Literatur

Neumann, Heinz-Dieter

doi:10.1007/978-3-662-08485-4_10

Cited by 1 publication

(1 citation statement)

References 4 publications

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“…CAT activity was measured by the biphasic partition assily of Neumann et al (1987) and P-galactosidase activity by the spectrophotometric method of Nielsen et al (1983).…”

Section: Tissue Culture and Transfection Of Primary Lens Culturesmentioning

confidence: 99%

Molecular analyses of carbonic anhydrase‐II expression and regulation in the developing chicken lens

Buono

Linser

Cuthbertson

et al. 1992

Developmental Dynamics

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The expression of carbonic anhydrase-I1 (CA-11) in the developing chicken lens was examined and compared with that in the retina of the chicken embryo. CA-I1 expression was measured by immunohistochemistry and radioimmunoassay during development, and CA-I1 mRNA was quantified by Northern blot and densitometric scanning and localized by in situ hybridization. A functional promoter of the chicken CA-I1 gene was identified by transfection of primary embryonic chicken lens epithelial cells and analyzed in deletion mutants. The results establish that CA-I1 makes u p about 0.1% of the total soluble protein of the embryonic chicken lens, an amount insufficient to make it a candidate for an enzyme crystallin in this species. Lens fiber differentiation coincided with a loss of CA-I1 mRNA and protein; by contrast, CA-I1 persisted in the epithelial cells of the embryonic and mature lens. This and previous studies showed that CA-I1 amounts to as much as 3% of the protein of the embryonic chicken retina and follows a different developmental time course of expression; like the lens, CA-I1 decreases until day 10 in the embryonic retina, but, unlike the lens, it increases thereafter and plateaus at hatching. Progressive deletions of the 5' flanking regions (from position -1314 to +32) of the CA-I1 gene fused to the bacterial chloramphenicol acetyltransferase (CAT) reporter gene resulted in a gradual loss of promoter activity, consistent with an additive effect of putative cis-regulatory elements found in many crystallin genes. These experiments provide the foundation for a molecular analysis of the developmental and differential regulation of the CA-I1 gene in lens and retina. o 1992 Wiley-Liss, Inc.

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“…CAT activity was measured by the biphasic partition assily of Neumann et al (1987) and P-galactosidase activity by the spectrophotometric method of Nielsen et al (1983).…”

Section: Tissue Culture and Transfection Of Primary Lens Culturesmentioning

confidence: 99%