Immunohistochemical Changes in the Testicular Excurrent Duct System of Healthy, Male Japanese Quail (Coturnix coturnix japonica) Observed at 4, 6–7, 12, and 52 Weeks of Age
Abstract
:1. Introduction
2. Results
2.1. Immunohistochemistry
2.1.1. Cytokeratin 5
2.1.2. Desmin
2.1.3. Fibronectin
2.1.4. Alpha-Smooth Muscle Actin (α-SMA)
2.1.5. Beta-Tubulin
2.1.6. Vimentin
2.2. TUNEL-Stained Sections
3. Discussion
3.1. Immunohistochemistry
3.1.1. Cytokeratin 5
3.1.2. Desmin and α-SMA
3.1.3. Fibronectin
3.1.4. Beta-Tubulin
3.1.5. Vimentin
3.2. TUNEL-Stained Sections
4. Materials and Methods
4.1. Animals and Tissue Preparation
4.2. Immunohistochemistry
4.3. Immunofluorescence for Beta-Tubulin-Cy3 Antibody
4.4. TdT dUTP Nick End Labeling (TUNEL) Assay
4.5. TUNEL-Positive Cell Counting
4.6. Statistical Analysis
5. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Antibody | Clone | Dilution | Antigen Retrieval | Manufactures | Manufacturer’s Recommended Species Reactivity |
---|---|---|---|---|---|
Cytokeratin 5 | Polyclonal rabbit ab53121 | 1:50 | Citrate buffer (pH 6) | Abcam, Cambridge, United Kingdom | Domestic rabbit, human, mouse, rat |
Desmin | Monoclonal mouse ab6322 | 1:300 | Citrate buffer (pH 6) | Abcam, Cambridge, United Kingdom | Human, mouse, rat |
Fibronectin | Polyclonal rabbit ab2413 | 1:250 | Tris-EDTA (pH 9) | Abcam, Cambridge, United Kingdom | Human, mouse |
Alpha-smooth muscle actin | Monoclonal mouse 1A4 | 1:50 | Citrate buffer (pH 6) | DakoCytomation, Glostrup, Denmark | Avian, chicken, goat, human, mouse, non-human primate, pig, rat, sheep, xenopus |
Beta-tubulin | Monoclonal mouse ab44928 | 1:500 | Tris-EDTA (pH 9) | Abcam, Cambridge, United Kingdom | Drosophila, human, melanogaster, mouse, rat, |
Vimentin | Monoclonal mouse 3B4 | 1:25 | Citrate buffer (pH 6) | DakoCytomation, Glostrup, Denmark | amphibian, bovine, canine, chicken, human, monkey |
Age Group | Epithelium | |||
---|---|---|---|---|
Rete Testis | Proximal Efferent Duct | Distal Efferent Duct | Epididymal Duct Unit | |
4-week-old | CK5(+++); SMA(+); T(+++); V(+++) | CK5(+++) a; SMA(+); T([+++] b and [++/+++] c); V(+++) | CK5(+++) a; SMA(+); T([+++] b and [++/+++] c); V(+++) | CK5(+++); SMA(+); T(+++); V(−) |
6–7-week-old | CK5(+++); T(++/+++); V(++/+++) | CK5(++/+++) a; T([+++] b and [++/+++] c); V(++) d | CK5(++/+++) a; T([+++] b and [++/+++] c); V(++) d | CK5(+++); T(++); V(−) |
12-week-old | CK5(+++); T(++/+++); V(++/+++) | CK5(++/+++) a; T([+++] b and [++/+++] c); V(++) d | CK5(++/+++) a; T([+++] b and [++/+++] c); V(++) d | CK5(+++); T(++); V(−) |
52-week-old | CK5(+/++); T(+/++); V(+/++) | CK5(+/++) a; T([+++] b and [+] c); V(+) e | CK5(+/++) a; T([+++] b and [+] c); V(−) | CK5(++); T(+); V(−) |
Age Group | Peri- and Interductal Tissues | |||
---|---|---|---|---|
Smooth Muscle Layer | Blood Vessel | Fibroblast | ||
Endothelial Cell | Tunica media | |||
4-week-old | CK5(+); D(+++); F(++/+++); SMA(+++); T(++/+++) | CK5(++); F(+++); T(+++); V(+++) | CK5(+); D(++); SMA(+++); T(++) | CK5(−); F(++/+++); T(++/+++); V(+++) |
6–7-week-old | CK5(−/+); D(++/+++); F(+/++); SMA(+++); T(++) | CK5(++); F(+++); T(++); V(++) | CK5(+); D(+); SMA(++); T(+) | CK5(−); F(+/++); T(++); V(++) |
12-week-old | CK5(−/+); D(++/+++); F(+/++); SMA(+++); T(++) | CK5(++); F(+++); T(++); V(++) | CK5(−/+); D(+); SMA(++); T(+) | CK5(−); F(+/++); T(++); V(++) |
52-week-old | CK5(−); D(+/++); T(+/++); F(+); SMA(+++) | CK5(+); F(++); T(+); V(+) | CK5(−); D(+); SMA(++); T(+) | CK5(−); F(+); T(−/+), V(−) |
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Ibrahim, M.I.A.; Williams, J.H.; Botha, C.J. Immunohistochemical Changes in the Testicular Excurrent Duct System of Healthy, Male Japanese Quail (Coturnix coturnix japonica) Observed at 4, 6–7, 12, and 52 Weeks of Age. Int. J. Mol. Sci. 2022, 23, 14028. https://doi.org/10.3390/ijms232214028
Ibrahim MIA, Williams JH, Botha CJ. Immunohistochemical Changes in the Testicular Excurrent Duct System of Healthy, Male Japanese Quail (Coturnix coturnix japonica) Observed at 4, 6–7, 12, and 52 Weeks of Age. International Journal of Molecular Sciences. 2022; 23(22):14028. https://doi.org/10.3390/ijms232214028
Chicago/Turabian StyleIbrahim, Mohammed I. A., June H. Williams, and Christo J. Botha. 2022. "Immunohistochemical Changes in the Testicular Excurrent Duct System of Healthy, Male Japanese Quail (Coturnix coturnix japonica) Observed at 4, 6–7, 12, and 52 Weeks of Age" International Journal of Molecular Sciences 23, no. 22: 14028. https://doi.org/10.3390/ijms232214028