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Robust Comparison of Protein Levels Across Tissues and Throughout Development Using Standardized Quantitative Western Blotting
Published on: April 9, 2019
Comparison of MMP-2 and TIMP-2 expressions in yak testes at different ages
Juanjuan Song1,2, Ligang Yuan1,2, Hua Wang1,2
1College of Veterinary Medicine, Gansu Agricultural University, Lanzhou, China.
Abstract:
This study aimed to investigate the distribution and expression of matrix metalloproteinase-2 (MMP-2) and tissue inhibitor of matrix metalloproteinase-2 (TIMP-2) in yak testes. The testes of healthy yaks at different ages: newborn [3 days], young [1 year], adult [4 years], and old [9 years] were collected for microscopic analyses using hematoxylin and eosin staining, immunohistochemistry and immunofluorescence, as well as western blot to compare the expression of MMP-2 and TIMP-2. Furthermore, the levels of MMP-2mRNA and TIMP-2mRNA was detected by real-time quantitative polymerase chain reaction (qPCR). The results of immunohistochemistry and immunofluorescence demonstrated that MMP-2 and TIMP-2 were mainly located in gonocytes of newborn, Sertoli cells of young, spermatozoa of adult and Leydig cells of old. The protein levels of MMP-2 and TIMP-2 exhibited a downward from newborn to adult, but increased again in old yaks. The analysis of qPCR showed that MMP-2 was higher in young compared with newborn or adult(**p < .01), but a lower expression was detected in adult compared with old yak testicular tissues (*p < .05). Compared with adults, TIMP-2 was significantly higher in newborn and young yaks (**p < .01), and slightly higher in old yaks (*p < .05). Hence, The location of MMP-2 and TIMP-2 in gonocytes were associated with the development of newborn yak testes. The expression of MMP-2 and TIMP-2 in Sertoli cells at young and adult yaks suggested that they provided a clue for the regulation of spermatogenesis. The positive labeling of MMP-2 and TIMP-2 in Leydig cells in old yaks suggested that both may be involved in the interstitial metabolism of the testes during this period. This study revealed the possible role of MMP-2 and TIMP-2 in testicular functionality of yaks at different ages.
Insights
This study examined matrix metalloproteinase-2 (MMP-2) and tissue inhibitor of matrix metalloproteinase-2 (TIMP-2) in yak testes across ages. Findings reveal their varied distribution and expression, suggesting roles in testicular development and function.
Area of Science:
- Reproductive Biology
- Molecular Biology
- Animal Science
Background:
- Matrix metalloproteinases (MMPs) and their inhibitors (TIMPs) are crucial for tissue remodeling.
- Understanding MMP-2 and TIMP-2 roles in yak testicular physiology is limited.
- Yak reproductive health is vital for agriculture and conservation.
Purpose of the Study:
- To investigate the localization and expression patterns of MMP-2 and TIMP-2 in yak testes.
- To correlate MMP-2 and TIMP-2 expression with testicular development and aging in yaks.
- To elucidate the functional significance of MMP-2 and TIMP-2 in yak testicular functionality.
Main Methods:
- Tissue collection from yaks of different age groups (newborn, young, adult, old).
- Histological analysis (H&E staining), immunohistochemistry, and immunofluorescence.
- Protein expression analysis via Western blot.
- mRNA expression analysis using real-time quantitative PCR (qPCR).
Main Results:
- MMP-2 and TIMP-2 were localized in gonocytes (newborn), Sertoli cells (young), spermatozoa (adult), and Leydig cells (old).
- Protein levels decreased from newborn to adult, then increased in old yaks.
- qPCR revealed distinct mRNA expression patterns for MMP-2 and TIMP-2 across age groups, with significant differences noted.
Conclusions:
- MMP-2 and TIMP-2 localization in gonocytes is linked to newborn yak testicular development.
- Expression in Sertoli cells suggests a role in regulating spermatogenesis in young and adult yaks.
- MMP-2 and TIMP-2 in Leydig cells may influence interstitial metabolism in aging yaks, highlighting their importance in testicular function.

