Biochemical and Biophysical Research Communications
Male specific expression of a cytochrome P450 (Cyp312a1) in Drosophila melanogaster
Section snippets
Materials and methods
Drosophila strains. Three parental strains of D. melanogaster, Oregon R, Canton S, and HKJ were used. HKJ is a field strain collected from Higashi Kurume-shi, Tokyo, Japan, in 2002 using a bait trap with grape peel. All strains were maintained at 25 °C on a standard medium based on the corn meal, glucose, and ebios.
Preparation of P450 cDNAs. cDNAs used for spotting on the cDNA array filter were prepared as follows. Sequences of amino acid and genomic DNA of all cytochrome P450s in D. melanogaster
cDNA array analysis
In total, we spotted cDNAs of 86 P450s and four internal controls. Each luminescence level was calculated based on the expression of the internal β-actin gene since the expression of this gene was least affected by synthesis of large amount of vitellogenin proteins in the ovary of female. Expression patterns are shown in Fig. 1. Cyp4g1 had the highest signal intensities, which was consistent with a previous study [21]. Of the 86 P450s, 17 isoforms were excluded from further analysis due to low
Discussions
Cyp312a1 was found to be expressed about 80-fold higher in adult male than female of Drosophila. This unique phenomenon is conserved among three strains of different origins, suggesting that this P450 is playing an important role, which is characteristic of male. In mammals, liver P450s involved with steroid hydroxylation have been well studied. Several P450s that metabolize sex hormones such as testosterone and estrogen are expressed in a sex-dependent manner [22]. On the contrary, insects do
Acknowledgements
We express our gratitude to Dr. Amanda Persad for the critical correction of the manuscript and to Dr. Noriaki Agui for his helpful comments on the manuscript. We also thank Kazue Ito for her technical support on the rearing of Drosophila melanogaster and to Kyoko Kasai for her help to collect HKJ strain. This work was supported by Grant-in-Aid for The Ministry of Education, Culture, Sports, Science and Technology (14560046).
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