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Membrane type-1 matrix metalloproteinases and tissue inhibitor of metalloproteinases-2 RNA levels mimic each other during Xenopus laevis metamorphosis.

Walsh LA, Carere DA, Cooper CA, Damjanovski S - PLoS ONE (2007)

Bottom Line: Here, using RT-PCR analysis, we examine the metamorphic RNA levels of two membrane-type MMPs (MT1-MMP, MT3-MMP), two TIMPs (TIMP-2, TIMP-3) and a potent gelatinase (Gel-A) that can be activated by the combinatory activity of a MT-MMP and a TIMP.In the metamorphic tail and intestine the RNA levels of TIMP-2 and MT1-MMP mirror each other, and closely resemble that of Gel-A as all three are elevated during periods of cell death and proliferation.Intriguingly, TIMP-3, which has been shown to have anti-apoptotic activity, is found at low levels in tissues during periods of apoptosis.

View Article: PubMed Central - PubMed

Affiliation: Department of Biology, University of Western Ontario, London, Ontario, Canada.

ABSTRACT
Matrix metalloproteinases (MMPs) and their endogenous inhibitors TIMPs (tissue inhibitors of MMPs), are two protein families that work together to remodel the extracellular matrix (ECM). TIMPs serve not only to inhibit MMP activity, but also aid in the activation of MMPs that are secreted as inactive zymogens. Xenopus laevis metamorphosis is an ideal model for studying MMP and TIMP expression levels because all tissues are remodeled under the control of one molecule, thyroid hormone. Here, using RT-PCR analysis, we examine the metamorphic RNA levels of two membrane-type MMPs (MT1-MMP, MT3-MMP), two TIMPs (TIMP-2, TIMP-3) and a potent gelatinase (Gel-A) that can be activated by the combinatory activity of a MT-MMP and a TIMP. In the metamorphic tail and intestine the RNA levels of TIMP-2 and MT1-MMP mirror each other, and closely resemble that of Gel-A as all three are elevated during periods of cell death and proliferation. Conversely, MT3-MMP and TIMP-3 do not have similar RNA level patterns nor do they mimic the RNA levels of the other genes examined. Intriguingly, TIMP-3, which has been shown to have anti-apoptotic activity, is found at low levels in tissues during periods of apoptosis.

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Related in: MedlinePlus

Representative RT-PCR Results of Similar MT1-MMP and TIMP-2 Levels of RNA During Natural and T3 Induced Metamorphosis.PCR products were agarose gel fractionated, stained, and photographed. MT1-MMP and TIMP-2 transcription patterns mimic each other at all stages and conditions. In the intestine, MT1-MMP and TIMP-2 transcripts are poorly detected at stage 56 prior to metamorphosis. Levels of both increase during stages 58, 61 and 63 and then drop to pre-metamorphic levels at stage 65. Conversely, in the tail, transcripts are detectable at stage 56, rise to stage 61, and then begin to drop at stage 63. T3 treatment induces a drop in levels after one day, but an increase after three days for both genes. EF1α RNA levels are shown for all comparable stages and treatments.
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pone-0001000-g003: Representative RT-PCR Results of Similar MT1-MMP and TIMP-2 Levels of RNA During Natural and T3 Induced Metamorphosis.PCR products were agarose gel fractionated, stained, and photographed. MT1-MMP and TIMP-2 transcription patterns mimic each other at all stages and conditions. In the intestine, MT1-MMP and TIMP-2 transcripts are poorly detected at stage 56 prior to metamorphosis. Levels of both increase during stages 58, 61 and 63 and then drop to pre-metamorphic levels at stage 65. Conversely, in the tail, transcripts are detectable at stage 56, rise to stage 61, and then begin to drop at stage 63. T3 treatment induces a drop in levels after one day, but an increase after three days for both genes. EF1α RNA levels are shown for all comparable stages and treatments.

Mentions: All five genes examined displayed increases in RNA levels as intestine metamorphosis commenced (Figure 2, stage 56 vs. 58). During intestine metamorphosis MT1-MMP, TIMP-2 and TIMP-3 RNA levels followed a general ‘M’ pattern, starting low, increasing, decreasing, increasing and then decreasing again as metamorphosis progressed (Figure 2). The first peak of RNA at about stage 58 corresponded to approximately when larval epithelial cells were beginning to undergo apoptosis (Figure 1). Several other MMPs (stomelysin-3, collagenase-3, collagenase-4 and MT1-MMP) have already been described to be expressed during intestine metamorphosis [3], [9], where MMP activation at this stage is involved in remodeling of the basement membrane resulting in the death of overlying cells [5]. Increased TIMP-2 RNA, in conjunction with MT1-MMP may also increase ECM remodeling by possibly activating Gel-A (whose RNA is also increased at this time). Indeed, when representative RT-PCR RNA levels of MT1-MMP and TIMP-2 are compared, they mirror each other in both the metamorphic intestine and tail (Figure 3).


Membrane type-1 matrix metalloproteinases and tissue inhibitor of metalloproteinases-2 RNA levels mimic each other during Xenopus laevis metamorphosis.

Walsh LA, Carere DA, Cooper CA, Damjanovski S - PLoS ONE (2007)

Representative RT-PCR Results of Similar MT1-MMP and TIMP-2 Levels of RNA During Natural and T3 Induced Metamorphosis.PCR products were agarose gel fractionated, stained, and photographed. MT1-MMP and TIMP-2 transcription patterns mimic each other at all stages and conditions. In the intestine, MT1-MMP and TIMP-2 transcripts are poorly detected at stage 56 prior to metamorphosis. Levels of both increase during stages 58, 61 and 63 and then drop to pre-metamorphic levels at stage 65. Conversely, in the tail, transcripts are detectable at stage 56, rise to stage 61, and then begin to drop at stage 63. T3 treatment induces a drop in levels after one day, but an increase after three days for both genes. EF1α RNA levels are shown for all comparable stages and treatments.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC1991586&req=5

pone-0001000-g003: Representative RT-PCR Results of Similar MT1-MMP and TIMP-2 Levels of RNA During Natural and T3 Induced Metamorphosis.PCR products were agarose gel fractionated, stained, and photographed. MT1-MMP and TIMP-2 transcription patterns mimic each other at all stages and conditions. In the intestine, MT1-MMP and TIMP-2 transcripts are poorly detected at stage 56 prior to metamorphosis. Levels of both increase during stages 58, 61 and 63 and then drop to pre-metamorphic levels at stage 65. Conversely, in the tail, transcripts are detectable at stage 56, rise to stage 61, and then begin to drop at stage 63. T3 treatment induces a drop in levels after one day, but an increase after three days for both genes. EF1α RNA levels are shown for all comparable stages and treatments.
Mentions: All five genes examined displayed increases in RNA levels as intestine metamorphosis commenced (Figure 2, stage 56 vs. 58). During intestine metamorphosis MT1-MMP, TIMP-2 and TIMP-3 RNA levels followed a general ‘M’ pattern, starting low, increasing, decreasing, increasing and then decreasing again as metamorphosis progressed (Figure 2). The first peak of RNA at about stage 58 corresponded to approximately when larval epithelial cells were beginning to undergo apoptosis (Figure 1). Several other MMPs (stomelysin-3, collagenase-3, collagenase-4 and MT1-MMP) have already been described to be expressed during intestine metamorphosis [3], [9], where MMP activation at this stage is involved in remodeling of the basement membrane resulting in the death of overlying cells [5]. Increased TIMP-2 RNA, in conjunction with MT1-MMP may also increase ECM remodeling by possibly activating Gel-A (whose RNA is also increased at this time). Indeed, when representative RT-PCR RNA levels of MT1-MMP and TIMP-2 are compared, they mirror each other in both the metamorphic intestine and tail (Figure 3).

Bottom Line: Here, using RT-PCR analysis, we examine the metamorphic RNA levels of two membrane-type MMPs (MT1-MMP, MT3-MMP), two TIMPs (TIMP-2, TIMP-3) and a potent gelatinase (Gel-A) that can be activated by the combinatory activity of a MT-MMP and a TIMP.In the metamorphic tail and intestine the RNA levels of TIMP-2 and MT1-MMP mirror each other, and closely resemble that of Gel-A as all three are elevated during periods of cell death and proliferation.Intriguingly, TIMP-3, which has been shown to have anti-apoptotic activity, is found at low levels in tissues during periods of apoptosis.

View Article: PubMed Central - PubMed

Affiliation: Department of Biology, University of Western Ontario, London, Ontario, Canada.

ABSTRACT
Matrix metalloproteinases (MMPs) and their endogenous inhibitors TIMPs (tissue inhibitors of MMPs), are two protein families that work together to remodel the extracellular matrix (ECM). TIMPs serve not only to inhibit MMP activity, but also aid in the activation of MMPs that are secreted as inactive zymogens. Xenopus laevis metamorphosis is an ideal model for studying MMP and TIMP expression levels because all tissues are remodeled under the control of one molecule, thyroid hormone. Here, using RT-PCR analysis, we examine the metamorphic RNA levels of two membrane-type MMPs (MT1-MMP, MT3-MMP), two TIMPs (TIMP-2, TIMP-3) and a potent gelatinase (Gel-A) that can be activated by the combinatory activity of a MT-MMP and a TIMP. In the metamorphic tail and intestine the RNA levels of TIMP-2 and MT1-MMP mirror each other, and closely resemble that of Gel-A as all three are elevated during periods of cell death and proliferation. Conversely, MT3-MMP and TIMP-3 do not have similar RNA level patterns nor do they mimic the RNA levels of the other genes examined. Intriguingly, TIMP-3, which has been shown to have anti-apoptotic activity, is found at low levels in tissues during periods of apoptosis.

Show MeSH
Related in: MedlinePlus