Progress 09/01/13 to 08/31/16
Outputs
Target Audience:Target audience reached during this reporting period includes formal classroom instruction, undergraduates completing research fellowships and the univerity scientific community. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?One Ph.D. studentworked onthis project as part of their dissertation. Four undergraduate students have been trained under this project as part of an undergraduate research fellowship. How have the results been disseminated to communities of interest?To date, results have been disseminated to the scientific community through an undergraduate research poster symposium. We are working on a manuscript for publication. What do you plan to do during the next reporting period to accomplish the goals?
Nothing Reported
Impacts
What was accomplished under these goals?
To date, we conducted experiments on capacitated, capacitated with mitochondrial inhibitor and control incubated bovine sperm (n=9 bulls). Associated FITC-PSA capacitation assay has been conducted to confirm the capacitations tatus of each sample. We conducted RNA-SEQ on each population but we were unable to obtain enough reads to conduct reliable bioinformatic analysis to compare the populations and test our hypothesis to identify if there were differences in the amount of transcripts in capacitated vs uncapacitated samples. We completed qPCR analysis of 12 candidate capacitation transcripts that were identified from previous publications. These results indicated variation in transcript amount after capacitation when compared to control and there was not a clear pattern of transcript increase or decrease with capacitation. We cloned 6 spermatozoal transcripts for production of RNA in situ hybridization probes to complete the localization studies in Aim 2 but have been unable to generate successful probes to date. Finally, due to the variation we found in the qPCR results on capaciated samples, we have tested different RNA isolation methods to determine if that was a source of variation. Use of Trizol vs. RNeasy RNA isolation methods yield different RNA concentration and purity that warrant further investigation into the contribution into variations in RNA-SEQ sperm profiles that were found in capacitated and non-capacitated samples.
Publications
- Type:
Other
Status:
Other
Year Published:
2015
Citation:
Toler C, Card C, Sartini BL. 2015. Comparison of RNA Isolation Methods from Bovine Sperm. Poster Presentation at the Coastal Fellow Undergraduate Scientific Poster Presentation, University of Rhode Island.
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Progress 09/01/14 to 08/31/15
Outputs
Target Audience:To date, results have been disseminated to the scientific research community by participation in an undergraduate research poster symposium. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?Three undergraduate students have been trained under this project as part of an undergraduate research fellowship. Two students presented results from their work in December 2013 and December 2014. The third student will present results in December 2015. How have the results been disseminated to communities of interest?To date, results have been disseminated to the scientific research community through an undergraduate research poster symposium. What do you plan to do during the next reporting period to accomplish the goals?In the next reporting period, bioinformatic analysis of the RNA-Seq experiments will be completed to identify further candidate transcripts for qPCR analysis, in situ localization (Aim 2) and protein expression (Aim 3).
Impacts
What was accomplished under these goals?
To date, we have conducted experiments on capacitated, capacitated with mitochondrial inhibitor and control incubated bovine sperm (N=9 bulls). Associated FITC-PSA capacitation assay has been conducted to confirm capacitation status of each sample. We are currently conducting bioinformatic analysis of RNA-Seq reads from the three experimental groups to identify if spermatozoal transcripts are decreased during capacitation providing information about the function of spermatozoal RNAs. We have also completed qPCR analysis of 12 candidate capacitation transcripts that were identified from previous publications. These results indicate variation in transcript amount after capacitation compared to control. More target transcripts, as identified from RNA-SEq experiments, need to be tested to determine if a pattern of transcript use is present. We have also cloned 6 spermatozoal transcripts for production of RNA in situ hybridization probes to complete the localizations studies described in Aim 2.
Publications
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Progress 09/01/13 to 08/31/14
Outputs
Target Audience: To date, results have been disseminated to the scientific research community by participation in an undergraduate research poster symposium. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided? Two undergraduate students have been trained under this project as part of an undergraduate research fellowship. One student presented a poster of the results in December 2013 and continued working on this project to train a new undergraduate student in the summer of 2014. The second undergraduate student will present results of their work on this project in a poster session in December 2014. How have the results been disseminated to communities of interest? To date, results have been disseminated to the scientific research community through undergraduate research poster symposium. What do you plan to do during the next reporting period to accomplish the goals? In the next reporting period, the RNA-Seq data will be analyzed to identify further candidate transcripts for qPCR analysis. Once targets have been identified, we will commence work on Aims 2 and 3.
Impacts
What was accomplished under these goals?
To date, we have conducted experiments on capacitated, capacitated with mitochondrial inhibitor and control incubated sperm (N=9 bulls). Associated FITC-PSA capacitation assay has been conducted to confirm capacitation status of each sample. Isolated RNA from each treatment has been submitted for RNA-Seq analysis to compare of the transcript profiles of the three treatments to identify candidate transcripts for further validation. We have also completed qPCR analysis on 9 candidate transcripts, identified from previous publications.
Publications
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