Project description:Most cartilaginous fishes live in seawater (SW), but a few exceptional elasmobranchs (sharks and rays) are euryhaline and can acclimate to freshwater (FW) environments. The plasma of elasmobranchs is high in NaCl and urea concentrations, which constrains osmotic water loss. However, these euryhaline elasmobranchs maintain high levels of plasma NaCl and urea even when acclimating to low salinity, resulting in a strong osmotic gradient from external environment to body fluid. The kidney consequently produces a large volume of dilute urine to cope with the water influx. In the present study, we investigated the molecular mechanisms of dilute urine production in the kidney of Japanese red stingray, Hemitrygon akajei, transferred from SW to low-salinity environments. We showed that red stingray maintained high plasma NaCl and urea levels by reabsorbing more osmolytes in the kidney when transferred to low salinity. RNA-seq and qPCR analyses were conducted to identify genes involved in NaCl and urea reabsorption under the low-salinity conditions, and the upregulated gene expressions of Na+-K+-Cl- cotransporter 2 (nkcc2) and Na+/K+-ATPase (nka) were found in the FW-acclimated individuals. These upregulations occurred in the early distal tubule (EDT) in the bundle zone of the kidney, which coils around the proximal and collecting tubules to form the highly convoluted structure of batoid nephron. Considering the previously proposed model for urea reabsorption, the upregulation of nkcc2 and nka not only causes the reabsorption of NaCl in the EDT, but potentially also supports enhanced urea reabsorption and eventually the production of dilute urine in FW-acclimated individuals. We propose advantageous characteristics of the batoid-type nephron that facilitate acclimation to a wide range of salinities, which might have allowed the batoids to expand their habitats.
Project description:Comparative transcriptome profile of genes differentially expressed in longissimus dorsi muscles between Japanese black (Wagyu) and Chinese Red Steppes cattle by RNA-seq
Project description:We performed a comparative genome-wide methylation analysis of longissimus dorsi muscles between the Japanese Black (Wagyu) and Chinese Red Steppes cattle, which exhibit significant differences in meat quality traits. This will allow us to better understand the correlation between DNA methylation variants and meat quality traits.
Project description:The mechanism driving the remarkable ability of the remaining kidney to enlarge and increase its function following the removal of its contralateral pair remains elusive. To explore the pathways driving compensatory renal hypertrophy, comprehensive RNA-seq transcriptional studies were undertaken in the kidneys of C57BL/6 mice undergoing hypertrophy at 24, 48, and 72 hours following nephrectomy, and these results were compared with mice undergoing sham operations. In addition, mass spectrometry was carried out at 24 hours to examine changes in protein expression. Single-nuclei RNA-Seq was used to delineate bulk RNA-seq data into cell types at 24 hours post-nephrectomy. HK-2 renal tubular cells were examined for their ability to undergo hypertrophy in the presence of IGF-1 via the activation of cholesterol biosynthesis pathways. Bulk RNA-seq revealed substantial time-dependent enhancement of cholesterol biosynthesis pathways, increases in mitochondrial gene expression, and cell cycle perturbations. Single-nuclei RNA-Seq at 24 hours post-nephrectomy showed that Sterol Binding Protein 2 (SREBP2) activity increases in medullary thick ascending limb cells and, to a lesser extent, in proximal tubular cells, consistent with the role of promoting cholesterol synthesis. Furthermore, SREBP2 was found to regulate cell size following IGF-1 stimulation in HK-2 cells. There are early, cell-specific alterations in gene expression of cholesterol biosynthesis pathways, mitochondrial genes, and the cell cycle in kidneys undergoing compensatory hypertrophy. SREBP2 activity in the medullary thick ascending limb and, to a lesser extent, in proximal tubules may play a previously undescribed role in promoting cholesterol metabolism in the mechanism underlying compensatory renal hypertrophy.
Project description:Transcriptomic analysis of cancer samples helps to identify the mechanism and molecular markers of cancer. However, transcriptomic analyses of pancreatic ductal adenocarcinoma from the Japanese population are lacking. We performed RNA sequencing of flesh or frozen pancreatic ductal adenocarcinoma tissues and adjacent normal pancreatic tissue from 12 Japanese patients to identify genes critical for the clinical pathology of pancreatic cancer among the Japanese population.