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）蛋白在发育性腺的性别依赖性结构的建立中起着重要作用。基质金属蛋白酶（MMP）是ECM调节的主要参与者。我们的假设是，EMC的MMP依赖性调节对于建立性腺的正确睾丸或卵巢结构至关重要。我们在体外培养了发育中的小鼠性腺在存在MMP抑制剂（α-2-巨球蛋白，亮肽素，磷酰胺）或MMP激活剂APMA（4-氨基苯基乙酸汞）的情况下。这些抑制剂和活化剂在不同程度上抑制/活化基质金属蛋白酶，但是抑制/活化的确切机制仍然未知。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，而磷酰胺对性腺的结构影响最大。α-2-巨球蛋白引起ECM略有增加，并且不破坏性腺结构。Leupeptin导致ECM的强积累，导致在睾丸和卵巢中形成类似于睾丸索的结构，并导致凋亡增加和生殖细胞完全丧失。磷酰胺引起最强的ECM积累，ECM分离单个细胞并完全阻止睾丸和卵巢中的细胞间粘附。由于形态异常，经磷酰胺处理的性腺的性别在形态上无法识别。APMA-MMP的激活剂导致ECM丢失，从而导致细胞粘附，细胞分散和性腺异常形态丧失。这些结果表明，ECM的积累是依赖MMPs的，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。经磷酰胺处理的性腺的性别在形态上无法识别。APMA-MMP的激活剂导致ECM丢失，从而导致细胞粘附，细胞分散和性腺异常形态丧失。这些结果表明，ECM的积累是依赖MMPs的，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。经磷酰胺处理的性腺的性别在形态上无法识别。APMA-MMP的激活剂导致ECM丢失，从而导致细胞粘附，细胞分散和性腺异常形态丧失。这些结果表明，ECM的积累是依赖MMPs的，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。