The importance of the lipoxygenase-hepoxilin pathway in the mammalian epidermal barrier

This review covers the background to discovery of the two key lipoxygenases (LOX) involved in epidermal barrier function, 12R-LOX and eLOX3, and our current views on their functioning. In the outer epidermis, their consecutive actions oxidize linoleic acid esterified in ω-hydroxy-ceramide to a hepoxilin-related derivative. The relevant background to hepoxilin and trioxilin biochemistry is briefly reviewed. We outline the evidence that linoleate in the ceramide is the natural substrate of the two LOX enzymes and our proposal for its importance in construction of the epidermal water barrier. Our hypothesis is that the oxidation promotes hydrolysis of the oxidized linoleate moiety from the ceramide. The resulting free ω-hydroxyl of the ω-hydroxyceramide is covalently bound to proteins on the surface of the corneocytes to form the corneocyte lipid envelope, a key barrier component. Understanding the role of the LOX enzymes and their hepoxilin products should provide rational approaches to ameliorative therapy for a number of the congenital ichthyoses involving compromised barrier function. This article is part of a Special Issue entitled The Important Role of Lipids in the Epidermis and their Role in the Formation and Maintenance of the Cutaneous Barrier. Guest Editors: Kenneth R. Feingold and Peter Elias.     

Comparing molecular measures for detecting inbreeding depression

Correlations between heterozygosity and components of fitness have been investigated in natural populations for over 20 years. Positive correlations between a trait of interest and heterozygosity (usually measured at allozyme loci) are generally recognized as evidence of inbreeding depression. More recently, molecular markers such as microsatellites have been employed for the same purpose. A typical study might use around five to ten markers. In this paper we use a panel of 71 microsatellite loci to: (1) Compare the efficacy of heterozygosity and a related microsatellite‐specific variable, mean d², in detecting inbreeding depression; (2) Examine the statistical power of heterozygosity to detect such associations. We performed our analyses in a wild population of red deer (Cervus elaphus) in which inbreeding depression in juvenile traits had previously been detected using a panel of nine markers. We conclude that heterozygosity‐based measures outperform mean d²‐based measures, but that power to detect heterozygosity‐fitness associations is nonetheless low when ten or fewer markers are typed.     

Epistasis,complex traits,and mapping genes

Using a three-locus model wherein two loci regulate a third, candidate locus, I examine physiological epistasis from the gene's eye view of the regulated locus. I show that, depending upon genetic background at the regulatory loci, an allele at the candidate locus can be dominant, additive, recessive, neutral, over-dominant, or under-dominant in its effects on fitness. This kind of variation in allelic effect caused by variation in genetic background from population to population, from time to time in the same population, or sample to sample makes finding and mapping the genes underlying a complex phenotype difficult. The rate of evolution of such genes can also be slowed, especially in genetically subdivided metapopulations with migration. Nevertheless, understanding how variation in genetic background causes variation in allelic effects permits the genetic architecture of such complex traits to be dissected into the interacting component genes. While some backgrounds diminish allelic effects and make finding and mapping genes difficult, other backgrounds enhance allelic effects and facilitate gene mapping.     

EVOLUTION OF THE MAGNITUDE AND TIMING OF INBREEDING DEPRESSION IN PLANTS

Estimates of inbreeding depression obtained from the literature were used to evaluate the association between inbreeding depression and the degree of self-fertilization in natural plant populations. Theoretical models predict that the magnitude of inbreeding depression will decrease with inbreeding as deleterious recessive alleles are expressed and purged through selection. If selection acts differentially among life history stages and deleterious effects are uncorrelated among stages, then the timing of inbreeding depression may also evolve with inbreeding. Estimates of cumulative inbreeding depression and stage-specific inbreeding depression (four stages: seed production of parent, germination, juvenile survival, and growth/reproduction) were compiled for 79 populations (using means of replicates, N = 62) comprising 54 species from 23 families of vascular plants. Where available, data on the mating system also were collected and used as a measure of inbreeding history. A significant negative correlation was found between cumulative inbreeding depression and the primary selfing rate for the combined sample of angiosperms (N = 35) and gymnosperms (N = 9); the correlation was significant for angiosperms but not gymnosperms examined separately. The average inbreeding depression in predominantly selfing species (δ = 0.23) was significantly less (43%) than that in predominantly outcrossing species (δ = 0.53). These results support the theoretical prediction that selfing reduces the magnitude of inbreeding depression. Most self-fertilizing species expressed the majority of their inbreeding depression late in the life cycle, at the stage of growth/reproduction (14 of 18 species), whereas outcrossing species expressed much of their inbreeding depression either early, at seed production (17 of 40 species), or late (19 species). For species with four life stages examined, selfing and outcrossing species differed in the magnitude of inbreeding depression at the stage of seed production (selfing δ = 0.05, N = 11; outcrossing δ = 0.32, N = 31), germination (selfing δ = 0.02, outcrossing δ = 0.12), and survival to reproduction (selfing δ = 0.04, outcrossing δ = 0.15), but not at growth and reproduction (selfing δ = 0.21, outcrossing δ = 0.27); inbreeding depression in selfers relative to outcrossers increased from early to late life stages. These results support the hypothesis that most early acting inbreeding depression is due to recessive lethals and can be purged through inbreeding, whereas much of the late-acting inbreeding depression is due to weakly deleterious mutations and is very difficult to purge, even under extreme inbreeding.     

Thrips tabaci and tomato spotted wilt virus: inheritance of vector competence

Populations of onion thrips, Thrips tabaci Lindeman (Thysanoptera: Thripidae), were shown to differ significantly in their ability to transmit an isolate of tomato spotted wilt virus (Tospovirus: Bunyaviridae) (TSWV) collected from potato [Solanum tuberosum L. (Solanaceae)]. To gain an understanding of the basis for this variation, we generated reciprocal crosses between an efficient and an inefficient transmitting population. The resulting F₁ progeny and progeny from the parental populations were tested for their ability to transmit TSWV. Our results indicate that the ability to transmit TSWV efficiently by T. tabaci is inherited as a recessive trait.     

Inheritance of vector competence by the thrips Ceratothripoides claratris (Shumsher) (Thysanoptera: Thripidae)

The thrips Ceratothripoides claratris is an efficient vector of the Capsicum chlorosis virus (CaCV). Transmission studies with a natural population of C. claratris found in a greenhouse ‘GH’ and a ‘colony’ derived from this ‘GH’ population by selection and inbreeding resulted in lowering the percentage of viruliferous individuals within the ‘colony’. After passing through approximately 20 generations, the ‘colony’ lost the ability to transmit the CaCV. When either viruliferous or non‐viruliferous virgin females reproduced parthenogenetically, 81% of F1 arrhenotokous males inherited their viruliferous status from their mothers, whilst, no viruliferous offspring arose from non‐viruliferous virgin mothers. Crosses between viruliferous and non‐viruliferous individuals suggest that the competence of the thrips C. claratris as a vector for this virus is probably a heritable trait controlled by a recessive allele.     

Serine proteinase activation of latent human skin collagenase

Latent and active collagenase were demonstrated following direct extraction from normal skin homogenates with 0.1M calcium chloride at 60 degrees C. 83% of the collagenase activity was in latent form and could be maximally activated with trypsin. Partial activation of the latent enzyme could also be demonstrated by incubation of the skin extract without added trypsin. This endogenous activation was inhibited by the addition of soya bean trypsin inhibitor, trasylol, di-isopropylphosphofluoridate and phenylmethanesulphonylfluoride, none of which inhibited collagenase directly. This suggests that the skin extracts contain a collagenase activating enzyme with the inhibition profile of a serine proteinase. A chymotryptic proteinase with a similar inhibition profile was extracted from normal human skin and partially purified. This enzyme activated fibroblast procollagenase derived from tissue culture of normal skin. The procollagenase was also partially activated by plasmin and chymotrypsin. This is the first demonstration of a collagenase activating enzyme in human skin and raises the possibility that collagenase activation by this mechanism may be responsible for collagen degradation in some disease processes.     

Orange, yellow and white-cream: inheritance of carotenoid-based colour in sunflower pollen

Inheritance of pollen colour was studied in sunflower (Helianthus annuus L.) using three distinct pollen colour morphs: orange, yellow and white‐cream. Orange is the most common colour of sunflower pollen, while the yellow morph is less frequent. These two types were observed in the inbred lines F11 and EF2L, respectively. White‐cream pollen is a rare phenotype in nature, and was identified in a mutant, named white‐cream pollen, recovered in the R₂ generation of an in vitro regenerated plant. The F11 inbred line was used as starting material for in vitro regeneration. The carotenoid content of these three pollen morphs differed, and was extremely reduced in white‐cream pollen. The phenotype of F₁ populations obtained by reciprocal crosses revealed that the orange trait was dominant over both white‐cream and yellow. Segregation of F₂ populations of both crosses, orange × yellow and orange × white‐cream, approached a 3:1 ratio, indicating the possibility of simple genetic control. By contrast, a complementation cross between the two lines with white‐cream and yellow pollen produced F₁ plants with orange pollen. The F₂ populations of this cross‐segregated as nine orange: four white‐cream: four yellow. A model conforming to the involvement of two unlinked genes, here designated Y and O, can explain these results. Accessions with yellow pollen would have the genotype YYoo, the white‐cream pollen mutant would have yyOO and the accession with orange pollen would have YYOO. Within F₂ populations of the cross white‐cream × yellow a new genotype, yyoo, with white‐cream pollen was scored. The results of the cross yyoo × YYoo produced only F₁ plants with yellow pollen, supporting a recessive epistatic model of inheritance between two loci. In this model, yy is epistatic on O and o. In F₂ populations, the distributions of phenotypic classes suggested that the genetic control of carotenoid content is governed by major genes, with large effects segregating in a background of polygenic variation. These three pollen morphs can provide insight into the sequence in which genes act, as well into the biochemical pathway controlling carotenoid biosynthesis in anthers and the transfer of these different pigments into pollenkitt.     

Absence of mutations in GJB2 (Connexin-26) gene in an ethnic group of southwest Iran

BACKGROUND:

The common GJB2 gene mutation (35delG) has been previously reported from Iranian patients that were affected with nonsyndromic autosomal recessive deafness. We, therefore, for the first time, investigated the prevalence and frequency of the GJB2 gene mutation in the Iranian deaf population with Arabian origins.

MATERIALS AND METHODs:

We amplified and sequenced the entire coding sequence of the GJB2 gene from 61 deaf patients and 26 control subjects.

RESULT:

None of the analyzed samples revealed deafness-associated mutation.

CONCLUSION:

This finding differs from several reports from Iran as we have focused on the GJB2 gene that possesses various mutations as the cause of congenital recessive deafness.