Coloration is one of the most variable characters in animals and provides rich material for studying the developmental genetic basis of pigment patterns. In the silkworm, more than 100 gene mutation systems are related to aberrant color patterns. The melanism (mln) is a rare body color mutant that exhibits an easily distinguishable phenotype in both larval and adult silkworms. By positional cloning, we identified the candidate gene of the mln locus, Bm-iAANAT, whose homologous gene (Dat) converts dopamine into N-acetyldopamine, a precursor for N-acetyldopamine sclerotin in Drosophila. In the mln mutant, two types of abnormal Bm-iAANAT transcripts were identified, whose expression levels are markedly lower than the wild type (WT). Moreover, dopamine content was approximately twice as high in the sclerified tissues (head, thoracic legs, and anal plate) of the mutant as in WT, resulting in phenotypic differences between the two. Quantitative reverse transcription PCR analyses showed that other genes involved in the melanin metabolism pathway were regulated by the aberrant Bm-iAANAT activity in mln mutant in different ways and degrees. We therefore propose that greater accumulation of dopamine results from the functional deficiency of BmiAANAT in the mutant, causing a darker pattern in the sclerified regions than in the WT. In summary, our results indicate that Bm-iAANAT is responsible for the color pattern of the silkworm mutant, mln. To our knowledge, this is the first report showing a role for arylalkylamine-N-acetyltransferases in color pattern mutation in Lepidoptera.Coloration is one of the most variable traits in insects, and pigmentation is known to play a role in mimicry, sexual selection, thermoregulation, and other adaptive processes in many insect groups (1). More recently, there has been much interest among biologists in the molecular mechanisms underlying the great diversity of insect colors and color patterns.Melanin, the dark pigment found in melanophores, is an important class of insect pigments (2-4). The genes responsible for pigment metabolism have been systematically studied in many insects, including Drosophila melanogaster, Manduca sexta, and Papilio xuthus (2, 3, 5-8). The melanin is derived from tyrosine (10, 11). Tyrosine hydroxylase and dopa decarboxylase convert tyrosine into dopa and dopa into dopamine, respectively (2). Dopa serves as the precursor of dopa melanin (which is black) and is converted to dopa melanin by the activities of the Yellow, Yellow-f 1 and Yellow-f 2 proteins (12). Furthermore, dopamine serves as the precursor of dopamine melanin (black or brown in color and a major pigment of the insect cuticle), the change being caused by the catalytic action of phenol oxidases (1,2,3,5,6,(12)(13)(14). The current study confirmed that the dopamine content was elevated in melanized insects or in the melanized regions of insects (5, 15). Alternatively, dopamine can reversibly convert to N--alanyldopamine (NBAD, 3 yellowish) by NBAD synthetase (EBONY) and NBAD hydrolase. NBAD deposi...
RNA-based fluorescent probes are currently limited by their low selectivity toward RNA versus DNA, and low specificity to different RNA structures. Poor membrane permeability is another defect of existing fluorogenic RNA probes for intracellular imaging. In this work, a naphthalimide derivative, probe 1, was developed for the rapid and selective detection of intracellular rRNA (rRNA). Probe 1 exhibited a 32-fold fluorescent enhancement in response to rRNA binding and showed desirable selectivity for rRNA versus DNA and other nucleic acids in phosphate buffer at pH 7.2. Importantly, probe 1 displayed excellent permeability of the nucleolus, could be taken up in 1 min by four different cell lines, and may be the fastest nucleolus dye. The excellent selectivity of probe 1 toward rRNA is attributed to the specific interaction between the complicated 3D structures of rRNA, which was confirmed by quantum calculations using molecular docking simulations. An appropriate lipophilic balance in 1 with the hydrophilic amine group and hydrophobic naphthalimide, as well as its high water solubility, guarantees the high permeability of 1 in cell membranes and nucleolus pores, compared to other analogues (e.g., probes 2–8 in this work). Furthermore, enlarged confocal laser micro images of nucleoli and RNase digestion tests revealed that 1 remained highly selective toward rRNA, even for intracellular imaging. As a live cell probe, 1 also exhibited better photostability than the commercial RNA dye, SYTO RNA select.
Under 980 nm excitation, upconversion (UC) emissions in the UV range of 270-320 nm were observed in nanocrystals Y(0.795-x)Gd(x)Yb(0.2)Tm(0.005)F(3) (x=0, 0.1, 0.2, 0.5, and 0.795), which were synthesized through a hydrothermal method. These UC emissions can be assigned to the transitions of (6)I(J), (6)P(J)-->(8)S(7/2)(Gd(3+)), and (3)P(0)/(1)I(6)-->(3)H(6) (Tm(3+)). The energy transfer from Tm(3+) to Gd(3+) plays a crucial role in populating the excited states of Gd(3+). The shortest wavelength of upconverted emission converted from the infrared region was demonstrated here.
Ultraviolet upconversion emissions of 246.2 and 252.8 nm from (6)D(J) levels of Gd(3+) ions were observed in GdF(3): 10% Yb(3+), 0.7% Tm(3+) nanocrystals under 980 nm excitation from a laser diode. The (6)D(J) levels of Gd(3+) ions can be efficiently populated by energy transfer processes of Yb-->Tm-->Gd and Yb-->Gd. A six-photon upconversion process was confirmed by the dependence of 252.8 nm emission intensity on the pumping power. The upconversion mechanism in the six-photon process was discussed based on excited state absorption of Gd(3+) ions, cross relaxation energy transfer between two excited Gd(3+) ions, and energy transfer between Gd(3+) and Yb(3+) or Tm(3+) ions.
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