Temporal modulation of the canonical clockwork in the suprachiasmatic nucleus and olfactory bulb by the mammary pheromone 2MB2 in pre-visual rabbits.

During the early stages of development, the olfactory system plays a vital role in the survival of altricial mammals. One remarkable example is the Oryctolagus cuniculus, whose mother-young interaction greatly depends on the 2-methylbut-2-enal (2MB2) pheromone that triggers nipple search and grasping behaviors. Olfactory stimulation with 2MB2 regulates the expression of the core body temperature and locomotor activity rhythms in rabbit pups, indicating the modulation of the circadian system by this volatile cue. To address this issue, in the present study, we determined the effect of stimulation with pulses of 2MB2 on the molecular circadian clockwork in the suprachiasmatic nucleus (SCN) and in the main olfactory bulb (MOB). For this purpose, 7-day-old rabbits were stimulated with distilled water (CON), with ethyl isobutyrate (ETHYL) or with the pheromone (2MB2) at different times of the cycle, and 1h later, the expression of the activity marker C-FOS and of the clock proteins PER1, CRY1 and BMAL1 was evaluated in the SCN and in the three layers of the MOB. The clock proteins were abundantly expressed in both structures; nevertheless these showed diurnal rhythmicity only in the MOB, confirming that central pacemakers exhibit a heterochronical development of the molecular clockwork. C-FOS expression in the SCN and in the MOB was modulated by exposure to ETHYL and to 2MB2 only when these stimulants were presented at ZT00 and at ZT18. In contrast, the clock proteins were essentially modulated by 2MB2 at ZT00 and at ZT06 in both structures. In addition, the PER1 and CRY1 proteins exhibited differential responses to stimulation in the three layers of the MOB. For the first time, we report a modulatory and time-dependent effect of the mammary pheromone 2MB2 on the expression of the core clock proteins in the SCN and in the MOB in rabbits during pre-visual stages of development.

Differential maturation of the molecular clockwork in the olfactory bulb and suprachiasmatic nucleus of the rabbit.

Recent studies suggest that the main olfactory bulb (OB) represents a functional circadian pacemaker. In many altricial mammals, during pre-visual stages of development the olfactory system plays a vital role in their survival. One remarkable example is the European rabbit; the newborns are normally raised in a dark nursery burrow, and the lactating female briefly visits her young approximately once every 24 h. Under these conditions, newborn rabbits depend on the circadian system to anticipate the arrival of the lactating doe as well as on pheromonal cues on the mother's ventrum to locate nipples and suckle efficiently. To investigate the development of the rabbit's circadian system, we characterized the 24-h pattern of expression of clock genes in the OB and suprachiasmatic nucleus (SCN) of pre-visual week-old rabbits and compared this with the pattern of expression in visual juvenile rabbits several weeks after weaning. We report for the first time that Per1, Cry1, and Bmal1 are expressed in the OB of newborn and juvenile rabbits. In addition, the diurnal pattern of clock gene expression develops earlier in the OB than in the SCN of newborn rabbits. Given the early maturation of the molecular clockwork and the biological relevance of this structure during development, it is possible that the OB plays an important role in temporal regulation during pre-visual life in rabbits.