Quantitative fluorescence of 5‐FU–treated fetal rat limbs using confocal laser scanning microscopy and Lysotracker Red

Abstract

Recent studies have revealed that an orphan receptor gene of the steroid/thyroid hormone nuclear receptor superfamily, the Nurr1 gene, is essential for the neurogenesis and differentiation of dopaminergic neurons in the midbrain of mice. Transgenic mice lacking the Nurr1 gene soon die after birth and are devoid of dopaminergic neurons in the midbrain. Heterozygous mice survive postnatally without obvious locomotor deficits; however, they have increased vulnerability to dopaminergic neurotoxin 1‐methyl‐4‐phenyl‐1,2,3,6‐tetrahydropyridine (MPTP). In view of the importance of dopamine neurotransmission in brain function, we were interested to know if the human homologous gene of murine Nurr1, the NR4A2 gene, may play a role in the pathogenesis of schizophrenia. We systematically sequenced all the exons of the human NR4A2 gene to search for molecular variants in a cohort of Chinese schizophrenic patients from Taiwan. Two molecular variants were identified: a G‐insertion in intron 6 (designated IVS6 + 17∼ + 18insG), and a G‐deletion in the untranslated exon 1 (designated c.‐469delG). The IVS6 + 17∼ + 18insG is a polymorphic one; further case control study, however, did not reveal association of this polymorphism with schizophrenia. The c.‐469delG is a rare variant found in two unrelated patients among 177 schizophrenic patients, but not in 130 nonpsychotic controls. The result suggests that the c.‐469delG and possibly other variants of the NR4A2 gene may be of relevance to the complex factors involved in the pathogenesis of schizophrenia. © 2001 Wiley‐Liss, Inc.

Autophagy and endocytosis in the amnion

“…According to published procedures (Price et al, 2003), AE cells on glass slides were stained with LTR (5 lM; Molecular Probes, Eugene, OR, USA) for a period of 30 min at 37°C and washed with PBS. The coverslip was mounted with a drop of PBS on a glass slide.…”

rSac3, a novel Sac domain phosphoinositide phosphatase, promotes neurite outgrowth in PC12 cells

“…No colocalization of rSac3 with the early endosome marker early endosomal antigen 1 (Supplementary information Figure S1D-F) [41], the late endosome marker mannose 6-phosphate receptor Figure S1G-I) [7] or the lipid fraft marker caveolin-1 [42] could be detected. We also used different subcellular compartment markers, such as the mitochondria marker MitoTracker [43] and the lysosome marker LysoTracker [44], to determine the subcellular localization of rSac3, but could not detect any double-positive signal. Similar results were obtained by transfecting EGFP-tagged rSac3 into COS-7 cells (data not shown).…”

Dynamic patterns of apoptosis in the developing chicken heart

“…LTR labels acidic compartments within the apoptotic cell itself as well as in the acidic compartments of healthy cells that are engulfing apoptotic debris (Zucker et al, 2000). This technique has been used previously to study apoptosis in the embryonic rat and mouse (Zucker et al, 1998;Dunty et al, 2001Dunty et al, , 2002Price et al, 2003). While the staining patterns observed using LTR resembled those detected with other apoptosis assays in cardiac tissue, LTR allowed us to detect regions of cell death at earlier stages than was detected by terminal deoxynucleotidyl transferase mediated dUTP nick end labeling (TUNEL) staining in sections.…”