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Biomedical subjects

K A Raphael

Publications and source records attributed to K A Raphael.

10 recordsLinked to original sources

The scarlet eye colour gene of the tephritid fruit fly: Bactrocera tryoni and the nature of two eye colour mutations.

A homologue of the Drosophila melanogaster eye-colour gene, scarlet (st), has been isolated from the genome of the tephritid fruit fly, Bactrocera tryoni. The comparison of the B. tryoni and D. melanogaster scarlet gene shows 71.2% and 79.3% sequence identity at the DNA and the derived amino acid level, respectively. Two allelic eye-colour mutations of B. tryoni, orange-eyes and lemon-eyes, have been recovered and found to be colocalized with the st gene. The st gene sequence in the two mutant strains has been examined for DNA sequence changes and expression levels.

Alleles↗

The period gene in two species of tephritid fruit fly differentiated by mating behaviour.

The period gene is important for the generation and maintenance of biological rhythms. It served as an ideal candidate for the investigation of the mating time isolation between two sibling Queensland fruit fly species, Bactrocera tryoni and Bactrocera neohumeralis. We have isolated the homologues of the period gene in the two species, and show that their putative amino acid sequences are identical. No length polymorphism was detected in the Thr-Gly repeat region. per mRNA expression, assayed in light-dark diurnal conditions, displayed circadian oscillation in both the head and abdomen of B. tryoni and B. neohumeralis, with the same cycling phase. An alternatively spliced intron was identified in the 3' untranslated region. The effect of temperature on the splicing and mRNA expression was examined.

3' Untranslated Regions↗

Growth factor expression in skin during wool follicle development.

A variety of growth factors are likely to be involved in initiation and morphogenesis of wool follicles. To enable direct comparisons of the expression of different growth factors, reverse transcriptase-polymerase chain reactions (RT-PCR) were developed for ovine and murine TGF alpha, TGF beta 1, TGF beta 2, TGF beta 3, IGF1, IGF2, and FGF-2, which could all be carried out on a single cDNA sample. These RT-PCR were used with 16 sheep RNA samples from different foetal stages, neonatal sheep and mouse skin. The mRNAs for these growth factors were detected throughout gestation in sheep skin, except for TGF beta 1 mRNA which was not expressed in 51-day-old skin, but was expressed in 54-day and older samples. Since the first microscopically visible changes of follicle initiation occur around 62 days gestation, these results suggest that TGF beta 1 expression may be a signal for follicle initiation.

Animals↗

myc protooncogenes of wool and hair growth.

The growth of hard keratin fibers such as wool and hair is dependent on the proliferation of cells in the follicle bulb. If the cells leaving the bulb could be induced to undergo an extra division, then fiber growth should increase. The cellular division within the follicle is complex and probably involves one or more growth factors, which act by altering the expression of transcription factors and other nuclear proteins. We propose that the expression of the myc protooncogenes is a central part of this mechanism. In support of this hypothesis we have detected the mRNAs for TGF-beta 1, basic FGF, TGF-alpha, and c-myc in plucked wool follicles using PCR amplification. We have also shown that the TGF-beta 1, TGF-beta 2, TGF-beta 3, EGF, TGF-alpha, basic FGF, N-myc, and c-myc genes are expressed in mouse skin, and we looked for changes during the hair cycle. The PCR data suggest that in whole skin the levels of mRNA for TGF-beta 1, TGF-beta 2, TGF-alpha, and c-myc do not change. In Quackenbush mice the levels for N-myc, TGF-beta 3, and basic FGF mRNA appear to be lower at the end of the hair cycle. We have confirmed in CBA/C57 black mice that lower levels of N-myc mRNA are detected when hair growth ceases in catagen and telogen. To test our hypothesis further and to assess its practical application, we are making transgenic mice in which the N-myc gene is overexpressed in the hair follicle by way of a wool keratin promoter. The transgene consists of 3.3 kb of 5' sequence from an ovine type 1 IF gene, the murine N-myc genomic coding sequence, and an SV40 polyadenylation signal. The native keratin type 1 IF gene is expressed exclusively in the wool follicle, as shown by in situ hybridization. However, in mice the injection of the transgene has resulted in high embryonic mortality and some embryos with large body size and head malformations. Since these mice were not transgenic, this is likely to be an effect of transient expression of the transgene during embryogenesis. The two transgenic mice produced so far have a normal phenotype.

Aging↗

The site of action of the naked locus (N) in the mouse as determined by dermal-epidermal recombinations.

The dermal-epidermal recombination technique was used to determine the site of action of the naked (N) locus in the skin of the mouse. The skin of athymic (nude) mice was used as a host site for growth of recombined epidermis and dermis from 13- and 14-day N/+ and +/+ embryos. Grafts that contained mutant epidermis lost their hair by 26 days after grafting (at the end of the first hair cycle) and again after 47 days (at the end of the second hair cycle); grafts that contained normal epidermis retained hair throughout the experiment. It was concluded that the N locus acts in the epidermis.

Animals↗