Antibodies to Dopamine: Radioimmunological Study of Specificity in Relation to Immunocytochemistry

Abstract: Two classes of anti-3,4-dihydroxyphenylethylamine (dopamine) antibodies were raised in rabbits using dopamine conjugated to albumin either via formaldehyde or via glutaraldehyde. Each was usable for immunohistochemical detection of dopamine neurons provided that the tissue was fixed by the homologous cross-linking agent. However, anti-dopamine-glutaraldehyde antibodies turned out to be of more general use because of the better fixative properties of glutaraldehyde which fixed dopamine in rat and in teleost, whereas formaldehyde only worked in lower vertebrates (such as goldfish) and not in rat brain. The specificity of antidopamine-glutaraldehyde antibodies was firmly established by competition experiments in equilibrium dialysis, using an immunoreactive tritiated derivative synthesized by coupling dopamine to N -α-acetyl-L-lysine N -methylamide via glutaraldehyde. Specificity studies in vitro and immunohistological results demonstrating the specific staining of dopaminergic neurons were found to correlate well.     

α-Ketoglutarate and Malate Uptake and Metabolism by Synaptosomes: Further Evidence for an Astrocyte-to-Neuron Metabolic Shuttle

This study was undertaken to provide further evidence relevant to the hypothesis that astrocytes supply one or more citric acid cycle intermediates to synaptic terminals, thereby serving an anaplerotic function necessitated by the synthesis and release of amino acid neurotransmitters. In our experiments, two populations of synaptosomes obtained from the brain of rats were separated from myelin and mitochondria by using Percoll to generate continuous density gradients. Both synaptosomal populations readily accumulated 14C-labelled alpha-ketoglutarate and L-malate by high-affinity transport systems. Hofstee plots of uptake velocity as a function of substrate concentration were highly nonlinear, indicating that uptake was mediated by two or more carriers, or was subject to negative cooperativity. At least one carrier was selective for alpha-ketoglutarate and another for malate, whereas a third carrier appeared to be present which transported both substrates. At low concentrations (approximately 1 microM), alpha-ketoglutarate transport was almost totally Na+-dependent, whereas malate uptake exhibited little Na+-dependency. The transport of alpha-ketoglutarate was associated with a net influx, and therefore was not due to a homoexchange process. alpha-Ketoglutarate and malate were metabolized rapidly to glutamate and aspartate, respectively, by both synaptosomal preparations; however, in all cases, label accumulated in gamma-aminobutyric acid rather slowly. The incorporation of label into glutamine from alpha-ketoglutarate was much greater in the high-density synaptosomes that in low-density synaptosomes, an indication that the former contained a higher proportion of astrogliasomes.(ABSTRACT TRUNCATED AT 250 WORDS)     

A proton-deuterium exchange study of three types ofDesulfovibrio hydrogenases

Summary Hydrogenases are among the main enzymes involved in bacterial anaerobic corrosion of metals. The study of their mode of action is important for a full comprehension of this phenomenon. The three types ofDesulfovibrio hydrogenases [(Fe), (NiFe), (NiFeSe)] present different patterns in the pH dependence of their activity. The periplasmic enzyme fromDesulfovibrio salexigens and the cytoplasmic enzyme fromDesulfovibrio baculatus both have pH optima at 7.5 for H₂ uptake and 4.0 for H₂ evolution and H⁺–D₂ exchange reaction (measured by membrane-inlet mass-spectrometry). The H₂ to HD ratio at pH above 5.0 is higher than 1.0. The periplasmic hydrogenase fromD. gigas presents the same pH optimum (8.0) for the H⁺–D₂ exchange as for H₂ consumption. In contrast, the enzyme fromD. vulgaris has the highest activity in H₂ production and in the exchange at pH 5.0. Both hydrogenases have a H₂-to-HD ratio below 1.0.     

The relationship between hydrogen metabolism,sulfate reduction and nitrogen fixation in sulfate reducers

Summary Hydrogenase and nitrogenase activities of sulfate-reducing bacteria allow their adaptation to different nutritional habits even under adverse conditions. These exceptional capabilities of adaptation are important factors in the understanding of their predominant role in problems related to anaerobic metal corrosion. Although the D₂–H⁺ exchange reaction indicated thatDesulfovibrio desulfuricans strain Berre-Sol andDesulfovibrio gigas hydrogenases were reversible, the predominant activity in vivo was hydrogen uptake. Hydrogen production was restricted to some particular conditions such as sulfate or nitrogen starvation. Under diazotrophic conditions, a transient hydrogen evolution was followed by uptake when dinitrogen was effectively fixed. In contrast, hydrogen evolution proceeded when acetylene was substituted as the nitrogenase substrate. Hydrogen can thus serve as an electron donor in sulfate reduction and nitrogen metabolism.     

The reserve proteins in the cells of mature cotyledons ofLupinus albus var.Lucky

Summary The nuclear DNA content of cotyledonary cells of two lupin seeds (L₁ and L₂) with markedly different total protein content, were investigated by scanning cytophotometry. Both seeds had polyloid nuclei with DNA levels varying between 8 C and 64 C, the majority being either 16 C or 32 C. The highest DNA levels were found in the abaxial and central cotyledonary zones of both seeds; seed L₂ had a higher ploidy level than L₁. It is shown that the volume of condensed chromatin (chromocenters) increased proportionally with the DNA content of the nucleus. A comparison was made between the distribution of protein, previously determined byLe Gal andRey (1986) and the DNA throughout the cotyledon. The L₂ seed, which has the highest total protein and the highest protein content per cell, also exhibited the greatest DNA content per cell. For both seeds, the r-value for association of DNA and protein content per cell was highly significant (0.98).     

Postnatal development of a demyelinating disease in avian spinal cord chimeras

Xenogeneic spinal cord chimeras were constructed by grafting fragments of quail neural primordium into chick embryos at 2 days of incubation. Hatched birds displayed normal motor behavior for about 5 to 7 weeks, whereupon they developed a neurological syndrome; in the grafted spinal cord the pathological signs of the disease were very similar to those of the active plaques of multiple sclerosis and of the lesions of experimental allergic encephalomyelitis and neuritis, including Ia expression by brain capillary endothelia, rupture of the blood-brain barrier, leukocytic infiltration in the nervous tissue, and demyelination. In the animals at the most advanced stage of the disease an autoimmune attack occurred on the host's nervous system with the same histopathological signs.     

Leishmaniasis in Bolivia. II. The involvement of Psychodopygus yucumensis and Psychodopygus llanosmartinsi in the selvatic transmission cycle of Leishmania braziliensis braziliensis in a lowland subandean region

An epidemiological survey of the vectors of cutaneous leishmaniasis ("espúndia" type) was carried out in the Alto Beni region of Bolivia, an area of Andean foothills at the Eastern limit of the Amazonian lowlands. The climate is typical wet tropical (15 degrees S latitude). Anthropophilic phlebotomine sandfly species were sampled at 20 sites, all forested. The importance of species from the Psychodopygus group, already suspected as a vector in the transmission of Leishmania from the braziliensis complex, was confirmed by: the aggressiveness and diversity of the species encountered (83% of catches, nine species), the discovery of a new anthropophilic species, P. yucumensis and the isolation of a strain of Leishmania braziliensis braziliensis indistinguishable from human strains from the same area, from two species, P. llanosmartinsi and P. yucumensis.     

Detailed analysis of the mouse H-2Kb promoter: enhancer-like sequences and their role in the regulation of class I gene expression

Sequencing and deletion analyses of the H-2Kb promoter have suggested that several regions may be important for expression and regulation of this gene. Two of these regions are conserved inside the promoter of several genes coding for classical transplantation antigens, but not in the promoter of class I genes located in the Qa region. They display enhancer-like activity in cells that express H-2 genes, but show some tissue specificity in that they function very poorly in undifferentiated embryonal carcinoma cells in which H-2 genes are not expressed. They also have been shown not to be the target of the adenovirus-12 induced repression of class I gene expression recently demonstrated by Schrier et al. The promoter of the beta 2-microglobulin gene also contains a sequence with enhancer-like activity, but shares no homology with the H-2Kb promoter region.     

Central injections of arginine vasopressin prolong extinction of active avoidance

Behavioral and physiological effects of arginine vasopressin (AVP) were examined following intracerebroventricular (ICV) injection in the rat. ICV injections prolonged extinction of active avoidance at doses of 1.0 and 10.0 ng/rat and this effect was blocked by peripheral injection of the vasopressor antagonist of vasopressin [dPtyr(Me)AVP] at a dose of 30 micrograms/kg (SC). However, 1.0 ng of AVP ICV failed to alter systemic blood pressure and also failed to produce taste aversions in a one or two bottle test. Results suggest that central AVP has a central action independent of systemic changes in blood pressure, but that the receptor mediating this action is functionally similar to the AVP V1 (vasopressor) receptor.     

Characterization and expression of collagen-like genes in Drosophila melanogaster

We have used a cloned chicken collagen cDNA sequence to help identify hypothetic members of the collagen gene family from Drosophila melanogaster. Several experimental evidences have been obtained which indicate that the Drosophila genome contains numerous collagen-like sequences. We have characterized in more detail ten distinct DNA sequences that hybridized strongly to the heterologous collagen probe. By in situ hybridization we have shown that these sequences are dispersed throughout the Drosophila genome. Two of them are shown to originate from the previously described DCg 1 and DCg 2 collagen genes. In other respects, we show that in addition to DCg 1 and DCg 2, at least five putative collagen genes are expressed during the Drosophila lifetime. These genes are unique, and some of them are seen to be transcribed into different size classes of mRNAs. Additionally, the data presented so far demonstrate that the expression of these genes is regulated temporally and/or quantitatively during the Drosophila life cycle.