title: Cell type-restricted expression of erythrocyte tropomodulin Isoform41 in exon 1 knockout/LacZ knock-in heterozygous mice.
abstract: Full-length erythrocyte tropomodulin (E-Tmod or Tmod1) isoform of 41 kDa is an actin nucleation protein and caps the pointed end of tropomyosin-coated actin filaments. It participates in the length control of short actin protofilaments in the erythrocyte membrane skeletal network as well as the organization of microfilaments in non-erythroid cells. Recently we discovered and characterized a truncated isoform of 29 kDa, which lacks the N-terminal sequence encoded by exons 1 and 2 required for nucleation and capping. Thus, it is important to study the expression pattern of solely the E-Tmod41 isoform in tissues. We utilized our exon 1 knockout (KO) mouse model with a knock-in lacZ reporter gene which reports the expression of E-Tmod41, but not E-Tmod29. Because this homozygous isoform-specific KO is an embryonic lethal mutation, we used heterozygous mice. X-gal staining localized specific signals at the single cell level and revealed a timed expression during embryonic development and restricted expression in adult mice. Our results showed that E-Tmod41 expressing cells include developing and young erythroid cells, developing somites, young fiber cells in the lens, certain subtype(s) of tubular cells in the kidney, smooth muscle cells in various tissues, and horizontal cells in the retina. A comparison with previous studies revealed that most if not all tissues known to express E-Tmod contained lacZ-expressing cells. Interestingly, some tubular cells were lacZ-positive while others in the same renal tubule were not, indicating heterogeneity within the tubular cells. Combined with double immunocytochemistry, we further localized E-Tmod41 to dendritic spines of horizontal cells. These timed and cell-type restricted expressions of E-Tmod41 suggest a role of actin nucleation and/or short actin protofilaments in these cell types and sub-cellular structures.
authors.affiliation: Department of Bioengineering, University of California, 9500 Gilman Drive, La Jolla, San Diego, CA 92093-0412, United States.
authors.forename: Weijuan
authors.initials: W
authors.lastname: Yao
authors.affiliation: Department of Bioengineering, University of California, 9500 Gilman Drive, La Jolla, San Diego, CA 92093-0412, United States.
authors.forename: Xin
authors.initials: X
authors.lastname: Chu
authors.affiliation: Department of Bioengineering, University of California, 9500 Gilman Drive, La Jolla, San Diego, CA 92093-0412, United States. Electronic address: amysung@bioeng.ucsd.edu.
authors.forename: Lanping Amy
authors.initials: LA
authors.lastname: Sung
mesh_terms: D000818:Animals; D003713:Dendritic Cells / Q000378:metabolism; D004912:Erythrocytes / Q000166:cytology; D005091:Exons; D007763:Lac Operon / Q000235:genetics*; D051379:Mice; D018345:Mice, Knockout; D018482:Muscle, Skeletal / Q000378:metabolism; D009130:Muscle, Smooth / Q000378:metabolism; D009928:Organ Specificity; D020033:Protein Isoforms / Q000378:metabolism; D012160:Retina / Q000378:metabolism; D051348:Tropomodulin / Q000235:genetics* / Q000378:metabolism*
chemical_list: D020033:Protein Isoforms; D051348:Tropomodulin
keywords: Actin; Dendritic spine; E-Tmod; Horizontal cells; Renal cortex; Smooth muscle cells