Figure 1

Behavioral and neuropathological examinations of Atg7^flox/flox:TH-Cre mice. (A) Schematic representation of the genetic cross between mice carrying the floxed Atg7 allele (Atg7 F/F) and knock-in mice carrying Cre inserted at the 3’UTR of the TH gene (TH-Cre) to generate Atg7 F/F:TH-Cre (or Atg7^flox/flox:TH-Cre) mice. (B) Runway test of Atg7^flox/flox mice (a) and Atg7^flox/flox:TH-Cre mice. (b) In contrast to the Atg7^flox/flox mice, which exhibited well-coordinated movement and almost no slips of the forepaw or hindpaw from the beam, Atg7^flox/flox:TH-Cre mice could hardly move on the beam and the hindpaw slipped frequently (over 110 weeks of age). In this figure, the hindpaw of the Atg7^flox/flox:TH-Cre mice has slipped off the beam. (c) The number of hindlimb slips was recorded as 85-week-old and 120-week-old mice cross the challenging beam. Data show means ± SE (error bars), and statistical significance was evaluated using Student’s t-test. **P < 0.01. (C) In the accelerating rotarod assay, rotation was accelerated from 3 rpm to 35 rpm over 5 min, and fall latency was recorded, in mice from 95 to 120 weeks of age. Red line, Atg7^flox/flox:TH-Cre mice; blue line, Atg7^flox/flox mice. Data show means ± SE (error bars), and statistical significance was evaluated using Student’s t-test. **P < 0.01. (D) Histological analyses of locus coeruleus (LC) in a 9-month-old Atg7^flox/flox:TH-Cre mouse. Paraffin sections were stained with hematoxylin–eosin (HE) (a) and immunostained for TH (b), ubiquitin (c), and p62 (d). In the each staining (a-d), many inclusions are apparent (arrow). Scale bars: 10 µm. (E) Immunofluorescence labeling of p62 (green) or ubiquitin (red) in the LC of an Atg7^flox/flox:TH-Cre mouse (a–c) and an Atg7^flox/flox mouse (d–f)., Scale bars: 20 µm.