Balanced effect of PACAP and FasL on granule cell death during cerebellar development: a morphological, functional and behavioural characterization.

It is now established that the development of the CNS requires equilibrium between cell survival and apoptosis. Pituitary adenylate cyclase-activating polypeptide (PACAP) exerts a powerful protective effect on cerebellar granule cells by inhibiting the caspase 3. In contrast, Fas ligand (FasL) plays an essential role during ontogenesis in eliminating supernumerary neurons by apoptosis. To determine if PACAP and FasL interact during cerebellar development, we characterized the effects of these factors on cerebellar morphogenesis and caspase 3 activity in PACAP+/+ and PACAP-/- mice. First, we demonstrated in vivo that PACAP is able to reverse the diminution of internal granule cell layer thickness induced by FasL in PACAP+/+ and PACAP-/- mice. Second, ex vivo and immunohistochemical studies revealed that interaction between FasL and PACAP occurs through the caspase 3 activity. Third, behavioural study showed a significant difference for the PACAP + FasL group in the righting reflex test at P8 which does not persist at P60. Finally, a time course study revealed that the pro-apoptotic effect of FasL characterized at P8 was followed by a progressive compensatory mechanism in caspase 3 activity and bromodeoxyuridine incorporation. These data suggest that PACAP and FasL interact during cerebellar development to control apoptosis of granule cells and may affect some motor cerebellar functions.

Free versus forced exposure to an elevated plus-maze: evidence for new behavioral interpretations during test and retest.

RATIONALE: The rodent elevated plus-maze is based on an approach/avoidance conflict between secure closed arms and aversive open arms that can be measured to assess anxiety. Despite this apparent simplicity, several discrepancies emerge from the interpretation of an animal's behavior in the maze, especially when considering the one-trial tolerance effect. OBJECTIVES AND METHODS: In order to bring new elements of interpretation, we compared the behavior of rats exposed to the standard version of the test (forced exposure) to the behavior of rats that were allowed to freely explore the apparatus. We also compared the effects of testing/retesting and chlordiazepoxide in these two situations. RESULTS: Our results confirm that open-arm avoidance is a natural tendency and therefore that it is not learned during initial exposure to the maze. In addition, comparison of the two situations suggests that some of the open-arm entries during a forced confrontation with the maze are better interpreted as attempts to avoid the whole situation, rather than as indications of a low level of anxiety. Finally, the one-trial tolerance effect was partially reduced in the free-exposure situation. CONCLUSIONS: Our results contradict the hypothesis that there is acquisition of a phobic-like response to open arms during trial 1. Rather, they are discussed in line with the hypotheses by Rodgers and Shepherd (Psychopharmacology (Berl) 113:237-242, 1993) and Bertoglio and Carobrez (Behav Brain Res 108:197-203, 2000) concerning the acquisition of spatial information about the whole apparatus, leading on trial 2 to an unbalanced approach/avoidance conflict and to the inefficiency of anxiolytic drugs.

Effects of harmaline on anxiety-related behavior in mice.

Harmaline (HA) is a beta-carboline commonly known to provoke motor alterations through activation of cells in the inferior olive. In addition, this pharmacological agent also induces cognitive disturbances such as motor and spatial learning impairments. In order to complete and extend these data, we examined the effects of this drug on state anxiety in mice, employing elevated plus maze test. We report here that lower doses of harmaline (5-10 mg/kg) have anxiogenic since higher doses (20 mg/kg) have anxiolytic-like properties. Overall pattern of our behavioral results provides evidence that harmaline also acts on emotional reactivity in mice by influencing their decision making when placed in an anxiogenic situation.

Further evidences that risk assessment and object exploration behaviours are useful to evaluate emotional reactivity in rodents.

In this study, we tested the ability of risk assessment and exploration behaviours to emphasise PH effects. Indeed, postnatal handling (PH) decreases emotional reactivity in rats but inconsistent behavioural results can be observed and may be due to false negative (i.e. existing effects are not detected). Risk assessment behaviours were measured in the elevated plus maze, in the free exploration paradigm and in the open field. In addition, we measured object exploration behaviours towards familiar/new objects in the open field. PH increased general activity in the elevated plus maze and in the free exploration paradigm and risk assessment behaviours allowed demonstrating that these effects were specific to emotional reactivity. In the open field, PH increased object exploration as early as first exposition while general activity was unaffected. PH also decreased behavioural inhibition in response to the introduction of a novel object. On the whole, our results show that risk assessment and object exploration behaviours are valuable tools to measure more precisely emotional reactivity in rodents. This reinforces the idea that these behaviours should be used more frequently in order to avoid false negative when emotional reactivity changes are expected in unconditioned conflict tests.

The emotional reactivity increase at mid-pregnancy is attenuated in female rats handled during their infancy.

In rats, while pregnancy has been reported to increase emotional reactivity, postnatal handling, consisting in a short daily maternal separation during early life, has been shown to reduce it in several behavioural tests. In the present study, we investigated the potential effects of handling in pregnant rats confronted to various stressful situations. For that purpose, groups of DA/Han female rats were tested either in the elevated plus maze test, in the probe burying test or in a cat exposure test. At the age of 10-14 weeks, control and handled females were mated. Pregnant females were isolated (day 0 of gestation) as well as some non-pregnant females (day 0 of isolation). On day 12, pregnant and non-pregnant females were subjected to one of the three emotional reactivity tests. Our results show that pregnant females (control and handled) were more reactive than non-pregnant ones in two of the three testing situations, namely the elevated plus maze and the probe burying test. In particular, pregnant females spent less time in the open arms of the elevated plus maze than their non-pregnant counterparts and their number of close sniffing towards the probe was decreased in the burying test. Comparatively, beneficial effects of postnatal handling were obvious in the three experiments, both for non-pregnant and pregnant females. We conclude (1) that pregnancy increases emotional reactivity at day 12 of pregnancy, particularly in tests involving low or mildly stressful conditions, and (2) that postnatal handling can partially reverse those effects in all three tests, even in the more stressful situation represented by the cat exposure.

The positive effects of postnatal handling on defensive burying are more obvious in a situation that enlarges the potential coping responses.

A recent study reported few effects of postnatal handling in the defensive burying test. Since the importance of testing conditions has been emphasised in this paradigm, the lack of handling effects in this test could be attributed to the testing situation. Our experiment was carried out to test whether postnatal handling in DA/HAN strain rats have positive effects in two situations of the defensive burying test: a no-hide situation, in which avoidance of the probe was not possible, and a hide situation, in which animals were allowed to avoid the probe by sheltering in a hiding compartment. Our results showed no difference between control and handled rats defensive reactions in the no-hide situation. However, the general coping style of handled rats was different, mostly by a higher level of exploration of the apparatus and of the probe. In the hide situation, the time spent burying the probe was significantly lower in handled rats and they also spent less time in the hiding compartment. Those results, along with approach/avoidance behaviours directed towards the probe demonstrate that postnatal handling do have some positive effects in the defensive burying test. In addition, our results also point out that testing situations in which complex coping strategies are available are appropriate to test the effects of early stimulation such as handling.