Cell‐Free Translation of Free and Membrane‐Bound Polysomes and Polyadenylated mRNA from Rabbit Brain Following Administration of <i>d</i>‐Lysergic Acid Diethylamide <i>In Vivo</i>

Heikkila, John J.; Cosgrove, James W.; Brown, Ian R.

doi:10.1111/j.1471-4159.1981.tb01722.x

Cited by 30 publications

(14 citation statements)

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“…Free polysomes were prepared as described (11) Ranu and London (12). The lysate was treated with micrococcal nuclease and utilized in a cell-free translation system essentially as described by Pelham 5-,l aliquots were removed and diluted into 1 ml of ice-cold distilled H20.…”

Section: Methodsmentioning

confidence: 99%

Heat shock protein in mammalian brain and other organs after a physiologically relevant increase in body temperature induced by D-lysergic acid diethylamide.

Cosgrove¹,

Brown²

1983

Proc. Natl. Acad. Sci. U.S.A.

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A physiologically relevant increase in body temperature from 39.7 to 42.50C, which was generated after the intravenous injection of D-lysergic acid diethylamide (LSD), caused the induction of synthesis of a 74,000-dalton heat shock protein in the brain, heart, and kidney of the young adult rabbit. A marked increase in the relative labeling of a 74,000-dalton protein was noted after analysis of both in vivo labeled proteins and cell-free translation products of isolated polysomes. A temporal decrease in the synthesis of this protein was noted as LSD-induced hyperthermia subsided. The 74,000-dalton protein, which is induced in various organs of the intact animal at a body temperature similar to that attained during fever reactions, may play a role in homeostatic control mechanisms.Increase of ambient temperature has been reported to induce the synthesis of a specific set of heat shock proteins in a wide range oftissue culture systems and unicellular organisms (1-9). The exact number and molecular weight ofthe induced proteins is dependent on the particular system under investigation; however, the induction of several proteins in the molecular mass range of 68,000-74,000 daltons and 95,000 daltons is common to most systems (1,3,5).Understanding the functional role that these proteins may play in mammalian cells is particularly important given the suggestion raised by Kelley and Schlesinger (3) that these proteins may be involved in the response of the body to fever reactions. An intriguing question that requires examination is whether heat shock proteins are induced within organs of the intact mammal during physiologically relevant increases in body temperature. In this report we demonstrate that an increase in body temperature similar to that attained during fever reactions induces the synthesis ofa 74-kilodalton (kDal) protein in the brain, heart, and kidney of the young adult rabbit. 0.20C prior to treatment were selected for experimentation. Animals that were injected with LSD had rectal temperatures of 42.5 ± 0.30C 1 hr after drug administration. It has been shown (10) that rectal temperature in rabbits is an accurate representation of the body temperature of organs and that rectal temperature and organ temperature increase in parallel during hyperthermia. MATERIALS AND METHODSIn Vivo Labeling of Proteins. Proteins in the cerebral hemisphere of the brain were labeled by injection of 0.5 mCi (1 Ci = 3.7 x 10'°becquerels) of [3S]methionine (New England Nuclear) 20 min after the drug injection. The isotope was administered into the lateral ventricles of the brain through stereotaxically implanted stainless steel cannulae. The animals were pulsed for 1 hr and then dispatched by cervical dislocation. The cerebral hemispheres were dissected out and homogenized in 2 vol of 0.32 M sucrose/50 mM Hepes/KOH, pH 7.5/140 mM potassium acetate/5 mM magnesium acetate/2.5 mM dithiothreitol. The homogenate was centrifuged at 10,000 rpm for 10 min in a Sorvall SS34 rotor at 40C to obtain a postmitochondrial supernatant....

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Section: Methodsmentioning

confidence: 99%

Heat shock protein in mammalian brain and other organs after a physiologically relevant increase in body temperature induced by D-lysergic acid diethylamide.

Cosgrove¹,

Brown²

1983

Proc. Natl. Acad. Sci. U.S.A.

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“…Although cultured cells have been used in most of the studies of mammalian stress proteins, they have been induced in rats by heat (Currie and White, 1981) and arterial ligation (Currie and White, 1981;Hammond et al, 1982), in rabbits by d-lysergic acid diethylamide-stimulated hyperthermia (Heikkila et al, 1981), and in humans following major surgery (Anderson et al, 1982). Therefore, it is likely that stress proteins have roles in responses of animals to trauma.…”

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confidence: 99%

The 73 kilodalton heat shock cognate protein purified from rat brain contains nonesterified palmitic and stearic acids

Guidon

Hightower

1986

Journal Cellular Physiology

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A protein related to the 71 kilodalton inducible rat heat shock protein was purified to electrophoretic homogeneity in milligram amounts from brain tissue of nonheat-stressed rats. The protein has been designated as a stress cognate protein based on previous studies and data presented herein that this protein cross-reacted with a monoclonal antibody originally raised against the Drosophila 70 kilodalton heat shock protein. The purified protein had an apparent molecular mass of 73 kilodaltons when analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis and an apparent mass of 150 kilodaltons as determined by nondissociative gel chromatography, suggesting that the purified protein is a homodimer. The purified protein had isoelectric points of 5.0 under nondissociative conditions and 5.6 when exposed to protein denaturants, suggesting loss of bound anionic molecules and/or net exposure of basic residues upon denaturation. Chloroform/methanol extraction of the purified protein and subsequent analyses by thin layer and gas-liquid chromatography resulted in the identification of palmitic and stearic acids noncovalently bound to the protein. Approximately four molecules of fatty acids were bound per dimer with palmitic and stearic acids present in a one-to-one ratio. The purified protein did not bind exogenously added radioactive palmitate, indicating that the fatty acid-binding sites of the cognate protein were fully occupied and that the associated fatty acids were too tightly bound to exchange readily. The possible significance of the fatty acids associated with the 73 kilodalton stress cognate protein is discussed.

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“…For example, it has been suggested that the synthesis of heat shock proteins is correlated to the phenomenon of acquired thermotolerance (4)(5)(6)(7). The induction of heat shock proteins is relevant to the intact mammal, since increases in body temperature similar to those attained during fever induce synthesis of a heat shock protein of 70-74 kDa in the mammalian brain and other organs (8)(9)(10)(11)(12)(13)(14)(15)(16). Because these proteins may be involved in the response of the body to fever reactions, it is important to increase our understanding of heat shock proteins in intact Mammalian organs.…”

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confidence: 99%

“…Aliquots were lyophilized to dryness and solubilized in 9.5 M urea/2% (vol/vol) Nonidet P-40/0.8% (wt/vol) ampholytes, pH 3-10/5% 2-mercaptoethanol. Isoelectric focusing (first dimension) was performed in 10-mm cyclindrical gels containing 4% acrylamide, 9 M urea and 2% ampholytes, pH 3-10. Cathode and anode buffers were 20 mM NaOH (degassed) and 10 mM H3PO4, respectively.…”

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Axonal transport of a heat shock protein in the rabbit visual system.

Clark¹,

Brown²

1985

Proc. Natl. Acad. Sci. U.S.A.

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Intraocular injection of [35S~methionine wasused to demonstrate the pronounced induction of a 74-kDa heat shock protein in the rabbit retina after a 3YC increase in body temperature was generated by intravenous administration of D-lysergic acid diethylamide. Two-dimensional polyacrylamide gel electrophoresis and fluorography revealed that the induced heat shock protein underwent axonal transport from retinal ganglion cells into the optic nerve and subsequently down the contralateral optic tract to synaptic termini in the visual projection area. Since the heat shock protein took more than 8 days to move down the optic nerve to the superior colliculus, it is transported by slow rather than by fast axonal transport.

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Cell‐Free Translation of Free and Membrane‐Bound Polysomes and Polyadenylated mRNA from Rabbit Brain Following Administration of d‐Lysergic Acid Diethylamide In Vivo

Cited by 30 publications

References 36 publications

Heat shock protein in mammalian brain and other organs after a physiologically relevant increase in body temperature induced by D-lysergic acid diethylamide.

Heat shock protein in mammalian brain and other organs after a physiologically relevant increase in body temperature induced by D-lysergic acid diethylamide.

The 73 kilodalton heat shock cognate protein purified from rat brain contains nonesterified palmitic and stearic acids

Axonal transport of a heat shock protein in the rabbit visual system.

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