The Infona portal uses cookies, i.e. strings of text saved by a browser on the user's device. The portal can access those files and use them to remember the user's data, such as their chosen settings (screen view, interface language, etc.), or their login data. By using the Infona portal the user accepts automatic saving and using this information for portal operation purposes. More information on the subject can be found in the Privacy Policy and Terms of Service. By closing this window the user confirms that they have read the information on cookie usage, and they accept the privacy policy and the way cookies are used by the portal. You can change the cookie settings in your browser.
DNA-damaging agents typically react with DNA to form a plethora of adducts within the genome. Historically, it has been difficult to ascertain the genetic effects of individual DNA adducts by studying randomly modified DNAs. Over the past decade techniques that allow the precise evaluation of the biological effects of single DNA adducts have been developed. This paper describes the methodology required...
We describe here the methodology developed in our laboratory to study the frequency and repair of cyclobutane pyrimidine dimers induced by ultraviolet light (254 nm) in the individual strands of specific DNA sequences. We have used this technique to establish the direct correlation between transcription by RNA polymerase II and the preferential repair of the transcribed strand of an active gene in...
The ability of mismatch binding proteins, in particular the Escherichia coli MutS protein, to bind DNA containing mismatched base pairs or one to four unpaired bases makes them ideally suited to use in mutation/polymorphism detection assays. Several methods, including nuclease protection, band shift, and filter binding assays, have been employed with varying degrees of success. New assays, including...
Excision repair of DNA is a major cellular mechanism to correct DNA lesions produced by a variety of physical and chemical agents. The availability of a cell-free system is expected to provide a powerful tool with which to study the complex biochemical pathways of excision repair in eukaryotes. Here we describe such a cell-free system in the yeast Saccharomyces cerevisiae that supports both nucleotide...
Nucleotide excision repair of damaged DNA can be carried out in a cell-free environment, making the process amenable to biochemical study and manipulation. In this article, methods for a versatile repair synthesis assay are described. Protocols are given for preparing plasmid DNA substrates, preparing whole cell extracts, performing repair synthesis reactions, and analyzing the data obtained. Methods...
Clonal neuronal cell lines of rat hippocampal origin have been developed by two alternative strategies using retroviral transduction of a temperature-sensitive simian virus 40 large tumor antigen (T ts ). Initially, T ts was introduced into dividing E17 rat embryonal hippocampal cells. In method 1, cell lines were generated directly from these progenitor cells by shifting...
The availability of in vitro cell culture systems demonstrating differentiated, mature phenotypes is crucial for the understanding of cellular functions and their regulation. Complex, heterogeneous systems such as the central nervous system had been refractory to this type of analysis. Recently, however, technical advances in molecular biology and a better understanding of the spectrum of growth factors...
The generation of cell lines in the sympathoadrenal lineage has greatly facilitated our understanding of how precursor cells that do not respond to NGF give rise to mature NGF-dependent neurons. The neuronal developmental pathway in this lineage has been worked out by studying both primary precursor cells in culture and the v-myc-immortalized MAH cell line. MAH cells were established by retroviral...
P19 is a pluripotent stem cell line which can be induced to differentiate along either mesodermal or ectodermal lineages. When treated with retinoic acid, P19 cultures differentiate into cell types similar to those derived from neuroectoderm. Lineage-specific markers have shown neurons and astroglia to predominate in these cultures. By manipulating the growth conditions, differentiated cultures can...
The PC12 cell line is one of the most widely used of all neuronal cell lines. Over 2200 studies using PC12 cells have been published since the first in 1974. It is so widely used because it possesses properties that make it applicable to a wide variety of neurobiological problems. This article is a brief overview of some neurobiological issues that have been approached in recent years using the PC12...
The neuroendocrine hypothalamus has been the object of intensive study in vivo and in tissue slices. However, using these models it is difficult to approach questions at the molecular and cellular level and to differentiate between direct effects and those mediated by other neurons. By using the regulatory domain of the rat gonadotropin-releasing hormone (GnRH) gene to target expression of the oncogene...
Thyroid C-cell lines, such as the CA77, 44-2C, and TT lines, are derived from calcitonin-secreting endocrine cells of neural crest origin. An intriguing property of C-cells is that they possess neuronal properties. These include neurite extension and expression of neuronal markers including neurofilaments, the neuropeptide calcitonin gene-related peptide (CGRP), and Ca 2+ and Na +...
Two neuronal cell lines, RN33B and RN46A, were derived from E12 rat medullary raphe neuroepithelial cells by infection with a retrovirus encoding the temperature-sensitive mutant of SV40 large T antigen (T-ag). At permissive temperature (33 o C), both cell lines express nuclear T-ag immunoreactivity, low levels of neuron-specific enolase (NSE), and low (NF-L) and medium (NFM), but not high...
The ND series of cell lines was isolated by fusing a proliferating neuroblastoma with primary dorsal root ganglion (DRG) neurons and selecting for the drug resistance of the DRG cells. These cell lines retain a variety of sensory neuron properties that are not found in the parental neuroblastoma and, in addition, one of these lines (ND7) can be induced to differentiate to a nondividing phenotype bearing...
The B-cell compartment is composed of a complex mixture of distinct subsets. Three anatomic sites that are particularly rich in B-cell subpopulations are the bone marrow, spleen, and peritoneal cavity. Our laboratory has found CD23, the low-affinity lgE Fc receptor (FcϵRII), to be of value in discriminating a number of murine B-cell subsets, especially when used in combination with other B-cell markers...
The B-cell immune system is functionally compartmentalized into B1- and B2-cell subsets. Both developmental and Ag selection processes collaborate in the establishment of these populations. The earliest-detected hematopoletic progenitors in the mouse embryo (8.5 d.p.c. paraaortic splanchnopleura; 12-13 d.p.c. omentum) preferentially undergo differentiation processes leading to B1 lymphocytes. This...
B1 cells are a subset of B lymphocytes, found in many species, that differ from conventional B cells (B2) in phenotype and function. B1 cells produce large amounts of low-affinity IgM, appear to be self-renewing, respond primarily to T-independent antigens, and, in the peritoneal cavity, do not generate lymphoid follicles. While B2 lymphocyte-T lymphocyte interactions result in proliferation, differentiation,...
Set the date range to filter the displayed results. You can set a starting date, ending date or both. You can enter the dates manually or choose them from the calendar.