Nicotinic receptor distribution in the human thalamus: autoradiographical localization of [3H]nicotine and [125I] alpha-bungarotoxin binding.

The thalamus plays a major role in relaying and transforming information that is relayed to the cortex and in turn modulates cortical outputs. The reticular nucleus projects to the other thalamic nuclei, modulating and integrating their activity. The distribution of high affinity nicotine and alpha-bungarotoxin (alpha BTX) receptors in the human thalamus has been investigated by radioligand autoradiography in post mortem human tissue. [3H]nicotine binding in the human thalamus was high in most thalamic nuclei, especially in the lateral dorsal, the medial geniculate, lateral geniculate and anterior nuclei. The distribution of [125I] alpha BTX binding was quite distinct from [3H]nicotine binding. [125I] alpha BTX binding was generally lower (< 0.26-11.62 fmol/mg protein compared with 6.68-36.17 fmol/mg protein for nicotine binding) and concentrated in the reticular nucleus, with discrete groups of cells displaying higher binding in the latter. These results indicate differences between the distribution of nicotinic receptors in humans and those previously reported in mice and monkeys. Changes in high affinity nicotine and alpha BTX receptors in the thalamus may contribute to symptoms observed in neuropathological conditions associated with disorders of perception and movement such as Dementia with Lewy Bodies, Alzheimer's Disease and Schizophrenia.

Selenium homeostasis in the central nervous system of the rat.

These experiments have investigated selenium movement between blood and CNS in anaesthetised rats. Each animal was surgically anaesthetised and the left femoral blood vessels cannulated for blood withdrawal and solute infusion. Each rat received 75-selenium as sodium selenite infused in normal saline and experiments lasted between 5 minutes and 5 hours during which blood samples were periodically taken. At termination, the CNS was removed, regionally dissected and analysed with the plasma samples for 75-Se radioactivity by gamma-counting. Data were analysed by graphical analysis. Results showed unidirectional uptake of 75-Se into the CNS and regional differences were not found except for the hypothalamus. On average the CNS influx rate constant (Kin) was about 7 +/- 1 x 10(-5) ml/min/g. Data suggest that the 75-Se most likely entered the CNS as a free ionic form.

Uptake of 75-Selenium into the central nervous system of the rat.

These experiments have investigated selenium movement between blood and the CNS in anaesthetized rats. Each animal was anaesthetized and the left femoral blood vessels cannulated for blood withdrawal and solute infusion. Each rat received 75-Se as sodium selenite infused in normal saline and experiments lasted between 5 minutes and 5 hours during which blood samples were periodically taken. At termination, the CNS was removed, dissected and analysed with the plasma samples for 75-Se radioactivity by gamma-counting. Data were analyzed by multiple-time uptake analysis. Results showed unidirectional uptake of 75-Se into the CNS and some regional differences were found. On average the CNS influx rate constant (Kin) was about 7 +/- 1 x 10(-5) ml/min/g. This indicates that the 75-Se most likely entered the CNS in a protein-bound form.

Germanium-68 as a possible marker for silicon transport in rat brain.

Silicon (Si) is an essential trace element normally present in brain and cerebrospinal fluid, although the mechanism by which it enters and distributes in brain is largely unknown. Due to the short radioactive half-life of 31Si (156 min) we have investigated the use of 68Germanium (68-Ge, half-life 282 days) as a possible marker for Si transport in rat brain over longer periods than are possible with 31Si. Adult male anaesthetised rats were given a bolus of 68Ge I.V. and arterial blood samples taken during experiments that lasted between 5 min and 3 days. At termination, the brain was removed and analysed for radioactivity as were the plasma samples. Data were analyzed by Graphical Analysis which showed that the blood-brain barrier permeability to 68Ge (Kin approximately 10(-4) ml/min/g) is similar to that for many non protein-bound electrolytes in plasma and that 68Ge fluxes across cerebral capillaries are bidirectional. The autoradiographic distribution of 68Ge in brain was homogenous. Our results are in agreement with those of previous studies using 31Si or 68Ge, which suggest 68Ge may be a useful marker for Si when investigating the role of this element in conditions such as neurodegenerative diseases.

Brain iron homeostasis.

The anatomical and cellular distribution of non-haem iron, ferritin, transferrin, and the transferrin receptor have been studied in postmortem human brain and these studies, together with data on the uptake and transport of labeled iron, by the rat brain, have been used to elucidate the role of iron and other metal ions in certain neurological disorders. High levels of non-haem iron, mainly in the form of ferritin, are found in the extrapyramidal system, associated predominantly with glial cells. In contrast to non-haem iron, the density of transferrin receptors is highest in cortical and brainstem structures and appears to relate to the iron requirement of neurones for mitochondrial respiratory activity. Transferrin is synthesized within the brain by oligodendrocytes and the choroid plexus, and is present in neurones, consistent with receptor mediated uptake. The uptake of iron into the brain appears to be by a two-stage process involving initial deposition of iron in the brain capillary endothelium by serum transferrin, and subsequent transfer of iron to brain-derived transferrin and transport within the brain to sites with a high transferrin receptor density. A second, as yet unidentified mechanism, may be involved in the transfer of iron from neurones possessing transferrin receptors to sites of storage in glial cells in the extrapyramidal system. The distribution of iron and the transferrin receptor may be of relevance to iron-induced free radical formation and selective neuronal vulnerability in neurodegenerative disorders.

Blood-brain exchange routes and distribution of 65Zn in rat brain.

Zinc is essential for normal development and function of the CNS although much is to be learned about brain Zn homeostasis. In these experiments adult male Wistar rats within the weight range 500-600 g were used. Ventriculo-cisternal perfusion was performed to allow the measurement of 65Zn fluxes between blood and csf across the choroid plexuses. Blood-brain or blood-cerebrospinal fluid barrier permeability to 65Zn has been determined by graphical analysis in experiments that lasted between 5 and 180 minutes. Cerebral capillary permeability to 65Zn was found to be low with a K(in) of about 5 x 10(-4)ml/min/g. Choroid plexus permeability to 65Zn was about 12 fold greater, although Zn influx to brain via this route was less than 5% that across cerebral capillaries. The autoradiographic distribution of 65Zn in brain showed regional variation with lowest levels in white matter and high levels in the dentate gyrus and hippocampus.

Uptake and distribution of iron and transferrin in the adult rat brain.

Brain uptake of iron-59 and iodine-125-labelled transferrin from blood in the adult rat has been investigated using graphical analysis to determine the blood-brain barrier permeability to these tracers in experiments that lasted between 5 min and 8 days. The blood-brain barrier permeability (K(in)) to 59Fe was 89 x 10(-5) ml/min/g compared to the value of 7 x 10(-5) ml/min/g for 125I-transferrin, which is similar to that of albumin, a plasma marker. The autoradiographic distribution of these tracers in brain was also studied to determine any regional variation in brain uptake after the tracers had been administered either systemically or applied in vitro. No regional uptake was seen for 125I-transferrin even after 24 h of circulation. In contrast, 59Fe showed selective regional uptake by the choroid plexus and extra-blood-brain barrier structures 4 h after administration. After 24 h of circulation, 59Fe distribution in brain was similar to the transferrin receptor distribution, as determined in vitro, but was unlike the distribution of nonhaem iron determined histochemically. The data suggest that brain iron uptake does not involve any significant transcytotic pathway of transferrin-bound iron into brain. It is proposed that the uptake of iron into brain involves the entry of iron-loaded transferrin to the cerebral capillaries, deposition of iron within the endothelial cells, followed by recycling of apotransferrin to the circulation. The deposited iron is then delivered to brain-derived transferrin for extracellular transport within the brain, and subsequently taken up via transferrin receptors on neurones and glia for usage or storage.

65Zn uptake from blood into brain in the rat.

Zinc is essential for the normal development and function of the CNS, although little is known about brain zinc homeostasis. Therefore, in this investigation we have studied 65Zn uptake by brain from blood and have measured the blood-brain barrier permeability to 65Zn in the anaesthetised rat in vivo. Adult male Wistar rats within the weight range 500-600 g were used. 65ZnCl2 and 125I-albumin, the latter serving as a vascular marker, were injected intravenously in a bolus of normal saline. Sequential arterial blood samples were taken during experiments that lasted between 5 min and 5 h, after which the whole brain was removed, dissected, and analysed for radioisotope activity. Data have been analysed by graphical analysis, which suggests that after 30 min of circulation, 65Zn uptake by brain from blood is unidirectional with an influx rate constant, Kin, of approximately 5 X 10(-4) ml/min/g. At circulation times of less than 30 min, 65Zn fluxes between blood and brain are bidirectional, where influx has a K value of greater than 5 X 10(-4) ml/min/g. In addition to the blood space, the brain appears to contain a rapidly exchanging compartment(s) for 65Zn of approximately 4 ml/100 g, which is not CSF.

65zinc uptake from blood into brain and other tissues in the rat.

Zinc is essential for normal growth, development and brain function although little is known about brain zinc homeostasis. Therefore, in this investigation we have studied 65Zn uptake from blood into brain and other tissues and have measured the blood-brain barrier permeability to 65Zn in the anaesthetized rat in vivo. Adult male Wistar rats within the weight range 500-600 g were used. 65ZnCl2 and [125I]albumin, the latter serving as a vascular marker, were injected in a bolus of normal saline I.V. Sequential arterial blood samples were taken during experiments that lasted between 5 min and 5 hr. At termination, samples from the liver, spleen, pancreas, lung, heart, muscle, kidney, bone, testis, ileum, blood cells, csf, and whole brain were taken and analysed for radio-isotope activity. Data have been analysed by Graphical Analysis which suggests 65Zn uptake from blood by all tissues sampled was unidirectional during this experimental period except brain, where at circulation times less than 30 min, 65Zn fluxes were bidirectional. In addition to the blood space, the brain appears to contain a rapidly exchanging compartment(s) for 65Zn of about 4 ml/100g which is not csf.

Gallium-67 as a potential marker for aluminium transport in rat brain: implications for Alzheimer's disease.

Evidence of a link between aluminium and Alzheimer's disease, parkinsonism-dementia of Guam, and dialysis encephalopathy raises questions regarding the role of this element in the pathogenesis of these conditions. Therefore, we have investigated the use of gallium-67 (67Ga) as a marker for brain uptake of aluminium. The binding of 67Ga to plasma proteins has been studied, and the blood-brain barrier permeability and autoradiographic distribution of this isotope in rat brain determined in vivo. The autoradiographic distribution of 125I-Fe-transferrin receptors in rat brain has also been determined in vitro. Results show that 67Ga was bound to plasma transferrin, entered the brain with a blood-brain barrier permeability of 2.48 x 10(-6) ml/min/g, and showed a marked regional distribution that was very similar to that of 125I-Fe-transferrin receptors. Our data suggest that the vulnerability of the hippocampus, amygdala, and cerebral cortex in conditions such as those mentioned above may be partly due to an increased uptake and deposition of aluminium in these regions by the iron transport system.

Bulk flow of cerebrospinal fluid into brain in response to acute hyperosmolality.

Brain volume is regulated during acute hyperosmolality based, in part, on the tissue gain of Na, Cl, and K. This study evaluates the contribution of bulk flow of cerebrospinal fluid (CSF) into brain to the volume regulatory gain of electrolytes. Artificial CSF containing radiolabeled albumin and diethylenetriamine penta-acetic acid (DTPA) was perfused for 60 min through the ventricles and/or subarachnoid space of anesthetized rats, and tracer clearances from CSF to brain were measured as a function of plasma osmolality. Osmolality was elevated after 30 min of perfusion by intraperitoneal injection of hypertonic NaCl or mannitol. Albumin and DTPA clearances increased with osmolality at the same rate, despite a sevenfold difference in diffusion coefficient, consistent with osmotic stimulation of a bulk flow component of tracer influx into brain. The volume shift estimated on the basis of this data is 114 microliters CSF/g dry wt brain for a 60-mosmol/kg increase in osmolality. Results indicate that CSF is a major source of the volume regulatory gain of Na and Cl, but not of K.

Effect of ventricular tonicity upon cerebrospinal fluid production in rabbits.

Ventriculo-cisternal perfusion in rabbits has been employed to examine steady-state relations between ventricular sodium and water fluxes and ventricular osmolality. These fluxes have been determined in individual rabbits when the ventricular fluid was either similar to normal cerebrospinal fluid (c.s.f.) or when its osmolality was changed to one value within the range of about 150-300 mosmol/l. The ventricular osmolality was changed by perfusing the ventricles with sucrose solutions of different concentrations that were either ion free, contained a low concentration of sodium, or contained both sodium and furosemide to inhibit the active production of c.s.f. Results suggest that this experimental range of ventricular osmolality is without significant effect upon a constant active sodium-coupled water movement into the ventricles, whereas a passive osmotic water flux into the ventricles increases with ventricular osmolality.