Characterization and Location of Met5-Enkephalin-Arg6-Phe7 Stored in Various Rat Brain Regions

A specific antiserum against met⁵-enkepha-lin-arg⁶-phe⁷ was raised and used to study the distribution and characterization of met⁵-enkephalin-arg⁶-phe⁷-like immunoreactive material in rat brains by radioimmunoassay and immunohistochemical procedures. The antiserum appears to be directed to the COOH-terminus of the peptide, as it fails to cross-react with met⁵-enkeph-alin, met³-enkephalin-arg⁶, met⁵-enkephalin-arg⁶-arg⁷, met⁶-enkephalin-lys⁶, and leu-enkephalin. However, it cross-reacts with phe-met-arg-phe by about 10% and with phe-met-arg-phe-NH₂ to an insignificant degree. The highest content of met⁵-enkephalin-arg⁶-phe⁷ was found in the striatum, which contains a dense network of immunoreactive varicose fibers and terminals, as well as immunoreac tive cell bodies. The met⁵-enkephalin-arg⁶-phe⁷ in striatum can be released in a Ca²⁺-dependent manner by a depolarizing concentration of KC1, raising the possibility of a neu-roregulatory role for met⁵-enkephalin-arg⁶-phe⁷. Characterization of the immunoreactive material by gel filtration and high pressure liquid chromatography revealed the presence of multiple forms of immunoreactive material in some brain regions.     

Opioid antinociception and positive reinforcement are mediated by different types of opioid receptors

Fentanyl (FEN) and diprenorphine's (DIPR) potentials for analgesia and reinforcement were assayed using rats. Analgesia was measured by the classic tail-flick test. The test germane to opioid reinforcement involved measuring pressing rates for direct electrical stimulation of the lateral hypothalamus and ventral tegmental area. FEN, as does morphine and heroin, produced strong analgesia and enhanced pressing rates for brain stimulation. DIPR produced no analgesia and antagonized FEN's analgesia. DIPR, at doses antagonizing FEN's analgesia, enhanced pressing for brain stimulation. DIPR's enhancement of pressing was antagonized by naloxone (100 micrograms/kg). When FEN and DIPR were given concurrently, pressing for brain stimulation was not reduced and was greater than after FEN alone was given. These data support a conclusion that different types of receptors are associated with opioid analgesia and reinforcement.     

Muscarinic Receptors Modulate Dopamine-Activated Adenylate Cyclase of Rat Striatum

We investigated the effect of acetylcholine (ACh) on the activation of adenylate cyclase by dopamine (DA) in a lysed synaptosomal preparation from rat striatum. ACh reduced both basal and the DA-activated adenylate cyclase with an apparent IC50 of approximately 1 microM. From a kinetic analysis it appeared that ACh reduced the Vmax for activation by DA but not the activation constant for DA. For most preparations the Vmax was reduced by 30-40%. The presence of atropine did not affect the activation of the enzyme by DA but it blocked the inhibition by ACh. Following 6-hydroxydopamine lesion of the nigrostriatal pathway, the enzyme became supersensitive to activation by DA and also more sensitive to inhibition by ACh. Inhibition of adenylate cyclase by ACh appeared to be rather specific for activation by DA, as ACh had no effect on activation of adenylate cyclase by the adenosine analogue N6-(L-2-phenylisopropyl)adenosine. These results indicate that some striatal muscarinic and dopaminergic receptors are probably coupled to the same adenylate cyclase domain. Moreover, they suggest a biochemical model for the dynamic balance of cholinergic and dopaminergic neurons that innervate the striatum.     

In vitro assessment of the toxicity of metal compounds

A review has been compiled illustrating the directions taken in examining the genotoxic effects of metals and their compounds centering only on those studies pertaining to effects of metals and their compounds on DNA structure and function, such as the induction of DNA strand breaks, production of DNA-protein crosslinks, induction of chromosomal aberrations, and sister chromatid exchanges. Although it is premature to declare a cause and effect relationship between the carcinogenic activity of metals and their ability to induce one or more lesions in DNA, strong evidence is emerging to suggest such a relationship. Low concentrations of metals induce the appearance of DNA lesions, such as strand breaks and crosslinks, or induce sister chromatid exchanges or DNA repair synthesis. Assays based upon these events constitute extremely sensitive probes for genotoxic effects of metals and their compounds. These effects of metals on DNA are consistent with the currently accepted mechanism of chemical carcinogenesis, allowing the acquisition and propagation of altered DNA function. The lack of complete information on the activity of metals in producing DNA lesions allow only preliminary conclusions to be drawn. Certain compounds containing potentially or actually carcinogenic elements, such as Ni, Be, As, Cr, Cd, and to a minor extent Pb, have yielded positive responses in one or more DNA lesion assays. At relatively nontoxic levels of Ni and Cr, considerable evidence suggests that multiple types of DNA lesions are induced.     

PROTEIN METABOLISM AND AMINO ACID ACCUMULATION IN THE RAT SUBMAXILLARY GLAND DURING REDUCED SYMPATHETIC ACTIVITY

Abstract— Unilateral sympathetic decentralization of the superior cervical ganglion of rats was performed 3 days prior to the experiments. A two-compartment kinetic model was proposed to describe the effect of decentralization on (1) the uptake of a nonphysiological amino acid from plasma to the submaxillary gland and (2) the incorporation of a physiological amino acid from precursor pool into protein. The calculations based on the model showed that the fractional rate constant for efflux of the nonphysiological amino acid, α-[3-¹⁴C] aminoisobutyric acid, was greater in the decentralized than in the normal gland. However, efflux rate was equal in the two glands because the extrapolated zero time value of the initial concentration was greater in the normal gland.
The labelled physiological amino acid, [¹⁴C]leucine, was used in initial experiments to assess turnover rate of the gland proteins but it was rapidly metabolized to many other radioactive compounds. Therefore, arginine[¹⁴C]guanido was employed-arginine being the only labelled amino acid found after injection. Since the steady state content of submaxillary gland proteins was not changed but the fractional rate constant of conversion of free arginine into protein (k_p) was greater in the decentralized gland (k_p= 0-40 h^_l) than in the normal (k_p= 0-27 h⁻¹), we can conclude that decentralization increases protein turnover rate; thus, assuming that arginine[¹⁴C]guanido is homogeneously distributed in the tissue pools of free arginine, the rate of protein turnover is greater in the sympathetically decentralized gland than in the normal.     

The relationship between the size of yellow water traps and catches of aphids

Yellow water traps are often used for sampling populations of flying aphids. This note suggests that the size and probably the shape of traps should be standardised, because trapping efficiency (nos. caught/unit area), and the relative attractiveness of traps to different species, depends on trap size. Aphidologists using water traps to compare mixed populations of flying aphids, should therefore compare catches from different traps with caution.Traps of three sizes were made up from individual trays, each 29×21.5×5cm, and painted Hansa yellow inside; 1 tray alone, 4 arranged in a 2×2 rectangle, and 9 in a 3×3 rectangle, gave trapping surfaces of approx. 625, 2500 and 5700 cm² respectively. The traps were half-filled with water plus a drop of detergent, placed on bare land and daily catches collected on 19 days in June and July 1966.