Biology subjects

Iizuka, T.

Publications and source records attributed to Iizuka, T..

7 recordsLinked to original sources

bioRxiv2025

An ATP-binding cassette subfamily C is crucial for flavonoid sequestration in the domestic silkworm, Bombyx mori

Some herbivorous insects have evolved sequestration mechanisms, the ability to take up and accumulate plant secondary metabolites for their own benefit. The domestic silkworm, Bombyx mori, and its wild ancestor, B. mandarina, take up flavonoids from mulberry leaves and accumulate the molecules as their glucosides in their tissues and cocoon shell. This sequestration enhances the cocoons protective property against ultraviolet or bacterial proliferation. Here, we show that an ATP-binding cassette transporter subfamily C (ABCC) gene, BmABCC4, plays a crucial role in the flavonoid sequestration in the silkworms. BmABCC4 is located at Green c, a cocoon color-associated locus predicted in 1941 and whose detailed position we previously identified. This transporter is expressed in the midgut and silk glands, and the expression is upregulated in the midgut in late period of final instar larva. Knockout of BmABCC4 significantly reduced the total flavonoid content in the tissues and cocoon shell. Our results suggest that BmABCC4 transports flavonoid glucosides from the midgut cells to hemolymph and from the silk gland cells to the silk gland lumen.

genetics↗

bioRxiv2023

A major endogenous glycosidase mediating quercetin uptake in Bombyx mori

Quercetin is a common plant flavonoid that is involved in herbivore-plant interactions. Mulberry silkworms (domestic silkworm, Bombyx mori, and wild silkworm, Bombyx mandarina) uptake quercetin from mulberry leaves and accumulate the metabolites in the cocoon, thereby improving its protective properties. Here we identified and characterized a glycosidase, named LPH-like quercetin glycoside hydrolase 1 (LQGH1), that initiates quercetin metabolism in the domestic silkworm. LQGH1 is expressed in the midgut where it mediates quercetin uptake by deglycosylating the three most common quercetin glycosides present in mulberry leaf: rutin, quercetin-3-O-malonyl-glucoside, quercetin-3-O-glucoside. Despite being located in an unequal crossing-over hotspot, LQGH1 is conserved in some species in clade Macroheterocera, including the wild silkworm, indicating the adaptive significance of quercetin uptake. LQGH1 is important also in breeding: defective mutations of LQGH1, which result in discoloration of the cocoon and increased silk yield, are homozygously conserved in 27 of the 32 Japanese white-cocoon domestic silkworm strains and 12 of the 30 Chinese ones we investigated.

genetics↗

bioRxiv2023

A novel monocarboxylate transporter involved in 3-hydroxykynurenine transport for ommochrome coloration

Ommochromes are widespread pigments in invertebrates utilized for screening pigments in compound eyes and for reddish coloration in epidermis and wings. Ommochromes are derived from 3-hydroxykynurenine (3OHK), which is incorporated into cells from hemolymph or synthesized from tryptophan within cells. While the synthetic pathway from tryptophan to 3OHK has been well characterized, the gene responsible for cellular uptake of 3OHK has been poorly understood. In the silkworm Bombyx mori, adult compound eyes and eggs contain a mixture of ommochrome pigments. By using positional cloning method, we found that a novel monocarboxylate transporter, 3-hydroxykynurenine transporter (3OHKT), is responsible for the recessive mutant maternal brown of Tsujita (b-t) of B. mori. In b-t mutant, the color of the eggs is light brown, whereas the color of the compound eyes is normal, and we identified a 2-kb deletion in 3OHKT gene. TALEN-mediated knockout of 3OHKT gene produced the same coloration phenotype as b-t mutant, and the complementation test between b-t mutant and 3OHKT knockout strain proved that 3OHKT is responsible for b-t phenotype. 3OHKT protein was localized in the cellular membrane, and LC-MS analysis indicated that the uptake of 3OHK from hemolymph into the ovary was suppressed in the b-t mutant. Moreover, we confirmed that 3OHKT gene is specifically expressed at the reddish region and the time of pigmentation in the pupal wing of nymphalid butterflies. RNA interference of 3OHKT prevented reddish pigmentation in wings, highlighting its general involvement in ommochrome-based pigmentation other than compound eyes. SignificanceOmmochromes are widely distributed pigments in invertebrates and are synthesized from intracellular tryptophan or 3-hydroxykynurenine (3OHK). Ommochrome-based red markings on butterfly wings are often used for sexual selection, warning colors and mimicry. Most genes involved in the ommochrome synthesis pathway have been elucidated from analyses of eye color mutants in Drosophila. However, this study reveals that the ommochrome synthesis pathway has a different genetic repertoire depending on the tissues, and that the novel monocarboxylate transporter identified in this study has a major role in ommochrome pigmentation other than in compound eyes. In particular, our results suggest that classical ommochrome-related genes are rarely involved in the wing pigmentation of the nymphalid butterflies.

genetics↗

bioRxiv2023

Direct link between convergent evolution at sequence level and phenotypic level of septal pore cap in Agaricomycotina

Several apparently homologous morphological characters are known to have independently evolved in different lineages multiple times. However, the genetic backgrounds of such morphological convergences are not well understood. To detect any correlated genes potentially responsible for morphological convergence at the phenotypic level, we focused on the morphology of the septal pore cap (SPC), which is involved in mycelias complex multicellularity in fungi. SPCs are classified into three morphological types: perforate, imperforate, and vesiculate. To understand what evolutionary events occurred at the sequence level during morphological convergence of perforate SPCs in Agaricomycotina, we examined sequence differences between species with different SPC types by comparative genomic analysis. If sequences from species with perforate SPCs formed a cluster, the associated gene might be involved in generating perforate SPCs in multiple lineages. Based on this assumption, we detected eight candidate genes, including an SPC-related gene, spc33. The results showed that some amino acid substitutions independently occurred in both lineages in which species with perforate SPCs emerged. From these results, we speculate that the amino acid substitutions in spc33 were critical in the emergence of perforate SPCs in multiple lineages. We also found that spc33 evolved just before imperforate SPC emergence based on homology search of spc33 and the species phylogeny. These findings illustrate the first step to clarifying the genetic basis of SPC morphological evolution. Our study contributes to both clarifying the genetic basis of morphological convergence and pioneering our understanding of fungal evolutionary morphology.

evolutionary biology↗

bioRxiv2022

Highly accurate genome assembly of an improved high-yielding silkworm strain, Nichi01

The silkworm (Bombyx mori) is an important lepidopteran model insect, and an industrial domestic animal traditionally used for silk production. Here, we report the genome assembly of an improved Japanese strain Nichi01, in which the cocoon yield is comparable to that of commercial silkworm strains. The integration of PacBio Sequel II long-read and ddRAD-seq-based high-density genetic linkage map achieved the highest quality genome assembly of silkworms to date; 22 of the 28 pseudomolecules contained telomeric repeats at both ends, and only four gaps were present in the assembly. A total of 452 Mbp of the assembly with an N50 of 16.614 Mbp covered 99.3% of the complete orthologs of the Arthropod core genes. Although the genome sequence of Nichi01 and that of the previously reported low-yielding tropical strain p50T assured their accuracy in most regions, we corrected several regions, misassembled in p50T, in our assembly. A total of 18,397 proteins were predicted using over 95 Gb of mRNA-seq derived from 10 different organs, covering 96.9% of the complete orthologs of the Arthropod core genes. The final assembly and annotation files are available in KAIKObase (https://kaikobase.dna.affrc.go.jp/index.html) along with a genome browser and BLAST searching service, which would facilitate further studies and the breeding of silkworms and other insects.

genomics↗

bioRxiv2021

Arousal-dependent auditory response in the brain of Bengalese finches as measured by gene expression

Songbirds use auditory feedback to maintain their own songs. Juveniles also memorize a tutor song and use memory as a template to make up their own songs through auditory feedback. A recent electrophysiological study revealed that HVC neurons respond to BOS playback only in low arousal, sleeping, or anesthetized conditions. One outstanding question is how does auditory suppression occur in the brain? Here, we determined how arousal affects auditory responses simultaneously in the whole brain and over the song neural circuit in Bengalese finches, using the immediate early gene egr-1 as a marker of neural activity. Our results showed that auditory responses in the low-arousal state were less susceptible to gating, which was also confirmed by gene expression, and that the suppression may be weaker than observed in previous zebra finch studies. This may be because the Bengalese finch is a domesticated species. In addition, our results suggest that information may flow from the MLd.I of the midbrain to higher auditory regions. Altogether, this study presents a new attempt to explore the auditory suppression network by simultaneously investigating the whole brain using molecular biology methods.

neuroscience↗

bioRxiv2019

Deep learning-based imaging classification identified cingulate island sign in dementia with Lewy bodies

The differentiation of dementia with Lewy bodies (DLB) from Alzheimers disease (AD) using brain perfusion single photon emission tomography is important but has been a challenge because these conditions have common features. The cingulate island sign (CIS) is the most recently identified specific feature of DLB for a differential diagnosis. The present study aimed to examine the usefulness of deep learning-based imaging classification for the diagnoses of DLB and AD. We also investigated whether CIS was focused by the deep convolutional neural network (CNN) during differentiation.\n\nBrain perfusion single photon emission tomography images were acquired from 80 patients each with DLB and with AD and 80 individuals with normal cognition (NL). The CNN was trained on brain surface perfusion images. Gradient-weighted class activation mapping (Grad-CAM) was applied to the CNN for visualization of the features that the trained CNN focused on.\n\nBinary classifications between DLB and NL, DLB and AD and AD and NL were 94.69%, 87.81% and 94.38% accurate, respectively. The CIS ratios closely correlated with softmax output scores for DLB-AD discrimination (DLB/AD scores). The Grad-CAM highlighted CIS in the DLB discrimination. Visualization of learning process by guided Grad-CAM revealed that CIS became more focused by the CNN as the training progressed. DLB/AD score was significantly associated with three core-features of DLB.\n\nDeep learning-based imaging classification was useful not only for objective and accurate differentiation of DLB from AD but also for predicting clinical features of DLB. The CIS was identified as a specific feature during DLB classification. The visualization of specific features and learning process could have important implications for the potential of deep learning to discover new imaging features.

neuroscience↗