Piprek Rafal P, Kloc Malgorzata, Kubiak Jacek Z
Department of Comparative Anatomy, Institute of Zoology and Biomedical Research, Jagiellonian University, Krakow, Poland.
The Houston Methodist Research Institute, Houston, TX, USA; Department of Surgery, The Houston Methodist Hospital, Houston, TX, USA; University of Texas, MD Anderson Cancer Center, Houston, TX, USA.
Differentiation. 2019 Mar-Apr;106:23-34. doi: 10.1016/j.diff.2019.01.006. Epub 2019 Mar 2.
The extracellular matrix (ECM) proteins play an important role in the establishment of the sex-dependent structure of developing gonads. The matrix metalloproteinases (MMPs) are the major players in the regulation of ECM. Our hypothesis was that the MMPs-dependent regulation of EMC is crucial for the establishment of the correct, either testis or ovary, structure of developing gonad. We cultured developing mouse gonads in vitro in the presence of the MMPs inhibitors (α-2-macroglobulin, leupeptin, phosphoramidon) or the MMPs activator, APMA (4-aminophenylmercuric acetate). These inhibitors and activator inhibit/activate, to a different degree, matrix metalloproteinases, but the exact mechanism of inhibition/activation remains unknown. We found that the MMP inhibitors increased accumulation of ECM in the developing gonads. The α-2-macroglobulin had the weakest, and the phosphoramidon the strongest effect on the ECM and the structure of the gonads. The α-2-macroglobulin caused a slight increase of ECM and did not disrupt the gonad structure. Leupeptin led to the strong accumulation of ECM, resulted in the formation of the structures resembling testis cords in both testes and ovaries, and caused increase of apoptosis and complete loss of germ cells. Phosphoramidon caused the strongest accumulation of ECM, which separated individual cells and completely prevented intercellular adhesion both in the testes and in the ovaries. As a result of aberrant morphology, the sex of the phosphoramidon-treated gonads was morphologically unrecognizable. The APMA - the activator of MMP caused ECM loss, which led to the loss of cell adhesion, cell dispersion and an aberrant morphology of the gonads. These results indicate that the ECM accumulation is MMPs-dependent and that the correct amount and distribution of ECM during gonad development plays a key role in the formation of the gonad structure.
细胞外基质（ECM）蛋白在发育中的性腺性别依赖性结构的建立中起着重要作用。基质金属蛋白酶（MMPs）是ECM调节的主要参与者。我们的假设是，MMPs依赖性的EMC调节对于发育中性腺正确的睾丸或卵巢结构的建立至关重要。我们在存在MMPs抑制剂（α-2-巨球蛋白、亮抑酶肽、磷酰胺素）或MMPs激活剂APMA（4-氨基苯基汞乙酸盐）的情况下，体外培养发育中的小鼠性腺。这些抑制剂和激活剂以不同程度抑制/激活基质金属蛋白酶，但其抑制/激活的确切机制仍不清楚。我们发现MMP抑制剂增加了发育中性腺中ECM的积累。α-2-巨球蛋白对ECM和性腺结构的影响最弱，而磷酰胺素的影响最强。α-2-巨球蛋白导致ECM略有增加，并未破坏性腺结构。亮抑酶肽导致ECM强烈积累，在睾丸和卵巢中均导致形成类似睾丸索的结构，并导致细胞凋亡增加和生殖细胞完全丧失。磷酰胺素导致ECM最强的积累，使单个细胞分离，并完全阻止睾丸和卵巢中的细胞间粘附。由于形态异常，经磷酰胺素处理的性腺的性别在形态上无法识别。MMP的激活剂APMA导致ECM丧失，从而导致细胞粘附丧失、细胞分散和性腺形态异常。这些结果表明，ECM积累是MMPs依赖性的，并且性腺发育过程中ECM的正确数量和分布在性腺结构形成中起关键作用。
