Generation of a conditionally null allele of the laminin alpha1 gene.

Laminins are heterotrimeric glycoproteins of the basement membranes. Laminin 1 (alpha1, beta1, gamma1) is the major laminin expressed during early mouse embryogenesis. To gain access to the physiological function of laminin alpha1 chain, we developed a conditionally null allele of its encoding gene (Lama1) using the cre/loxP system. Floxed-allele-carrying mice (Lama1(flox/flox)) display no overt phenotype. Lama1(flox/flox) mice were crossed with transgenic deleter mice (CMV-Cre) to generate Lama1-deficient mice (Lama1(Delta/Delta)). Lama1(Delta/Delta) embryos die during the early postimplantation period after embryonic day 6.5. They lack Reichert's membrane, an extraembryonic basement membrane in which laminin alpha1 is normally highly expressed. In parallel, Lama1(Delta/Delta) embryos display 1) parietal and visceral endoderm differentiation defects with altered expression of cytokeratin 19 and GATA4, respectively, and 2) an induction of apoptosis. This new mouse model is of particular interest as it will allow time- and tissue-specific inactivation of the Lama1 gene in various organs.

Oligodendrocytes express different isoforms of beta-amyloid precursor protein in chemically defined cell culture conditions: in situ hybridization and immunocytochemical detection.

The expression of beta-amyloid precursor protein (betaAPP) by astrocytes is well documented; however, data concerning oligodendrocytes remain controversial. The main goal of the present study was to determine whether or not oligodendrocytes in culture constitutively express the different betaAPP isoforms. Oligodendrocytes were cultured in a chemically defined medium that avoids putative effects of unknown serum factors on oligodendrocyte development. We have employed immunocytochemistry and in situ hybridization with antibodies and synthetic oligonucleotides recognizing, respectively, specific protein epitopes and mRNA transcripts of rat betaAPP isoforms. Oligodendrocytes, in both mixed primary cultures in the presence of serum or in secondary cultures in defined medium, were clearly labeled by antibodies directed to different betaAPP sequences. Antibodies against the serine protease inhibitor domain of betaAPP, also strongly labelled oligodendrocytes. Immunohistochemistry and in situ hybridization were combined to determine precisely the expression of different isoforms of betaAPP. In situ hybridization revealed the presence in oligodendrocytes of mRNA transcripts coding not only for betaAPP695 but also for betaAPP770 and betaAPP751. This indicates that betaAPP immunoreactivity found in oligodendrocytes corresponds to constitutive expression of betaAPP. Oligodendrocyte cultured in chemically defined medium are able to express not only betaAPP695 but also betaAPP770, betaAPP751 isoforms containing the Kunitz protease inhibitor domain. Although the role of betaAPP in the pathological processes of Alzheimer's disease (AD) remains unknown, possible disturbances of betaAPP processing and/or synthesis in oligodendrocytes may account for some myelin disorders observed in AD and other senile dementias.