Mammalian expression constructs based on the Rous sarcoma virus LTR in high yield plasmids.
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Avian Sarcoma Viruses↗
Biomedical subjects
T Kamalati
Publications and source records attributed to T Kamalati.
Heterogeneity in spinal radial glia demonstrated by intermediate filament expression and HRP labelling.
Considerable evidence indicates that radial glial cells play an active role in guiding growing neurites during development of the vertebrate CNS. In this paper we describe subpopulations of radial glial in the spinal cord of the axolotl. Amphibians maintain radial glia throughout life, and subpopulations are described using anatomical criteria following filling of individual cells with horseradish peroxidase and immunocytochemical staining with a range of intermediate filament antibodies. Radial glial cells in specific regions of the spinal cord stain with a range of antibodies specific to human keratins 8 and 18, and to glial fibrillary acid protein (GFAP). Some of these antibodies show selective localized to specific regions of individual glial cell processes. Immunoblotting analysis indicates that two keratins are present in the axolotl CNS corresponding to the two earliest embryonic keratins of vertebrates, keratin 8 and 18. Comparisons of molecular weight indicate that these may correspond to keratins identified in Xenopus laevis, the genes of which have been cloned. Axolotl GFAP is also identified in Western blots and may be present in two forms of differing molecular weight. These results are discussed in terms of the likely role of radial glial cells, and comparisons are drawn between the keratin and GFAP types seen in the axolotl spinal cord and of those in other vertebrate groups.
Ambystoma↗
Expression of markers of differentiation in a transformed human keratinocyte line induced by coculture with a fibroblast line.
SVK14, an SV40-transformed human keratinocyte line, has previously been reported to be almost completely unable to differentiate, and indeed, to express a set of keratins characteristic of simple epithelia rather than the stratifying epithelium from which they were derived. We have recently shown that IGF I stimulation of SVK14 results in expression of keratin 14, a marker of stratifying epithelia, as well as expression of markers which are characteristic of differentiation in normal human keratinocytes such as involucrin and keratin 10. To study further the capacity of SVK14 to differentiate, we have cocultured SVK14 with a variety of fibroblastic cell lines with a view to examining whether the cocultured partner can promote or interfere with their differentiation. We have observed that SVK14, when cocultured with Swiss 3T3, form organized structures through specific cell-cell interactions in which SVK14 express keratins 14 and 5 and involucrin, while maintaining T-antigen expression. These results are interesting since they show coculturing of a transformed human keratinocyte cell line and a particular fibroblast line can result in induction of characteristics of stratifying epithelia in a cell line with characteristics of simple epithelia. This may be analogous to the epithelial-mesenchymal interactions seen during epithelial development in the very early embryo.
Animals↗
IGF I induces differentiation in a transformed human keratinocyte line.
A comparison of normal epithelial cells with their transformed counterparts could lead to the definition of parameters related to growth and differentiation which are altered by viral transformation and which may be relevant to malignant changes in vivo. Using the SV40-transformed human keratinocyte line, SVK14, which exhibits characteristics of simple, nonkeratinizing epithelia, we have shown that IGF I stimulation of these cells results in extensive multilayering, increased cell size, accumulation of involucrin, modulation of keratin 18 and expression of keratins 14 and 10, whilst T-antigen expression is maintained in the multilayered cells. Since T-antigen expression is correlated directly with impairment of stratification and differentiation, it is interesting that treatment of SVK14 with a single growth factor. IGF I, results in molecular events characteristic of differentiating normal keratinocytes.
Antigens, Viral, Tumor↗