Yoshihiro Aikawa

The effect of walking on regional blood flow and acetylcholine in the hippocampus in conscious rats

Recent studies in our laboratory have demonstrated that stimulation of the septal complex (i.e., the medial septal nucleus and the nucleus of the diagonal band) increases extracellular acetylcholine (ACh) release and, consequently, results in an increase in regional cerebral blood flow in the hippocampus (Hpc CBF) via activation of the nicotinic ACh receptors (nAChRs) [Neurosci. Lett. 107 (1989) 135; Neurosci. Lett. 112 (1990a) 263]. The present study aimed to examine the effects of walking on Hpc CBF, measured by laser Doppler flowmetry, in conscious rats. Walking at a moderate speed (4 cm/s) on a treadmill for 30 s produced increases in Hpc CBF and mean arterial pressure (MAP), reaching 107 +/- 1% and 105 +/- 1% of the prewalking control values, respectively. Walking for 3 min produced an increase in ACh release in the extracellular space of the hippocampus. The increase in Hpc CBF during walking was attenuated by mecamylamine (abbreviated as MEC here; 2 mg/kg, i.v.), a nAChR antagonist permeable to the blood-brain barrier (BBB), but not by hexamethonium (denoted as C6 here; 20 mg/kg, i.v.), a nAChR antagonist impermeable to the BBB, while the walking-induced increase in MAP was abolished by either agent. The response of Hpc CBF and MAP were not altered by atropine (abbreviated as ATR here; 0.5 mg/kg, i.v.), a muscarinic AChR antagonist permeable to the BBB. The increase in Hpc CBF during walking was attenuated by N(omega)-nitro-L-arginine methyl ester (L-NAME, 3 and 30 mg/kg, i.v.), a nitric oxide synthase (NOS) inhibitor, and the reduced responses were reversed following the intravenous (i.v.) administration of a physiological precursor of NO, L-arginine (600 mg/kg). The results suggest that the increase in Hpc CBF during walking is independent of MAP and attributable at least to activation of the nAChRs by the cholinergic vasodilator nerves projecting to the hippocampus and to production of NO in the hippocampus.

Neural regulation of hepatic blood flow in rats: an in vivo study.

The aim of the present study was to elucidate the influence of the hepatic sympathetic and parasympathetic (vagal) nerves on the hepatic blood flow (HBF), both tonically and when stimulated, using urethane-anesthetized rats as an in vivo experimental model. HBF was measured at the surface of the lateral left lobe of the liver using laser Doppler flowmetry and the hydrogen gas clearance method. Denervation of the hepatic sympathetic nerves had no influence on the HBF, while electrical stimulation of the hepatic sympathetic nerves caused the HBF to decrease in a frequency-dependent manner. This decrease was shown to occur via alpha-adrenergic receptors. In contrast, neither denervation nor electrical stimulation of the hepatic vagal nerves elicited significant changes in the HBF. These results demonstrate that the sympathetic and vagal hepatic nerves have little or no tonic influence on the HBF of rats under urethane anesthesia, whereas the HBF decreases in response to activation of the hepatic sympathetic nerves.

Effect of Moxibustion Stimulation of Various Skin Areas on Cortical Cerebral Blood Flow in Anesthetized Rats

The effect of moxibustion stimulation of various skin areas (cheek, forepaw, upper arm, chest, back, lower leg, hindpaw and perineum) on cerebral blood flow (CBF) of the parietal cortex was examined in anesthetized rats after eliminating emotional influences. Moxibustion stimulation was performed by burning a moxa cone of about 4 mg weight placed on the shaved skin. CBF of the parietal cortex was measured using a laser Doppler flowmeter. Stimulation of the cheek, forepaw, upper arm and hindpaw produced significant increases in CBF, but stimulation of the other areas did not produce significant responses. Moxibustion stimulation of the forepaw and hindpaw produced an increase in the mean arterial pressure (MAP), while stimulation of the other areas did not. After spinal transection at the 2nd thoracic level, the MAP response to stimulation of the forepaw was abolished, whereas the CBF response to stimulation of the forepaw remained. The CBF response in spinalized rats was not affected by cutting cervical sympathetic and facial parasympathetic nerves, while it was almost abolished by intravenous administration of muscarinic and nicotinic cholinergic blocking agents. The CBF response was abolished by crushing the brachial plexus ipsilateral to the stimulated side. It is suggested that the increase in CBF, independent of MAP and emotional responses, elicited by moxibustion stimulation is a reflex response whose afferent pathway is composed of somatic afferent nerves, and whose efferent pathway involves intracerebral cholinergic nerves. A contribution of endogenous opioids in the present CBF responses was neglected, because naloxone did not influence the CBF responses.

Reflex choroidal blood flow responses of the eyeball following somatic sensory stimulation in rats

The effect of cutaneous mechanical stimulation on choroidal blood flow (ChBF) of the eyeball measured using a laser Doppler flowmeter was examined in anesthetized rats. Noxious pinching stimulation of a forepaw for 20 s produced increases in ChBF and mean arterial blood pressure (MAP), whereas brushing of a forelimb produced no changes in either parameter. After spinal transection at the fourth thoracic (T4) level, forepaw pinching stimulation did not produce any MAP changes in 9 of 11 spinalized rats. In these nine spinalized animals, pinching stimulation of a forepaw produced no significant responses in ChBF. After the cutting of cervical sympathetic trunks in five spinal rats, forepaw pinching showed no effect on MAP, but produced an increase in ChBF, which was abolished by an intravenous (i.v.) injection of 1-(2-trifluoromethylphenyl) imidazole (TRIM), a selective inhibitor of neuronal nitric oxide synthase (nNOS). In another four spinalized rats, whose cervical sympathetic trunks were intact and the superior salivary nucleus (SSN) was destroyed, forepaw pinching showed no effect on MAP, but produced a decrease in ChBF, which was abolished by an i.v. injection of phentolamine, an alpha-adrenoceptor antagonist. The present experiment shows that somatic afferent stimulation can produce reflex responses of the ChBF of the eyeball, either a vasodilative response using parasympathetic efferent fibers or a vasoconstrictive response using sympathetic efferent fibers, independent of systemic blood pressure. It was also shown that the somatically induced vasodilative response was due to a release of nitric oxide (NO) from parasympathetic nerves and the vasoconstrictive response was due to a release of noradrenaline from sympathetic nerves.

Ovarian blood flow is reflexively regulated by mechanical afferent stimulation of a hindlimb in nonpregnant anesthetized rats

The effects of mechanical afferent stimulation of either a hindpaw or a hind leg on ovarian blood flow and systemic arterial blood pressure were studied in anesthetized nonpregnant adult female rats. Ovarian blood flow at the left side was continuously measured using a laser Doppler flowmeter. A mechanical stimulus (by pinching, brushing or pressing) was delivered for 30 s to either a hindpaw or a hind leg, at the right or left side. Pinching of a hindpaw at the right or left side decreased ovarian blood flow 8+/-3% accompanied with an increase in blood pressure. Brushing or pressing of a hindpaw produced no effects on these two parameters measured. Pressing stimulation of a hind leg at the right or left side produced a decrease in ovarian blood flow 8+/-2% accompanied with a decrease in blood pressure. Pinching or brushing of a hind leg produced no effects on both parameters measured. All responses of ovarian blood flow and blood pressure mentioned above were abolished after severing the femoral and sciatic nerves at the same side in which stimulation was unilaterally delivered. The blood pressure response, either increased or decreased depending on the stimulus modality and location stimulated, was not influenced by severing of ovarian sympathetic nerves. However, the decrease response in ovarian blood flow following pinching of a hindpaw reversed to the increase response, while the decrease response in ovarian blood flow following pressing of a hind leg was slightly augmented. The activity of sympathetic nerves innervating the ovary was increased during pinching of a hindpaw of either side, and decreased during pressing of a hind leg of either side. Electrical stimulation of the distal part of the severed splanchnic nerve at the left side decreased the left ovarian blood flow, and this response was abolished by intravenous injection of phentolamine, the alpha-adrenoceptor antagonist. The present results demonstrate that ovarian blood flow increase or decrease passively to responses of an increase or a decrease in systemic arterial blood pressure following mechanical afferent stimulation of a hindlimb, and these passive responses of ovarian blood flow are modulated by reflexive activity of ovarian sympathetic vasoconstrictor nerves to attenuate blood pressure-dependent passive changes in ovarian blood flow.

Modulation of somatosensory-evoked cortical blood flow changes by GABAergic inhibition of the nucleus basalis of Meynert in urethane-anaesthetized rats

Vascular changes associated with brain functions are thought to be tightly coupled with neuronal activity through neuronal glucose consumption or the local release of vasoactive agents. In contrast, another view suggests that cortical blood flow is strongly regulated by the nucleus basalis of Meynert (NBM), independently of regional metabolism. Thus, although cortical regional cerebral blood flow (rCBF) variations induced by somatosensory stimulation are strongly linked to neuronal activity, they may also be partly controlled by the NBM. In the present study, cortical rCBF alterations in response to innocuous brushing of the hindlimb (HL) were investigated by laser speckle contrast imaging. The contribution of NBM to these changes was examined after injection of the GABAergic agonist muscimol into the right NBM, allowing comparison of somatosensory‐evoked cortical rCBF modifications before and after NBM inactivation. As expected, HL brushing elicited a robust rCBF increase in the contralateral parietal cortex (PC), over the representation of the HL. However, these alterations were decreased, by approximately 40%, in the hemisphere ipsilateral to muscimol inactivation of NBM, whereas vehicle injection did not produce any significant variation. The results demonstrate that cortical rCBF changes induced by somatosensory stimulation are partly regulated by NBM.

High-performance liquid chromatographic separation of carbohydrates on a stationary phase prepared from polystyrene-based resin and novel amines

New anion-exchange stationary phases On (n = 1, 2 and 3) with a dimethylamino terminal functional group, where n is the number of oxyethylene units [-(CH2CH2O)n-], were prepared by the reaction of chloromethylated porous styrene-divinylbenzene copolymer beads and amines [(CH3)2N-(CH2CH2O)nCH2CH2-N(CH3)2]. HPLC separations of monosaccharides (sorbitol, fucose, glucosamine, mannose, glucose, galactose, fructose, allose and altrose) and disaccharides (trehalose, lactose, cellobiose and maltose) were performed successfully on these stationary phases. The ether group of the stationary phases On was found to affect the separation of carbohydrates.

Number, size, conduction, and vasoconstrictor ability of unmyelinated fibers of the ovarian nerve in adult and aged rats.

The effect of aging on the number, size, conduction velocity, and vasoconstrictive function of unmyelinated fibers in ovarian nerve accompanying the ovarian artery was studied in adult (4-7mo) and aged (28-31mo) rats. Morphological observation by electron microscopy showed that the ovarian nerve contains mainly unmyelinated fibers with only a small percentage (less than 4%) of myelinated fibers in either age group. The number of unmyelinated fibers tended to decrease in aged rats (717±59) compared to adult rats (801±48), especially in fibers of smaller diameter, although this difference was not statistically significant. The maximum conduction velocity of unmyelinated fibers within the ovarian nerve was similar when compared between adult (1.05±0.04m/s) and aged (1.02±0.05m/s) rats. Under anesthesia, electrical stimulation of the distal portion of a severed ovarian nerve reduced ovarian blood flow, as measured by laser Doppler flowmetry, when the stimulus intensity was above the threshold for unmyelinated C fibers. Stimulation of the ovarian nerve with supra-maximum intensity (10V) at 2-20Hz frequencies produced frequency-dependent reductions in ovarian blood flow in both adult and aged rats. There were no significant differences in magnitude of the reduction in ovarian blood flow with comparable frequencies of electrical stimulation of the ovarian nerve between adult and aged rats. Collectively, these data indicate that unmyelinated C fibers in ovarian nerve are maintained in number, size, conduction ability, and vasoconstrictor function in aged rats.

HPLC Separation of All Aldopentoses and Aldohexoses on an Anion-Exchange Stationary Phase Prepared from Polystyrene-Based Copolymer and Diamine: The Effect of NaOH Eluent Concentration

To investigate the separations of all aldopentoses (ribose, arabinose, xylose and lyxose) and aldohexoses (glucose, galactose, allose, altrose, mannose, gulose, idose and talose) on the D6 stationary phase prepared by the reaction of chloromethylated styrene-divinylbenzene copolymer and N,N,N’,N’-tetramethyl-1,6-diaminohexane, we examined the effect of varying the concentration of the NaOH eluent on the elution orders. Separations of these aldoses were achieved using a 20 mM NaOH eluent. The elution behaviors of the aldoses were probably due to not only the individual pKa values, but also the chemical structures of the cyclic aldoses.

Hepatic blood flow responses to mechanical stimulation of the skin in anaesthetised rats

The aim of the present study was to investigate how hepatic blood flow (HBF) changes in response to mechanical stimulation of different areas of the skin in anaesthetised rats, by focusing on involvement of the hepatic sympathetic nerves in and contribution of systemic circulatory changes to the HBF responses. HBF was measured at the surface of the left lateral lobe using the laser Doppler flowmetry. Both innocuous and noxious mechanical stimuli were applied to skin areas of the abdomen and hindlimb. Innocuous mechanical stimulation (brushing) of the abdomen and hindlimb did not significantly change HBF, while noxious mechanical stimulation (pinching) of the abdomen and hindlimb did. The responses to pinching were dependent on the sites stimulated. Pinching of the abdomen decreased, while pinching of the hindlimb increased the HBF. The decrease of HBF in response to abdominal pinching remained after the spinal cord was transected at T1-2 level, but the response was diminished after hepatic sympathetic nerves were severed. On the other hand, the increase of HBF in response to hindlimb pinching was dependent on the increase in blood pressure, and was not influenced by the severance of hepatic sympathetic nerves, and the responses to hindlimb pinching were almost absent after the spinal cord was transected. Based on these results, we suggest that noxious mechanical stimulation of the skin produces changes of HBF, either as a reflex response via activation of the hepatic sympathetic nerves or as a passive response to systemic circulatory changes, depending on the sites stimulated.