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Morais da Silva S
Tomoregulin-1 (TMEFF1) was first identified as a gene implicated in pituitary secretion in Xenopus laevis. The predicted structure of TMEFF1 is that of a transmembrane protein with a highly conserved cytoplasmic tail, two follistatin domains and one modified EGF domain in its extracellular region. We report the cloning of the newt orthologue, and show that the expression of TMEFF1 is upregulated in the blastema during limb regeneration, and is also expressed in mouse embryonic limb development.
Fig. 3. In situ hybridization analysis of TMEFF1 expression during newt limb regeneration and during mouse development. Sagittal section of normal forelimb (a), or sagittal section through a forelimb stump at one day after amputation (b). Note the expression in some of the myofibre nuclei (black arrows). (c) Higher magnification view of a positive myofibre from the forelimb stump shown in (b). (d) Sagittal section through an early blastema, 8 days after amputation. Note the absence of signal in the wound epidermis (arrow). (e) Higher magnification of blastemal cells from the blastema shown in (d). (f) Sagittal section through a midbud blastema at 20 days after amputation. (g) Sagittal section through a palette stage blastema at 35 days after amputation. (h) Higher magnification of blastemal cells from the blastema shown in (g). Note that both positive and negative cells are present. (i) Whole-mount in situ hybridization of 12 and 13 dpc mouse embryos. Note the expression in the forelimb and hindlimb. (j) Higher magnification of the 12 dpc embryo forelimb (left) and hindlimb (right).
Fig. 4. Immunohistochemistry with antibodies to the follistatin modules. (a) Sagittal section of a newt blastema at 20 days after amputation. Note that the majority of the cells from the blastema are positive, and the absence of significant staining in the epidermis. (b) Higher magnification of blastemal cells shown in (a). Expression appears to be high at the periphery of the cells. (c) TMEFF1 protein expression in a transfected COS-7 cell after reaction of the live cells with antibody to the follistatin modules. Note that the signal is indicative of expression at the cell surface. (d) The same cell as (c) showing expression of GFP which was used to identify transfected cells.