Mordanting Plant Tissues

Selected current ideas on the mordanting of plant tissues are presented in the hope that they will be of practical assistance, especially to the beginner. The nature of the mordanting process and the results which may be expected following mordanting are briefly described. Specific technics are presented for the mordanting of natural dyes, synthetic dyes generally, the basic synthetic dyes and the acid synthetic dyes.     

INTERACTION OF IONIZING AND VISIBLE RADIATION IN MUTATION INDUCTION IN NEUROSPORA CRASSA

Klein , Richard M. (New York Bot. Gdn., New York, N.Y.), and Deana T. Klein. Interaction of ionizing and visible radiation in mutation induction in Neurospora crassa. Amer. Jour. Bot. 49(8): 870–874. 1962.—Conidia of the purple adenineless strain of N. crassa were irradiated with 25 kr of X rays and then exposed to far-red or red radiations or to far-red followed by red radiation. Far-red light, without effect on un-irradiated conidia, augmented the genetic damage caused by X rays as measured by survival (colony count), back mutation to adenine prototrophy, and the induction of mutants affecting colony morphology. Post-X-irradiation with red light ameliorated the severity of X-radiation as measured by survival and back mutation. The potentiation of X-ray-induced genetic damage by far-red light could be completely negated by subsequent exposure to red light.     

Phytochrome controlled acid RNase: An “attached” protein of ribosomes

The light-dependent increment in RNase activity (which is ribosome bound in cell extracts) is distributed as a gradient increasing from base to hook of lupin hypocotyls. No evidence was found of non-specific or of specific activation of pre-formed enzyme molecules following isolation, either before or after (latent activity) destruction of particles. The autodegradation capacity of ribosomes isolated from irradiated cells was almost double that of ribosomes from etiolated tissue. It is proposed that association between the bulk of the light-controlled RNase fraction and lupin ribosomes results from binding of soluble protein. It is not clear whether binding is specific or an artifact of isolation.     

Phosphoproteome Profiling of the Receptor Tyrosine Kinase MuSK Identifies Tyrosine Phosphorylation of Rab GTPases

Muscle-specific receptor tyrosine kinase (MuSK) agonist antibodies were developed 2 decades ago to explore the benefits of receptor activation at the neuromuscular junction. Unlike agrin, the endogenous agonist of MuSK, agonist antibodies function independently of its coreceptor low-density lipoprotein receptor–related protein 4 to delay the onset of muscle denervation in mouse models of ALS. Here, we performed dose–response and time-course experiments on myotubes to systematically compare site-specific phosphorylation downstream of each agonist. Remarkably, both agonists elicited similar intracellular responses at known and newly identified MuSK signaling components. Among these was inducible tyrosine phosphorylation of multiple Rab GTPases that was blocked by MuSK inhibition. Importantly, mutation of this site in Rab10 disrupts association with its effector proteins, molecule interacting with CasL 1/3. Together, these data provide in-depth characterization of MuSK signaling, describe two novel MuSK inhibitors, and expose phosphorylation of Rab GTPases downstream of receptor tyrosine kinase activation in myotubes.     

Improved Stability of Methanolic Wright's Stain with Additive Reagents

Additive reagents have been investigated to improve the stability of methanolic Wright's stain. The addition of ammonium halides, monoalkyiamine hydrochlorides, dialkylamine hydrochlorides or trialkylamine hydrochlorides to methanolic Wright's stain was found to enhance the stability of stain components in methanol. No change in performance is observed with these additives present. Random precipitation in the stain solution was still observed with the addition of ammonium halides and monoalkyiamine hydrochlorides. No precipitation was found in stain solutions containing hydrochlorides of most dialkylamines and trialkylamines. Of the compounds evaluated, 0.6% diethylamine hydrochloride added to methanolic stain solutions produced the most desirable overall results. Mechanisms of stabilization and precipitation in these stain solutions are proposed, Essentially, separation of the thiazine-eosinate ion pair through interaction with an appropriate additive increases stain stability. The solubilities of thiazine-eosinate or additive cation-eosinate ion pairs in methanol determine the formation of precipitate in such stain solutions.