A spontaneous mutation, enNK14, was a new allele of engrailed (en) in Drosophila melanogaster. Females of enNK14 have three spermathecae, instead of two in wild type, under a wide range of developmental temperatures, while the males show no abnormal phenotype. Spermathecae of the mutant female can accept inseminated sperms, albeit with a delay of at least an hour until full acceptance compared with wild type. The time course of decrease in the number of stored sperms was thoroughly similar between the mutant and wild type. enNK14 females produced fewer progeny than wild type females despite storing a larger number of sperms. The delay of sperm entry and lower fecundity suggested some functional defects in secretory products of the spermathecae. In addition, some spermathecae in the mutant were accompanied by a mass of brown pigments in the adipose tissue surrounding the capsule. Six contiguous amino acids, Ser340–Ala345, were replaced by one Thr in enNK14. In another mutant, enspt, Ser325 was also shown to be substituted by a Cys. These amino acid changes were located within a serine-rich region, in which Ser325, Ser340 and Thr341 were suggested as targets of Protein Kinase C by an in silico analysis. The splicing pattern of en mRNA did not differ between enNK14 and wild type in embryo, larva, pupa or adult. Our results suggest that en plays an important role in determining the number of spermathecae as well as in sperm storage function in the Drosophila female.
While the house mouse (Mus musculus), widely distributed in Eurasia, is known to have substantial coat color variation between and within local populations, in both primary and secondary distribution areas, including the Japanese archipelago, the evolutionary history of the color variation is poorly understood. To address the ventral fur color variation, we quantified the lightness of museum skin specimens, and found that the southern subspecies, M. m. castaneus (CAS), has high and low lightness in dry and rainy geographic regions, respectively. The northern subspecies, M. m. musculus (MUS), has low and high levels of lightness in the high and middle latitudes of northern Eurasia, respectively. We examined sequence variation of the agouti signaling protein gene (Asip), which is known to be responsible for the ventral fur color. We performed phylogenetic analyses with 196 haplotype sequences of Asip (~180 kb) generated by phasing the whole-genome data of 98 wild mice reported previously. Network and phylogenetic tree construction revealed clustering of haplotypes representing the two subspecies, MUS and CAS. A number of subclusters with geographic affinities appeared within the subspecies clusters, in which the essential results were consistent with those reconstructed with whole mitochondrial genome data, indicating that the phased haplotype genome sequences of the nuclear genome can be a useful tool for tracing the dispersal of geographical lineages. The results of phylogeographic analysis showed that CAS mice with darker ventral fur possessed similar Asip haplotypes across the geographic distribution, suggesting that these haplotypes are major causes of the historical introduction of Asip haplotypes for darker ventral fur in mice from northern India to the peripheral areas, including the Japanese archipelago. Similarly, MUS in East Asia, which has a white abdomen, formed an Asip haplogroup with that from northern Iran, also with a white abdomen.
Long non-coding RNAs have been proposed as biomarkers for the detection, prevention and screening of various malignancies. In this study, two lncRNAs (ANRIL and BANCR) were assessed for biomarker application in the early detection of colorectal cancer (CRC) through stool specimen testing, as a non-invasive and cost-effective methodology. A total of 40 stool samples were collected from patients referred to the hospital with colorectal cancer or adenomatous polyps as pre-cancerous lesions; patients were diagnosed using colonoscopy and pathology reports were available. Twenty control samples were also obtained from healthy subjects for comparison. RNA extraction and cDNA synthesis were followed by real-time PCR to evaluate lncRNA expression. The up-regulation of ANRIL in 20% of samples taken from polyp patients, combined with up-regulation in 65% of patients with CRC, confirmed the potential usefulness of ANRIL as a prognostic biomarker (AUC 0.95; P < 0.0001). BANCR relative expression analysis illustrated significant up-regulation in polyp (P < 0.04) and tumoural participants (P < 0.03) compared with normal control individuals. The expression patterns of ANRIL and BANCR in polyp cases were significantly correlated according to correlation analysis (r = 0.45, P < 0.045). ANRIL expression patterns in stool specimens of polyp and tumour cases supported the use of ANRIL as a prognostic biomarker for screening patients in the early stages of CRC. Up-regulation of BANCR in pre-cancerous lesions as well as down-regulation of ANRIL may also be a specific marker pair for easy, convenient and fast CRC prognosis.
Mycoheterotrophic plants can derive carbon from fungi rather than from photosynthesis. Habitat destruction and sensitivity to environmental perturbation may result in the loss of biodiversity including genetic variation of mycoheterotrophic plants. Burmannia nepalensis (Miers) Hook.f. (Burmanniaceae) is a mycoheterotrophic plant with a wide distribution across southern China and southern and eastern Asia. As part of our endeavor to reveal population genetic patterns of mycoheterotrophic plants, fifteen microsatellite loci were developed by RAD (restriction site-associated DNA) sequencing in 89 individuals from four populations of B. nepalensis. A total of 49 alleles were amplified. The number of alleles per locus ranged from two to six with an average of 3.3. The observed and expected heterozygosity per population varied from 0.000 to 1.000 and from 0.000 to 0.722, respectively. A transferability test showed that only one to five loci could be cross-amplified successfully in four other congeneric species of Burmannia. These markers can be used to reveal population genetic diversity in B. nepalensis, and will help to elucidate the evolutionary history and to enhance efforts for conservation of mycoheterotrophic plants.