Even when statistical models have extracted a temporal modulation from influences of location and speed (Lepage et al., 2012; MacDonald et al., 2011), it remains possible that temporal tuning occurs only when the animal is moving. In addition, in previous studies when animals remained in a relatively constant location, elapsed time was confounded with the distance the animal traveled Selleckchem IOX1 (the number of steps taken), allowing for the possibility that variations in firing rate reflect an integration of distance along an egocentrically defined path. Indeed,

several theoretical conceptions have proposed that path integration is the primary function of hippocampal networks (Etienne and Jeffery, 2004; McNaughton et al., 1991, 1996, 2006; O’Keefe and Burgess, 2005; Samsonovich and McNaughton, 1997). To fully understand the extent to which time and distance, as well as location, govern hippocampal neuronal

firing patterns, it is critical to disentangle these parameters. Here, we distinguished influences of location, time, and distance by recording from multiple hippocampal neurons as rats ran continuously in place at different learn more speeds on a treadmill placed in the stem of a figure-eight maze (Figure 1). On each trial, the rats entered the central stem of the maze from one of two directions (left or right), and then walked onto the treadmill where they received a small water reward. After a short delay, the treadmill accelerated to a speed randomly chosen from within a predetermined range, and the rats ran in place until the treadmill stopped automatically and another small water reward was delivered. Subsequently the animals finished the trial by turning in the direction opposite from their entry to the stem (spatial alternation) to arrive at a water port at the end of a goal arm. Our strategy in distinguishing behavior, location, time, and distance was to “clamp” the behavior and location

of the animal on the maze, and vary the treadmill speed to decouple the distance the rat traveled from the time spent on the treadmill. Multiple analyses showed that the activity of most hippocampal neurons that were active when the rat was on the treadmill aminophylline could not be attributed to residual variations in location, but were heavily influenced by time and distance. Most neurons were influenced to differing extents by both time and distance, but some were best characterized as representing time but not distance and others as representing distance and not time. During treadmill running, the rats’ heads were consistently facing forward, and 75% of the time spent on the treadmill could be accounted for by an area with a radius of approximately 3.3 cm (average area: 35 cm2; standard deviation: 15.9 cm2; range: 12 to 59 cm2). This indicates that the location of the rats’ heads were generally consistent despite fluctuations in position due to side-to-side, forward, and backward shifts on the treadmill.