The effects of putative amino acid neurotransmitters on somata isolated from neurons of the locust central nervous system

Changes in spike frequency, membrane potential and input resistance of somata freshly isolated from neurons in the metathoracic ganglia of adult locusts (Schistocerca gregaria) during bath and ionophoretic application of putative amino acid transmitters and analogues were studied using intracellular techniques. gamma-Aminobutyrate, glycine, taurine, cysteine and DL-ibotenate hyperpolarized the isolated soma, the response to kainic acid was depolarizing whereas L-glutamate and L-aspartate evoked a variety of potential changes. All of these compounds reduced the input resistance of the isolated soma. Ionophoretic studies showed that the receptors for L-glutamate and gamma-aminobutyrate are diffusely distributed over the somal surface.     

Locust nymphal neurones in culture: a new technique for studying the physiology and pharmacology of insect central neurones

A new technique for studying the physiology and pharmacology of locust central neurones is described. Somata isolated from neurones in the meso and metathoracic ganglia of third instar locusts (Schistocerca gregaria) were maintained for up to 4 weeks in co-culture (monolayer) with embryonic locust neurones. Most of the cultured cells became multipolar but a few were monopolar like their in vivo counterparts. They had diameters of 40-80 microns and "clean" (glial free) surface membranes. Cells 6-14 days in vitro were depolarized by acetylcholine and usually hyperpolarized by gamma-aminobutyrate, taurine and glycine. L-Glutamate and L-aspartate were inactive but further pharmacological studies are required to confirm this. Cultured larval neurones should provide excellent opportunities to study the molecular basis for drug-receptor interactions and voltage-sensitive membrane channels using the patch clamp technique.     

Primary structural variation among serologically indistinguishable DS antigens: the MB3-bearing molecule in DR4 cells differs from the MB3-bearing molecule in DR5 cells

HLA-DS molecules bearing the MB3 supertypic specificity have been isolated from two DR4 and two DR5 homozygous cell lines by using the monoclonal antibody IVD12 . Limited amino-terminal amino acid sequence analysis of these molecules demonstrates polymorphism of the HLA-DS subregion. Although the distribution of amino-terminal tyrosine residues in the alpha-chains of all IVD12 -reactive molecules was identical, amino-terminal amino acid sequence differences existed between DS beta-chains isolated from these two groups of cell lines bearing different DR specificities. These studies indicate that two DS molecules bearing the same serologic determinant ( MB3 ), although similar to one another, may be structurally distinct.     

HLA Haplotypes with C4B5; evidence for further allelic heterogeneity

Twenty-three individuals from various disease groups and normal controls were identified by immunofixation with anti-C4, C4-dependent lysis, determination of Rg (Rodgers) and Ch (Chido) phenotypes, and immunoblotting with C4-specific mouse monoclonal antibody. We found that one haplotype predominates with the C4B ^* 5 allele, HLA-A11, B22(55), Cw3, Bf ^* S, C4A ^* 4B ^* 5, which also carries the Ch ^{1,–2, 3} haplotype. The B5 allotype was also found with HLA-1360, HLA-1335 in Caucasoids, and HLA-B18 in non-Caucasoids; these carried the Ch ^{–1, –2, –3} haplotype. Our results are in accord with an earlier report of two B5 subtypes, B5Rg⁺ and B5Rg^– (Roos et al. 1984). The specificity of the mouse monoclonal antibodies IC4 and 21312 had been previously related to C4A and C4B, respectively, but our results suggest that they relate more closely to Rg and Ch determinants.     

Structural basis of the polymorphism of human complement components C4A and C4B: gene size, reactivity and antigenicity.

The human complement components C4A and C4B are highly homologous proteins, but they show markedly different, class-specific, chemical reactivities. They also differ serologically in that C4A generally expresses the Rodgers (Rg) blood group antigens while C4B generally expresses the Chido (Ch) blood group antigens. C4A 1 and C4B 5 are exceptional variants which possess their class-specific chemical reactivities, but express essentially the reversed antigenicities. The genes encoding the typical Rg-positive C4A 3a and Ch-positive C4B 3 allotypes and the interesting variants C4A 1 and C4B 5 have been cloned. Characterization of the cloned DNA has revealed that the genes encoding the A 3a, A 1 and B 3 allotypes are 22 kb long, but that encoding B 5 is only 16 kb long. Comparison of derived amino acid sequences of the polymorphic C4d fragment has shown that C4A and C4B can be defined by only four isotypic amino acid differences at position 1101-1106. Over this region C4A has the sequence PCPVLD while C4B has the sequence LSPVIH, and this presumably is the cause of their different chemical reactivities. Moreover, the probable locations of the two Rg and the six Ch antigenic determinants have been deduced. Our structural data on the C4A and C4B polymorphism pattern suggests a gene conversion-like mechanism is operating in mixing the generally discrete serological phenotypes between C4A and C4B.     

Heterogeneity of leukotriene receptors in guinea-pig trachea

The selective leukotriene (LT) antagonist FPL 55712 antagonized the contractile activity of synthetic LTD4 and E4 on guinea-pig trachea. Schild analysis of the antagonism provided evidence for two distinct receptors for LTD4: one with significantly higher affinity for FPL 55712 than the other. LTE4 appears to interact preferentially with the high affinity receptor.     

Revertants and Secondary arom-2 Mutants Induced in Non-Complementing Mutants in the arom Gene Cluster of Neurospora Crassa

Extensive genetical and biochemical studies have been performed with revertants and secondary arom-2 mutants induced in two different primary non-complementing mutants which map within the arom gene cluster of Neurospora crassa. These studies indicate that mutant M54 but not M25 can revert by super-suppressor mutations in unlinked genes, thus confirming previous evidence that M54 contains a nonsense codon. At least three new super suppressors of M54 have been detected. All four super suppressors (including one previously detected) when combined with M54 result in high levels of all five of the arom enzymic activities in the form of arom multienzyme complexes very similar to (but not necessarily identical with) that in wild type (WT).-Evidence has also been obtained that the two non-complementing mutants can yield revertants which appear to result from true back mutations and produce arom aggregates essentially indistinguishable from that of WT. In addition, M25, but not M54, when plated on quinic acid yields revertants (secondary mutants) some of which are phenotypically indistinguishable from arom-2 primary mutants and others of which, although also mapping within the arom-2 gene, exhibit unusual properties. Genetic evidence indicates that the M25 secondary mutants are localized within the arom-2 gene, but that they arise from mutational events more complex than ones resulting in single base pair changes in the M25 codon.-The recovery of secondary arom-2 mutants as revertants of non-complementing arom mutants provides strong evidence, independent of earlier recombination data, that non-complementing arom mutants are located within the arom-2 structural gene of the arom gene cluster. In addition, the occurrence and characteristics of these secondary arom-2 mutants provide strong evidence, independent of the results with nonsense suppressors, that the arom gene cluster is transcribed, beginning with the arom-2 gene, as a single polycistronic messenger ribonucleic acid (mRNA) molecule which is subsequently translated into the arom multienzyme complex.     

Distribution of β-thalassemia trait and erythrocyte glucose-6-phosphate dehydrogenase deficiency in the Markham river valley of New Guinea

Ten villages in or near the Markham Valley, northeastern New Guinea, have provided a sample of 476 males for the ascertainment of glucose-6-phosphate dehydrogenase (G6PD) deficiency and 810 individuals of both sexes for the investigation of the β-thalassemia trait. An extreme heterogeneity was found in the prevalence of both traits when the villages were analyzed separately (from 1.5% to 18.2% demonstrating G6PD deficiency and from 0 to 22.8% evincing β-thalassemia trait). The latter result counters an earlier report from the same region that β-thalassemia trait frequencies correlated negatively with altitude and (presumably) positively with endemicity of malaria. The present findings, based on a more representative sample, do not necessarily invalidate the relationship between malaria and β-thalassemia trait suggested by earlier studies, but they do make clear that correlation in New Guinea, at least, may be complicated by random genetic drift, although other possibilities are discussed. The heterogeneous distribution of G6PD deficiency in the present study confirms similar earlier findings. The importance of sample provenience and anthropological data in the interpretation of differences in the distribution of genetically determined traits is stressed.