Brain Of Various Species Research Articles

High-affinity 3H-imipramine binding: a new biological marker in depression.

High-affinity binding of the tricyclic antidepressant drug, 3H-imipramine, has been demonstrated in the brain of various species including man. These specific binding sites have many of the characteristics to be expected for the specific site of action of a drug and appear to be associated with the neuronal uptake mechanism for serotonin. Chronic administration of tricyclic antidepressant drugs or the prolonged application of other antidepressant therapies, such as electroshock and sleep-deprivation, resulted in decreases in the density of 3H-imipramine binding sites. Apparently identical 3H-imipramine binding sites have been found in blood platelets from a variety of species including man. Clinical studies have shown that untreated severely depressed patients have a lower density of binding sites in their platelets than control volunteers. Longitudinal studies of these patients indicate that the density of 3H-imipramine binding sites tends not to change during treatment with tricyclic antidepressant drugs and the subsequent recovery from depression. 3H-imipramine binding in brain and platelets is proposed as a new biological marker in depression and as a useful research tool in biochemical and clinical pharmacological studies in affective disorders.

High-affinity binding of 3H-imipramine in brain and platelets and its relevance to the biochemistry of affective disorders

Specific high-affinity binding of 3H-imipramine has been demonstrated in the brain of various species including man. These binding sites have many of the characteristics to be expected for a pharmacological receptor and appear to be associated with the neuronal uptake mechanism for serotonin. Different antidepressant treatments like chronic administration of tricyclic antidepressants, chronic electroshock or sleep-deprivation result in decreases in the density of 3H-imipramine binding sites in normal animals. 3H-imipramine binding sites have also been found in blood platelets from different species including man. These sites are identical to those described in the brain. Clinical studies have shown that untreated severely depressed patients have a lower density of 3H-imipramine binding sites in their platelets when compared with control volunteers of the same age and sex. Longitudinal studies indicate that the low density of 3H-imipramine binding sites does not change during treatment with tricyclic antidepressant drugs and the subsequent clinical recovery from depression. 3H-imipramine binding in brain and platelets is proposed as a useful research tool in biochemical and clinical studies in affective disorders.

Antibodies as a means of isolating and characterizing biologically active substances: presence of a non-peptide, morphine-like compound in the central nervous system.

Antibodies generated against small molecular weight substances such as drugs are being used to isolate and characterize biologically active agonists. A morphine-like compound can be extracted from brain of various species which has determinant groups that are recognized by specific morphine antibodies. It has a regional distribution which can be quantitated as immuno-equivalents. Immunological, chemical and chromatographic tests show great similatiries of the compound to morphine. This morphine-like compound has biological activity as it inhibits the electrically induced contractions both of the guinea pig ileum and mouse vas deferens but the inhibition is not reversed by naloxone or naltrexone.

A Procedure for the Bulk Preparation of Brain Microsomes With Low Speed Centrifugation

A method is described for the rapid isolation of microsomes from brains of various species. The method depends on the interaction of calcium with microsomes. The procedure is limited because calcium inhibits certain microsomal enzymes. Magnesium and manganese may be substituted for calcium. However, the substitutions decrease the reliability and efficiency of the procedure.

Uptake of (3H)5-hydroxytryptamine and (3H)noradrenaline by slices of rat brain incubated in plasma from patients treated with chlorimipramine, imipramine or amitroptyline.

Abstract Great interest has been focused in recent years on the purported role of monoamines in the depressive syndrome (for review see McClure, 1971). Monoamine systems in the brains of various species have been mapped out (Fuxe, 1965; Ungerstedt, 1971) and noradrenaline nerve terminals have been demonstrated in human cerebral cortex (Nyström, Olsson & Ungerstedt, 1972). Tricyclic antidepressants have been shown to be effective in about 60% of depressions treated (Bennet, 1967). These drugs affect the monoamine neuron by inhibiting the uptake of the released transmitters noradrenaline and/or 5-hydroxytryptamine (5-HT) through the membrane pump of the nerve terminals. Various tricyclic antidepressants have different profiles of potency in inhibiting the uptake into noradrenaline and 5-HT nerve terminals, each drug having its own pattern (Carlsson, Corrodi & others, 1969; Shaskan & Snyder, 1970; Lidbrink, Jonsson & Fuxe, 1971; Ross, Renyi & Ögren, 1972). We have previously studied the active uptake of [3H]noradrenaIine (3H-NA) and [3H]5-hydroxytryptamine (3H-5-HT) into their respective nerve terminals by using slices of rat cerebral cortex incubated in human plasma which was drawn from patients before and during treatment with antidepressant drugs (Hamberger & Tuck, 1973).

DDT intoxication in birds: Subchronic effects and brain residues

After 5 days on ad libitum diets containing 25 to 800 ppm of technical DDT, the concentrations of DDT, DDE, and DDD in the brains of farm-reared bobwhite ( Colinus virgianus) could be correlated with dietary concentrations. Weight losses were significant when dietary concentrations reached 400 ppm but there were no other toxic signs. A diet of 800 ppm DDT produced mild signs of intoxication and total brain residues of 5.6–22.3 ppm. A diet containing 1600 ppm DDT caused severe toxic signs culminating in convulsions and death. Analysis of the brain showed an average concentration of DDD + DDT to be 28.4 ppm. The average concentrations of DDD + DDT in the brain of various species were: house sparrows ( Passer domesticus), 43.2 ppm; wild bobwhite, 31.3 ppm; farm-reared bobwhite, 28.4 ppm; cardinals ( Richmondena cardinalis), 26.7 ppm; and blue jays ( Cyanocitta cristata), 23.0 ppm. In bobwhite, cardinals, and blue jays concentrations in the brain ranging from 20 to 25 ppm strongly implicate DDT as the lethal agent. A concentration of 32.6 ppm in the brain of house sparrows was indicative of DDT poisoning.

The effect of hypoxia on isonicotinic acid hydrazide-induced seizures in chicks during ontogenesis

Abstract : The concentration of gamma-aminobutyric acid (GABA) in the brain of various species is elevated by hypobaric hpoxia, or by hypoxia caused by the breathing at ambient pressure of a gas mixture of low oxygen content. Hypobaric hypoxia also delays the onset of semi-carbazide- and methionine sulphoximine-induced seizures and it has been suggested that the elevation in brain GABA level may be responsible for the anticonvulsant action. Although the above studies clearly indicate that both an elevation in brain GABA concentration and an anticonvulsant actio are associated with hypoxic conditions, the results do not necessarily indicate a causal relation between the first two phenomena. The present investigation was therefore initiated to obtain additional information about this.

Methods for the estimation of acetylcholinesterase activity in the plasma and brain of laboratory animals given carbamates or organophosphorus compounds.

This report describes the modifications necessary to adapt the pH Stat and Ellman methods for the estimation of cholinesterase activity in the plasma and brain of various species of laboratory animals. Both methods were evaluated in studies in which cholinesterase inhibitors, carbamates and organophosphorus compounds, were given to the animals.

Modifications of the michel delta pH method for the estimation of plasma, erythrocyte and brain cholinesterase activities of various species of laboratory animals.

This report describes the modifications necessary to adapt the Michel method for the estimation of cholinesterase activity in the blood and brain of various species of laboratory animals.

A comparative study of gangliosides from brains of various species.

AbstractA comparison is made of the sialic acid and hexose content of crude gangliosides from the brains of 13 species, including mammals, a bird, and amphibian, a reptile and fish. The sialic acid content is relatively constant from species to species. Gangliosides are less abundant in tissues other than the brain. Four species (cat, dog, pig and sheep) were selected for the determination of their major ganglioside subfractions. The ganglioside subfractions were isolated (after extraction and partition dialysis) using descending thin layer chromatography. The population of ganglioside molecules varied from species to species. From dog and sheep a mono‐, a di‐ and a trisialoganglioside were obtained; from cat a mono‐, a di‐ and two trisialogangliosides; and from pig a mono‐ and two disialogangliosides. Each ganglioside subfraction was found to contain glucose, galactose and galactosamine in the ratio of approximately 1∶2∶1. The fatty acid moieties consisted of more than 80% stearic acid with lesser amounts of arachidic, palmitic and behenic acid. Sphingosine analyses indicated ratios of sphingosine to icosisphingosine of 7∶3 for the monosialo‐, 1∶1 for the disialo‐ and 3∶7 for the trisialogangliosides.

A quantitative mapping of alkaline phosphatase in the brain of the rhesus monkey.

THE distribution of acid phosphatase in the central nervous system as demonstrated by histochemical methods has been investigated by LANDOW, KABAT and NEWMAN (1942), WISLOCKI and DEMPSEY (1948), and by SHIMIZU (1950) in comparative studies that included mice, rats, guinea pigs, rabbits, cats and chickens. Numerous papers deal with the cytological distribution of acid phosphatase in selected regions of the brain of various species, but little attention has been given to gradations of enzyme activity among those regions. The present paper reports a mapping of acid phosphatase activity in the brain of the rhesus monkey. A quantitative histochemical method was used, based upon spectrophotometric measurements of dye extracts from standard size tissue discs taken from brain sections that had been incubated with a substrate for acid phosphatase. Previous mappings of oxidative enzymes (FRIEDE, 1961), and particularly a mapping of lactate dehydrogenase in the brain of the rhesus monkey (FRIEDE and FLEMING, 1963), facilitate comparison of enzyme patterns. MATERIAL AND METHODS Histochemical method. Preliminary testing and standardization of the quantitative method for acid phosphatase activity were done on cat and monkey brains; a detailed account of methodology will be given elsewhere. For the present experiment three adult, healthy rhesus monkeys were killed under pentothal anaesthesia; the brains were removed immediately and placed in 10 % formalin at 4". After 8 hr fixation, the material was cut into slices, 3-5 mm thick. Twenty-four hours after removal of each brain, 30 p frozen sections were cut, rinsed, and stored in chilled distilled water until all material from the brain had been prepared. All sections were transferred to a 4-litre Erlenmeyer flask containing a small portion of the buffer component of the incubation medium. The substrate and diazo salt were mixed with the remaining portion of the buffer and filtered as quickly as possible; the filtrate was added immediately to the sections and buffer in the Erlenmeyer flask and incubation was started. The incubation medium was a mixture of 1 mg of sodium-a-naphthyl phosphate (Signia Chemical Co. St Louis, Missouri) and 1 mg of the stable diazotate of o-amino azotoluene (Fast Garnet GBC) (Dajac Laboratories, Philadelphia) in 1 ml of 0.2 M-Walpole buffer of pH 5 (PEARSE, 1960). For each monkey brain approximately 2 I. of reaction mixture were used. The sections were incubated in an Eherbach water-bath shaker with constant agitation for 1 hr at 38". The reaction was stopped by transferring all sections into a large quantity of 10% formalin. The stained sections were mounted temporarily on a microslide covered with three layers of parafilm. Discs were cut from selected areas of the sections with stainless steel tubes, 1.1 mm and 2.4 mm in diameter. Each disc was placed in a stoppered test tube containing a drop of distilled water. Care was taken not to transfer particles of parafilm into the tubes since these would cause turbidity and false readings. The azo dye was extracted with 1 ml of chloroform-ethanol (1 :l), v/v and the colour density was determined within 10-30 min at 555 mp in microcells in a Beckman DU spectrophotometer. For the data collected in Table I an average of ten samples were taken from the regions indicated for each monkey, and means and standard deviations were calculated. The dependability of this method was tested in various experiments (unpublished data) the results of which are only briefly summarized: tissue fixed in formalin after incubation was used since it was impractical to cut discs from loose sections During the required 24-hr fixation period, some loss of

A QUANTITATIVE MAPPING OF ACID PHOSPHATASE IN THE BRAIN OF THE RHESUS MONKEY.

THE distribution of acid phosphatase in the central nervous system as demonstrated by histochemical methods has been investigated by LANDOW, KABAT and NEWMAN (1942), WISLOCKI and DEMPSEY (1948), and by SHIMIZU (1950) in comparative studies that included mice, rats, guinea pigs, rabbits, cats and chickens. Numerous papers deal with the cytological distribution of acid phosphatase in selected regions of the brain of various species, but little attention has been given to gradations of enzyme activity among those regions. The present paper reports a mapping of acid phosphatase activity in the brain of the rhesus monkey. A quantitative histochemical method was used, based upon spectrophotometric measurements of dye extracts from standard size tissue discs taken from brain sections that had been incubated with a substrate for acid phosphatase. Previous mappings of oxidative enzymes (FRIEDE, 1961), and particularly a mapping of lactate dehydrogenase in the brain of the rhesus monkey (FRIEDE and FLEMING, 1963), facilitate comparison of enzyme patterns. MATERIAL AND METHODS Histochemical method. Preliminary testing and standardization of the quantitative method for acid phosphatase activity were done on cat and monkey brains; a detailed account of methodology will be given elsewhere. For the present experiment three adult, healthy rhesus monkeys were killed under pentothal anaesthesia; the brains were removed immediately and placed in 10 % formalin at 4. After 8 hr fixation, the material was cut into slices, 3-5 mm thick. Twenty-four hours after removal of each brain, 30 p frozen sections were cut, rinsed, and stored in chilled distilled water until all material from the brain had been prepared. All sections were transferred to a 4-litre Erlenmeyer flask containing a small portion of the buffer component of the incubation medium. The substrate and diazo salt were mixed with the remaining portion of the buffer and filtered as quickly as possible; the filtrate was added immediately to the sections and buffer in the Erlenmeyer flask and incubation was started. The incubation medium was a mixture of 1 mg of sodium-a-naphthyl phosphate (Signia Chemical Co. St Louis, Missouri) and 1 mg of the stable diazotate of o-amino azotoluene (Fast Garnet GBC) (Dajac Laboratories, Philadelphia) in 1 ml of 0.2 M-Walpole buffer of pH 5 (PEARSE, 1960). For each monkey brain approximately 2 I. of reaction mixture were used. The sections were incubated in an Eherbach water-bath shaker with constant agitation for 1 hr at 38. The reaction was stopped by transferring all sections into a large quantity of 10% formalin. The stained sections were mounted temporarily on a microslide covered with three layers of parafilm. Discs were cut from selected areas of the sections with stainless steel tubes, 1.1 mm and 2.4 mm in diameter. Each disc was placed in a stoppered test tube containing a drop of distilled water. Care was taken not to transfer particles of parafilm into the tubes since these would cause turbidity and false readings. The azo dye was extracted with 1 ml of chloroform-ethanol (1 :l), v/v and the colour density was determined within 10-30 min at 555 mp in microcells in a Beckman DU spectrophotometer. For the data collected in Table I an average of ten samples were taken from the regions indicated for each monkey, and means and standard deviations were calculated. The dependability of this method was tested in various experiments (unpublished data) the results of which are only briefly summarized: tissue fixed in formalin after incubation was used since it was impractical to cut discs from loose sections During the required 24-hr fixation period, some loss of