The effects of age on the circadian clock system have been extensively studied, mainly in two rodent species, the laboratory rat and the golden hamster. However, less information is available on how aging alters circadian rhythmicity in a commonly studied rodent animal model, the mouse. Therefore, in the present study we compared the rhythm of wheel-running activity in adult (6–9 mo) and old (19–22 mo) C57BL/6J mice maintained under different lighting conditions for a period of 4 mo. During this period, mice were subjected to phase advances and phase delays of the light-dark (LD) cycle and eventually to constant darkness (DD). In LD (12 h light, 12 h dark), old mice exhibited delayed activity onset relative to light offset and an increase in the variability of activity onset compared with adult mice. After a 4-h phase advance of the LD cycle, old mice took significantly longer to reentrain their activity rhythm when compared with adult animals. Old mice also demonstrated a decline in the number of wheel revolutions per day and a tendency toward a decrease in the length of the active phase. An increase in fragmentation of activity across the 24-h day was obvious in aging animals, with bouts of activity being shorter and longer rest periods intervening between them. No age difference was detected in the maximum intensity of wheel-running activity. In DD, the free-running period was significantly longer in old mice compared with adults. In view of the rapidly expanding importance of the laboratory mouse for molecular and genetic studies of the mammalian nervous system, the present results provide a basis at the phenotypic level to begin to apply genetic methods to the analysis of circadian rhythms and aging in mammals.

Opiate peptides are thought to modulate the pattern of LH release in female rats. We tested the hypothesis that changes in proopiomelanocortin (POMC) gene expression occur in proestrous (PRO) and ovariectomized (OVX) steroid-treated rats which may explain their unique patterns of LH secretion. Using in situ hybridization, we examined whether diurnal changes in POMC gene expression occur in the arcuate nucleus. Four groups of rats were used in this study. 1) PRO rats were used after exhibiting at least two consecutive 4-day estrous cycles; 2) OVX rats were killed 9 days after ovariectomy; 3) estradiol (E2)-treated rats were OVX for 7 days and then treated for 2 days; and 4) E2-progesterone (P4)-treated rats were treated with E2 as described above, and on day 9 at 1030 h, P4 was administered. Rats were killed at 2300, 0300, 1000, 1300, 1500, 1800, or 2300 h, beginning on the evening of diestrous day 2 or day 8 after ovariectomy. POMC gene expression exhibited a diurnal rhythm on PRO. Levels of mRNA rose during the morning, peaked between 0300-1000 h, and decreased by 2300 h. In E2-treated rats, which exhibited a LH surge similar in timing to the PRO surge, POMC mRNA levels exhibited a diurnal rhythm strikingly similar to that observed in PRO animals. OVX abolished the rhythm; however, average POMC mRNA levels across the 24-h period were not significantly different from those in PRO or E2-treated rats. P4 treatment increased POMC mRNA levels by 2300 h compared to those in all other experimental groups.(ABSTRACT TRUNCATED AT 250 WORDS)

Reproductive activity in the Djungarian hamster is controlled by seasonal variations in day length. Exposure to long days stimulates testis development, while exposure to short days induces testis regression. We recently found that testis regression after gonadotropin deprivation in rats is associated with increases in apoptosis. Here we sought to determine whether or not apoptosis is associated with the testis regression and/or recrudescence that occurs naturally in seasonally breeding mammals. Newborn male hamsters were maintained on long days (16L:8D) until 3 wk of age before being transferred to short days (8L:16D). Following decreases in serum FSH within 3 days of exposure to short days, testis weight decreased by 52% at Day 10, reaching a 70% decrease after 21 days. Analysis of testis cell DNA fragmentation showed a 4.9-fold increase of low-molecular-weight DNA as early as 5 days after transfer to short days; this was followed by a time-dependent decrease. The observed increases in testis cell apoptosis were correlated with decreases in serum testosterone, but decreases in Leydig cell LH receptor content were delayed. In a second study, 6-wk-old hamsters with regressed testes due to a 3-wk exposure to short days were transferred back to long days. After increases in serum FSH within 3 days of photostimulation, a 2-fold elevation in testis weight was found at Day 5. The increase in testis weight was associated with a 65% decrease of testis apoptosis within 5 days of photostimulation. Also, increases in serum testosterone and LH receptor content were observed after 5 and 10 days of exposure to long days, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)

Aging of the female reproductive system in rats is marked by discrete stages in the disappearance of regular estrous cyclicity. We determined at which point in the transition to acyclicity changes in the LH pulse generator could be detected by comparing pulsatile LH release in ovariectomized young rats to three groups of middle-aged rats that were chronologically matched, yet exhibited different stages of reproductive senescence. Since changes in LH pulse amplitudes can result from hypothalamic and/or pituitary factors, we also assessed pituitary responsiveness to exogenous GnRH. Young rats (2-3.5 months) that had shown regular 4- or 5-day estrous cycles and middle-aged rats (9.5-12 months) that exhibited either regular cycles, irregular cycles, or persistent estrus were bilaterally ovariectomized and used 4 weeks later. Rats were implanted with right atrial cannulae and were bled 2 days later at 5-min intervals for 3 h. To test pituitary responsiveness to GnRH, rats were bled at 10-min intervals for 3 h and received GnRH (25 ng/100 g BW, iv) after the first and second hours. The mean inter-peak interval increased in middle-aged irregularly cycling and persistent estrous rats. The frequency distribution of inter-peak intervals was already significantly different in middle-aged regularly cycling compared to that in young regularly cycling rats. Middle-aged rats displayed fewer short inter-peak intervals and a greater frequency of longer intervals between LH pulses. LH pulse duration increased gradually in parallel with increasing reproductive senescence. Pulse amplitude decreased in all groups of middle-aged rats regardless of their prior reproductive status. Mean LH concentrations were significantly lower in middle-aged than in young regularly cycling rats, and a further significant decline was detected in the middle-aged irregularly cycling and persistent estrous groups. Middle-aged regularly and irregularly cycling rats showed decreased pituitary responsiveness to GnRH compared to young regularly cycling rats, whereas middle-aged persistent estrous rats displayed an intermediate level of pituitary responsiveness to GnRH. These data are the first evidence of changing pulse generator function in middle-aged rats tha had previously exhibited no change in the regularity of their estrous cycles. We suggest that such changes may play a role in the age-related transition to acyclicity.