Yang M and Kurkinen M (1998), Cloning and developmental regulation of tissue ...

XB-ART-14966

Gene 1998 Apr 28;2111:95-100. doi: 10.1016/s0378-1119(98)00077-8.

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Cloning and developmental regulation of tissue inhibitor of metalloproteinases-3 (TIMP3) in Xenopus laevis early embryos.

Yang M , Kurkinen M .

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We cloned a cDNA encoding tissue inhibitor of metalloproteinases-3 (TIMP3) from the frog Xenopus laevis. Similar to TIMP3 from other species, Xenopus TIMP3 has 188 residues including 12 conserved cysteines and Asn184, a putative site for N-linked sugars. Xenopus TIMP3 is 84% identical with human TIMP3. As shown by Northern blotting and RT-PCR, Xenopus TIMP3 mRNA is maternally inherited in eggs and midblastula (stage 8) embryos, downregulated in gastrula and then upregulated in neurula and pretailbud embryos. In select adult tissues, TIMP3 mRNA is present in heart, muscle, liver, skin, intestine and ovaries. These results suggest that TIMP3 is involved in the regulation of expression of matrix metalloproteinases in Xenopus early development and adult tissue remodeling.

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Species referenced: Xenopus laevis
Genes referenced: rpl8 timp3

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	Fig. 1. Xenopus TIMP3. Sequence comparison of human, mouse, rat, chicken, bovine and Xenopus TIMP3. SP is signal peptide cleavage site. Primers show two conserved TIMP sequences that were used in degenerate primer designed and TIMP3 cloning. Consensus shows identical amino acids in all six sequences. Twelve conserved cysteines are shown in bold. In human TIMP3, boxes refer to point mutations that all generate cysteine and cause a rare dominantly inherited eye disease called Sorsby fundus dystrophy ( Felbor et al., 1997). Circle is a conserved Asn residue and potential site for N-linked surgars.
	Fig. 3. Adult expression of Xenopus TIMP3 mRNA. RT-PCR with specific primers for Xenopus TIMP3 using total RNA (0.5 μg) of select adult Xenopus tissues. PCR of 32 cycles. In a control, specific primers for the Xenopus ribosomal protein rpL8 were used. PCR of 27 cycles. Lane M shows DNA size markers.
	Fig. 3. Adult expression of Xenopus TIMP3 mRNA. RT-PCR with specific primers for Xenopus TIMP3 using total RNA (0.5 μg) of select adult Xenopus tissues. PCR of 32 cycles. In a control, specific primers for the Xenopus ribosomal protein rpL8 were used. PCR of 27 cycles. Lane M shows DNA size markers.