Maintenance of fertility in cryopreserved Indian gerbil (Tatera indica) spermatozoa

This study is the first attempt at sperm cryopreservation, as well as a further examination of frozen sperm fertility by the hamster test, applied to the maintenance of an Indian gerbil (Tatera indica) colony, which is a newly developing experimental animal.The osmotic tolerance of the spermatozoa was initially investigated by subjection to hypertonicity, up to 620 mOsm/kg, for 5 min at room temperature prior to freezing. Although the percentage of total motile sperm was not affected, that of progressive motile spermatozoa began to drop at 400 mOsm/kg, and a significant decrease was observed at 620 mOsm/kg (p < 0.01). According to these results, the osmolality of the solutions for the freezing experiment, in which 6–22% raffinose was present, was fixed at approximately 400 mOsm/kg. Sperm, suspended in a plastic straw, were frozen in liquid nitrogen vapor for 5 min, followed by immersion in liquid nitrogen. Motile sperm were recovered from all freezing conditions, and high survival was obtained when sperm were frozen in the presence of 14% and 18% raffinose, with a normalized motility higher than 40%. Fertility of cryopreserved Indian gerbil sperm was examined by the zona-free hamster test. Thawed sperm adhered to 88% of the zona-free hamster oocyte surface, and some oocytes were penetrated and exhibited swollen sperm heads or male pronuclei, which we used to define fertilization. Although the fertilization rate of cryopreserved sperm to zona-free hamster eggs was significantly lower than that of fresh sperm (6% vs. 30%, p < 0.01), we demonstrated that thawed Indian gerbil spermatozoa have the ability to maintain their fertility.     

Physiological ecology of Mediterranean Blue Tits (Parus caeruleus) I. A test for inter-population differences in resting metabolic rate and thermal conductance as a response to hot climates

Blue Tits (Parus caeruleus) are widely distributed throughout Europe, reaching the southern limit of their range on islands in the Mediterranean and in northern Africa. On Corsica, one population located at Pirio in the Fango Valley breeds roughly one month later than populations in adjacent valleys or on the continent, thus exposing nestlings to high ambient temperatures (T_a). We tested the hypothesis that nestlings and possibly adult Blue Tits at Pirio would exhibit a reduction in resting metabolic rate (MR) and an increase in thermal conductance as a physiological response to high T_a. We compared the thermoregulatory response and evaporative water loss for nestlings from Pirio in Corsica and one continental site (Vic-le-Fesq) and for adults from two Corsican (Pirio and Muro) and one continental site (La Rouvière). At 12–14 days of age, nestlings from Pirio showed two distinct thermoregulatory patterns. Nestlings under 8.0 g behaved as heterotherms, whereby MR was correlated only with body temperature. At body masses above 8.0 g nestlings progressively acquired the ability to regulate T_b and at masses >9.0 g they behaved as homeotherms. When considering homeothermic nestlings and adults, population of origin did not affect either thermal conductance or resting MR. For homeothermic nestlings, mass-specific resting MR (mW · g-¹) was 15.5 ± 2.6 and 17.5 ± 2.5 for nestlings from Vic-le-Fesq and Pirio, respectively. For adults, mass-specific resting MR (mW · g-¹) was 17.5 ± 2.0, 17.8 ± 1.6, and 17.9 ± 1.0 for birds from Pirio, Muro, and La Rouvière, respectively. Although there was a weak but positive effect of T_a on evaporative water loss for homeothermic nestlings, no such trend was evident for adults over the range of T_a tested in this study. We thus find no evidence to indicate that either nestlings or adults exhibit the exponential increase in evaporative water loss associated with the non-convective regulation of T_b within the range of T_a tested (roughly ≤35 °C). We conclude that there is no evidence for a specific physiological adaptation in the Pirio population. Measures of nestbox temperatures indicate that nestlings rarely experience temperatures in excess of 33 °C. We conclude that, although some years may be hot enough to impose a thermal stress, temperatures at Pirio are not high enough to consistently impose a selective pressure for physiological adaptations to heat.     

Recessive yellow in the Mongolian gerbil (Meriones unguiculatus)

A new autosomal recessive coat color mutant in the Mongolian gerbil (Meriones unguiculatus) is described: recessive yellow. On the dorsal side the mutant has a rich yellow to ginger color. Ventrally it shows the typical creamy white belly of a wild-type Mongolian gerbil. The dorsal yellow hairs have short black tips, and a light olive green base. A clear demarcation line between dorsal and ventral color is present. Crosses between recessive yellow animals and multiple homozygous recessive tester animals (a/a; c^chm/c^chm; g/g; p/p) resulted only in animals of an agouti (wild-type) phenotype, showing that the new allele is not allelic with any of the known coat color mutations in the Mongolian gerbil. Molecular studies showed that the new mutant is caused by a missence mutation at the extension (E) locus. On a non-agouti background (a/a; e/e) mutant animals look like a dark wild-type agouti. In contrast to wild-type agouti it shows yellow pigmentation and dark ticking at the ventral side, resulting in the absence of a demarcation line. Since black pigment is present in both the agouti and non-agouti variant (A/A; e/e and a/a; e/e), we conclude that recessive yellow in the Mongolian gerbil is non-epistatic to agouti. Additionally we describe a second mutation at the same locus leading to a similar phenotype, however without black pigment and diminishing yellow pigment during life. Fertility and viability of both new mutants are within normal range. The extension (E) gene is known to encode the melanocortin 1 receptor (MC1R). Interestingly, this is the only gene that is known to account for substantial variation in skin and hair color in humans. Many different mutations are known of which some are associated with higher skin cancer incidence.     

Morphological features of the epididymal epithelium of gerbil, Meriones unguiculatus

Principal cells of the ducts epididymis of the Mongolian gerbil showed ultrastructural characteristics of lining epithelium cells close related to processes of protein secretion, and transcytosis occurring between adjacent principal cells which were mainly verified in the initial segment. Principal cells also presented roles of fluid phase and adsorptive endocytoses, as well as autophagic and heterophagic lysosomal activities mainly observed in the caput epididymis. Columnar (principal) cells of the corpus epididymidis presented great number of variable vesicles and vacuoles distributed in all the cytoplasmic levels occurring a progressive coalescence pattern among them, which help to guarantee formation of cytoplasmic channels for fluid phase transport between the tubular lumen and epididymal interstitium. Clear cells were presented in the initial segment and predominately in the cauda epididymis epithelium of the gerbil and showed marked ultrastructural characteristics of endocytosis activities occurrence, perhaps directly related to the turnover of fluid phase of spermatozoa stored into the lumen of the distal tail. Other epididymal epithelium cells were verified and described such as basal, halo, apical and dark cells, but they did not presented special ultrastructural features.     

Thermoregulatory, behavioral and seizure modulatory effects of AVP in the gerbil

Recent reports suggest that arginine vasopressin (AVP) may be an endogenous antipyretic peptide and a mediator of febrile convulsions [10,12]. The spontaneously seizing Mongolian gerbil was used to investigate the thermoregulatory, behavioral and seizure modulatory effects of AVP. Injection of AVP (1.0 and 5.0 μg IV and 0.01–1.0 mg/kg SC) caused dose-related falls in body temperature. Stereotypic scratching, terminated by a body shake, was observed after AVP (1.0–5.0 μg IV). However, such behavior was not observed after subcutaneous injection of AVP. AVP did not potentiate seizure induction in the gerbils but rather reduced the seizure incidence. The data demonstrate that AVP can reduce body temperature and cause specific behaviors, but it does not appear to play a role in the pathogenesis of seizures in the seizure sensitive strain of Mongolian gerbil.     

Diel thermoregulation of the crawfish Procambarus Clarkii (crustacea, cambaridae)

1. 1. In a diel cycle Procambarus clarkii has two preferred temperatures: 24.0 ± 0.15 SEM °C during the day and 26.7 ± 0.13 SEM °C at night.

2. 2. The preferred temperatures are independent from the weight of the organisms.

3. 3. In the photophase the animals are dispersed, in the scotophase they congregate.

4. 4. The crawfish seem to feed during the thermal interphases.

5. 5. Animals in a constantly dark condition maintain a diel preferendum of temperature.

Reproduction and postnatal development of the bushveld gerbil Gerbilliscus (formerly Tatera) leucogaster

The reproduction and postnatal development of the bushveld gerbil Gerbilliscus (formerly Tatera) leucogaster was studied in the laboratory. Nineteen pairs produced 23 litters. Mean litter size was 3.5 and gestation was 21–22 days. Neonates weighed 3.7 g on average and were altricial. Development was slow, with eyes usually opening 16–18 days after birth, and weaning occurring by about 24 days of age. The earliest age of sexual maturity was 6.6 weeks in females and 9.9 weeks in males. A comparison with other studies of G. leucogaster, and with closely and distantly related similar-sized murid rodents, indicates that reproduction generally varies with geographic location, and that the slow postnatal development of G. leucogaster appears to be phylogenetically constrained.     

Rate of metabolism in the smallest simian primate,the pygmy marmoset (Cebuella pygmaea)

Rate of metabolism was measured with six adult pygmy marmosets (Cebuella pygmaea) at regulated ambient temperatures ranging between 20°C and 35°C. A novel combined nest box and metabolic chamber was designed to allow nighttime measurements on immobile animals in their home cage without disturbance. The basal rate of metabolism (BMR) was 98 ml O₂ h⁻¹, representing 74% of the value expected from the equation of McNab [Quarterly Review of Biology 63:25–54, 1988] relative to body mass. The thermoneutral zone was approximately 27–34°C. Below the lower critical temperature (27–28°C), thermal conductance (12.9 ml O₂ h⁻¹ °C⁻¹) was close to the predicted value. Body temperature ranged between 34.9°C and 35.5°C at night. When two animals rested together overnight in the nest box, the lower critical temperature was slightly lowered, and individual energy expenditure at 20–21°C was reduced by about 34%. The basal rate of metabolism of C. pygmaea is much lower than reported in an earlier study based on daytime measurements but agrees with values reported from a more recent study conducted at night with a classical metabolic chamber. In order to compare the BMR of C. pygmaea with that of other primates, 23 species were included in a comparative study taking into account both phylogeny and body mass (independent contrasts approach). The scaling exponent of BMR to body mass obtained was indistinguishable from that published for eutherian mammals in general. Cebuella and Callithrix exhibit the lowest basal rates known for simians. This trait may possibly be linked to the natural diet, which includes a large proportion of gums that are difficult to digest, but additional metabolic studies on primates are needed for further examination of its adaptive significance. Am. J. Primatol. 41:229–245, 1997. © 1997 Wiley-Liss, Inc.     

Thermoregulation by Diabrotica undecimpunctata howardi and potential effects on overwintering biology

In North Carolina, southern corn rootworm (SCR), Diabrotica undecimpunctata howardi Barber, adults of both sexes (diapausing and nondiapausing) elevate body temperature (Tb) above ambient air temperature (Ta) by basking in direct sunlight on clear fall and winter days when Ta<13°C (Tb-Ta range: 0.8–13.3°C). On cloudy days, SCR adults did not exhibit basking behavior and Tb was more highly correlated with Ta, ground temperature (Tg), and substrate temperature (Ts) than on clear days. Ts was the best predictor of Tb regardless of Ta and the presence or absence of adult basking behavior (fall basking SCR, Ts vs. Tb: R²=0.94, Ta vs. Tb: R²=0.41; fall nonbasking SCR, Ts vs. Tb: R²=0.85, Ta vs. Tb: R²=0.55).These results suggest that SCR thermoregulation at low Ta is ectothermic regulation by microhabitat selection. Elevation of Tb by thermoregulation was often of sufficient magnitude to affect SCR behavior (flight, feeding, mating) and preovipositional development rate. This could explain why gravid SCR females have been found in North Carolina by 6 January when Ta's were too low for development. The relationship of Tb to Ta on cold sunny days would be important to include in any predictive day-degree model of SCR postdiapause development or SCR endoparasite development in the field.

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Résumé En Caroline du Nord, les adultes diapausants ou non des deux sexes de Diabrotica undecimpunctata howardi (SCR) élèvent la température de leurs corps (Tb) au-dessus de la température de l'air ambiant (Ta) en lézardant en plein soleil par jours clairs du printemps et de l'hiver, quand Ta<13°C (Tb-Ta de 0.8 à 13.3°C). Par temps nuageux, les adultes ne lézardent pas, et Tb dépend beaucoup plus de Ta, de la température du sol (Tg), et de la température du substrat (Ts), que par temps clair. Ts fournit la meilleure idée de Ta que les adultes lézardent ou non; quand SCR lézarde au printemps la corrélation de Ts avec Tb est R²=0.94, de Ta avec Tb:R²=0.41; au printemps, quand ils ne lézardent pas; Ts avec TbR²=0.85 et Ta avec Tb:R²=0.55. Ces résultats suggerent que la thermorégulation de SCR aux basses Ta est une régulation ectothermique par sélection du microhabitat. L'augmentation de Tb par thermorégulation est souvent suffisante pour modifier le comportement de SCR (vol, prise de nourriture, accouplement) et un développement préponte. Ceci pourrait expliquer pourquoi des femelles gravides ont été trouvées un 6 janvier en Caroline du Nord quand Ta était trop basse pour permettre le développement. II paraît important d'inclure la relation entre Tb et Ta par temps nuageux froid, dans tout modèle prédictif, basé sur la notion de degrés-jours, sur le développement de SCR après la diapause ou le dévelopment d'endoparasites de SCR dans la nature.

     

Seasonal changes in thermal biology of Podarcis lilfordi (Squamata,Lacertidae) consistently depend on habitat traits

During spring and summer, we studied the thermal ecology of two populations of the Balearic lizard, Podarcis lilfordi, from two coastal islets of Menorca (Balearic Islands, Spain): Aire and Colom. We calculated the accuracy of thermoregulation, that is, the extent to which body temperatures are close to species' thermal optima, the thermal quality of the habitat as the proximity of operative temperatures to thermal optima and effectiveness of thermoregulation, as the extent to which accuracy is higher than thermal quality of the habitat. We found that seasonality affects thermoregulation differently, depending on the lizard population. Those effects are consistent for all thermal parameters under study. The effects of seasonality were significantly stronger in Aire than in Colom islet. Many factors may be responsible for this different effect of seasonality, from differences on physiological traits to differences in the environmental conditions of the two islets, as their resource availability, predator pressure or habitat structure. Identifying the factors that boost or inhibit those seasonal changes would be important to understand thermoregulation in lizards. Slight changes on two similar populations can lead to great differences in thermal ecology of conspecific ectotherms.     

Postprandial thermophily in the Danube crested newt, Triturus dobrogicus

(1) Preferred temperatures were compared between fed and fasted newts with respect to their locomotor activity (LA) in an aquatic thermal gradient (5–32.5°C). (2) Locations and LA of 17 newts in postabsorptive phase were recorded over 24 h. (3) Nine randomly chosen newts were fed (10% of their body mass) and behaviour of all newts was recorded over the next 24 h. (4) Fasted newts preferred similar water temperatures during periods of both LA and inactivity. (5) Newts preferred 2–3°C higher water temperatures after feeding irrespective of the behavioural state.     

The effects of thermal biology and refuge availability on the restricted distribution of an alpine lizard

Aim In an effort to disentangle the ecological processes that confine ectotherms to alpine environments, we studied the thermoregulatory and microhabitat selection behaviours of the rock lizard Iberolacerta cyreni, which is endemic to some mountains of central Spain, and of the wall lizard Podarcis muralis, which is a potential competitor of rock lizards. Location We chose three areas in the Sierra de Guadarrama (central Spain) that differed in their thermal quality [mean deviation of environmental operative temperatures from the lizards’ preferred thermal range (PTR)] and refuge availability: a pine forest (1770 m a.s.l.) in which P. muralis was the only species found, and two mixed shrub and rock sites (1770 and 1900 m a.s.l.) where both species were present. Methods In the field we collected data on refuge availability, sun exposure, body temperature (T_b) and operative temperature (T_e). Thus, we estimated the thermal habitat quality of the areas sampled and the thermoregulation accuracy and effectiveness of both species. Results The pine forest had the lowest thermal quality and refuge availability. The lower‐elevation shrub site offered the best thermal quality, but refuges were much scarcer than at the higher‐elevation site. Both species thermoregulated accurately, because mean deviations of body temperature (T_b) from PTR were considerably smaller than those of T_e. Podarcis muralis had higher T_b values than did I. cyreni, which had similar T_b values at both shrub sites, whereas P. muralis had lower T_b values at higher elevation. Overall, the thermoregulatory effectiveness (extent to which T_b values are closer to the PTR than are T_e values) of both species was similar, but whereas I. cyreni thermoregulated more efficiently at higher elevation, the opposite was true for P. muralis. At the lower‐elevation shrub site, I. cyreni remained closer to refuges than did P. muralis. Main conclusions Our results suggest that the pine forest belt might prevent the expansion of rock lizards towards lower elevations as a result of its low thermal quality and scarcity of refuges, that the thermoregulatory effectiveness of rock lizards in alpine environments depends more on refuge availability than on thermal habitat quality, and that competition with wall lizards is unlikely to explain either the distribution or the thermoregulatory effectiveness of rock lizards.     

The interstitial cells in the renal medulla of rat,rabbit, and gerbil in different states of diuresis

Summary The inner zone of the renal medulla of rats, gerbils, and rabbits was investigated to determine whether or not there are any characteristic ultrastructural differences between the interstitial cells of these species. The effects on the interstitial cells of water deprivation and water loading were also investigated.In all three species, the Type 1 interstitial cells, the lipid containing cells, were abundant and their distribution and topographical relations as well as their general ultrastructure were similar. The previously reported significantly higher frequency in desert rats could not be confirmed. Although the lipid droplets of the interstitial cells were smaller in gerbils and rabbits when compared to rats, their fine structure was similar. Their electron dense outer zone was sometimes associated with a granular material and/or a lamellar material with a periodicity of about 40 Å resembling phospholipid myelin figures.Water-loaded rats showed a considerable increase in the number of lipid droplets when compared to dehydrated or untreated animals. In contrast, the interstitial cells of waterloaded gerbils and rabbits were depleted of lipid droplets.We are indebted to Professor A.B. Maunsbach for valuable discussions and criticism and to Mrss. Hanne Weiling and Birthe Overgaard for competent technical assistance. The gerbils used in this study were a gift from Leo AB, Helsingborg, Sweden. This study was supported by the Danish Medical Research Council (J. nos. 512-1067, 512-1545, 512-3633)     

蒙新区子午沙鼠种群的遗传多样性和遗传结构

为了探讨蒙新干旱区景观和环境因素对子午沙鼠种群遗传结构的影响,我们利用8 个多态性的微卫星位点,对子午沙鼠14 个局域种群的160 个个体进行了种群遗传多样性和遗传结构的分析。微卫星结果显示种群具有高的遗传多样性,微卫星位点的等位基因数(A)为22. 50 ±3.02 (19 ～ 28),多态信息含量(PIC)为0.912 ±0. 02 (0.872 ～ 0. 929);种群观测杂合度(HO )为0.68 ± 0. 19 (0. 52 ～0.85),期望杂合度(HE )为0.79 ± 0. 08(0. 71 ～ 0.85)。分子变异分析(AMOVA)结果显示,所得到的3 个组之间已发生显著的遗传分化(P < 0. 001)。
Mantel 检测结果表明,子午沙鼠种群遗传结构与地理距离无显著的相关性(P = 0.270 0),而在多元线性模型中,蒙新干旱区的海拔高度是影响种群遗传结构的关键因素。     

Central thermoregulatory effects of lactate in the toad Bufo paracnemis

Hypoxia induces a regulated decrease in body temperature (Tb; anapyrexia) in organisms ranging from protozoans to mammals, but very little is known about the mechanisms involved. Several candidates have been suggested to mediate hypoxia-induced anapyrexia, among them lactate, which is a classical companion of hypoxic stress in vertebrates. The present study was designed to assess the central thermoregulatory effects of lactate in Bujo paracnemis. Toads equipped with a temperature probe were tested over a thermal gradient (10–40°C). Lactate injected systemically (4.0 mmol kg⁻¹) caused a significant reduction of Tb from 24.6±2.1 to 17.4±3.9°C. To assess the role of central thermoregulatory mechanisms, a lower dose (0.4 mmol kg⁻¹) of lactate was injected into the fourth cerebral ventricle or systemically. Intracerebroventricular injection of lactate caused a similar decrease in Tb, whereas systemic injection caused no change. The data indicate that lactate may play a role in hypoxia-induced anapyrexia in central rather than peripheral sites.     

Incubation regimes of cold-climate reptiles: the thermal consequences of nest-site choice, viviparity and maternal basking

Cold-climate reptiles show three kinds of adaptation to provide warmer incubation regimes for their developing embryos: maternal selection of hot nest sites; prolonged uterine retention of eggs; and increased maternal basking during pregnancy. These traits may evolve sequentially as an oviparous lineage invades colder climates. To compare the thermal consequences of these adaptations, I measured microhabitat temperatures of potential nest sites and actual nests of oviparous scincid lizards ( Bassiana duperreyi ), and body temperatures of pregnant and non-pregnant viviparous scincid lizards ( Eulamprus heatwolei ). These comparisons were made at a site where both species occur, but close to the upper elevational limit for oviparous reptiles in south-eastern Australia. Viviparity and maternal basking effort had less effect on mean incubation temperature than did maternal nest-site selection. Eggs retained in utero experienced bimodal rather than unimodal diel thermal distributions, but similar mean incubation temperatures. Often the published literature emphasizes the ability of heliothermic (basking) reptiles to maintain high body temperatures despite unfavourable ambient weather conditions; this putative ability is central to many hypotheses on selective forces for the evolution of viviparity. In cold climates, however, opportunities for maternal thermoregulation to elevate mean body temperatures (and thus, incubation temperatures) above ambient levels may be severely limited. Hence, at least over the broad elevational range in which oviparous and viviparous species live in sympatry, maternal selection of 'hot' nests may be as effective as is viviparity in providing favourable incubation regimes.  © 2004 The Linnean Society of London, Biological Journal of the Linnean Society , 2004, 83 , 145–155.     

Intraventricular blood vessels associated with the deep pineal gland of the Mongolian gerbil,Meriones unguiculatus

Summary Intraventricular blood vessels and choroidal-like cells were studied using scanning electron microscopy and correlative light microscopy. The intraventricular blood vessels were covered on their ependymal surface with a layer of cells essentially identical to the ependyma of the choroid plexus in the gerbil. Similar choroidal-like cells were seen either singly or in clusters associated with the cerebrospinal fluid-contacting pinealocytes of the suprapineal recess. Processes of the cerebrospinal fluid-contacting pinealocytes were seen extending to and making contact with the choroidal-like cells. The intraventricular blood vessels appeared to be derived from the choroid plexus, and typically took one of three courses in and around the surface of the deep pineal: (1) the vessels or their equivalent were located in the suprapineal recess with no indication of penetration into the substance of the deep pineal; (2) the vessels coursed from the suprapineal recess around the anterior surface of the habenular commissure to enter the ventral surface of the deep pineal; or (3) the vessels entered the parenchyma of the deep pineal from its dorsal surface and could be seen coursing through the substance of the gland. The close association between the choroidal-like cells and the intraventricular blood vessels with the deep pineal gland add morphological support for the possibility of interaction between the cerebrospinal fluid, or perhaps the choroid plexus, and the deep pineal gland.