Insights Into Pigmentary Phenomena Provided by Grafting and Chimera Formation in the Axolotl

The expression of pigmentation patterns in axolotl pigmentary mutants was observed following three types of experimental manipulations including chimera formation, reciprocal neural crest grafts, and grafts of gonadal primordia. Three pigmentary genes were utilized including the wild type (D), white (d), and albino (a). In chimeras between white and albino embryos, melanoblasts from the white half crossed the graft interface to differentiate in albino skin. Neural crest grafts from white embryos to albinos provided melanophores of white origin that were capable of differentiation in albino skin. Grafts of gonadal primordia from albino to white embryos provided albino germ cells that formed unpigmented ovocytes together with dark ovocytes: white ovocytes from the albino grafted ovary, and dark ovocytes from the host ovary. The donor albino white ectoderm included in the graft was able to support the differentiation of melanophores, iridophores, and xanthophores that invaded the graft ectoderm from the neural crest of the white host. It was concluded that manifestation of the white or wild phenotypes may be related to the possible presence or absence of inhibiting or stimulating pigmentary factors in the skin. This possibility was discussed in the light of recent discoveries of such factors as Agouti Signaling Protein (ASP) from mammalian skin.     

Inference of macromolecular assemblies from crystalline state

We discuss basic physical-chemical principles underlying the formation of stable macromolecular complexes, which in many cases are likely to be the biological units performing a certain physiological function. We also consider available theoretical approaches to the calculation of macromolecular affinity and entropy of complexation. The latter is shown to play an important role and make a major effect on complex size and symmetry. We develop a new method, based on chemical thermodynamics, for automatic detection of macromolecular assemblies in the Protein Data Bank (PDB) entries that are the results of X-ray diffraction experiments. As found, biological units may be recovered at 80-90% success rate, which makes X-ray crystallography an important source of experimental data on macromolecular complexes and protein-protein interactions. The method is implemented as a public WWW service.     

Single QTL mapping and nucleotide-level resolution of a physiologic trait in wine Saccharomyces cerevisiae strains

Natural Saccharomyces cerevisiae yeast strains exhibit very large genotypic and phenotypic diversity. However, the link between phenotype variation and genetic determinism is still difficult to identify, especially in wild populations. Using genome hybridization on DNA microarrays, it is now possible to identify single-feature polymorphisms among divergent yeast strains. This tool offers the possibility of applying quantitative genetics to wild yeast strains. In this instance, we studied the genetic basis for variations in acetic acid production using progeny derived from two strains from grape must isolates. The trait was quantified during alcoholic fermentation of the two strains and 108 segregants derived from their crossing. A genetic map of 2212 markers was generated using oligonucleotide microarrays, and a major quantitative trait locus (QTL) was mapped with high significance. Further investigations showed that this QTL was due to a nonsynonymous single-nucleotide polymorphism that targeted the catalytic core of asparaginase type I (ASP1) and abolished its activity. This QTL was only effective when asparagine was used as a major nitrogen source. Our results link nitrogen assimilation and CO(2) production rate to acetic acid production, as well as, on a broader scale, illustrating the specific problem of quantitative genetics when working with nonlaboratory microorganisms.     

Effect of quinolinic acid on the distribution of hepatic metabolites

The intracellular distribution of hepatic metabolites in normal and quinolinic acid (QA)-treated rats has been calculated. QA, an inhibitor of gluconeogenesis, raises the total cell content of malate, aspartate, α-ketoglutarate and citrate. The calculated mitochondrial content of all four metabolites was raised, as was the mitochondrial/cytosolic gradient, and the cytosolic content of oxaloacetate and α-ketoglutarate decreased. The fall of cytosolic oxaloacetate in QA-treated rats suggests a control at PEPCK by substrate limitation. It is suggested that QA may have an action on the translocation of polycarboxylate anions across the mi tochondrial membrane.     

低能氩离子注入对甜瓜幼苗抗氧化系统的影响

就低能氩离子注入对甜瓜幼苗抗氧化系统的影响进行研究,使用不同剂量的离子注入甜瓜种子,结果表明,不同剂量离子注入后伽师瓜、皇后品种的超氧化物歧化酶(SOD)分别在5×1016Ar /cm2、4×1016Ar /cm2剂量时酶活性最大,之后下降。抗坏血酸过氧化物酶(ASP)的活性的变化趋势与超氧化物歧化酶相似,分别在6×1016Ar /cm2、4×1016Ar /cm2剂量时酶活性最大。抗坏血酸和类黄酮的含量都有不同程度的上升,说明离子注入能诱导甜瓜幼苗抗氧化系统的生物合成。     

朱砂叶螨危害对豇豆幼苗叶片活性氧代谢相关酶的影响

朱砂叶螨危害豇豆幼苗后,叶绿体内与活性氧代谢有关的酶SOD、ASP活性及同工酶均受到不同程度的影响。(1)受害2～8d,SOD活性与对照相比均升高,差异显著。不同虫口密度之间在4d时SOD活性差异显著;(2)在危害期内,随着危害时间的延长ASP活性显著升高,且不同虫口密度之间差异极显著,虫口密度越大,ASP活性越高;(3)SOD同工酶谱带显示,随着危害程度的加强,一些分子量较大的同工酶谱带亦加强。     

4-(4-(dimethylamino)phenyl)-1-methylpyridinium (APP+) is a fluorescent substrate for the human serotonin transporter

Monoamine transporters terminate synaptic neurotransmission and are molecular targets for antidepressants and psychostimulants. Fluorescent reporters can monitor real-time transport and are amenable for high-throughput screening. However, until now, their use has mostly been successful to study the catecholamine transporters but not the serotonin (5HT) transporter. Here, we use fluorescence microscopy, electrophysiology, pharmacology, and molecular modeling to compare fluorescent analogs of 1-methyl-4-phenylpyridinium (MPP(+)) as reporters for the human serotonin transporter (hSERT) in single cells. The fluorescent substrate 4-(4-(dimethylamino)phenyl)-1-methylpyridinium (APP(+)) exhibits superior fluorescence uptake in hSERT-expressing HEK293 cells than other MPP(+) analogs tested. APP(+) uptake is Na(+)- and Cl(-)-dependent, displaced by 5HT, and inhibited by fluoxetine, suggesting APP(+) specifically monitors hSERT activity. ASP(+), which was previously used to study catecholamine transporters, is 10 times less potent than APP(+) at inhibiting 5HT uptake and has minimal hSERT-mediated uptake. Furthermore, in hSERT-expressing oocytes voltage-clamped to -60 mV, APP(+) induced fluoxetine-sensitive hSERT-mediated inward currents, indicating APP(+) is a substrate, whereas ASP(+) induced hSERT-mediated outward currents and counteracted 5HT-induced hSERT currents, indicating ASP(+) possesses activity as an inhibitor. Extra-precise ligand receptor docking of APP(+) and ASP(+) in an hSERT homology model showed both ASP(+) and APP(+) docked favorably within the active region; accordingly, comparable concentrations are required to elicit their opposite electrophysiological responses. We conclude APP(+) is better suited than ASP(+) to study hSERT transport fluorometrically.