Dataset Information

Identification of Circulating miRNAs in a Mouse Model of Nerve Allograft Transplantation under FK506 Immunosuppression by Illumina Small RNA Deep Sequencing.

ABSTRACT:

Background

This study aimed to establish the expression profile of circulating microRNAs (miRNAs) during nerve allotransplantation in the presence and absence of FK506 immunosuppression.

Methods

A 1?cm BALB/c donor sciatic nerve graft was transplanted into the sciatic nerve gaps created in recipient C57BL/6 mice with or without daily FK506 immunosuppression [1?mg/(kg·d)]. At 3, 7, and 14?d after nerve allotransplantation, serum samples were collected for miRNA expression analysis by Illumina small RNA deep sequencing.

Results

Sequence analysis showed that the dominant size of circulating small RNAs after nerve allotransplantation was 22 nucleotides, followed by 23-nucleotide sequences. Nine upregulated circulating miRNAs (let-7e-5p, miR-101a-3p, miR-151-5p, miR-181a-5p, miR-204-5p, miR-340-5p, miR-381-3p, miR-411-5p, miR-9-5p, and miR-219-2-3p) were identified at 3?d, but none was identified at 7 or 14?d. Among them, miR-9-5p had the highest fold-change of >50-fold, followed by miR-340-5p with 38.8-fold. The presence of these nine miRNAs was not significant at 7 and 14?d after nerve allotransplantation with or without immunosuppression, showing that these miRNAs are not ideal biomarkers for monitoring rejection of deep-buried nerve allografts, a response usually observed later.

Conclusions

We identified nine upregulated circulating miRNAs, which may have a biological function, particularly during the early stages after nerve allotransplantation under FK506 immunosuppression.

SUBMITTER: Wu SC

PROVIDER: S-EPMC4578739 | biostudies-literature | 2015

REPOSITORIES: biostudies-literature

ACCESS DATA

Publications

Identification of Circulating miRNAs in a Mouse Model of Nerve Allograft Transplantation under FK506 Immunosuppression by Illumina Small RNA Deep Sequencing.

Wu Shao-Chun SC Rau Cheng-Shyuan CS Yang Johnson Chia-Shen JC Lu Tsu-Hsiang TH Wu Yi-Chan YC Chen Yi-Chun YC Tzeng Siou-Ling SL Wu Chia-Jung CJ Lin Chia-Wei CW Hsieh Ching-Hua CH

Disease markers 20150908

<h4>Background</h4>This study aimed to establish the expression profile of circulating microRNAs (miRNAs) during nerve allotransplantation in the presence and absence of FK506 immunosuppression.<h4>Methods</h4>A 1 cm BALB/c donor sciatic nerve graft was transplanted into the sciatic nerve gaps created in recipient C57BL/6 mice with or without daily FK506 immunosuppression [1 mg/(kg·d)]. At 3, 7, and 14 d after nerve allotransplantation, serum samples were collected for miRNA expression analysis ...[more]

PMID: 26435568

Similar Datasets

Project description:This study aimed to establish the expression profile of circulating microRNAs (miRNAs) during nerve allotransplantation. Experimental design Male Balb/c and C57BL/6 mice (age, 10M-bM-^@M-^S12 weeks; weight, 30M-bM-^@M-^S35 g) were purchased from BioLasco (Yi-Lan, Taiwan). The Balb/c mice served as sciatic nerve allograft donors. The C57BL/6 mice were randomized into the group of nerve allograft transplantation with FK506 treatment and the group without FK506 treatment. These species of mice were selected on the basis of disparity at the MHC locus and prior experience with reciprocal rejection of grafts between these murine strains [18]. The mice were anesthetized by intraperitoneal injection of an anesthetic cocktail consisting of 0.1 mg/g ketamine and 0.01 mg/g xylazine. The anesthetized mice were restrained in a supine position on a heated pad to maintain the body temperature at 37M-BM-0C. Under aseptic conditions, with sterile povidone/iodine preparation and 70% ethanol and sterile instruments and drapes, the skin over the proximal right hindlimb was incised, and the underlying biceps femoris muscle was bluntly dissected to expose the sciatic nerve. An established mouse sciatic nerve allotransplantation model was used. In brief, 1-cm Balb/c donor sciatic nerve grafts were transplanted in reverse orientation into 0.5-cm sciatic nerve gaps created in the recipient Balb/c mice. Tension-free repair was then performed under an operating microscope with three 11M-bM-^@M-^S0 nylon (Ethicon Inc., Somerville, NJ) interrupted epineurial sutures under 40X magnification. The muscle was closed with 5M-bM-^@M-^S0 vicryl sutures, and the skin with interrupted 5M-bM-^@M-^S0 nylon sutures. The animals were monitored to ensure appropriate feeding and diet after surgery. FK506 was administered subcutaneously at 1 mg/(kg-d) throughout the experimental course. At 3, 7, and 14 d after initial surgery (n = 3 animals per group at each time point), the whole blood samples were collected at the indicated times in tubes containing anticoagulant. (F)3d, (F)7d and (F)14d were used to indicate the serum samples of allotransplantation mice with FK506 immunosuppression for 3, 7, and 14 d, respectively. (N)3d, (N)7d and (N)14d were used to indicate the serum samples of allotransplantation mice without FK506 immunosuppression and sacrificed at postoperative 3, 7, and 14 d, respectively. After the whole blood samples were incubated at room temperature for 15 min, they were centrifuged at 3,000 M-CM-^Wg for 10 min, white blood cells were slowly removed from the corresponding layers, and the serum was extracted and stored at M-bM-^@M-^S80M-BM-0C before processing for RNA analyses. Small RNA libraries generated from the serum of mice of above groups were analyzed using an Illumina HiSeq2000 Sequencer.

Dataset Information

Identification of Circulating miRNAs in a Mouse Model of Nerve Allograft Transplantation under FK506 Immunosuppression by Illumina Small RNA Deep Sequencing.

Background

Methods

Results

Conclusions

Publications

Identification of Circulating miRNAs in a Mouse Model of Nerve Allograft Transplantation under FK506 Immunosuppression by Illumina Small RNA Deep Sequencing.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets