Source: TEXAS A&M UNIVERSITY submitted to NRP
LASER CAPTURE MICRODISSECTION (LCM) SYSTEM FOR ANIMAL RESEARCH
Sponsoring Institution
National Institute of Food and Agriculture
Project Status
ACTIVE
Funding Source
Reporting Frequency
Annual
Accession No.
1029311
Grant No.
2022-70410-38426
Cumulative Award Amt.
$211,000.00
Proposal No.
2022-06403
Multistate No.
(N/A)
Project Start Date
Sep 1, 2022
Project End Date
Aug 31, 2026
Grant Year
2022
Program Code
[EGP]- Equipment Grants Program
Recipient Organization
TEXAS A&M UNIVERSITY
750 AGRONOMY RD STE 2701
COLLEGE STATION,TX 77843-0001
Performing Department
(N/A)
Non Technical Summary
Several molecular approaches are currently available for investigating epigenetic, transcriptomic, proteomic, and metabolic alterations in biological samples. However, to elucidate physiological and disease processes, it is critical that these studies provide spatial information by analyzing samples obtained from well-defined organ substructures and/or homogenous populations of cells. This is particularly relevant for tissues with great cellular heterogeneity, such as the brain, uterus, placenta, and other reproductive organs. Laser capture microdissection (LCM) allows the identification and precise removal of specific tissue substructures and cell types withoutcompromising the integrity of samples. LCM provides several advantages compared to other techniques for cell purification, particularly with respect to precision, simplicity, and versatility, since samples can be used for analysis of DNA, RNA, protein, and metabolite content. Theacquisition of a LCM system will allow a large group of federally-funded investigators to generate valuable knowledge related to the spatial pattern of gene and protein expression in reproductive, endocrine, and metabolic tissues from different animals models that benefit agricultural and biomedical research. The LCM system will support ongoing projects, future collaborations and grant applications related to animal physiology, reproduction, and genetics at Texas A&M University, Texas A&M AgriLife Research, and Prairie View A&MUniversity. This project aligns with the following USDA Strategic Goals: Maximize the Ability of American Agricultural Producers to Prosper by Feeding and Clothing the World; Promote American Agriculture Products and Exports; Facilitate Rural Prosperity and Economic Development; and Provide all Americans Access to a Safe, Nutritious, and Secure Food Supply.
Animal Health Component
40%
Research Effort Categories
Basic
60%
Applied
40%
Developmental
(N/A)
Classification

Knowledge Area (KA)Subject of Investigation (SOI)Field of Science (FOS)Percent
3013610102040%
3013310102040%
3013510102020%
Goals / Objectives
The primary objective of of this project is to acquirea LCM system that will allow a large group of federally-funded investigators (USDAand NIH) to generate valuable knowledge related to the spatial pattern of gene and protein expression in reproductive, endocrine, and metabolic tissues from different animals models that benefit agricultural and biomedical research.
Project Methods
All collaborators will be informed of the project timeline and progress throughout the project. Additionally, once the equipment is available for use, emails will be sent to all faculty members in the College of Agriculture and Life Sciences, College of Veterinary Medicine & Biomedical Sciences, College of Medicine, and College of Science at TAMU to enhance collaborations and recruit additional users. Information regarding training, standard operating procedures, and costs to use the equipment will be shared with all potential users. During the first year of use, Dr. Cardoso and members of his laboratory will provide monthly training sessions for new users (PIs and lab members). Dr. Cardoso has previous experience performing laser capture microdissection. Afterwards, training will be offered upon request as new users are identified. We anticipate that 15 to 20 investigators will use the LCM system during the first year of use. This projection is based on the number of investigators who have demonstrated interest in gaining access to the instrument. All users will be requested toproperly acknowledge the public funding (USDA NIFA) in all published articles, manuscripts, dissertations, posters, presentations, inventions, patents, and press releases.

Progress 09/01/23 to 08/31/24

Outputs
Target Audience:Scientific community • Graduate students • Undergraduate students • Livestock producers • Veterinarians and industry professionals Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?Several graduate students (approximately 20 students) and research staff members received training regarding principles of laser capture microdissection and how to properly use the equipment purchased. How have the results been disseminated to communities of interest?Two peer-reviewed manuscripts have been published • Abstracts at scientific meetings (2024Society for the Study of Reproduction, 2024American Society of Animal Science Annual Meeting, and 2023 International Ruminant Reproduction Symposium) • Presentations at conventions attended by beef cattle producers, veterinarians, and industry professionals (2023Texas A&M Beef Cattle Short Course, and Texas A&M AgriLife Extension programs across Texas). What do you plan to do during the next reporting period to accomplish the goals?The LCM system will continue to be available to potential users and new users will receive training as needed. Moreover, new hires will be informed periodically (twice a year) about the opportunity to use this shared equipment. It is expected that several publications acknowledgingthe support from USDA-AFRI-NIFA will be generated in the next reporting period.

Impacts
What was accomplished under these goals? The Laser Capture Microdissection (LCM) System (Manufacturer: Applied Biosystems, ThermoFisher Scientific; Model: Arcturus Cellect Fluorescence) was successfully purchased and installed in January 2023 as planned. Training was provided by ThermoFisher Scientific technicians to all potential users listed in the proposal. Two in-person sessions were offered and approximately 28 researchers (PIs, research staff, and graduate students) received training. The equipment has been available and used by multiple investigators since then. Approximately 8 PIs have used the equipment so far.

Publications

  • Type: Conference Papers and Presentations Status: Accepted Year Published: 2023 Citation: May 2023 - International Ruminant Reproduction Symposium, Galway, Ireland. Impact of pre- and postnatal nutrition on number of kisspeptin- and neurokinin 3 receptor-immunopositive neurons in the arcuate nucleus of sexually mature heifers. Garza V, West SM, Gurule SC, King LN, Doyle KE, Williams GL, Cardoso RC.
  • Type: Conference Papers and Presentations Status: Accepted Year Published: 2023 Citation: May 2023 - International Ruminant Reproduction Symposium, Galway, Ireland. Effects of prenatal and postnatal nutrition on alpha-melanocyte-stimulating hormone neuronal projections to kisspeptin neurons in the arcuate nucleus of beef heifers. West SM, ONeil MMO, Welsh Jr T, Williams GL, Cardoso RC.
  • Type: Peer Reviewed Journal Articles Status: Published Year Published: 2023 Citation: Garza V, West SM, Cardoso RC. Review: Gestational and postnatal nutritional regulation of puberty and subsequent reproductive performance in heifers. Animal, 17: 100782, 2023.
  • Type: Peer Reviewed Journal Articles Status: Published Year Published: 2024 Citation: West SM, Garza V, Cardoso RC. Puberty in beef heifers: effects of prenatal and postnatal nutrition on the development of the neuroendocrine axis. Animal Reproduction, 2024


Progress 09/01/22 to 08/31/23

Outputs
Target Audience:Scientific community • Graduate students • Undergraduate students • Livestock producers • Veterinarians and industry professionals Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?Several graduate students (approximately 15 students) and research staff members received training regarding principles of laser capture microdissection and how to properly use the equipment purchased. How have the results been disseminated to communities of interest?• A peer-reviewed manuscript published in Animal (Garza et al., 2023). • Abstracts at scientific meetings (2023 Society for the Study of Reproduction, 2023 American Society of Animal Science Annual Meeting, and 2023 International Ruminant Reproduction Symposium) • Presentations at conventions attended by beef cattle producers, veterinarians, and industry professionals (2022 Texas A&M Beef Cattle Short Course, and Texas A&M AgriLife Extension programs across Texas). What do you plan to do during the next reporting period to accomplish the goals?The LCM system will continue to be available to potential users and new users will receive training as needed. Moreover, new hires will be informed periodically (twice a year) about the opportunity to use this shared equipment.

Impacts
What was accomplished under these goals? TheLaser Capture Microdissection (LCM) System (Manufacturer: Applied Biosystems, ThermoFisher Scientific; Model: Arcturus Cellect Fluorescence) was successfully purchased and installed in January 2023 as planned. Training was provided by ThermoFisher Scientific technicians to all potential users listed in the proposal. Two in-person sessions were offered and approximately 28 researchers (PIs, research staff, and graduate students) received training. The equipment has been available and used by multiple investigators since then.

Publications

  • Type: Journal Articles Status: Published Year Published: 2023 Citation: Garza, Viviana, Sarah M. West, and Rodolfo C. Cardoso. "Gestational and postnatal nutritional effects on the neuroendocrine control of puberty and subsequent reproductive performance in heifers." animal 17 (2023): 100782.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2023 Citation: May 2023 - International Ruminant Reproduction Symposium, Galway, Ireland. Impact of pre- and postnatal nutrition on number of kisspeptin- and neurokinin 3 receptor-immunopositive neurons in the arcuate nucleus of sexually mature heifers. Garza V, West SM, Gurule SC, King LN, Doyle KE, Williams GL, Cardoso RC.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2023 Citation: May 2023 - International Ruminant Reproduction Symposium, Galway, Ireland. Effects of prenatal and postnatal nutrition on alpha-melanocyte-stimulating hormone neuronal projections to kisspeptin neurons in the arcuate nucleus of beef heifers. West SM, ONeil MMO, Welsh Jr T, Williams GL, Cardoso RC.
  • Type: Conference Papers and Presentations Status: Accepted Year Published: 2022 Citation: September 2022  European Federation of Animal Science (EAAP) Annual Meeting, Porto, Portugal. Nutritional Programming of Puberty in Bos indicus-influenced Heifers. Cardoso RC, West SM, Garza V, Williams GL.