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M P Gerkema

Publications and source records attributed to M P Gerkema.

11 recordsLinked to original sources

Being circadian or not: vasopressin release in cultured SCN mirrors behavior in adult voles.

We studied vasopressin (AVP) release patterns from organotypic suprachiasmatic nucleus (SCN) cultures obtained from circadian rhythmic and non-rhythmic voles. All eight SCN cultures made from non-rhythmic voles did not produce any circadian pattern in AVP release, while five out of six SCN cultures of rhythmic voles produced significant (circadian) peak values. The total amount of AVP released was 2-fold higher in SCN cultures from rhythmic vole. These data confirm our previously formulated AVP release deficit hypothesis for nonrhythmic voles, and suggest that AVP in the vole SCN plays an important role in mediating output of its circadian clock, regulating circadian organization of locomotor behavior.

Animals↗

Photic entrainment of circadian activity patterns in the tropical labrid fish Halichoeres chrysus.

Yellow wrasses (Halichoeres chrysus) show clear daily activity patterns. The fish hide in the substrate at (subjective) night, during the distinct rest phase. Initial entrainment in a 12h:12h light-dark (12:12 LD) cycle (mean period 24.02h, SD 0.27h, n = 16) was followed by a free run (mean period 24.42h, SD 1.33h) after transition into constant dim light conditions. Light pulses of a comparable intensity as used in the light part of the LD cycles did not result in significant phase shifts of the free-running rhythm in constant darkness. Application of much brighter 3h light pulses resulted in a phase-response curve (PRC) for a fish species, with pronounced phase advances during late subjective night. The PRCs differed from those mainly obtained in other vertebrate taxa by the absence of significant phase delays in the early subjective night. At that circadian phase, significant tonic effects of the light pulses caused a shortening of the circadian period length. Entrainment to skeleton photoperiods of 1:11 LD was observed in five of six wrasses exposed, also after a 3h phase advance of this LD cycle. Subsequently, a 1:11.25 LD cycle resulted in entrainment in four of the six fish. It is suggested that the expression of the circadian system in fish can be interpreted as a functional response to a weak natural zeitgeber, as present in the marine environment. This response allows photic entrainment as described here in the yellow wrasse.

Animals↗

The suprachiasmatic nucleus in organotypic slice cultures of the common vole (Microtus arvalis): comparison of development with rat and hamster and the effect of age.

The intrinsic properties of the suprachiasmatic nucleus (SCN), the site of the main circadian pacemaker in mammals, have recently been studied in vitro by means of organotypic slice culturing. So far, only neonatal rats and mice have been used for such developmental and functional analyses of the isolated pacemaker. Here, the authors present a comparative developmental study of the SCN of voles, rats, and hamsters in organotypic slice cultures. In contrast to strictly circadian organization of behavior in rats and hamsters, common voles (Microtus arvalis) are characterized by large variability in the strength of circadian organization of behavior. It is not known to what extent this variability is reflected in the intrinsic features of the SCN. Cultures were prepared from rat, hamster, and vole pups (6 to 9 days old) for the purpose of species comparison. In addition, the authors studied the relation between age and development in cultures from pup (7 to 10 days old), juvenile (15 to 16 days old), and young adult (1 to 2 months old) voles. In contrast to the situation in rat and hamster, the most striking feature in neonatal voles is the variability in shape of the final, fully developed culture and its poor resemblance with the in vivo SCN. The SCN of adult voles, however, could be cultured successfully while retaining its morphological organization seen in situ. Phase-contrast microscopy and immunocytochemical staining for vasopressin and glial fibrillary acidic protein revealed that cultures of pup and juvenile voles still have potential for neurogenesis and morphological reorganization. Young voles, therefore, can serve as a model to study the developmental establishment of a functional circadian pacemaker, while adult voles allow the study of intrinsic pacemaker properties in relation to previously recorded behavior of the donor and aging-related pacemaker dysfunction.

Aging↗

Severe loss of vasopressin-immunoreactive cells in the suprachiasmatic nucleus of aging voles coincides with reduced circadian organization of running wheel activity.

Aging leads to a decrease in circadian organization of behavior. Whether this general observation is related to the finding that in older subjects the arginine-vasopressin (AVP) system in the suprachiasmatic nucleus (SCN) has deteriorated is an unsolved question. Here we assessed circadian organization of running wheel behavior and numbers of AVP cells in the SCN of old voles (n=12, 11. 5 months of age) and compared the results with data from young voles (n=16, 4.5 months of age). A third of the young voles, but three-quarter of the old voles lost circadian rhythmicity. Analysis of daily onset to onset periodicity of running wheel activity at the age of 5 and 10 months in individual voles revealed a significant loss of precision of circadian rhythmicity at the higher age. The number of AVP cells in the SCN of old voles decreased substantially, over 78% compared to young voles in general. AVP cell numbers, however, cannot be directly correlated with the state of rhythmicity in old voles; in one of the three circadian rhythmic old voles the SCN contained the least AVP cells. This study does not support the idea of a causal relationship between aging induced reduction in AVP cells in the SCN and the presence of circadian rhythmicity in behavior.

Aging↗

Lack of circadian patterns in vasoactive intestinal polypeptide release and variability in vasopressin release in vole suprachiasmatic nuclei in vitro.

Organotypic hypothalamic cultures of neonatal rats comprising the suprachiasmatic nuclei (SCN) produce stable 20 h release patterns of vasoactive intestinal polypeptide (VIP) and arginine-vasopressin (AVP). Compared with rats, voles show variably expressed circadian activity patterns. In this study we measured neuropeptidergic release patterns in organotypic SCN cultures of neonatal common voles (Microtus arvalis, n = 6). Slices were prepared at postnatal day 6. After 14 days of incubation, 2 h samples of medium were collected during 50 h. None of the vole SCN slices showed a circadian modulation in VIP release. Peaks in AVP occurred, 20 h apart from each other, in four of six vole SCN slices. These findings contrast with the concurrent release patterns of VIP and AVP in rat SCN slices. The results suggest an independent role of both neuropeptides in the oscillatory output pathways of the circadian pacemaker in the common vole.

Animals↗

Organotypic suprachiasmatic nuclei cultures of adult voles reflect locomotor behavior: differences in number of vasopressin cells.

This study is the first to demonstrate organotypic culturing of adult suprachiasmatic nuclei (SCN). This approach was used to obtain organotypic SCN cultures from adult vole brain with a previously determined state of behavioral circadian rhythmicity. We examined vasopressin (AVP) immunoreactivity in these organotypic slice cultures. AVP is one of the major neuropeptides produced by the SCN, the main mammalian circadian pacemaker. AVP immunoreactivity in the SCN of adult common voles in vivo has been shown to correlate with the variability in expression of circadian wheel-running behavior. Here, cultures prepared from circadian rhythmic and nonrhythmic voles were processed immunocytochemically for AVP. Whereas in all cultures AVP could be observed, AVP immunoreactivity differed considerably between vole SCN cultures. SCN cultures from rhythmic voles contained significantly lower numbers of AVP immunoreactive (AVPir) cells per surface area than cultures from nonrhythmic voles. The correlation between timing of behavior and AVP immunoreactivity in vitro is similar to the correlation found earlier in vivo. Apparently, such correlation depends on intrinsic AVP regulation mechanisms of SCN tissue, and not on neural or hormonal input from the environment, as present in intact brain.

Animals↗

Concurrent decrease of vasopressin and protein kinase Calpha immunoreactivity during the light phase in the vole suprachiasmatic nucleus.

Vasopressin (AVP) is a major neuropeptide in the suprachiasmatic nucleus, the mammalian hypothalamic circadian pacemaker. Protein kinase Calpha is a putatively coupled intracellular messenger. Mean numbers of AVP- and protein kinase Calpha-immunoreactive neurons were determined in the suprachiasmatic nucleus of common voles, entrained to a 12:12 h light-dark (LD) cycle, at the beginning of the light period (zeitgeber time zero) and 6 h later (zeitgeber time six). At zeitgeber time zero, mean numbers of AVP- and protein kinase Calpha- immunoreactive neurons were 2194 and 9897, respectively. Both numbers decreased significantly with about 40% at zeitgeber time six. This concurrent decrease was most pronounced in the dorsomedial aspect of the suprachiasmatic nucleus. These findings are consistent with the findings of a peak of AVP release in rats during the early light phase.

Animals↗

Expression of circadian rhythmicity correlates with the number of arginine-vasopressin-immunoreactive cells in the suprachiasmatic nucleus of common voles, Microtus arvalis.

We have studied the number of arginine-vasopressin (AVP)-immunoreactive (IR) cells in the suprachiasmatic nuclei (SCN) in common voles, Microtus arvalis, with a strong (n = 18), weak (n = 10) or absent (n = 9) expression of circadian rhythmicity. Spontaneous expression of rhythmicity was assessed from records of wheel running activity and general activity, measured with passive infra-red detection during 4 weeks of continuous low light (LL) conditions. Subsequently, 20 voles were perfused in LL. After additional exposure to a 12:12 h light-dark (LD) cycle during 14 days, the other 17 voles were perfused in the early morning. AVP-positive neurons were visualized immunocytochemically with a polyclonal IgG antibody. AVP-stained cell somata were present predominantly in the dorsomedial SCN, whereas terminals, contacting both AVP positive and negative neurons, were distributed throughout the SCN. In LL conditions, AVP-staining intensity of neurons was low in all 20 voles. Also the number of AVP-IR cells in the SCN in brain sections obtained in LL conditions was low and not different for the three categories of rhythmicity. In LD conditions, the staining intensity of AVP positive SCN neurons was low in rhythmic animals, high in non-rhythmic animals not expressing, and intermediate in animals weakly expressing circadian rhythmicity. The number of AVP positive cells in LD conditions showed a negative correlation with the propensity of animals to lose circadian rhythmicity under LL conditions. Differences in AVP-IR were found solely in the dorsomedial SCN, and neither in the rostral and caudal aspects of the SCN nor in the area adjacent to the SCN.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Phase control of ultradian feeding rhythms in the common vole (Microtus arvalis): the roles of light and the circadian system.

In their ultradian (2- to 3-hr) feeding rhythm, common voles show intraindividual synchrony from day to day, as well as interindividual synchrony between members of the population, even at remote distances. This study addresses the question of how resetting of the ultradian rhythm, a prerequisite for such synchronization, is achieved. Common voles were subjected to short light-dark cycles (1 hr darkness with light varying between 0.7 and 2.5 hr); to T cycles (long light-dark cycles in the circadian range--16 hr darkness and 3-13 hr light); to light pulses (15 min) during different circadian and ultradian phases; and to addition of D2O to the drinking water (25%). Short light-dark cycles and D2O were also applied to voles without circadian rhythmicity, after lesions of the suprachiasmatic nuclei. In these experiments, four hypotheses on synchronization of ultradian rhythmicity were tested: (I) synchronization by a direct response to light; (II) synchronization via the circadian system with multiple triggers, here called "cogs," each controlling a single ultradian feeding bout; and (III and IV) synchronization via the circadian system with a single "cog," which resets an ultradian oscillator and either (III) originates directly from the circadian pacemaker, or (IV) is mediated via the overt circadian activity rhythm. Short light-dark cycles failed to entrain ultradian rhythms, either in circadian-rhythmic or in non-circadian-rhythmic voles; light pulses did not cause phase shifts; and in extreme T cycles no stable phase relationship with light could be demonstrated. Thus, Hypothesis I was rejected. Changes in the circadian period (tau) were generated as aftereffects of light pulses, by entrainment in various T cycles, and by the addition of D2O to the drinking water. These changes in tau did not lead to parallel, let alone proportional, changes in the ultradian period. This excluded Hypothesis II. Both in T-cycle experiments and in the D2O experiments with circadian-rhythmic voles, the phase of ultradian feeding bouts was locked to the end of circadian activity rather than to the most prominent marker of the pacemaker, the onset of circadian activity. This was not expected under Hypothesis III, but was consistent with entrainment via activity (Hypothesis IV). On the basis of these experiments, we conclude that the most likely mechanism of ultradian entrainment is that of a light-insensitive ultradian oscillator, reset every dawn by the termination of the activity phase controlled by the circadian pacemaker, which is itself entrained by the light-dark cycle. Neither in circadian-rhythmic nor in non-circadian-rhythmic voles was the period of the feeding rhythm lengthened by administration of D2O. This insensitivity to deuterium is exceptional among biological rhythms.

Activity Cycles↗

Ongoing ultradian activity rhythms in the common vole, Microtus arvalis, during deprivations of food, water and rest.

The timing mechanism underlying ultradian (2-3 h) activity patterns in the common vole, Microtus arvalis, was studied using behavioral deprivation experiments. These were aimed at distinguishing between a homeostatic control mechanism, in which the rhythmic behaviour itself is part of the causal loop, and a clock mechanism, independent of the behaviour. In 175 experiments, deprivation of food during 3 ultradian cycles in (subjective) daytime did not result in significant changes in the ultradian periodicity of attempts to obtain the food, compared with ad lib, access to food and water. A minor, but significant increase in ultradian activity time (alpha') occurred in the course of the deprivation, but this was compensated by a shorter ultradian rest (rho'). These results were obtained both in intact animals (n = 24), which showed ultradian and circadian rhythmicity in behaviour, and in animals (n = 21) with electrolytic lesions aimed at the suprachiasmatic nuclei (SCN), which lacked the circadian modulation of behaviour. Simultaneous deprivation of water and food in 8 voles without circadian rhythmicity during 40 experiments also did not lead to any change in the ultradian periodicity of feeding attempts. Rest deprivation was studied in 5 SCN lesioned voles, by forcing running wheel activity to continue following spontaneous running. Thus, the experimental activity bout alpha' was artificially lengthened to 2-9 h in 67 experiments. The onset of the subsequent rest episodes occurred independent of the duration of the preceding alpha'. The duration of rho' was dependent on the preceding, experimental alpha' in a periodic fashion.(ABSTRACT TRUNCATED AT 250 WORDS)

Activity Cycles↗

Differential elimination of circadian and ultradian rhythmicity by hypothalamic lesions in the common vole, Microtus arvalis.

Effects of hypothalamic lesions on the ultradian and circadian organization of wheel running and feeding were studied in the common vole, Microtus arvalis. Circadian organization broke down within 30 days in continuous darkness in 24% of intact voles (n = 135). Ultradian rhythmicity of feeding (period 2-3 hr) persisted in constant conditions in all intact voles. Following lesions of the suprachiasmatic nuclei (SCN), circadian rhythmicity disappeared when lesions were complete (n = 8) or more extensive than 25% of the total SCN volume (n = 5). Absence of circadian rhythmicity was also found in animals with substantial lesions in the diencephalic paraventricular area (PVA) and in the retrochiasmatic area (RCA) and/or adjacent arcuate nucleus (Arc). Complete loss of ultradian and circadian organization occurred in eight voles with damage to the RCA and/or Arc. In three of these, the SCN was intact. The SCN is a likely candidate for a circadian pacemaker in voles (as in other rodents), while the loss of circadian rhythmicity following PVA and RCA/Arc lesions may be due to destruction of efferent pathways from the SCN. The RCA/Arc area is apparently necessary for the expression of ultradian rhythms. The intact SCN is neither necessary nor sufficient for the generation of ultradian rhythmicity.

Activity Cycles↗