Calmodulin,a calmodulin acceptor protein,and calcimedins: Unique antibody localizations in hamster sperm

A calmodulin acceptor protein has been identified in isolated hamster caudal sperm by immunofluoresence and Western transfer techniques. The protein shows a localization in sperm heads identical to calmodulin. Fluorescence of both calmodulin and the acceptor protein are lost by treatment with MgCl₂, conditions which release the acrosome. These results are consistent with the proposed function of calmodulin in a sperm function.     

Localization of choline acetyltransferase in laminae of the rat olfactory tubercle

Abstract: We report the distribution of choline acetyltransferase (ChAT) activity in the laminae of the rat olfactory tubercle. Within its posterior medial portion, the tubercle contains three parallel histological laminae that can be separated by cutting tangential sections from frozen tissue. ChAT was measured in homogenates of consecutive sections (16 μm) cut parallel to these laminae. The distribution of ChAT activity, as a function of tubercle depth, showed a broad peak centered at 500 μm from the ventral surface of the brain. Enzyme activity measured at this depth (85 pmol acetylcholine formed/μg protein/h) was 2 1/2 times greater than that measured in the outermost, plexiform, layer. Stereotaxic injections of kainic acid (1 μg in 1 μ1) made directly into the tubercle were used to eliminate intrinsic neurons. Three days after injection, histological examination revealed the almost total absence of neuronal cell bodies and the proliferation of glial cells. The greatest decreases in ChAT activity (50%) were seen at depths of 300–600 μm whereas no loss of activity occurred in the plexiform layer.     

The New Face of the Lipid Droplet: Lipid Droplet Proteins

The lipid droplet (LD) is an organelle with vital functions found in nearly all organisms. LD proteomic research has provided fundamentally important insights into this organelle's functions. The review provides a summary of LD proteomic studies conducted across diverse organisms and cell and tissue types. The accumulated proteomic data are reviewed for evidence of a protein targeting mechanism for the organelle. The hypotheses for several specific localization mechanisms based on what is known about targeting mechanisms for other organelles and vesicles are provided. Although the nature of the mechanism is not known, the functional data demonstrate that the targeting mechanism and, indeed, the organelle itself, is conserved from prokaryotes to eukaryotes. It is hoped that the review will help inspire further research leading to novel discoveries in the field.     

酿酒酵母全基因组SNARE蛋白的亚细胞定位研究

在真核细胞中,内质网、高尔基体、质膜等膜结构间的蛋白质运输主要通过囊泡出芽和融合实现。SNARE蛋白家族在介导囊泡与目的膜结构融合过程中发挥关键作用。在模式生物酿酒酵母中,对全基因组SNARE蛋白的系统研究仍有不足。此研究构建了一套用于标记酿酒酵母基因组全部24种SNARE的工具质粒。该系列质粒既能呈现出良好的定位特征,又避免了过度表达造成的定位异常。通过与细胞器标记共定位验证了SNARE蛋白的亚细胞定位。结果发现3种SNARE的定位与之前报道不符:Bos1定位于早高尔基体,Snc1和Bet1定位于晚高尔基体/早内体。另外,Sec9定位于芽尖和芽颈,这是首次观察到Sec9在活酵母细胞中的定位。这项工作首次全面的检验了酵母SNARE家族蛋白的亚细胞定位,为后续SNARE蛋白功能研究提供了新线索,并为相关研究提供了一套工具质粒。     

Subcellular cadmium distribution and antioxidant enzymatic activities in the leaves of four Hylotelephium spectabile populations exhibit differences in phytoextraction potential

Hylotelephium spectabile with high tolerance to cadmium (Cd) might be a potential candidate for phytoremediation. However, the mechanisms for Cd accumulation and tolerance in H. spectabile are poorly understood. Four H. spectabile populations, namely HB1, HB2, JS, and LN, were selected to investigate their Cd extraction potential and the underlying mechanism of Cd accumulation, focusing on subcellular distribution and antioxidant enzymes. The Cd concentration, bioconcentration factor and transfer factor of the LN was significantly higher than other populations, particularly with increasing Cd exposure, and no obvious growth inhibition observed. Segregation of excessive Cd to Cd-rich granule in LN was much higher than other populations which reveal one possible mechanism of Cd accumulation. A significant increase in superoxide dismutase (SOD) and catalase (CAT) activities with increasing Cd stress suggested SOD and CAT contribute to the Cd tolerance of H. spectabile. LN displayed significantly higher and constant peroxidase (POD) activities than other populations, which indicated that an effective mechanism existed in the LN to cope with Cd stress. Therefore, the subcellular distribution and antioxidant enzymes might play important roles in Cd accumulation and tolerance of H. spectabile. LN possessed high Cd extraction potential, and further studies under field conditions are warranted.     

Characteristics of Cd accumulation and distribution in two sweet potato cultivars

In this study, we compared the chemical forms and subcellular distribution of Cd in high-Cd (X16) and low-Cd (N88) sweet potato cultivars through hydroponic experiments and examined the Cd distribution in their roots by histochemical staining. The results showed that inorganic and pectate/protein-integrated Cd predominated in the leaves, and Cd concentrations were significantly higher in X16 than in N88. However, in the roots, Cd was mostly integrated with pectate and protein, and Cd concentration was higher in N88 than in X16. It was mainly stored through vacuolar sequestration and cell wall binding. In the leaves and stems, Cd concentrations in all subcellular fractions were higher in X16 than in N88; the opposite was observed in the roots. In X16, Cd was mostly accumulated in the root stele, and its Cd translocation factor was higher than that of N88. Overall, the subcellular fractions of X16 roots retained less Cd than N88 roots, and more Cd entered the root stele of X16 and subsequently moved to the shoots. The higher amounts of inorganic, water-soluble, and pectate/protein-integrated Cd with high mobility in the shoots of X16 than in N88 might facilitate Cd remobilization to other tissues, but this needs to be further studied.     

Transcriptional Heterogeneity of Beta Cells in the Intact Pancreas

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A novel de novo PDE4D gene mutation identified in a Chinese patient with acrodysostosis

Acrodysostosis is an extremely rare disorder at birth, that is, characterized by skeletal dysplasia with short stature and midfacial hypoplasia, which has been reported to be caused by PDE4D and PRKAR1A gene mutations. Here, a Chinese boy with acrodysostosis, ventricular septal defect, and pulmonary hypertension was recruited for our study, and his clinical and biochemical characteristics were analyzed. A novel de novo heterozygous missense mutation (NM_001104631: c.2030A>C, p.Tyr677Ser) of the PDE4D gene was detected by whole exome sequencing and confirmed by Sanger sequencing. The c.2030A>C (p.Tyr677Ser) variant was located in exon 15 of the PDE4D gene, predicted to be damaging by a functional prediction program and shown to be highly conserved among many species. Further functional analysis showed that the p.Tyr677Ser substitution changes the function of the PDE4D protein, affects its subcellular localization in transfected cells, increases PDE4 activity in the regulation of cAMP signaling and affects cell proliferation. Our study identified a novel de novo PDE4D mutation in acrodysostosis of Chinese origin that not only contributes a deeper appreciation of the phenotypic characteristics of patients with PDE4D mutations but also expands the spectrum of PDE4D mutations.