The visual pigment and visual cycle of the lobster,Homarus

Summary The visual pigment of the American lobster,Homarus americanus, has been studied in individual isolated rhabdoms by microspectrophotometry. Lobster rhodopsin has _max at 515 nm and is converted by light to a stable metarhodopsin with _max at 490 nm. These figures are in good agreement with corresponding values obtained by Wald and Hubbard (1957) in digitonin extracts. Photoregeneration of rhodopsin to metarhodopsin is also observed. The absorbance spectrum of lobster metarhodopsin is invariant with pH in the range 5.4–9, indicating that even after isomerization of the chromophore fromcis totrans, the binding site of the chromophore remains sequestered from the solvent environment. Total axial density of the lobster rhabdom to unpolarized light is about 0.7.As described for several other Crustacea, aldehyde fixation renders the metarhodopsin susceptible to photobleaching, a process that is faster at alkaline than at neutral or acid pH. Small amounts of a photoproduct with _max at 370 nm are occasionally seen. A slower dark bleaching of lobster rhabdoms (_1/2–2 h) also occurs, frequently through intermediates with absorption similar to metarhodopsin.The molar extinction coefficient of metarhodopsin is about 1.2 times greater than that of rhodopsin, each measured at their respective _max. Isomerization of the chromophore fromcis totrans is accompanied by a change in the orientation of the absorption vector of about 3°. The absorption vector of metarhodopsin is either tilted more steeply into the membrane or is less tightly oriented with respect to the microvillar axes.When living lobsters are kept at room temperature, light adaptation does not result in an accumulation of metarhodopsin. At 4 °C, however, the same adapting lights cause a reduction of rhodopsin and an increase in metarhodopsin. There is thus a temperature-sensitive regeneration mechanism that supplements photoregeneration. Following 1 ms, 0.1 joule xenon flashes that convert about 70% of the rhodopsin to metarhodopsin in vivo, dark regeneration occurs in the living eye with half-times of about 25 and 55 min at 22 °C and 15 °C respectively.This work was supported by USPHS research grant EY 00222 to Yale University. S.N.B. was aided by NIH Postdoctoral Fellowship EY 52378.     

Dark adaptation,sensitivity, and rhodopsin level in the eye of the lobster,Homarus

Summary Dark adaptation of living lobsters was measured by recording the ERG at several temperatures in the range 5–20 °C following adapting flashes that convert about 70% of the rhodopsin to metarhodopsin. Recovery of log threshold is rapid, and at 10–20° is nearly complete in 10 min. Only at 5 °C is dark adaptation significantly slowed. Comparison of dark adaptation with data on regeneration of pigment (Bruno et al., 1977) is consistent with the hypothesis that as rhodopsin concentration rises and falls, its only effect on sensitivity is to alter the probability of quantum catch. This interpretation is further bolstered by observations on winter lobsters that have a 70% deficiency of rhodopsin without the concomitant increase in metarhodopsin that accompanies light adaptation. No effect of metarhodopsin on sensitivity was detected. These experiments support the growing body of evidence indicating that the relationship between rhodopsin concentration and log threshold is fundamentally different in the rhabdomeric photoreceptors of invertebrates and the rods and cones of vertebrates.This work was supported by USPHS research grant EY 00222 to Yale University. S.N.B. was aided by NIH Postdoctoral Fellowship EY 52378, by funds made available through the Unidel Foundation, and by a grant from the University of Delaware Research Foundation.     

Identification of protein X of Escherichia coli as the recA+/tif+ gene product.

Summary Protein X, molecular weight 40,000, has been separated from the other proteins of E. coliby a two-dimensional gel electrophoretic technique which separates proteins according to isoelectric point (pI) in the firstdimension and according to molecular weight in the second. When protein X is induced in wild-type cells by mitomycin C treatmentit has a pI6.0. However, when protein X is induced in a tif-1 mutant, either by temperatureshift-up to 42° or by mitomycin C treatment at 30°, it has a pI6.2. The low level of protein X which is present inuninduced tif mutants at 30° also has a pI6.2. These results suggest thattif-1 is a mis-sense mutation in the gene coding for protein X. Since transduction andcomplementation studies indicate that tif-1 is a mutation of therecA ⁺ gene (Castellazzi, Morand, George and Buttin, 1977) it follows that protein X is the recA ⁺ gene product.A model has been formulated to account for the relationship between protein X synthesis and the recA ⁺ and lexA ⁺ genes. In this model, a repressor coded by lexA ⁺ binds to the operator of the recA ⁺ gene from whence it can normally only be removed by the combined action of an inducer and protein X, the recA ⁺ product. Thus, protein X controls its own synthesis. The tif-1 mutation leads to a temperature sensitive form of protein X which, at 42°, can spontaneously remove the repressor without the intervention of the inducer.     

Discriminative effects of morphine administered intracerabrally in the rat

Rats were trained in a two-choice discrete trial avoidance paradigm to discriminate between saline and 3.0 mg/kg of morphine administered S.C. The microinjection of 0.3–3.0 μg of morphine into the lateral ventricle produced discriminative effects equivalent to those of the systemic training dose as measured by responding on the morphine-appropriate choice lever. Discriminative effects equivalent to those of the morphine training dose were not consistently produced by administration of morphine into the periaqueductal gray, lateral septum or dorsomedial thalamus in doses as high as 10 μg. However, the discriminative effects of systematically administered morphine were blocked by 10–30 μg of naloxone administered intracerebrally at all of the brain sites tested. Thus, the primary site at which morphine acts to produce discriminative effects in the rat is central, although the specific brain areas mediating these effects remain unidentified. The actions of naloxone could be the result of diffusion of the drug into the ventricular system or into the systemic circulation.     

Chemical induction of β-carotene biosynthesis

Marsh white seedless grapefruit were treated with the 2-diethylaminoethanol esters of the following acids: benzoic, phenylacetic, hydrocinnamic, 4-phenylbutyric, 5-phenylvaleric, valeric, hexanoic, heptanoic, octanoic, nonanoic, 5-chlorovaleric, cyclohexanecarboxylic, phenoxyacetic, p-chlorophenoxyacetic, 3-phenoxypropionic, cinnamic and p-chlorocinnamic. Several of these esters, in particular the hexanoate, 4-phenylbutyrate and cinnamate, caused the accumulation of large amounts of β-carotene. The effects of the hexanoate and of 2-phenoxytriethylamine, which causes only lycopene accumulation, were studied as functions of time. The hexanoate caused the rapid accumulation of lycopene during the first day. The amount of lycopene then began to decrease and that of β-carotene increased until, after 14 days, β-carotene was the major pigment. 2-Phenoxytriethylamine caused rapid lycopene accumulation during the first day and a slow steady increase afterwards. Thus, the mode of action of the β-carotene inducers may be similar to that of the lycopene inducers except that the former are probably rapidly hydrolysed by the esterase(s) in the flavedo, so that they no longer inhibit the cyclase(s), and β-carotene is accumulated at the expanse of lycopene.     

Effects of insecticides on acetylene reduction by Azotobacter vinelandii and soybean nodules

Summary Nine organophosphate and carbamate insecticides were tested for effects on ability of Azotobacter vinelandii to reduce acetylene. Only Gardona^R, at higher concentrations, was significantly inhibitory. The same pesticides were tested with soybeans (Glycine max L.). Some minor phytotoxic effects were noted, but there was no inhibition of the ability of the excised nodules of the plants to reduce acetylene.Published with the approval of the Director of the North Dakota Agricultural Experiment Station as Journal Article No. 726. Portion of a thesis presented by the senior author in partial fulfillment of the requirements for the M.S. degree in bacteriology at North Dakota State University.Published with the approval of the Director of the North Dakota Agricultural Experiment Station as Journal Article No. 726. Portion of a thesis presented by the senior author in partial fulfillment of the requirements for the M.S. degree in bacteriology at North Dakota State University.     

C-13 nuclear magnetic resonance spectroscopy of humic substances

Summary Evidence is presented from a preliminary study showing that C-13 N.M.R. spectroscopy is a useful new tool for the elucidation of the structure of soil organic matter.     

Inhibition of rosette formation by antithymocyte globulin

Summary The peripheral blood leukocytes from 29 patients with adenocarcinoma of the colon were studied sequentially for the T-cell level, the rosette-inhibition titer of antithymocyte globulin and the blastogenic response to PHA and Con A to evaluate the T-cell immunocompetence. The level of E-rosette-forming cells and the blastogenic response did not reflect the immunocompetence of the T-cell population. Fluctuations in the rosette-inhibition titer of antithymocyte globulin (ATG) were observed; an increased ATG titer indicated an unfavorable clinical course, while a decreased ATG titer was observed with patients who had a favorable clinical course. The rosette-inhibition titer with antithymocyte globulin was observed to be an important indicator of competent T cells in patients with colorectal cancer.Supported by the Physicians' Medical Education and Research Foundation and General Research Supports FundsAbbreviations used that are not spelled out in the text are: AET 2-aminoethylisothiouronium bromide; ATG Antithymocyte globulin; ALG Antilymphocyte globulin; E-RFC Erythrocyte rosette-forming cells; PHA Phytohemagglutinin; Con A Concanavalin A; E Sheep erythrocyte