Na+-dependent transport of basic, zwitterionic, and bicyclic amino acids by a broad-scope system in mouse blastocysts

Mouse blastocysts which had been activated from diapause in utero appeared to take up amino acids via a Na+-dependent transport system with novel characteristics. In contrast to other cell types, uptake of 3-aminoendobicyclo [3,2,1]octane-3-carboxylic acid (BCO) by blastocysts was largely Na+ dependent. Moreover, L-alanine and BCO met standard criteria for mutual competitive inhibition of the Na+-dependent transport of each other. The Ki for each of these amino acids as an inhibitor of transport of the other had a value similar to the value of its Km for transport. In addition, both 2-aminoendobicyclo [2,2,1]heptane-2-carboxylic acid (Ki approximately 1.0 mM) and L-valine (Ki approximately 0.10 mM) appeared to inhibit Na+-dependent transport of alanine and BCO competitively. Finally, alanine and L-lysine appeared to compete for the same Na+-dependent transport sites in blastocysts. For these reasons, we conclude that lysine, alanine, and BCO are transported by a common Na+-dependent system in blastocysts. In addition, the apparent interaction of the system with other basic amino acids, such as 1-dimethylpiperidine-4-amino-4-carboxylic acid, which has a nondissociable positive charge on its side chain, and L-arginine and L-homoarginine, whose cationic forms are highly predominant at neutral pH, suggests that the cationic forms of basic amino acids are transported by the wide-scope system.     

Neurokinin A-like immunoreactivity in rat primary sensory neurons; coexistence with substance P

Summary Rat spinal cord, dorsal root ganglia and skin were investigated employing immunohistochemical technique with specific antisera to neurokinin A and substance P. Neurokinin A-like immunoreactivity was detected in the spinal dorsal horn and skin with a similar distribution pattern as that of substance P-like immunoreactivity. After dorsal root transection a parallell decrease of neurokinin A and substance P-like immunoreactivity was observed in the dorsal horn. Using colchicine pretreatment a population of neurokinin A positive cell bodies was seen in the dorsal root ganglia, and by comparison of consecutive sections of the same cells stained for substance P it was revealed that these neurons also display substance P-like immunoreactivity. However, substance P-, but not neurokinin A-, immunoreactive cells were also observed. It is concluded that neurokinin A- and substance P-like immunoreactivity coexist in a population of rat primary sensory neurons.     

Asexual propagation and reproductive strategies in aquatic Oligochaeta

A great variety of asexual reproductive modes are known among aquatic oligochaetes. The main types of these modes are shortly described. Based upon observations on natural populations, the possible genetical and ecological implications of asexual reproduction are discussed. The following points are emphasized: (1) The often expressed expectations of a strong predominance of one particularly adaptive genotype is not born out. (2) In most cases, a number of genetically distinct clones are present in each population, and they show a strong differential distribution in heterogeneous environments, indicating an effective exploitation of the available resources. (3) Most cases of asexual propagation are reproductive strategies of their own and not escape mechanisms. (4) The mechanisms underlying asexual propagation are complex and involve many aspects of the life history. The great variety of types among aquatic oligochaetes offer particularly useful models for the study of these problems.     

Organic ion transport during seven decades. The amino acids

The amino acids are ions of various charge combinations, and one can argue that historically they were the first ions for which the ongoing problem of membrane transport was presented; also that among transported ions these may undergo a highly detailed molecular recognition. Furthermore, the distribution of charge on the amino acid molecule determines by what route or routes it is conducted across the biological membrane, with what directional and structural specificity, and therefore what regulation is imposed, and where. Cases where a presumably charged chemical group behaves as if it were somehow absent from the amino acid have been observed to fall into several categories: Straightforward cases where the pH has been low enough or high enough to remove the charge by protonation or deprotonation, even in free solution. Cases where that protonation or deprotonation is facilitated at the binding site, and perhaps by the total transport process. The cystine molecule can apparently thus be rendered either a tripolar anion or a tripolar cation for transport. Cases where an otherwise co-transported Na+ is omitted to redress charge, or where a Na+ serves as a surrogate for a missing charged group on the amino acid molecule. A case where the protonation occurs reversibly at the receptor site rather than on the amino acid molecule.     

Trypanosoma brucei-induced blockage of expulsion of Echinostoma revolutum and of homologous E. revolutum resistance in mice

Experiments were designed to study the effect of Trypanosoma brucei on the expulsion of Echinostoma revolutum and on the development and maintenance of homologous E. revolutum resistance in the mouse. T. brucei given 3, 2, and 1 wk before and 1 wk after infection with E. revolutum completely inhibited the expulsion of the E. revolutum worm burden for a period of at least 6 wk following infection, and T. brucei given 2 or 3 wk after infection with E. revolutum conferred a significant delay in the expulsion of the E. revolutum worm burden. T. brucei given 1 wk before and 1 wk after a primary E. revolutum infection blocked completely the resistance of the mouse to a homologous E. revolutum challenge given 2 wk after the primary infection. A similar blockage of resistance to a homologous challenge was experienced by mice given T. brucei 3 wk after the primary E. revolutum infection and challenged following another 2 wk. The mechanisms underlying the T. brucei-induced interference with the expulsion of E. revolutum and with the development and maintenance of homologous E. revolutum resistance in mice are presumably immunologically mediated.     

Tachykinin multiplicity in rat central nervous system as studied using antisera raised against substance P and neurokinin A

Antisera were raised in rabbits against the tachykinins neurokinin A (NKA) and substance P (SP). All NKA-antisera tested cross-reacted markedly with NKB, kassinin and eledoisin in radioimmunoassay (RIA), but virtually not with SP and physalaemin. Also when used for immunohistochemistry, one of the NKA-antisera was found to be virtually without cross-reactivity with SP. The most specific SP-antiserum did not cross-react with NKA but to some extent with NKB at the immunohistochemical level. Using these two antisera, the same distribution pattern of immunoreactivity was seen in both the rat substantia nigra and dorsal spinal cord. In neutral extracts of the substantia nigra, all NKA-antisera used for RIA detected a major component which eluted at the position of NKA in reverse phase high performance liquid chromatography, while no or only little immunoreactivity was detected at the position of NKB. A major component of substance P-like immunoreactivity (SPLI) co-eluting with SP and one or two minor SPLI-components were also detected in these extracts. An SP-antiserum, which cross-reacted markedly with physalaemin, detected an additional rather prominent component. In neutral water extracts of dorsal spinal cord the component detected with the NKA-antisera at the position of NKB, as well as one of the SPLI-components not eluting in the position of SP, were much more prominent than in the corresponding extracts of substantia nigra. In acetic acid extracts of both tissues, only one major SPLI-component co-eluting with SP could be detected, while only very small amounts of immunoreactivity eluting at the position of NKA and NKB (dorsal spinal cord only) could be detected using the NKA-antisera. The present results illustrate the importance of the extraction method used in immunochemical studies and demonstrate that the relative proportions of various tachykinins are markedly different in the rat substantia nigra and dorsal spinal cord.     

Effect of dextran and dextran modifications on the thermal and proteolytic stability of conjugated bovine testis beta-galactosidase and human serum albumin

In order to study carbohydrate-induced protein stabilization bovine testis beta-galactosidase and human serum albumin were conjugated with dextran, partially acetylated dextran and partially methylated dextran. The conjugates and the free proteins were compared with respect to thermal stability at 50 degrees C and resistance to proteolytic digestion by subtilopeptidase A. Both beta-galactosidase and serum albumin were stabilized by conjugation with polysaccharide. However, higher stability was achieved by conjugating the proteins with the hydrophilic polysaccharides, dextran and acetylated dextran, than by conjugation with the hydrophobic polysaccharide, methylated dextran. The results are discussed in relation to possible explanations of carbohydrate-induced protein stabilization.     

A subcortical, pigment-containing structure in Xenopus eggs with contractile properties

An accumulation of insoluble, finely granular material has been observed under the pigmented surface of Xenopus eggs by a specialized "dry fracture" technique and scanning electron microscopy. Cortical granules and pigment granules can be recognized with the techniques and can be seen to be embedded in the material. Thin sections show that the region also contains mitochondria and membranous vesicles or reticula. Yolk platelets are largely excluded from the heaviest accumulations of the material. The substance is most dense just under the cortex and grades off gradually into the more diffuse, yolk-containing network of the endoplasm. The accumulation of material is much thicker in the animal hemisphere of the egg than in the vegetal hemisphere, and the pigment embedded in it defines the pigmented area of the animal hemisphere. In the pigmented area the material excludes yolk for a thickness of 3-7+ microns from the surface. In the vegetal hemisphere there is no such accumulation and yolk platelets can be found almost touching the plasmalemma. Cortical contractions have been experimentally induced in eggs. Their relative strength correlates with the relative thickness of the finely granular, subcortical material. During contraction the material accumulates to much greater thicknesses, excluding yolk from thicknesses of 15-30+ microns from the surface. The contracting entity is, or is in, the finely granular material. Injection of cytochalasins into the eggs inhibits cleavage furrow operation but does not inhibit the induced cortical contractions. The thus do not seem to be dependent on actin microfilamentogenesis as is the operation of the contractile ring of the cleavage furrow. The differential sensitivity to cytochalasins of the contractile ring and the system responding in the induced cortical contractions, suggests a two-component system for cortical contractions in the egg. A model is presented which accommodates the available data.