Differential G-protein-coupled receptor phosphorylation provides evidence for a signaling bar code

G-protein-coupled receptors are hyper-phosphorylated in a process that controls receptor coupling to downstream signaling pathways. The pattern of receptor phosphorylation has been proposed to generate a "bar code" that can be varied in a tissue-specific manner to direct physiologically relevant receptor signaling. If such a mechanism existed, receptors would be expected to be phosphorylated in a cell/tissue-specific manner. Using tryptic phosphopeptide maps, mass spectrometry, and phospho-specific antibodies, it was determined here that the prototypical G(q/11)-coupled M(3)-muscarinic receptor was indeed differentially phosphorylated in various cell and tissue types supporting a role for differential receptor phosphorylation in directing tissue-specific signaling. Furthermore, the phosphorylation profile of the M(3)-muscarinic receptor was also dependent on the stimulus. Full and partial agonists to the M(3)-muscarinic receptor were observed to direct phosphorylation preferentially to specific sites. This hitherto unappreciated property of ligands raises the possibility that one mechanism underlying ligand bias/functional selectivity, a process where ligands direct receptors to preferred signaling pathways, may be centered on the capacity of ligands to promote receptor phosphorylation at specific sites.     

Impaired striatum-dependent behavior in GASP-1-knock-out mice

G protein-coupled receptor (GPCR) associated sorting protein-1 (GASP-1) is suspected to play a key role in recycling and degradation of several GPCRs. In a previous study, we have shown that GASP-1-knock-out (GASP-1-KO) mice displayed deficits in acquiring a cocaine self-administration task, associated with an exacerbated down-regulation of striatal dopaminergic and cholinergic receptors. Among several possibilities, GASP-1 deficiency could have impaired memory processes underlying the acquisition of the operant conditioning task. Therefore, the present study investigated cognitive performances of GASP-1-KO mice and their wild-type littermates (WT) in a broad variety of memory tasks. Consistent with a deficit in procedural memory, GASP-1-KO mice showed delayed acquisition of a food-reinforced bar-press task. During water-maze training in hidden- or visible-platform paradigms, mutant and WT mice acquired the tasks at the same rate. However, GASP-1 mice exhibited persistent thigmotaxic swimming, longer distance to the platform, and reduced swim speed. There was no deficit in several tasks requiring simple behavioral responses (Barnes maze, object recognition and passive avoidance tasks). Thus, the ability to acquire and/or express complex responses seems affected in GASP-1-deficient mice. Hippocampal functions were preserved, as the retention of an acquired memory in spatial tasks remained unaffected. The pattern of behavioral deficits observed in GASP-1-KO mice is coherent with current knowledge on the role of striatal GPCRs in acquisition/expression of skilled behavior and in motivation. Together with the previous findings, the so far established phenotype of GASP-1-KO mice makes them a potentially exciting tool to study striatal functions.     

Untersuchungen zur Bildung der Vomerspange beiSalamandra salamandra (L.)

Zusammenfassung BeiSalamandra salamandra wurde die Entwicklung der Vomerspangen anhand von Exstirpations- und Transplantationsversuchen vor und während der Metamorphose untersucht. Die Vomerspange bildet sich aus Bindegewebe, das am Caudalrand des Larvenvomer liegt. Exstirpierte Vomerteile werden nicht regeneriert. Ein kurzer caudaler Abschnitt der larvalen Vomerzahnleiste genügt für die vollständige Ausbildung der Zahnleiste der Vomerspange. Wird die Spitze der auswachsenden Vomerspange reseziert, bleibt die Spange unvollständig. Die Bildung des Definitivvomer bleibt davon unberührt. In die Subcutis der Rumpfhaut transplantiertes Gewebe der sich bildenden Vomerspange bleibt zwar erhalten, wächst aber nicht weiter. In den Spangenbildungsbereich eingefügte Rumpfhaut verhindert das Auswachsen der Vomerspange.

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Investigations on the development of the vomerine bar inSalamandra salamandra (L.)

Summary The development of the vomerine bar inSalamandra salamandra was studied by amputation and transplantation procedures before and during metamorphosis. The bar of the vomer arises from the connective tissue near the caudal margin of the larval vomer. The larval vomer does not regenerate after amputation. A short caudal part of its dental lamina is sufficient for the outgrowing of the complete dental lamina of the vomerine bar. If the tip of the outgrowing vomerine bar is removed, the bar will remain incomplete, which procedure has no influence on the development of the definitive vomer. The material of the developing vomerine bar transplanted to the subepithelial connective tissue of the skin remains intact, but does not continue to grow. Fragments of the skin transplanted to the region from which the bar of the vomer arises prevent the development of the bar.

     

Neuromuscular onset succession of high level gymnasts during dynamic leg acceleration phases on high bar

In several athletic disciplines there is evidence that for generating the most effective acceleration of a specific body part the transfer of momentum should run in a “whip-like” consecutive succession of body parts towards the segment which shall be accelerated most effectively (e.g. the arm in throwing disciplines). This study investigated the question how this relates to the succession of neuromuscular activation to induce such “whip like” leg acceleration in sports like gymnastics with changed conditions concerning the body position and momentary rotational axis of movements (e.g. performing giant swings on high bar). The study demonstrates that during different long hang elements, performed by 12 high level gymnasts, the succession of the neuromuscular activation runs primarily from the bar (punctum fixum) towards the legs (punctum mobile). This demonstrates that the frequently used teaching instruction, first to accelerate the legs for a successful realization of such movements, according to a high level kinematic output, is contradictory to the neuromuscular input patterns, being used in high level athletes, realizing these skills with high efficiency.Based on these findings new approaches could be developed for more direct and more adequate teaching methods regarding to an earlier optimization and facilitation of fundamental movement requirements.     

番茄红素β-环化酶基因的玉米转化及 后代遗传分析

利用农杆菌介导法将番茄红素β-环化酶基因(Lycb)转入由玉米自交系天塔五号植株,分析基因在T0转化及后代的遗传情况,结果表明,在27株T0转基因植株中,PCR初步检测后8株呈阳性;将T1代转基因植株以株系为单位用200mg/L草铵膦抗性筛选后,收获抗性植株种子。T2代转基因植株进一步进行PCR、RT-PCR和田间草铵膦涂抹检测,结果表明,PCR、RT-PCR为阳性的6个株系植株均具有草铵膦抗性。选取6株阳性植株提取叶片总类胡萝卜素,经HPLC分析其β-胡萝卜素含量显著高于野生型,表明目的基因Lycb成功的转入玉米,并得到了稳定遗传。     

利用脉冲电泳技术转化玉米获得转基因植株的研究

本研究采用脉冲电泳技术将外源Bar基因和B-t基因转化玉米种胚并直接成苗,通过除草剂抗性初步筛选,再进行PCR检测、Southem杂交验证,探讨了脉冲电泳介导外源基因直接转化玉米种胚的有效性,结果表明：脉冲电泳技术介导玉米转基因是有效可行的,其T0代植株PCR阳性率与T1代植株转化率分别为11．36％和1．04％。在外源基因的转化中,处于不同启动子下的B-t基因和Bar基因共转化频率为90．9％。T1代PCR表现为阳性的植株Southem杂交结果显示脉冲电泳法介导的外源基因在受体中多为单拷贝,且已遗传给后代,并获得稳定表达。     

黄花菜不同外植体形成的愈伤组织再生苗观察

研究了黄花菜不同外植体愈伤组织的形成与植株再生。结果表明,出愈率是花柄>花茎>叶片。愈伤组织在MS+6-BA 2mg/L的培养基上出现致密愈伤组织颗粒,经增殖→分割→增殖的程序逐渐形成“球状体”似的愈伤组织。叶片、花茎形成的球状体经20代继代培养,再生能力没有减退,苗形态正常,根尖细胞染色体数目为2n=2x=22。该球状体经秋水仙素处理获得多种形态的四倍体苗和少数重复加倍现象。花柄球状体再生苗中出现了叶片花瓣状或类似花蕾状的形态异常苗,其频率与花柄所处的花期及花蕾大小有密切关系。再生异常苗的球状体继代培养,或者由异常苗叶片重新形成的球状体仍然再生异常苗。花柄球状体经秋水仙素处理,获得的变异株其染色体数目类型较多。因而不宜用花柄作黄花菜快速繁殖的外植体。