ポスター
C. 感覚系と運動系
C. Sensory and Motor Systems |
|---|
| 2022年7月2日 11:00～12:00 沖縄コンベンションセンター 展示棟 ポスター会場1
3P-001
筋肉および神経系で発現するアミノ酸輸送体SLC46の低温耐性への関与
Amino acid transporter SLC46 expressed in muscle and neurons is involved in cold tolerance of C. elegans
*山城 芹奈(1)、水野 賢美(1)、本村 晴佳(1)、太田 茜(1)、久原 篤(1,2)
1. 甲南大学、2. PRIME, AMED
*Serina Yamashiro(1), Satomi Mizuno(1), Haruka Motomura(1), Akane Ohta(1), Atsushi Kuhara(1,2)
1. Konan University, 2. PRIME, AMED
Keyword: C.elegans, cold tolerance, SLC46A3, pharyngeal muscle
Organisms can survive by adapting to changes in environmental temperature, which is a dynamic environmental factor. C. elegans exhibits a cold tolerance in which wild-type worms reared at 25℃ die after 48 hours of cold stimulation at 2℃, whereas more than 90% of the wild-type reared at 15℃ can survive at 2℃.In order to identify novel molecules that control cold tolerance, we attempted to isolate novel cold-tolerant mutants by forward genetics. We treated wild-type strains with mutagen EMS and isolated a cold intolerant mutant, KHR018, which showed abnormalities such that KHR018 died at 2°C after being raised at 15°C. Decoding whole genome and genetic analysis revealed that a responsible gene for defective cold tolerance of KHR018 was C46C11.2, which encodes a homolog of human "amino acid transporter" SLC46A3. This identified gene C46C11.2 was named as slcr-46.1. Human SLC46A3 localizes to the lysosomal membrane, but its function in vivo is largely unknown.To identify the subcellular localization of the SLCR-46.1 protein in C. elegans, we introduced a fusion gene consisting of the full length of slcr-46.1 ORF fused with GFP gene into wild-type, and found that SLCR-46.1::GFP was observed in the intestine, pharyngeal muscle, and epidermis. In another experiment, slcr-46.1 promoter:: slcr-46.1 cDNA::gfp in wild-type was found in some granules of the intestine and in two pairs of neurons in the head. These two pairs of neurons were identified as thermosensory neuron ASG and oxygen-sensing neuron BAG, by co-staining with the NeuroPAL, a strain with a stereotyped fluorescent color map to identify all neurons (Yemini et al., Cell, 2021). SLCR-46.1::GFP protein was co-localized to the lysosomal protein LMP-1.To identify functional cells of SLCR-46.1 in the cold tolerance, we performed rescue experiments using tissue-specific promoters. Expressing slcr-46.1cDNA::GFP in pharyngeal muscle of slcr-46.1 mutant by the myo-2 promoter restored abnormal cold tolerance of slcr-46.1 mutant. This suggests that SLCR-46.1 in the pharyngeal muscle is essential for the normal cold tolerance in wild-tyep. We are attempting to investigate whether SLCR-46.1 localizes into lysosomes of pharyngeal muscle, and if so, to analyze how muscular SLCR-46.1 controls cold tolerance. | | | |
|
|
