the subfornical organ (SFO) of the rat brain has been studied by means of non-radioactive in situ hybridization combined with immunocytochemistry for glial fibrillary acidic protein (GFAP) and Neutral red staining. The AT 1 receptor mRNA expression is shown to be within putative nerve cells without any association with the glial fibrillary acidic protein (GFAP)-immunoreactive (IR) cells. In contrast the angiotensinogen cRNA expression is associated predominantly with GFAP-IR cells.

The distribution of angiotensin II AT1 receptor subtype mRNA in the rat brain.

The present study demonstrates the existence and regional distribution of angiotensin II AT1 receptor subtype mRNA expression in the rat brain by the use of in situ hybridization and RNase protection assay. Substantial expression levels in the brain have only been detected in certain distinct areas, such as the subfornical organ, the parvocellular part of the paraventricular hypothalamic nucleus, and the median preoptic nucleus. The results give further evidence for the involvement of the angiotensin II AT1 receptor subtype in the classical functions of central angiotensin II, like blood pressure control, body fluid homeostasis and in corticotropin-releasing factor (CRF) secretion.

The semi-quantitative distribution and cellular localization of angiotensinogen mRNA in the rat brain.

The present study describes the regional distribution and cellular localization of angiotensinogen-mRNA in the rat brain as investigated by means of in situ hybridization also in combination with immunocytochemistry for glial fibrillary acidic protein. The angiotensinogen gene expression seemed to be restricted to astroglia and showed marked regional differences. In some areas angiotensinogen-mRNA was present in almost all astrocytes with a strong signal (e.

High blood pressure in transgenic mice carrying the rat angiotensinogen gene.

Transgenic mice were generated by injecting the entire rat angiotensinogen gene into the germline of NMRI mice. The resulting transgenic animals were characterized with respect to hemodynamics, parameters of the renin angiotension system, and expression of the transgene. The transgenic line TGM(rAOGEN)123 developed hypertension with a mean arterial blood pressure of 158 mmHg in males and 132 mmHg in females.

The brain renin-angiotensin system: localization and general significance.

This report summarizes the present data about the existence of components of the renin-angiotensin system in the rat brain. Angiotensinogen mRNA, mas proto-oncogene mRNA, angiotensin II (Ang II), and Ang II receptors have been mapped in the brain by using in situ hybridization, immunocytochemistry, and receptor autoradiography. These markers turned out to be widely distributed throughout the brain and to be not only restricted to areas related to cardiovascular control, but also to be present in functionally different areas, suggesting also other functions of angiotensin peptides.