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Research note: Turkey (Meleagris gallopavo) semen CD14+ macrophages - management, key features, and classification -
Ewa M Drzewiecka1, Anna Szóstek-Mioduchowska2, Joanna Wiśniewska3
1Team of Gamete Biology, Institute of Animal Reproduction and Food Research, Polish Academy of Sciences in Olsztyn, Trylińskiego 18, 10-683 Olsztyn, Poland.
Abstract:
Semen macrophages (Mф) play a role in mediating innate immune responses and supporting adaptive immunity within the male reproductive tract. Knowledge about the semen Mф in avian species remains limited. This study aimed to isolate turkey semen Mф and optimize protocols for their characterization based on specific Mф markers, specifically cluster of differentiation 14 (CD14), inducible nitric oxide synthase (iNOS), and interleukin 10 (IL10), for further classification into CD14+ type 1 macrophages (CD14+iNOS+, akin CD14+Mф1) and CD14+ type 2 macrophages (CD14+IL10+, akin CD14+Mф2) subpopulations. In the current study, immunofluorescence (IF) staining and fluorescence-activated cell sorting (FACS) were employed to identify these markers, and the morphological features of CD14+Mф were examined. Our results demonstrated that turkey semen CD14+Mф are spherical, may contain one or more nuclei, and often exhibit large vacuoles. Based on FACS results, CD14+iNOS+(akin CD14+Mф1) accounted for 48.5 ± 12.45 % of CD14+ cells, whereas CD14+IL10+ (akin CD14+Mф2) represented 42.48 ± 14.62 % of them. This study provides a foundational framework for future investigations into the functional roles of turkey CD14+Mф in semen.
Insights
This study isolated and characterized turkey semen macrophages (Mф), identifying two subpopulations: CD14+ type 1 Mф (CD14+iNOS+) and CD14+ type 2 Mф (CD14+IL10+). These findings establish a basis for understanding avian reproductive immunology.
Area of Science:
- Immunology
- Veterinary Science
- Cell Biology
Background:
- Semen macrophages (Mф) are crucial for immune responses in the male reproductive tract.
- Limited knowledge exists regarding avian semen Mф characteristics and functions.
Purpose of the Study:
- To isolate turkey semen Mф and optimize protocols for their characterization.
- To classify Mф into CD14+ type 1 (CD14+iNOS+) and CD14+ type 2 (CD14+IL10+) subpopulations using specific markers.
Main Methods:
- Immunofluorescence (IF) staining to identify macrophage markers.
- Fluorescence-activated cell sorting (FACS) for quantitative analysis and subpopulation classification.
- Morphological examination of isolated turkey semen Mф.
Main Results:
- Turkey semen CD14+ Mф were identified as spherical cells, potentially multinucleated, with large vacuoles.
- CD14+iNOS+ (akin CD14+Mф1) constituted 48.5 ± 12.45% of CD14+ cells.
- CD14+IL10+ (akin CD14+Mф2) comprised 42.48 ± 14.62% of CD14+ cells.
Conclusions:
- This study provides the first detailed characterization of turkey semen macrophages.
- Established protocols enable future functional studies of these Mф subpopulations in avian reproduction.
- The findings contribute to a foundational understanding of male reproductive tract immunity in poultry.
