Fatty acid composition of brown adipose tissue in genetically heat-tolerant FOK rats.

The phospholipid fatty acid composition of brown adipose tissue (BAT) was examined in inbred heat-tolerant FOK rats and compared with that in conventional Wistar rats not previously exposed to heat. The FOK rats showed higher unsaturation states, as indicated by higher levels of polyunsaturated fatty acids and a higher unsaturation index and polyunsaturated fatty acids/saturated fatty acids ratio. This higher level of unsaturation was characterized by the higher amount of polyunsaturated fatty acids such as linoleic acid, arachidonic acid and docosahexaenoic acid. It may be concluded that the increased docosahexaenoic acid level in BAT phospholipids brings about the hyperplasia of BAT, causing an enhancement of its in vivo thermogenic activity as well as the systemic non-shivering thermogenesis observed in heat-tolerant FOK rats.

Thermoregulatory responses of the inbred heat-tolerant FOK rat to cold.

The responses of inbred heat-tolerant FOK rats to cold were compared with those of Wistar King A/H (WKAH) and Std:Wistar (WSTR) strains. The fall of colonic temperature during cold exposure was unexpectedly smaller in FOK than in other groups, but the onset of shivering was delayed in FOK. Norepinephrine (NE)-induced in vivo oxygen consumption and the mitochondrial uncoupling protein 1 level of brown adipose tissue (BAT) were not different among the groups, but the cold-induced increases in in vivo oxygen consumption as well as plasma glycerol and free fatty acids were higher in FOK than in other groups. In vitro NE-induced oxygen consumption of BAT was less in FOK than WSTR, but not WKAH. The magnitude of the NE-induced increase in blood flow through BAT was higher in FOK than in other groups. These results suggest that FOK paradoxically have a high capacity for nonshivering thermogenesis in spite of their high capacity for heat tolerance, probably due to an increased lipid utilization and improved circulation of BAT.

Thermoregulatory responses to acute heat loads in the FOK rat.

FOK is an inbred rat strain with a genotypic adaptation to hot environments. The present study investigated the mechanism of the high heat tolerance of the FOK rat. Male FOK and WKAH rats were used. They were loosely restrained and placed individually in a direct calorimeter with an ambient temperature of 24 degrees C. Their hypothalamic temperature, evaporative and non-evaporative heat loss and heat production were measured. After thermal equilibrium had been attained, the rats were warmed for 30 min with a chronically implanted intraperitoneal electric heater (internal heating). At least 90 min after the heating, the jacket water temperature surrounding the calorimeter chamber was gradually raised from 24 degrees C to 36 degrees C in 80 min (external warming). During the internal heating, changes in the thermoregulatory parameters did not differ between the groups. During the external warming, the evaporative heat loss of the FOK rat was significantly greater than that of the WKAH rat, while changes in nonevaporative heat loss and heat production did not differ between the groups. The results suggest that in the FOK rat, the improved heat tolerance is attributable to an enhanced evaporative heat loss response, but not to a facilitation of nonevaporative heat loss or of metabolic depression.

Differences in thermal salivation between the FOK rat (a model of genotypic heat adaptation) and three other rat strains.

Rats secrete saliva in response to heat. In the present study, details of thermal salivation were investigated using the FOK rat in comparison with Sprague-Dawley (SD), Donryu, and ACI rats. The FOK rat is a strain inbred for genotypic heat adaptation and endures heat for long periods. Conscious rats of all four strains were exposed to 42.5 degrees C. The order of heat endurance times at this temperature was FOK >> SD > Donryu = ACI. FOK rats spread their saliva over their entire ventral surface, their faces, and their outside legs. This saliva area was wider than those made by the other three strains. SD rats spread in an area wider than those of the Donryu and ACI rats. Saliva spreading in the FOK rats continued for 4.0-4.5 h, far longer than in the other strains. Under ketamine anesthesia and exposure to 40 degrees C, the FOK rats secreted saliva at 1390+/-235 microL/100 g of body weight during a 60-min observation period. This was the highest rate among the four rat strains (p < 0.0001). The body temperature increase rate in anesthetized FOK and SD rats was lower than in the other two strains, suggesting a minor contribution of unknown factors. Ligation of the submandibular gland ducts abolished the thermal salivation of the FOK rats, whereas ligation of the parotid duct had no effect. The submandibular, sublingual, and lachrymal glands in the FOK rats were 1.3-1.5, 1.25-1.4, and 1.3-1.5 times heavier, respectively, than those in the other three strains, whereas the parotid gland of the FOK rats was not enlarged. These findings indicate that the rats' saliva spreading and ET values are significantly correlated. A potentiated and long-lasting salivation from the submandibular gland was acquired during development of genotypic heat adaptation. This salivation is actuated in response to heat. The pronounced thermal salivation is probably attributable to adaptive changes in the superior salivatory nucleus-chorda tympani-submandibular gland pathway.

Thermoeffector thresholds and preferred ambient temperatures of the FOK rat.

The FOK is an inbred rat strain with a genotypic adaptation to hot environments. The present study compared the thermoeffector thresholds and preferred ambient temperatures (Tpref) of the FOK rat with those of other rat strains. Male FOK, WKAH, and Donryu rats were used. First, they were loosely restrained and placed individually in a metabolic chamber with an ambient temperature of 26.0 degrees C. Their hypothalamic temperature (T(hy)), tail skin temperature (Tsk), and heat production (M) were measured. After thermal equilibrium had been attained, the rats were gradually warmed and then cooled using an intravenous thermode. The threshold T(hy) values for tail skin vasodilation and cold-induced thermogenesis were defined as the points at which sharp increases in Tsk and M occurred, respectively. The two thresholds of the FOK rat were lower than those of the WKAH and Donryu rats. In a second set of experiments, the FOK and WKAH rats were placed individually in a thermocline. Their intra-abdominal temperatures (T(ab)) were measured by a biotelemetry system, and the rats' Tpref values were estimated with the thermal gradient. Mean T(ab) and Tpref over a 24-h period for the FOK rat were significantly lower than those of the WKAH rat. The results suggest that in the FOK rat the control ranges of autonomic and behavioral thermoregulation are lower than those of the other rat strains examined. This contributes to the maintenance of core temperature at low levels.

Heat production by brown adipocytes of FOK rats genetically resistant to a hot environment.

The FOK rat was established as an inbred strain with genotypic adaptation to a hot environment. To investigate the mechanism of the heat resistance of FOK rats, heat production by isolated brown adipocytes in FOK and WKAH rats was measured at 37.0 and 39.9 degrees C. Basal heat production by brown adipocytes of FOK rats was not significantly different from that of WKAH rats. Isoproterenol increased the heat production in a dose-dependent manner in both strains, while 10(-9) M isoproterenol caused a significant increase in WKAH rats and a negligible change in FOK rats. These results indicate that the reason why FOK rats are resistant to the severe heat is partly due to the low heat production by brown adipocytes.

Genetic development of an inbred rat strain with increased resistance adaptation to a hot environment.

The purpose of this study was to demonstrate resistance adaptation based on genotypic adaptation and to develop an inbred rat strain with genotypic resistance adaptation to a hot environment. Survival time (ST) at an ambient temperature (Ta) of 42.5 degrees C was determined without appreciable thermal damage. Rats with the longest ST were sibmated over 30 generations and designated FOK. The ST, evaporative water loss, and body water economy increased rapidly during the first 10-15 generations, followed by a more gradual increase. The FOK rat resisted a Ta of 42.5 degrees C for > 5 h; this ability was hereditary. Body size progressively decreased during the early generations. The ST was longer (P < 0.01) and evaporative water loss was significantly higher (P < 0.01) in the FOK rat compared with three other strains. Tail lengths in the FOK rat were slightly shorter than those of three controls (P < 0.01). The heat loss system in the FOK rat may depend on the ability to mobilize and evaporate body fluids efficiently. FOK rats can be used for phenotypic comparisons with other strains, as well as in molecular genetic studies on thermoregulation, osmoregulation, and resistance adaptation to heat using recombinant inbred and recombinant congenic lines.

Sialyl cholesterol enhances the development of grafted neurons and motor recovery.

Trophic actions of alpha-sialyl cholesterol (SC) and its sialidase-tolerant derivative, alpha-(3 beta-hydroxysialyl) cholesterol (SCt), were carried out on the development of midbrain neurons both in vitro and in vivo transplantation studies. Low to moderate concentrations of SC (0.01 to 0.05 micrograms/ml) facilitated neurite extension but had no effects on cell survival of primary cultured midbrain neurons. However, high concentration of SC (0.1 micrograms/ml) disturbed both neurite genesis and cell survival. SCt had a similar effect on midbrain neurons. At higher concentrations, SC and SCt induced concentration-dependent morphological changes in astrocytes from flat to fibrous. The effect on astrocytes was stronger in SCt than SC. At highest concentration tested (20 micrograms/ml), the proliferation of astrocytes was completely blocked, cells became detached and finally died. This effect of SC and SCt was partially blocked by simultaneous application of aFGF. Following dopaminergic cell grafting in vivo, SC and SCt had biphasic effects: a low dose (0.2 mg/kg, SC) enhanced motor recovery at 4 and 6 weeks after transplantation, while the highest dose (20 mg/kg, SC) disturbed motor recovery at all periods tested. These effects on motor recovery were paralleled by an effect on neurite genesis as studied by tyrosine hydroxylase immunostaining. Thus, at low concentrations, SC and SCt are neurotrophic agents that stimulate the development and differentiation of dopaminergic neurons.

Overshoot and high equilibrium in body temperature responses of rats to ambient heat: relation to thermoregulatory ability and improvement in estimation of survival time in a hot environment.

Physiological significance of body temperature responses to ambient heat (BTRAH) with high overshoot or a high equilibrium phase was studied in relation to thermoregulatory ability. Subsequently, an attempt was made to predict survival time (ST) by measuring the period required to attain a colonic temperature (Tco) of 42.0 degrees C (t42.0), whereas ST hitherto having been calculated on the basis of a period until Tco attains 42.5 degrees C (t42.5). Tco of rat was monitored continuously with a Cu-Co thermocouple during exposure to an ambient temperature (Ta) of 42.5 degrees C. The lower the overshoot temperature (Tos) or the equilibrium temperature (Teq), the longer the rats survived in a hot environment. The present findings further suggest that these two types of BTRAH are immature variations of the typical triphasic BTRAH which is characteristic of heat resistant individuals. A new regression line of ST (Y) as a function of the t42.0 (X) was obtained for most rats as follows: Y = 0.963X + 43.85 (male); and Y = 0.973X + 39.10 (female). This equation enabled to calculate ST without thermal death. However, the former approach based on t42.5 must be applied yet in small number of rats which showed the irregular BTRAHs with Tos or Teq higher than 42.0 degrees C. Delayed influence on survival and fecundity at a Ta of 24 degrees C were not found during one year following this hyperthermia up to 42.5 degrees C.

Long-term survival of grafted cells, dopamine synthesis/release, synaptic connections, and functional recovery after transplantation of fetal nigral cells in rats with unilateral 6-OHDA lesions in the nigrostriatal dopamine pathway.

In animal models of hemi-Parkinson's disease, survival of grafted nigral cells, their synaptic connections, dopamine (DA) synthesis/release, and recovery from motor disturbances were investigated, and these were compared among 3 groups of animals raised for 3 months, 1 year and 2 years after the transplantation. Fetal nigral DAergic cell suspensions were transplanted in the ipsilateral caudate nucleus of rats with unilateral 6-OHDA lesions in the nigrostriatal DA pathway. Motor disturbances, assessed by methamphetamine-induced rotation, recovered partly in the 2nd week, significantly in the 4th week after the grafting, and remained stable thereafter. Many tyrosine hydroxylase (TH)-positive cells were detected along the grafting tracks. The number of TH-positive cells was similar in the 3 groups of animals. These TH-positive cells made synaptic connections in the host caudate. By in vivo microdialysis measurement, extracellular DA, dihydroxyphenylacetic acid (DOPAC) and homovanillic acid (HVA) around the grafted sites recovered to 30-100% of those of controls. No significant differences were observed in the concentration of DA, DOPAC and HVA among 3 groups of animals. They also responded to methamphetamine loading though the magnitudes were smaller. Using a TH cDNA probe, TH-positive cells were found to express TH mRNA in in situ hybridization-autoradiographic analysis. Data indicate that grafted fetal DAergic cells survive, synthesize and release DA, make synaptic connections in the host brain and ameliorate motor disturbances for over 2 years. There were no differences in these parameters among the 3 groups of animals, and no untoward side effects were observed even at 2 years after the grafting. Thus it was confirmed that the grafting of neuronal cells into the brain is a promising approach to restore disturbed function.

tGS ganglioside induces peculiar morphological features in grafted dopaminergic cells and promotes motor recovery in rats with unilateral lesions in the nigrostriatal dopamine pathway.

A cell suspension of substantia nigra from fetal rats was introduced into the ipsilateral caudate nucleus of rats with unilateral lesions in the nigrostriatal dopamine pathway, and effects of bovine total ganglioside (tGS) and monosialoganglioside (GM1) treatment on the morphological features of the transplanted cells and recovery from motor imbalance (rotation induced by methamphetamine) were investigated. Gangliosides (30 mg/kg) were administered intraperitoneally once a day for 2 weeks after transplantation to test animals while control animals received saline alone. tGS animals showed definite motor recovery in the 2nd week (P less than 0.05) while control and GM1 animals exhibited slight recovery only. At 6 weeks after transplantation, motor imbalance disappeared in all 3 groups. Tyrosine hydroxylase (TH) immunocytochemical staining revealed that in the 2nd week TH-positive cells in tGS animals had more primary dendrites and more large neurites (meganeurites) than did controls. TH-positive cells of all 3 groups often had spiny processes at that time. In the 20th week, TH-positive cells became more multigonal and had wider dendritic fields in all groups, and had less meganeurites and spines. Motor recovery of each animal was dependent on the number of TH-positive cells and no significant difference was observed in the number of TH-positive cells among the three groups. tGS treatment for 2 weeks without grafting induced immunohistologically no axonal sprouting in the substantia nigra, medial forebrain bundle, accumbens and caudate nucleus when the chemical lesions were complete. Data suggest that tGS induces hypertrophy but not hyperplasia of the transplanted nigral cells, and increases the morphological plasticity. This might be the basis for promotion of recovery in motor function after transplantation.

Thermal salivation in rats, anesthetized with barbiturates, chloralose, urethane and ketamine.

1. The relationship between thermal salivation (TS) and thermoregulation was studied in anesthetized rats. 2. Of the 6 anesthetics used, ketamine-anesthetized rats secreted the largest amount of saliva. Salivation, however, was thermal and not induced by ketamine itself. 3. Ketamine-anesthetized rats readily secreted saliva at core temperatures less than 40 degrees C but TS was remarkably enhanced by hyperthermia of 40-42.5 degrees C. 4. The equilibrium phase in the triphasic heat response of core temperature was a consequence of equilibrium between heat gain and heat loss by salivation.

Estimation of rat thermoregulatory ability based on body temperature response to heat.

A new simple but general estimation method for survival time in a hot environment is presented in this study. Even in heat-tolerant rats showing a triphasic heat response, an accurate estimation of survival time (ST) is possible. Rat groups, which included some heat-tolerant individuals, were exposed to 42.5 degrees C, 40% rh. Colonic temperature (Tco) was measured continuously by copper-constantan thermocouple. The ST (Y) of male and female rats were expressed as a linear function of time (X) until the Tco of 42.5 degrees C was reached: Y = 0.976X + 30.6 and Y = 0.968X + 31.6, respectively. A Tco of 42.5 degrees C at rest was just below the maximum survivable body temperature and above the steady-state equilibrium Tco levels during the second phase of the triphasic heat-response curve. Heat-tolerant individuals showed lower equilibrium temperatures than heat-intolerant rats. All 140 rats survived the Tco of 42.5 degrees C and lived for more than 8 wk, thus enabling them to be used for future experiments on thermoregulation. The heat survivors were able to reproduce, and their genetically controlled offspring could be used for thermoregulatory experiments.