Serotonin Levels in the Rabbit Retina Are Elevated Following Intraocular Injection of Forskolin

Analysis of the mammalian retina for serotonin immunoreactivity suggests an absence of the amine. However, following an intraocular injection of forskolin (1 microM) into a rabbit eye 1 h before analysis of the retina, serotonin immunoreactivity is associated with a subpopulation of amacrine cells. These cells correspond in size and position to the "indoleamine-accumulating cells" of the retina. Biochemical experiments show that forskolin treatment produces an increase in levels of endogenous serotonin and 5-hydroxytryptophan but has no effect on the uptake of serotonin or tryptophan or the metabolism of 5-hydroxytryptophan. These results suggest that the "indoleamine-accumulating cells" in the retina are "serotonergic cells" and that the level of amine is elevated sufficiently for localisation following forskolin treatment. It would appear that forskolin either directly or indirectly activates tryptophan hydroxylase.     

Mutation-Selection Balance at a Modifier-of-Imprinting Locus

We propose a pair of population genetic models for a modifier-of-imprinting locus for which different genotypes imprint different proportions of an imprintable target locus in their gametes. The two models examine the situations in which imprinting is advantageous or disadvantageous, and we discuss three cases for which the modifier is respectively partially dominant, dominant, or recessive. The models predict the stable equilibrium frequencies of the mutant modifier and functionally diploid individuals in a large population in terms of up to four parameters: the mutation rate at the modifier locus, V; the selection coefficient against the disadvantageous phenotype, s; the proportion of unimprinted eggs produced by homozygotes for the mutant modifier, θ, and, in the partially dominant models, the dominance parameter, k. The equilibrium frequency of the mutant phenotypes is shown to be approximately twice that of standard Mendelian models: 2V/s or 4V/s when the modifier is recessive or dominant, respectively. Mathematical equivalences between these and nonimprinting models are noted.     

The scl gene product is required for the generation of all hematopoietic lineages in the adult mouse.

Homozygosity for a null mutation in the scl gene causes mid-gestational embryonic lethality in the mouse due to failure of development of primitive hematopoiesis. Whilst this observation established the role of the scl gene product in primitive hematopoiesis, the death of the scl null embryos precluded analysis of the role of scl in later hematopoietic development. To address this question, we created embryonic stem cell lines with a homozygous null mutation of the scl gene (scl-/-) and used these lines to derive chimeric mice. Analysis of the chimeric mice demonstrates that the scl-/- embryonic stem cells make a substantial contribution to all non-hematopoietic tissues but do not contribute to any hematopoietic lineage. These observations reveal a crucial role for the scl gene product in definitive hematopoiesis. In addition, in vitro differentiation assays with scl-/- embryonic stem cells showed that the scl gene product was also required for formation of hematopoietic cells in this system.     

The development of phloem anastomoses between vascular bundles and their role in xylem regeneration after wounding in Cucurbita and Dahlia

The differentiation of phloem anastomoses linking the longitudinal vascular bundles has been studied in stem internodes of Cucurbita maxima Duchesne, C. pepo L. and Dahlia pinnata Cav. These anastomoses comprise naturally occurring regenerative sieve tubes which redifferentiate from interfascicular parenchyma cells in the young internodes. In all three species, severing a vascular bundle in a young internode resulted in regeneration of xylem to form a curved by-pass immediately around the wound. The numerous phloem anastomoses in these young internodes were not involved in this process, the regenerated vessels originating from interfascicular parenchyma alone. Conversely, in mature internodes of Dahlia, the regenerated vessels originated from initials of the interfascicular cambia, and their phloem anastomoses did not influence the pattern of xylogenesis. On the other hand, in old internodes of Cucurbita, in which an interfascicular cambium was not yet developed, the parenchyma cells between the bundles had lost the ability to redifferentiate into vessel elements, and instead, regenerated vessels were produced in the phloem anastomoses. Thus, the wounded region of the vascular bundle was not bypassed via the shortest, curved pathway, but by more circuitous routes further away from the wound. Some of the regenerated vessels produced in the phloem anastomoses were extremely wide, and presumably efficient conductors of water. It is proposed that the dense network of phloem anastomoses developed during evolution as a mechanism of adaptation to possible damage in mature internodes by providing flexible alternative pathways for efficient xylem regeneration in plants with limited or no interfascicular cambium.This paper is dedicated to the memory of the late Isaac Blachmann (deceased 19 November 1995), father-in-law of the senior author, for encouragement and advice throughout the yearsThis research was supported by an International Scientific Exchange Award to R.A. from the Israel Academy of Sciences and The Royal Society.     

Gene technology-based antimetabolite design: the use of an in vitro protein expression system to facilitate antimetabolite design for virally-induced human diseases and malignant conditions

A precondition for the chemotherapeutic treatment of a variety of virally-induced human diseases and malignant conditions is a highly selective interaction of the drug molecule to be used with it's biological target. To ensure the development of novel, effective drugs, it is essential that the biological target is well characterised with regard to it's structure and activity. Such characterisation relies upon adequate amounts of pure target being available. One of the most important enzymatic importers for antimetabolites is the enzyme thymidine kinase. In this article an in vitro protein expression system is described which facilitates the production of milligram amounts of pure and biologically active thymidine kinase, from a number of important biological sources. Results have shown that the in vitro produced enzyme has the exact biochemical propeties of the in vivo enzyme. Thus the in vitro protein expression system is an ideal vechicle to facilitate an in depth investigation of the enzyme's biological properties.     

Effect of agitation speed on the synthesis of Mucor miehei acid protease

Different experiments using Mucor miehei CBS 370.65 were carried out to study the effect of agitation speed on the production of the mold acid protease. The experiments were conducted in shake flasks at a fixed substrate concentration of 58 g l⁻¹ of total carbohydrates and at shaker speeds from 80 to 380 rev min⁻¹. Enzyme production was found to be directly proportional to the shaker speeds, with the highest concentration of enzyme of 1,400 Soxhlet Rennet units (SU) ml⁻¹ obtained at 380 rev min⁻¹. The yield of product to substrate at 380 rev min⁻¹ was determined to be 27,081.0 SU g⁻¹ substrate and the productivity of the process was 221 SU g⁻¹ h⁻¹. Enzyme production was partially growth associated, and glucose supported both cell growth and enzyme production. Product formation and cell concentration were directly related to the rate of substrate consumption. The rate of product formation decreased when product started to accumulate, suggesting that the process was affected by feedback repression.     

Genetic and environmental variation in anther,pollen and pistil dimensions in sesame

Measurements of anther (length, width, depth), pollen grain (percent fertility, polar diameter, equatorial diameter, polar diameter/equatorial diameter ratio, volume) and pistil (stigma length, style length, ovary length, total pistil length, stigma width, style width, ovary width) were taken on 12 diverse sesame (Sesamum indicum L.) genotypes on each of four collection dates in 1994. Highly significant differences among genotype means were obtained for all characters except polar diameter. Highly significant differences among environment (collection date) means were found for ten of the 15 characters measured. Highly significant genotype x environment interactions were obtained for all characters except anther length. For the anther characters measured, relatively high repeatability values were found, ranging from 99.8% for length to 87.6% for depth. For the pollen grain characters measured, the repeatability values ranged from 67.6% for percent fertility to 23.1% for polar diameter. For the pistil characters measured, the repeatability values ranged from 94.0% for style width to 49.6% for total pistil length. These results indicate that genotype and environment influence anther, pollen grain and pistil characters. Variation in some of these morphological aspects could influence the consistency and interpretation of male transmission studies on both the applied and evolutionary levels.