细胞功能测定

This chapter provides an overview of the polyamine field and introduces the 32 other chapters that make up this volume. These chapters provide a wide range of methods, advice, and background relevant to studies of the function of polyamines, the regulation of their content, their role in disease, a ...

In addition to regulating gene transcription, polyamines also potently modulate gene expression posttranscriptionally. Posttranscriptional gene regulation, which includes processes such as mRNA transport, turnover, and translation, involves specific mRNA sequen ...

Unusual long polyamines such as caldopentamine and caldohexamine, and branched polyamines such as tetrakis(3-aminopropyl)ammonium and N 4-aminopropylspermidine were often found in cells of extreme thermophiles and hyperthermophiles belonging to both Bacteria and Arc ...

Polyamine blockade of inwardly rectifying potassium (Kir) channels underlies their steep voltage-�dependence observed in native cells. The structural determinants of polyamine blockade and the structure-activity profile of endogenous polyamines requires speciali ...

Polyamines are intimately linked to essential cellular processes that are required for cell growth and �proliferation, and abundant evidence links polyamine metabolism to tumor susceptibility and progression. Intensive efforts over the past 2 decades have yielded numerous mo ...

Depletion of pancreatic acinar cell polyamines in response to activation of polyamine catabolism is associated with the development of acute pancreatitis in experimental rodent models. The disease is characterized by general hallmarks seen also in human pancreatitis, such as acc ...

Embyroid body (EB) formation is a key step in many embryonic stem cell (ESC) differentiation protocols. The EB mimics the structure of the developing embryo, thereby providing a means of obtaining any cell lineage. Traditionally, the two methods of EB formation are suspension and hanging drop. ...

Mesenchymal stromal/stem cells (MSCs) were originally isolated from bone marrow (BM), but are now known to be present in all fetal and adult tissues. These multipotent cells can be differentiated into at least three downstream mesenchymal lineages that include bone, cartilage, and fat. How ...

Proteomics aims to identify and classify the proteins present in a particular cell or tissue. However, we know that proteins rarely function alone and knowledge of which proteins interact with which other proteins is vital if we wish to understand how cells work. The budding yeast, Saccharomyc ...

Electron microscopic analysis of biological material requires optimal preparation of the samples. This is necessary to prevent degeneration processes and changes of the material during microscopic observation. Both would lead to artefacts. In this chapter, we present methods to p ...

Flow cytometry has found wide applications in areas like haematology and immunology, but also presents great potential in microbiology. The susceptibility of clinical isolates of Candida and Cryptococcus to antifungal compounds can be assayed by flow cytometry using fluorescent ...

Fungi are capable of undergoing apoptotic-like cell death, and display many of the characteristic features of apoptosis observed in multicellular organisms. These features include nuclear condensation, DNA fragmentation, translocation of phosphatidylserine from the cy ...

Due to the large number of microtubules in the wild-type strain, total internal reflection fluorescence (TIRF) microscopy was used to study cortical microtubule dynamics in leading hyphae of Neurospora crassa expressing β-tubulin-GFP. Detection of plus-end dynamics of individu ...

Fluorescence in situ hybridization (FISH) is a powerful technology for studying eukaryotic chromosomes and genomes from the combined view of cytogenetics and molecular biology, but its use in filamentous fungi has been limited. In this chapter, we describe protocols to perform three ba ...

The cytoskeleton provides the basic architectural organization and shape of the eukaryotic cell, and plays a key role in segregation of the genetic material. A method to visualize the actin and microtubule cytoskeleton in the fungus Ustilago maydis by indirect immunofluorescence is de ...

Critical cellular processes such as DNA replication, DNA damage repair, and transcription are mediated and regulated by DNA-binding proteins. Many efforts have been invested therefore in developing methods that monitor the dynamics of protein-DNA association. As older techniqu ...

As ubiquitous metabolic and signaling intermediaries, kinases regulate innumerable aspects of fungal growth and development. At its simplest, the enzymatic function of a kinase is to transfer a phosphate from a donor molecule (such as adenosine triphosphate) to an acceptor molecule, ...

Biochemical approaches are powerful tools for investigating mechanisms of biological processes. Here, we describe several biochemical approaches that have been successfully in our laboratory to study the filamentous fungus Neurospora crassa. These approaches include pr ...

Digital image analysis has been used to distinguish and quantify leaf color changes arising from a variety of factors. Its use to assess the percentage of leaf area with color differences caused by plant disease symptoms, such as necrosis, chlorosis, or sporulation, can provide a rigorous and qu ...

In host-pathogen interactions, identification of pathogen genes expressed during plant infection poses a challenge, even though these genes may be strongly induced by signals from the host. Here, we describe the application of a PCR-based differential screening method to plant-fun ...