Expression of laminin isoforms in mouse myogenic cells in vitro and in vivo.
نویسندگان
چکیده
The expression of laminin-1 (previously EHS laminin) and laminin-2 (previously merosin) isoforms by myogenic cells was examined in vitro and in vivo. No laminin alpha 2 chainspecific antibodies react with mouse tissues, 50 rat monoclonal antibodies were raised against the mouse laminin alpha 2 chain: their characterization is described here. Myoblasts and myotubes from myogenic cell lines and primary myogenic cultures express laminin beta 1 and gamma 1 chains and form a complex with a 380 kDa alpha chain identified as laminin alpha 2 by immunofluorescence, immunoprecipitation and PCR. PCR from C2C12 myoblasts and myotubes for the laminin alpha 2 chain gene (LamA2) provided cDNA sequences which were used to investigate the in vivo expression of mouse LamA2 mRNA in embryonic tissues by in situ hybridization. Comparisons were made with specific probes for the laminin alpha 1 chain gene (LamA1). LamA2 but not LamA1 mRNA was expressed in myogenic tissues of 14- and 17-day-old mouse embryos, while the laminin alpha 2 polypeptide was localized in adjacent basement membranes in the muscle fibres. In situ hybridization also revealed strong expression of the LamA2 mRNA in the dermis, indicating that laminin alpha 2 is expressed other than by myogenic cells in vivo. Immunofluorescence studies localized laminin alpha 2 in basement membranes of basal keratinocytes and the epithelial cells of hair follicles, providing new insight into basement membrane assembly during embryogenesis. In vitro cell attachment assays revealed that C2C12 and primary myoblasts adhere to laminin-1 and -2 isoforms in a similar manner except that myoblast spreading was significantly faster on laminin-2. Taken together, the data suggest that laminins 1 and 2 play distinct roles in myogenesis.
منابع مشابه
Evalauation of Laminin Expression during Mouse Lens Development
Introduction: Among the components of the extracellular matrix (ECM) and basement membrane (BM), laminitis heterotrimeric glycoprotein (laminin) and collagen type IV are the most important. In a previous study we have examined the role of collagen type IV in the developing lens capsule. The present study aims to determine the appearance and distribution of laminin in the BM and ECM of lenses ...
متن کاملUp-Regulation of Integrinsn α2β1 and α3β1 Expression in Human Foreskin Fibroblast Cells after In-Vitro Infection with Herpes Simplex Virus Type 1
The interaction of Herpes Simplex Virus type 1 (HSV-1) with human fetal foreskin fibroblast (HFFF) cell was studied using a recent isolate of HSV-1 which was propagated in Hep-2 cells. HFFF cells were challenged with HSV-1 with a multiplicity of infection (MOI) of 1 virus/cell for 24 hours. Flow cytometric analysis demonstrated that HSV-1 challenged HFFF cells expressed increased levels of α2β1...
متن کاملIn vitro versus In vivo: Development-, Apoptosis-, and Implantation-Related Gene Expression in Mouse Blastocyst
Background: While mammalian embryos can adapt to their environments, their sensitivity overshadows their adaptability in suboptimal in vitro conditions. Therefore, the environment in which the gametes are fertilized or to which the embryo is exposed can greatly affect the quality of the embryo and consequently its implantation potential. Objectives:</stro...
متن کاملP-67: Quantitative Expression of Pluripotency Specific-Genes in Mouse Blastocysts Produced by In Vitro Fertilization
Background: The efficiency of in vitro fertilization (IVF) is still low to be developed to blastocyst stage probably because of environmental conditions. It is likely that in vitro environment can not exactly mimic in vivo environment due to differences in media, metabolic content, atmospheric composition, temperature and pH. Therefore it may affect embryo quality by changing in embryo gene exp...
متن کاملThe Effect of Laminin and Gelatin Extracellular Matrix on Short-Term Cultivation of Neonate Mouse Spermatogonial Stem Cells
Purpose: To compare the effect of laminin and gelatin on short-term culture of spermatogonial stem cells (SSCs) from neonatal mouse testes.Materials and Methods: Cell suspension containing SSCs were isolated from testes of 6 day-old mice and cultured in the presence of Glial-derived neuroterophic factor (GDNF), Epidermal Growth Factor (EGF) and Basic Fibroblastic Growth Factor (bFGF) on laminin...
متن کاملذخیره در منابع من
با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید
برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید
ثبت ناماگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید
ورودعنوان ژورنال:
- Journal of cell science
دوره 108 ( Pt 12) شماره
صفحات -
تاریخ انتشار 1995