Toshiko Hara

Improvement of spatial cognition with dietary docosahexaenoic acid is associated with an increase in Fos expression in rat CA1 hippocampus

1. Twenty 5‐week‐old male Wistar rats were divided into two groups: one group was fed a fish oil‐deficient diet and the other group was fed the same diet supplemented with per orally administered docosahexaenoic acid (DHA) for 12 weeks.

Aging attenuates acquired heat tolerance and hypothalamic neurogenesis in rats

This study investigated age‐dependent changes in heat exposure‐induced hypothalamic neurogenesis and acquired heat tolerance in rats. We previously reported that neuronal progenitor cell proliferation and neural differentiation are enhanced in the hypothalamus of long‐term heat‐acclimated (HA) rats. Male Wistar rats, 5 weeks (Young), 10–11 months (Adult), or 22–25 months (Old) old, were subjected to an ambient temperature of 32°C for 40–50 days (HA rats). Rats underwent a heat tolerance test. In HA rats, increases in abdominal temperature (Tab) in the the Young, Adult, and Old groups were significantly smaller than those in their respective controls. However, the increase in Tab of HA rats became greater with advancing age. The number of hypothalamic bromodeoxyuridine (BrdU)‐immunopositive cells double stained with a mature neuron marker, neuronal nuclei (NeuN), of HA rats was significantly higher in the Young group than that in the control group. In Young HA, BrdU/NeuN‐immunopositive cells of the preoptic area/anterior hypothalamus appeared to be the highest among regions examined. Large numbers of newborn neurons were also located in the ventromedial and dorsomedial nuclei, as well as the posterior hypothalamic area, whereas heat exposure did not increase such numbers in the Adult and Old groups. Aging may interfere with heat exposure‐induced hypothalamic neurogenesis and acquired heat tolerance in rats. J. Comp. Neurol. 523:1190–1201, 2015.

β-amyloid infusion into lateral ventricle alters behavioral thermoregulation and attenuates acquired heat tolerance in rats

We investigated behavioral thermoregulatory function and acquired heat tolerance of β-amyloid (Aβ)-infused rats. Male Wistar rats were anesthetized and implanted in the intraperitoneal cavity with a temperature transmitter. Aβ peptide (4.9–5.5 nmol) was dissolved in a solvent of 35% acetonitrile and 0.1% trifluoroacetic acid (pH 2.0). The solvent was used as the vehicle. An osmotic pump contained 234 ± 13.9 μl of Aβ solution was subcutaneously implanted in the back and was cannulated into the left cerebral ventricle. Moreover, 0.5 µg of AlCl3 was injected into the right cerebral ventricle with a micro syringe pump (Aβ-infused rats). The solvent-infused rats were used as control rats (CN rats). After 2 weeks, rats were placed in a thermal gradient and their intra-abdominal temperature (Tab) and their ambient temperatures (Ta) selected (Ts) were measured for 3 consecutive days. In an additional study, rats were kept at a Ta of 32°C for 4 weeks to attain heat acclimation. Then, rats were subjected to a heat tolerance test, i.e. they were exposed to a Ta of 36°C for 160 min. Although there were clear day-night variations of Ts and Tab in CN rats, patterns were significantly abolished in Aβ-infused rats. Moreover, heat tolerance obtained by heat acclimation was attenuated in Aβ-infused rats. These results suggest that Aβ-infusion in the lateral ventricle modifies behavioral thermoregulation and lowers an ability to acclimate to heat in rats.

Neural progenitor cell proliferation in the hypothalamus is involved in acquired heat tolerance in long-term heat-acclimated rats

Constant exposure to moderate heat facilitates progenitor cell proliferation and neuronal differentiation in the hypothalamus of heat-acclimated (HA) rats. In this study, we investigated neural phenotype and responsiveness to heat in HA rats’ hypothalamic newborn cells. Additionally, the effect of hypothalamic neurogenesis on heat acclimation in rats was evaluated. Male Wistar rats (5 weeks old) were housed at an ambient temperature (Ta) of 32°C for 6 days (STHA) or 40 days (LTHA), while control (CN) rats were kept at a Ta of 24°C for 6 days (STCN) or 40 days (LTCN). Bromodeoxyuridine (BrdU) was intraperitoneally injected daily for five consecutive days (50 mg/kg/day) after commencing heat exposure. The number of hypothalamic BrdU-immunopositive (BrdU+) cells in STHA and LTHA rats was determined immunohistochemically in brain samples and found to be significantly greater than those in respective CN groups. In LTHA rats, approximately 32.6% of BrdU+ cells in the preoptic area (POA) of the anterior hypothalamus were stained by GAD67, a GABAergic neuron marker, and 15.2% of BrdU+ cells were stained by the glutamate transporter, a glutamatergic neuron marker. In addition, 63.2% of BrdU+ cells in the POA were immunolabeled with c-Fos. Intracerebral administration of the mitosis inhibitor, cytosine arabinoside (AraC), interfered with the proliferation of neural progenitor cells and acquired heat tolerance in LTHA rats, whereas the selected ambient temperature was not changed. These results demonstrate that heat exposure generates heat responsive neurons in the POA, suggesting a pivotal role in autonomic thermoregulation in long-term heat-acclimated rats.

Daily voluntary exercise enhances pilocarpine‐induced saliva secretion and aquaporin 1 expression in rat submandibular glands

Saliva—a water‐based fluid containing electrolytes, immunoglobulins, and enzymes—has many functions, including the protection and hydration of mucosal structures within the oral cavity and the initiation of digestion. Aquaporins (AQPs) are proteins that act as water channels through membranes. We have previously reported upregulation of the expression levels of AQP 1 and 5 in the submandibular glands (SMGs) in heat‐acclimated rats. In this study, we investigated pilocarpine‐induced saliva secretion and AQP expression in rats after voluntary exercise. Male, 10‐week‐old Wistar rats were initially maintained at an ambient temperature of 24 °C for 10 days and were then kept for 40 days in cages either with a running wheel (exercise rats, n = 6) or with a locked wheel [control rats (CN), n = 6]. After the training period, the rats were anesthetized and pilocarpine, an M3 muscarinic receptor agonist, was intraperitoneally injected (0.5 mg·kg−1) to stimulate saliva secretion. Saliva was collected, and the SMGs were sampled and subjected to western blot, RT‐PCR, and immunohistochemical analyses. Pilocarpine induced a greater amount of saliva in the exercised rats than in the CN. Expression levels of AQP1 mRNA and protein were significantly higher in SMGs of exercised rats than in those of the CN, but the expression of AQP5 was not affected by voluntary exercise. Voluntary exercise increased the expression of vascular endothelial growth factor (VEGF) and cluster of differentiation 31 (CD31), a marker for endothelial cells, in the SMGs. Voluntary exercise promoted pilocarpine‐induced saliva secretion, probably via an increase in the expression level of AQP1 due to VEGF‐induced CD31‐positive angiogenesis in the SMG.