Minisequencing with acyclonucleoside triphosphates tethered to lanthanide(III) chelates.

Four acyclic nucleoside triphosphates (derivatives of cytosine, thymine, 7-deazaadenine, and 7-deazaguanine) labeled with nonluminescent europium, terbium, dysprosium, and samarium chelates of 2,2',2'',2'''-[[4-(4-isothiocyanatophenyl)ethyl]pyridine-2,6-diyl]bis(methylenenitrilo)]tetrakis(acetic acid) were applied to minisequencing using two mutations (Delta F 508 and 1717-1 G to A) of cystic fibrosis as a model system. When synthetic targets were used, all four alleles involved could be analyzed in a single reaction using four terminating substrates labeled with four different lanthanide(III) chelates and DELFIA technology for detection. Blood spot samples without DNA isolations were used for PCR amplification and genotyping the target mutations by minisequencing.

Transgenic mice expressing p450 aromatase as a model for male infertility associated with chronic inflammation in the testis.

Our previous studies have shown that transgenic male mice expressing human P450 aromatase (AROM+) are infertile. In the present study, we followed the testis phenotype up to 15 months of age in these mice. The testes of the old AROM+ mice showed Leydig cell hypertrophy and hyperplasia, as indicated by the staining for steroidogenic enzymes and androgen and estrogen receptors.

Stressor-dependent regulation of the heat shock response in zebrafish, Danio rerio.

Heat shock transcription factors (HSFs) regulate expression of heat shock proteins (Hsps). We have previously shown that in zebrafish a unique isoform, zHSF1b, disappears concomitant with heat shock-induced Hsp70 expression. To characterize the role of zHSF1a and zHSF1b isoforms in the regulation of the stress response in vivo, we have carried out cadmium (10-100 microM) and copper (10-30 microM) exposures in order to specify whether the disappearance of HSF1b is specific for heat stress.