Subject(s)
HSP70 Heat-Shock Proteins/metabolism , Sleep/physiology , Animals , Body Temperature , Electroencephalography , Electromyography , Electrooculography , Genetic Vectors , HSP70 Heat-Shock Proteins/genetics , Immunohistochemistry , Lentivirus/genetics , Male , Microscopy, Confocal , Monitoring, Ambulatory , Motor Cortex/metabolism , Preoptic Area/metabolism , RNA, Small Interfering , Rats, Wistar , Telemetry , Time Factors , WakefulnessABSTRACT
For the first time, the biodistribution of recombinant heat shock protein in rhHsp70 rats with grafted intracranial C6 glioma was evaluated. It was assessed using the fluorescent antibody accumulation chaperone rhHsp70 conjugated with fluorochrome Alexa Fluor 555 in tumor cells by intratumoral or intravenous administration. Assessment of the distribution and accumulation of labeled protein was carried out on the model of subcutaneous B16/F10 melanoma in C57BL/6 mice with the use of single-photon emission computer tomography. After 60 minutes after intravenous administration rhHsp70-I123 (20 MBq, 5 mg chaperone) accumulation of the drug mainly in the liver and tumor tissue was showed. The coefficient of the differential accumulation of the labeled protein KDN(tumor/background) was 3.14. It was turned out that comparing the level of fixation of rhHsp70-I123 in the liver and the tumor KDN(tumor/ liver) = 0.76. After 24 hours from the time of injection of rhHsp70-I123 it was observed increase the level of fixation of the labeled protein in the liver and melanoma: KDN(tumor/background) = 3.43; KDN(tumor/liver = 0.78.
Subject(s)
Brain Neoplasms/metabolism , Glioma/metabolism , HSP70 Heat-Shock Proteins/metabolism , Liver/metabolism , Melanoma, Experimental/metabolism , Skin Neoplasms/metabolism , Animals , Fluorescent Dyes , HSP70 Heat-Shock Proteins/administration & dosage , HSP70 Heat-Shock Proteins/pharmacokinetics , Injections, Intralesional , Injections, Intravenous , Iodine Radioisotopes , Male , Mice , Mice, Inbred C57BL , Neoplasms, Experimental/metabolism , Rats , Rats, Wistar , Time Factors , Tissue Distribution , Tomography, Emission-Computed, Single-PhotonABSTRACT
Data obtained for the last 12 years and modern hypotheses on key function of sleep and the role of Heat Shock Protein 70 kDa (HSP70) molecular chaperones family in sleep modulation are insufficient to determine assotiation of sleep quantity to the level of chaperones in the basic "center" of sleep in the ventrolateral preoptic area (VLPA) of the hypothalamus. In the present study, to reduce the content of Hdj1 major co-chaperone of Hsp70 in the VLPA we employed a novel approach based on lentiviral construction containing specific Hdj1-shRNA. The immunoblotting data showed that in 6 weeks after infection the level of Hdj1 in VLPA was reduced by 80% that was accompanied by a considerable increase in the quantity of slow-wave sleep and a marked decrease in the level of anxiety; earlier we found that elevation of Hsp70 level in the rat brain resulted in similar changes. It is suggested that the increase in quantity of slow wave sleep and the decrease in the level of anxiety can be related to a sustained disorder in the integration between molecular systems based on chaperones Hdj1 and Hsp70 and to a compensatory increase in the Hsp70 chaperone activity/level in VLPA.
