Temporal and spatial expression of tissue inhibitors of metalloproteinases 1 and 2 TIMP-1 and -2 in the bovine corpus luteumReportar como inadecuado




Temporal and spatial expression of tissue inhibitors of metalloproteinases 1 and 2 TIMP-1 and -2 in the bovine corpus luteum - Descarga este documento en PDF. Documentación en PDF para descargar gratis. Disponible también para leer online.

Reproductive Biology and Endocrinology

, 1:85

First Online: 07 November 2003Received: 03 October 2003Accepted: 07 November 2003

Abstract

The matrix metalloproteinases MMPs and their endogenous inhibitors, tissue inhibitors of metalloproteinases TIMPs, may mediate the dramatic structural and functional changes in the corpus luteum CL over the course of its life span. In addition to regulating MMP activity, TIMPs are also involved in a variety of cellular processes, including cell proliferation and steroidogenesis. In a series of initial studies, we determined that matrix metalloproteinase inhibitory activity was present in protein extracts from early 4 days old, estrus = day 0, mid 10–12 days old and late 16 days old CL n = 3 for each stage. Reverse zymography revealed four metalloproteinase inhibitory protein bands with relative molecular masses that are consistent with those reported for TIMP-1 to -4. In order to gain a better understanding of TIMPs and their role in luteal function, we further characterized this inhibitory activity with a particular focus on the temporal and spatial expression of TIMP-1 and TIMP-2 in the bovine CL. Northern blotting revealed that the TIMP-1 transcript 0.9 kb was expressed at a higher p < 0.05 level in early and mid cycle CL than in the late stage. In contrast, two TIMP-2 mRNA species, one major 1 kb species and one minor 3.5 kb species, were significantly p < 0.05 increased in the mid and late cycle CL than in the early. Western blotting analyses demonstrated no differences in TIMP-1 29 kDa protein levels between early and mid stages, while its levels decreased p < 0.05 from the mid to late stage CL. Conversely, TIMP-2 22 kDa protein was detected at a low level in the early CL, but significantly p < 0.05 increased in the mid and late stages. Immunohistochemistry revealed that both TIMP-1 and -2 were localized to large luteal cells from all three ages of CL. TIMP-1 was also localized in capillary smooth muscle cells, while TIMP-2 was restricted to the endothelial cells in the capillary compartment. In conclusion, the different temporal expression patterns of TIMP-1 and TIMP-2 suggest that TIMP-1 may be important for luteal formation and development, while TIMP-2 may play significant roles during luteal development and maintenance. Furthermore, the distinct localization of these two inhibitors in the vascular compartment indicates that they may serve diverse physiological functions during different stages of luteal angiogenesis.

Electronic supplementary materialThe online version of this article doi:10.1186-1477-7827-1-85 contains supplementary material, which is available to authorized users.

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Autor: Bo Zhang - Marsha A Moses - Paul CW Tsang

Fuente: https://link.springer.com/



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