Assessment of micro-RNA profiles in small extracellular vesicles isolated from bovine colostrum with different immunoglobulin G concentrations.

1. Assessment of microRNA profiles in small
extracellular vesicles isolated from bovine
colostrum with different immunoglobulin G
concentrations.
Authors:
T. Ma, W. Li, Y. Chen, E. Cobo, C. Windeyer, L. Gamsjäger, Q. Diao, Y. Tu, L. Guan.
Published: July 26, 2022.
Journal Club
Course instructor: Dr. Nichole Anderson
Student: Rafaela Santos

2. Micro RNA in bovine colostrum
INTRODUCTION
Colostrum
Why is it important?
What is colostrum composition?
What are colostrum main functions?

3. Micro RNA in bovine colostrum
INTRODUCTION
What is micro RNA? What are its functions?
Why we should have a better understanding
of it?
Image credit: Sigma aldrich

4. Micro RNA in bovine colostrum
MATERIALS
This study was approved by the university of Calgary veterinary sciences animal
care committee and conducted in accordance with guidelines established by the
Canadian council on animal care.
•Colostrum samples were collected from
•6 beef heifers (first parity) and
•2 beef cows (third or higher parity) from
Mi RNA expression analysis challenges:
Source preparation: lack of standardized protocols
Detection techniques: sensitivity at single-base level
Normalization methods: universal normalizer

5. Micro RNA in bovine colostrum
METHOD
Defining the best purification + RNA isolation method for this experiment:
Image source: Micro RNA in bovine colostrum

6. Micro RNA in bovine colostrum
METHOD
Best results: purification kit + RNA isolation kit = Qiagen
1 - radial immunodiffusion
2- the 8 s-EV associated mi RNA libraries were constructed using Qiagen kit.
In brief, 100 ng of total RNA was ligated with adapters sequentially to the 3′ and
5′ ends of mi RNA, cDNA synthesis, followed by amplification of 14 cycles.
Image source: https://www.qiagen.com/us/products/

7. Micro RNA in bovine colostrum
RESULTS
In total, 527 mi RNA (CPM >1) were identified in 8 samples, which 389 mi RNA identified by both methods.
The profiles of the top 50 miRNA were the same and were detected in all 8 samples using mi rdeep2 and sRNA-
bench, which accounted for 91.7% ± 0.44% and 94.3% ± 0.45% of total mi RNA abundance, respectively,
suggesting that both approaches identified a “core” mi RNA in bovine colostrum.
Image source: Micro RNA in bovine colostrum

8. Micro RNA in bovine colostrum
RESULTS
The study speculate that mi RNA regulating the genes involved in the PI3K/AKR, signaling pathway could be key
in affecting physiology in the neonatal calf. The mir-30a, abundant in our colostrum samples, regulates insulin
sensitivity (mclean et al., 2015) and is- let functions (jiang et al., 2017) by signaling PI3K/AKT. Moreover, mir-30a-
5p could regulate glucose metabolism through PI3K/akt signaling in neonatal calves, mainly in young calves that
often experience glucose dysbalances (e.G., Postprandial hyperglycemia, hyperinsulinemia, and insulin resistance)
(bach et al., 2013; pantophlet et al., 2016).
Image source: Micro RNA in bovine colostrum

9. Micro RNA in bovine colostrum
RESULTS
Another key role of colostrum in homeostasis:
The establishment of the microbiome in the small intestine (malmuthuge et al., 2015), including the colonization
of beneficial bacteria such as bifidobacterium (song et al., 2019) and lactobacillus (ma et al., 2019b).
Image source: Micro RNA in bovine colostrum

10. Micro RNA in bovine colostrum
RESULTS
Bovine colostrum categorization:
- High
- Low
However, no mi RNA was uniquely identified in all samples from one group except for mi R-758, which was only
expressed in all low-igg samples using mirdeep2 (average CPM = 19). We showed similar mi RNA profiles in
colostrum with high and low-igg concentrations using both approaches.
Image source: Micro RNA in bovine colostrum

11. Micro RNA in bovine colostrum
CONCLUSION
miR CURY Exosome Cell/Urine/ CSF and miRNeasy Mini methods are suitable for sEV-associated mi RNA
examination in bovine colostrum. Both miRDeep2 and sRNAbench identified a similar core mi RNA, which may
play a role in regulating health and immune development functions in newborn calves beyond the known
passive immunity associated with IgG.
Image source: https://www.proteinsynthesis.org/what-is-the-role-of-mrna-in-protein-synthesis/

12. Micro RNA in bovine colostrum
ARTICLE FEEDBACK -
Image source: Micro RNA in bovine colostrum

13. Micro RNA in bovine colostrum
ARTICLE FEEDBACK +
Image source: https://dairy.extension.wisc.edu/
* Updated: Published: July 26, 2022.
* Relevant
* Short article

14. Micro RNA in bovine colostrum
REFERENCES
T. Ma, W. Li, Y. Chen, E. Cobo, C. Windeyer, L. Gamsjäger, Q. Diao, Y. Tu, L. Guan. 2022. Assessment of
microRNA profiles in small extracellular vesicles isolated from bovine colostrum with different
immunoglobulin G concentrations (original research) American Dairy Science Association. https:
https://doi.org/10.3168/jdsc.2022-0225
DOUBLE STRANDED RNA
CREDIT IMAGES :HTTPS://GIPHY.COM/SEARCH/RNA

15. QUESTIONS?
• Rafaela Santo
• rafsanto@ttu.edu