In vivo adenoviral vector-mediated gene transfer into balloon-injured rat carotid arteries

Homozygotes for the dsy1 desynaptic mutant of maize show massive failure of chiasma maintenance during diplotene and diakinesis. Although some chiasmata persist until anaphase I in most microsporocytes expressing this mutant, homozygotes are completely or nearly completely sterile, owing apparently to disjunctive irregularities. Pachytene synaptic errors and some synaptic failure also are found, but recombination nodules are common in homologously synapsed regions, and equational separation of a heterozygous knob into univalents or open arms at diakinesis clearly demonstrates that chiasma failure occurs following crossing-over. A wider than normal synaptonemal complex central region and uniform apparent weakness of central region cross connections to spreading procedures strongly suggest the presence of a genetic lesion in a synaptonemal complex central region component. The dsy1 mutant may provide an especially important source of material for molecular studies on the nature of chiasma maintenance mechanism.     

Intraamniotic administration of an adenoviral vector for gene transfer to fetal sheep and mouse tissues

Replication-deficient adenoviruses have been used to transfer various genes of interest to mammalian tissues in vivo. Effective gene therapy for inborn genetic defects presenting with significant morbidity and mortality at birth will require correction of the defect prenatally. To test the hypothesis that intra-amniotically administered adenovirus transfers gene expression to fetal tissues, replication-deficient human type 5 adenovirus carrying the lacZ gene which encodes nuclear-targeted bacterial beta-galactosidase (Av1LacZ4) was instilled into the amniotic cavity of fetal sheep (10(10) to 1.5 x 10(11) pfu) and fetal mice (10(9) pfu) at 0.8 term gestation. Amniotic membranes and gastrointestinal and respiratory tract tissues were harvested after 3 d, bacterial beta-galactosidase activity was determined by 5-bromo-4-chloro-3-indoyl-beta-D-galactopyranoside (X-gal) enzyme-histochemistry, and tissue integrity was assessed in sections stained with hematoxylin and eosin. Bacterial beta-galactosidase activity was abundant in amniotic membranes and present in lower levels in esophagus, stomach, and small intestine as well as in conducting airways and pulmonary alveoli. To determine whether gene transfer by intraamniotic injection of adenovirus was dose-dependent, Av1Luc1, an adenoviral vector carrying the gene for luciferase (10(5)-10(9) pfu), was injected intraamniotically into fetal mice at 0.8 term gestation. Luciferase activity measured after 3 d in tissue homogenates of Av1Luc1-treated fetal mice revealed a linear dose response in amniotic membranes and gastrointestinal and respiratory tract organs. Intraamniotic administration of an adenoviral gene vector leads to expression of the transferred gene in amniotic membranes as well as in fetal gastrointestinal and respiratory tract tissues in a dose-dependent manner.