SD Voles Research Articles

Disentangling the effects of obesity and high-fat diet on glucose homeostasis using a photoperiod induced obesity model implicates ectopic fat deposition as a key factor

ObjectiveObesity in laboratory rodents is generally induced by feeding them a high fat diet (HFD). This model does not permit separation of the impact of the HFD from the resultant obesity on metabolic defects such as impaired glucose homeostasis. In Brandt's voles we have previously shown that exposure to long photoperiod (LD: 16L: 8D) induces obesity even when they are fed a low fat diet. We show here that these voles are largely resistant to HFD. This model therefore permits some separation of the effects of HFD and obesity on glucose homeostasis. The objective was therefore to use this model to assess if glucose homeostasis is more related to diet or obesity MethodsMale voles, which were 35 days old and born in LD, were exposed to SD and a low fat diet for 70 days. We then randomly separated the animals into 4 groups for another 63 days: SL (short day and low fat diet: n = 19) group; SH (short day and high-fat diet, n = 20) group; LL (long day and low-fat diet, n = 20) group; LH (long day and high-fat diet, n = 18) group. Glucose tolerance tests (GTT) were performed after treatment for 56 days, and body compositions of the voles were quantified at the end by dissection. ResultsConsistent with our previous work LD voles were more obese than SD voles. Although total body weight was independent of dietary fat content, HFD did have an effect on fat storage. Photoperiod induced obesity had no effect on glucose homeostasis, and the fat content in both the liver and muscle. In contrast, HFD induced adiposity was linked with elevated fat deposition in muscle (but not in liver) and led to impaired glucose tolerance. ConclusionsThe contrasting effects of diet and photoperiod were consistent with the predictions of the ‘lipotoxicity hypothesis’. This may contribute to our understanding of why some human individuals are able to be obese yet remain metabolically healthy.

Telocytes are localized to testis of the bank vole (Myodes glareolus) and are affected by lighting conditions and G-coupled membrane estrogen receptor (GPER) signaling

We aim to explore the presence of a novel cell type, telocytes (TCs), in the bank vole testis interstitium following G-coupled membrane estrogen receptor (GPER) signaling withdrawal. In addition, the involvement of interstitial cells in lipid homeostasis was investigated.Bank voles (actively reproducing or regressed) were administered with GPER antagonist (G-15; 50 μg/kg bw) injections. To examine TC distribution, ultrastructure, function, and their connotation in the interstitial tissue lipid balance, electron microscopic observations were implemented. Immunohistochemistry and Western blot for the TC marker, CD34, and lipid balance molecules: leptin, adiponectin, and perilipin were performed. Photoperiod-regulated testis steroidogenic function was estimated via serum melatonin level and intratesticular cholesterol concentrations in immunoenzymatic assays.We demonstrate the presence of TCs in bank vole testis interstitium. Distinctive TC morphology: small cell bodies with very long, slender prolongations, constituting a three-dimensional network around the interstitial cells was seen. Ultrastructurally, scarce mitochondria, a few cisternae of the endoplasmic reticulum, and lipid droplets indicated possible TC implications in lipid homeostasis. Changes in CD34 expression in TCs were seen in relation to GPER disturbances. In GPER-blocked testis, single TCs were present in the LD interstitium when in SD ones they were occasionally absent. Moreover, in TCs of SD voles, a lack of lipid droplets was revealed, likely reflecting attenuated TC function during regression. However, melatonin levels decreased in GPER-blocked LD and SD. Concomitantly, leptin, adiponectin, and perilipin expressions together with cholesterol content varied after blockage.Based on our results we suggest TCs are an important component of the bank vole testis interstitium as they are implicated in ultramorphology maintenance, protein interactions, and lipid homeostasis.

Are aryl hydrocarbon receptor and G-protein–coupled receptor 30 involved in the regulation of seasonal testis activity in photosensitive rodent—the bank vole (Myodes glareolus)?

Within the reproductive system both aryl hydrocarbon receptor (AHR) and G-protein–coupled receptor 30 (GPR30) contribute to estrogen signaling and controlling of reproductive physiology. The specific question is whether and how AHR and GPR30 are involved in regulation of testis function in seasonally breeding rodents. Bank vole testes were obtained from animals reared under 18 hours light:6 hours dark (LD) and 6 hours light:18 hours dark (SD) conditions. Aryl hydrocarbon receptor and GPR30 expression were analyzed by quantitative reverse transcriptase-polymerase chain reaction, Western blot, and immunohistochemistry and/or immunofluorescent staining. In addition, the activity of enzymes involved in the intracellular signal transduction; extracellular signal-regulated kinase (ERK), protein kinase (PKA), matrix metalloproteinase 9 (MMP 9) and the concentrations of cyclic adenosine monophosphate (cAMP), cyclic guanosine monophosphate (cGMP), and calcium (Ca2+) were examined by immunohistochemical, immunoenzymatic, and colorimetric assays, respectively. Aryl hydrocarbon receptor and GPR30 were expressed in testes of actively reproducing voles and regressed ones although their expression at the messenger RNA and AHR also at protein level appeared to be photoperiod-dependent. A specific cellular localization and expression of AHR and GPR30 correlated with the expression of ERK, PKA, and MMP 9. Moreover, we found robust differences in the levels of cAMP, cGMP, and Ca2+ in testicular homogenates between LD and SD voles. In the testes of LD voles, the levels of second messengers were always higher compared to SD. In vole testis, AHR and GPR30 can induce signaling pathways that involve ERK, PKA, MMP 9 and cAMP, cGMP, Ca2+. In addition, in AHR, signaling the engagement of both photoperiod and estrogens, whereas in GPR30, signaling only estrogens is reported. It is likely that in vole, because of a differential activity of signaling molecules, signal transduction via AHR rather than through GPR30 plays a role in regulation of seasonal changes of testis physiology.

Corticotropin-releasing factor receptor densities vary with photoperiod and sociality

Life in social groups relies on prosocial behaviors as well as on reduction of antisocial behaviors such as aggression and territoriality. The mechanisms supporting variation in behaviors that give rise to group living (sociality) are largely unknown. Female meadow voles exhibit natural seasonal variation in sociality: females are aggressive and territorial in summer, while in winter they share burrows and nest in mixed-sex groups. This behavioral shift is paralleled in the lab by day length-dependent variation in partner preference formation and social huddling. We exploit natural variation in meadow vole sociality in order to examine changes in neural pathways that coincide with environmental and behavioral variations.Mounting evidence suggests that the corticotropin-releasing factor system, encompassing multiple peptides and two receptor subtypes (CRF1 and CRF2), may play an important role in regulating social behaviors. We report day-length dependent variation in CRF1 and CRF2 receptor binding in female meadow voles, and relate these findings to previously collected oxytocin receptor (OTR) binding data and behavioral data for the same individuals. CRF1 receptor binding was greater in summer-like long day lengths (LD), particularly in the hippocampus, while CRF2 receptor binding was greater in winter-like short day lengths (SD) in the cingulate cortex and hippocampus. OTR varied with day length in the bed nucleus of the stria terminalis, nucleus accumbens, and hippocampus. SD voles huddled more extensively than LD voles, and greater huddling time was associated with more CRF1 receptor binding and less CRF2 receptor binding in subregions of the lateral septum. CRF2 receptor associations with behavior mirrored those of OTR in the lateral septum. Finally, estradiol treatment affected density of CRF receptors in multiple brain regions. CRF receptors and their ligands are promising candidates for enhancing understanding of the regulation of non-sexual social behavior between group living peers.

Effects of photoperiod on energy intake, thermogenesis and body mass in Eothenomys miletus in Hengduan Mountain region

The present study was designed to examine whether photoperiod alone was effective to induce seasonal changes in physiology in voles ( Eothenomys.) from the Hengduan Mountain region. Eothenomys miletus were randomly assigned into either long photoperiod (LD; 16L: 8D) or short photoperiod (SD; 8L: 16D) for 4 weeks at constant temperature (25 °C). At the end of acclimation, SD voles showed lower body mass and body fat coupled with higher energy intake than LD voles. SD greatly enhanced the thermogenic capacity of E. miletus, as indicated by an elevated nonshivering thermogenesis (NST), mitochondrial protein in brown adipose tissue (BAT); basal metabolic rate (BMR) was also raised. Although no variations in serum leptin levels were found between SD and LD voles, serum leptin levels were positively correlated with body mass and body fat mass, and negatively correlated with energy intake and UCP1 content in BAT, respectively. To summarize, SD alone is effective in inducing higher thermogenic capacities and energy intake coupled with lower body mass and body fat mass in root voles. Leptin is potentially involved in the photoperiod induced body mass regulation and thermogenesis in E. miletus. Our study shows that SD alone is effective.

Diet-induced obesity in the short-day-lean Brandt's vole

To test the hypothesis that mammals that show decrease in body mass under short-day condition should be resistant to high-fat induced obesity, we traced the changes of energy balance in a wild rodent, Brandt's voles ( Lasiopodomys brandtii), which were acclimated to either long day (16 L: 8D, LD) or short day (8 L: 16D, SD) and fed either low-fat diet (LFD) or high-fat diet (HFD) in each photoperiodic manipulation. We found that Brandt's vole was not resistant to high-fat diet-induced obesity and SD, not HFD, induced the elevation in basal metabolic rate, the maximal rate of oxygen consumption after norepinephrine injection, and uncoupling protein 1 content in brown adipose tissue. HFD caused the increase in apparent digestibility and body fat mass, and the decrease in energy intake in both LD and SD voles. The enhancement of energy absorption associated with small intestine tissue recruitment can compensate the lower energy intake, which may contribute to the high-fat diet-induced body fat deposition. Thus, a decrease in body-weight gain but has no resistance to high-fat induced obesity implies an evolutionary and adaptive mechanism which is a benefit for their winter survival.

Day length and estradiol affect same-sex affiliative behavior in the female meadow vole

Non-sexual social bonding between adult mammals is poorly understood, despite its importance in many species. Female meadow voles are territorial and nest alone in long summer day lengths when circulating estradiol concentrations are high, but cohabit in groups in short winter photoperiods when estradiol secretion is low. The influence of day length and estradiol on same-sex huddling behavior was assessed in adult female pairs housed together in long day lengths (LDs) or short day lengths (SDs) from weaning. The behavior of intact, ovariectomized, and estradiol-treated ovariectomized females from each photoperiod was assessed during 3 h partner-preference tests. Intact SD voles, unlike intact LD voles, spent the majority of the test in proximity to their cage-mates. Estradiol treatment of SD voles significantly reduced time spent huddling with the partner. Neither ovariectomy nor estradiol treatment significantly affected the amount of time LD females spent in contact with their partners. Low estradiol availability is therefore a necessary but not sufficient condition for maintenance of high levels of huddling. These results establish that ovarian hormones interact with photoperiod to affect same-sex social behavior.

Effects of photoperiod history on body mass and energy metabolism in Brandt's voles (Lasiopodomys brandtii)

Many small mammals respond to seasonal changes in photoperiod via alterations in morphology, physiology and behaviour. In the present study, we tested the hypothesis that the preweaning (from embryo to weaning) photoperiod experience can affect subsequent development in terms of body mass and thermogenesis. Brandt's voles (Lasiopodomys brandtii) were gestated and reared to weaning under either a short (SD, 8 h:16 h L:D) or a long photoperiod (LD, 16 h:8 h L:D) at a constant ambient temperature (23 degrees C). At weaning, male juveniles were either maintained in their initial photoperiod or transferred to the alternative photoperiod for 8 weeks. Postweaning SD voles had a lower body mass but higher thermogenic capacity compared with LD voles. At the same time, preweaning photoperiod conditions had long-lasting effects on thermogenic capacity later in life. Serum leptin concentration was positively correlated with body mass and body fat mass, whereas it was negatively correlated with energy intake and uncoupling protein 1 content in brown adipose tissue. Our results suggest that postweaning development in terms of body mass and thermogenesis is predominantly influenced by the postweaning photoperiod, while the preweaning photoperiod experience could chronically modify thermogenesis but not body mass. Furthermore, serum leptin, acting as a potential adipostatic signal, may be involved in the regulation of both energy intake and energy expenditure.

Regulation of body mass and adiposity in the field vole, Microtus agrestis: a model of leptin resistance

Adult mammals are typically highly resistant to perturbations in their energy balance. In obese humans, however, this control appears to be lost. Apart from a few exceptional cases, this loss of control occurs despite appropriate levels of circulating leptin -- suggesting that elevated adiposity may be a consequence of failure to respond to the leptin signal: leptin resistance. When cold-acclimated male field voles (Microtus agrestis) are transferred from short (SD, 8 h light) to long (LD, 16 h light) photoperiods, they increase dramatically in body mass and fatness for about 4 weeks. After this period, their mass stabilizes at a new plateau about 25% higher than animals maintained in SD. The increase in adiposity is not caused by significant increases in food intake, but reflects an increase in digestive efficiency. Measures of circulating leptin reveal that the increased adiposity is matched by increased circulating leptin. By infusing voles with exogenous leptin, we have demonstrated that SD voles are leptin sensitive (reducing both body mass and food intake), whereas LD animals are leptin resistant. Voles may therefore be a useful model for understanding the process of leptin resistance. The change in leptin sensitivity in voles was not associated with changes in the levels of gene expression of the orexogenic or anorexogenic neuropeptides, such as neuropeptide Y, agouti-related peptide, POMC and cocaine- and amphetamine-regulated transcript, measured in the hypothalamic arcuate nucleus (ARC). During the phase that body mass was increasing, however, there was a transient increase in the ARC expression of suppressor of cytokine signalling-3 (SOCS3). These data suggest that the changes in the expression of SOCS3 in the ARC may be involved in leptin resistance. However, the mechanism by which these changes may be linked to alterations in digestive efficiency that underpin the changes in adiposity, or how the differences are signalled by changes in photoperiod, remains unclear.

Thermoregulatory and Endocrine Responses to Light Pulses in Short‐Day Acclimated Social Voles (Microtus socialis)

In mammals, nocturnal light pulses (NLP) have been demonstrated to affect physiology and behavior. However, the impact of NLP as a stressor has been less broadly examined. The purpose of this study was to examine the effect of NLP (three 15 min 450 lux light pulses) during each scotophase on both thermoregulation and endocrine stress responses under short‐day (SD; 8L:16D) acclimation. Voles were acclimated to either SD (SD voles) or SD+NLP (NLP voles). Resistance to cold was estimated by measurements of body temperature (Tb) during cold exposure (5°C). Daily rhythms of energy expenditure (calculated from oxygen consumption), urine production, and urinary adrenaline and serum cortisol levels were measured. Tb values of SD voles were generally unaffected by the cold stimulus, whereas in NLP voles, resistance to cold was markedly lowered. While SD‐ and NLP voles showed similar ultradian characteristics in energy expenditure with a period of 3.5 h, mean energy expenditure levels were lowest for voles exposed to NLP‐treatment. In SD voles, but not in NLP voles, urine production rates showed clear time variations and were consistently highest for SD voles, with significant differences during the scotophase. Both mean total urinary adrenaline and serum cortisol levels were significantly elevated in NLP‐treated voles compared with the control group. Taken together, the results suggest that NLP negatively affects winter acclimatization of thermoregulatory mechanisms of M. socialis, probably by mimicking summer acclimatization, and consequently the thermoregulatory mechanisms respond inappropriately to ambient conditions. One important finding of this study is that NLP may act as a stressor and correspondingly impose a major threat to the physiological homeostasis of M. socialis, such that over‐winter survival might be compromised.

Altered Expression of SOCS3 in the Hypothalamic Arcuate Nucleus during Seasonal Body Mass Changes in the Field Vole, Microtus agrestis

We have previously shown that cold-acclimated (8 degrees C) male field voles (Microtus agrestis) transferred from short day (SD, 8 h light) to long day (LD, 16 h light) photoperiod exhibit an increase in body mass lasting 4 weeks, after which they stabilise at a new plateau approximately 7.5 g (24.8%) higher than animals maintained in SD. By infusing voles with exogenous leptin, we have also demonstrated that SD voles respond to the hormone by reducing body mass and food intake, whereas LD animals increasing body mass are resistant to leptin treatment. In the present study, we investigated whether seasonal changes in body mass could be linked to modulation of the leptin signal by suppressor of cytokine signalling-3 (SOCS3). We used in situ hybridisation to examine hypothalamic arcuate nucleus (ARC) expression of SOCS3, neuropeptide Y (NPY), agouti-related peptide (AgRP), pro-opiomelanocortin (POMC) and cocaine- and amphetamine-regulated transcript (CART) genes in 90 voles exposed to either SD or LD for up to 11 weeks. LD voles increasing body mass had significantly higher levels of SOCS3 mRNA than SD or LD voles with a stable body mass. There were no associated changes in expression of NPY, AgRP, POMC and CART genes. These results suggest that voles that regulate body mass at either the lower (SD) or upper (LD) plateau remain sensitive to leptin action, whereas SOCS3-mediated leptin resistance is a short-term mechanism that enables animals to move between the stable body mass plateaus. Our data provide evidence that expression of SOCS3 in the ARC is involved in the modulation of the strength of the leptin signal to facilitate seasonal cycles in body mass and adiposity.

Photoperiodic regulation in energy intake, thermogenesis and body mass in root voles (Microtus oeconomus)

The present study was designed to examine whether photoperiod alone was effective to induce seasonal regulations in physiology in root voles (Microtus oeconomus) from the Qinghai-Tibetan plateau noted for its extreme cold environment. Root voles were randomly assigned into either long photoperiod (LD; 16L:8D) or short photoperiod (SD; 8L:16D) for 4 weeks at constant temperature (20 degrees C). At the end of acclimation, SD voles showed lower body mass and body fat coupled with higher energy intake than LD voles. SD greatly enhanced thermogenic capacities in root voles, as indicated by elevated basal metabolic rate (BMR), nonshivering thermogenesis (NST), mitochondrial protein content and uncoupling protein-1 (UCP1) content in brown adipose tissue (BAT). Although no variations in serum leptin levels were found between SD and LD voles, serum leptin levels were positively correlated with body mass and body fat mass, and negatively correlated with energy intake and UCP1 content in BAT, respectively. To summarize, SD alone is effective in inducing higher thermogenic capacities and energy intake coupled with lower body mass and body fat mass in root voles. Leptin is potentially involved in the photoperiod induced body mass regulation and thermogenesis in root voles.

Short photoperiod influences energy intake and serum leptin level in Brandt’s voles ( Microtus brandtii)

Photoperiod cues play important roles in the regulation of seasonal variations in body mass (BM) and energy balance for many small mammals. The present study was designed to examine the effects of photoperiod acclimation on BM, energy intake, and serum leptin levels in Brandt’s voles ( Microtus (Lasiopodomys) brandtii). After 4 weeks of acclimation to either long (LD; light:dark, 16:8) or short (SD; 8:16) photoperiod, SD voles had lower BM, body fat mass, and dry mass of liver and kidneys, but higher digestible energy intake in comparison to LD voles. SD voles also showed a lower level of serum leptin than did LD voles. Furthermore, the level of serum leptin was correlated positively with body fat mass and negatively with gross energy intake. Together, these data suggest that Brandt’s voles employ a strategy of minimizing body growth, increasing energy intake, and mobilizing fat deposition in response to cues associated with short photoperiod. Furthermore, leptin seems to be involved in the regulation of BM and energy balance mediated by photoperiod.

Photoperiod regulates leptin sensitivity in field voles, Microtus agrestis

We have previously shown that cold-acclimated (8 degrees C) male field voles (Microtus agrestis) transferred from short (SD, 8:16 h L:D) to long photoperiod (LD, 16:8 h L:D) exhibit increases in body mass, adiposity and food intake. To assess whether these increases were associated with decreased leptin sensitivity, we infused LD and SD voles with physiological doses of murine leptin (or saline) delivered peripherally for 7 days via mini-osmotic pumps. Measurements were made of body mass (weight-reducing effect of leptin), food intake (anorectic effect of leptin) and gene expression of uncoupling protein 1 (UCP1) in brown adipose tissue (BAT) (thermogenic effect of leptin). The SD animals were sensitive to the weight-reducing effects of leptin (mean body mass decrease of 1.2 g over 7 days) and appetite-reducing effect of leptin (mean food intake decrease of 2.5 g over 7 days), whereas LD voles were resistant to the hormone treatment. The switch from a leptin-sensitive to leptin-resistant state appears to act as a desensitisation mechanism that allows voles transferred from SD to LD to ignore elevated leptin levels generated by increased body fat and accumulate adipose tissue without stimulating compensatory changes opposing the weight gain. Neither SD nor LD voles responded to infusion of leptin by changes in BAT UCP1 gene expression, suggesting dissociation of anorectic and thermogenic effects of leptin, possibly related to chronic cold exposure. Our results indicate that cold-acclimated voles show photoperiod-regulated changes in leptin sensitivity and may provide an attractive model for elucidating molecular mechanisms of leptin resistance.

Influence of photoperiod and sex on locomotor behavior of meadow voles ( Microtus pennsylvanicus) in an automated light–dark ‘anxiety’ test

This study examined the influence of photoperiod on affective behavior (anxiety) of adult male and female meadow voles (Microtus pennsylvanicus), maintained in either a long or short day photoperiod, when tested in an automated (VersaMax) light-dark test. The light-dark test is based on an innate aversion of rodents to novel, brightly illuminated spaces and has been used with laboratory raised species, such as mice, to assess anxiety and/or fear related behaviors. Male and female meadow voles, housed either in a long day (LD: 16 h light) or short day (SD: 8 h light) photoperiod, were tested in the light-dark apparatus for 30 min on 3 consecutive days. All animals spent significantly (p < 0.001) less time in the brightly lit chamber (900 lux) than in the dark chamber. LD voles, especially females, spent significantly less time in the brightly lit area than did SD voles. Both horizontal and vertical movements occurred less frequently per unit time in the dark area relative to the light, but only in the LD voles. LD female voles were the least active group in the dark area on the first test day but the most active group in the light area, despite spending the least amount of time in this area on the second and third test days. The present results show that LD voles exhibit more anxiety related behaviors in this test situation than do SD voles. LD females avoided the brightly lit area the most, particularly when the apparatus was novel. Thus, both photoperiod and sex influence situation-based anxiety in this species. These findings suggest that meadow voles are an excellent animal model in which to examine the role of gonadal hormones, and their modulation of defence related neural systems, in the induction of anxiety.

Effect of photoperiod on body mass, food intake and body composition in the field vole,Microtus agrestis

Many small mammals respond to seasonal changes in photoperiod by altering body mass and adiposity. These animals may provide valuable models for understanding the regulation of energy balance. Here, we present data on the field vole (Microtus agrestis) - a previously uncharacterised example of photoperiod-induced changes in body mass. We examined the effect of increased day length on body mass, food intake, apparent digestive efficiency, body composition, de novo lipogenesis and fatty acid composition of adipose tissue in cold-acclimated (8 degrees C) male field voles by transferring them from a short (SD, 8 h:16 h L:D) to long day photoperiod (LD, 16 h:8 h L:D). During the first 4 weeks of exposure to LD, voles underwent a substantial increase in body mass, after which the average difference between body masses of LD and SD voles stabilized at 7.5 g. This 24.8% increase in body mass reflected significant increases in absolute amounts of all body components, including dry fat mass, dry lean mass and body water mass. After correcting body composition and organ morphology data for the differences in body mass, only gonads (testes and seminal vesicles) were enlarged due to photoperiod treatment. To meet energetic demands of deposition and maintenance of extra tissue, voles adjusted their food intake to an increasing body mass and improved their apparent digestive efficiency. Consequently, although mass-corrected food intake did not differ between the photoperiod groups, the LD voles undergoing body mass increase assimilated on average 8.4 kJ day(-1) more than animals maintained in SD. The majority (73-77%) of the fat accumulated as adipose tissue had dietary origin. The rate of de novo lipogenesis and fatty acid composition of adipose tissue were not affected by photoperiod. The most important characteristics of the photoperiodic regulation of energy balance in the field vole are the clear delineation between phases where animals regulate body mass at two different levels and the rate at which animals are able to switch between different levels of energy homeostasis. Our data indicate that the field vole may provide an attractive novel animal model for investigation of the regulation of body mass and energy homeostasis at both organism and molecular levels.