ABSTRACT
Baculoviruses serve as a stress factor that can activate both death-inducing and cytoprotective pathways in infected cells. In this report, induction of heat shock proteins (HSPs) of the 70-kDa family (HSP/HSC70) in Sf-9 cells after infection with AcMNPV was monitored by Western blot analysis. Two-dimensional electrophoresis in polyacrylamide gel revealed changes in the cellular pattern of HSP/HSC70s and synthesis of a new member of the HSP/HSC70 family in the infected cells. Although infection with AcMNPV moderately increased the HSP/HSC70 content in cells under standard conditions, the infection potentiated the response to heat shock boosting the HSP/HSC70s content in infected cells several-fold in comparison with uninfected cells. Addition of KNK437, a known inhibitor of inducible HSPs, decreased the rate of viral DNA synthesis in infected cells more than one order of magnitude and markedly suppressed the release of budded viruses indicating the importance of the heat shock response for baculovirus replication.
Subject(s)
Heat-Shock Response , Nucleopolyhedroviruses/physiology , Spodoptera/physiology , Spodoptera/virology , Animals , HSP72 Heat-Shock Proteins/genetics , HSP72 Heat-Shock Proteins/metabolism , Insect Proteins/genetics , Insect Proteins/metabolism , Nucleopolyhedroviruses/genetics , Spodoptera/genetics , Virus ReplicationABSTRACT
The expression pattern and distribution of proteasome immune subunits LMP7 and LMP2 in the developing rat spleen and liver as well as the periarterial lymphoid sheath formation were investigated. LMP7 and LMP2 were detected by immunoblotting in the spleen on the 21st embryonic day and during the first postnatal days in equal amounts. Their levels increased by the 8th and 18th postnatal days. Double immunofluorescent labeling the spleen cells revealed LMP7 and LMP2 in T and B lymphocytes localized in the red pulp in embryogenesis. Few T lymphocytes were discovered in periarterial zones on the 8th postnatal day. T lymphocytes filled these zones and formed lymphoid sheaths by the 18-19th day. In the liver, LMP7 and LMP2 were revealed by the 17-19th postnatal day. Immunofluorescent analysis showed their presence in hepatocytes at this period. The data suggest that T cell-mediated immune response in relation to hepatocytes is possible beginning from 18th to 19th postnatal day.
Subject(s)
Cysteine Endopeptidases/immunology , Cysteine Endopeptidases/metabolism , Lymphoid Tissue/embryology , Lymphoid Tissue/immunology , Multienzyme Complexes/immunology , Multienzyme Complexes/metabolism , T-Lymphocytes/enzymology , T-Lymphocytes/immunology , Animals , Embryonic Development/immunology , Female , Immunity, Cellular , Liver/embryology , Liver/enzymology , Liver/growth & development , Liver/immunology , Lymphoid Tissue/enzymology , Lymphoid Tissue/growth & development , Pregnancy , Proteasome Endopeptidase Complex , Rats , Rats, Wistar , Spleen/embryology , Spleen/enzymology , Spleen/growth & development , Spleen/immunology , Tissue DistributionABSTRACT
This study aimed to analyze the aqueous humor (AH) and the vitreous body (VB) of the eye of the adult frog Rana temporaria L. as a representative species of amphibians, which lead a semi-terrestrial life. The presence of collagen, albumin, uric acid and electron donors was shown in both media; however, there are slight differences in their concentrations. To determine collagen, a spectral-fluorescent probe, cyanine dye, was used. The presence of collagen in AH of the frog was found at the first time. The total content of electron donors (ascorbic and uric acids, tryptophan, and tyrosine) in VB and HA was roughly estimated at approximately 1.5x10(-4) mol/L. Both VB and AH absorb light in similar UV regions. The total protein and albumin contents in AH were found to be somewhat higher than those in VB. The uric acid content was at an equally low level in both intraocular media. It is supposed that the similarity of VB and AH compositions shown in this work is due to some exchange between VB and AH contents in the course of accommodation. The role of intraocular fluids in physiological functions of the eye and in protecting the retina against UV light is discussed.
Subject(s)
Aqueous Humor/metabolism , Rana temporaria/metabolism , Vitreous Body/metabolism , Albumins/metabolism , Animals , Collagen/metabolism , Electron Transport , Eye Proteins/metabolism , Fluorescent Dyes , Uric Acid/metabolismABSTRACT
The aim of this work was to develop a quick method for analysis of macromolecules of the extracellular matrix. Of great interest are soluble components of the extracellular matrix, in particular, carrier proteins, whose variation dynamics can characterize the studied tissue in its development, adult stage, and aging. We suggest the method of analysis of the extracellular matrix to reveal the presence of albumin and collagen by using an anionic cyanine dye as a spectral and fluorescence probe. The method was applied for the analysis of the human vitreous body in the course of its development. Albumin was detected by the appearance of the trans monomer absorption and fluorescence bands in the dye spectra, and collagen was detected by the absorption and fluorescence bands of J aggregates. Hyaluronic acid present in the vitreous body does not interfere with the results of the analysis. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blot analysis confirmed the presence of albumin in the vitreous body. We suppose that albumin as a protein carrying biologically active macromolecules plays an important role in the processes of differentiation and functional establishment of ocular tissues in the course of their prenatal development.
