Diffusion Tensor Imaging MRI With Spin-Echo Sequence and Long-Duration Measurement for Evaluation of Renal Fibrosis in a Rat Fibrosis Model.

BACKGROUND: Renal fibrosis (RF) is a well-known marker for chronic kidney disease (CKD) progression, including chronic renal injury after renal transplantation. However, invasive biopsy is an available examination for evaluation of RF. Diffusion MRI was once recognized as a promising option for RF. However, it is now controversial for RF evaluation in a unilateral ureteral obstruction (UUO) model. METHODS: To seek an optimal imaging method applicable for RF in UUO model kidneys, we attempted a series of MRI methods, including proton density-weighted imaging, T1-weighted imaging, T2-weighted imaging, T2*-weighted imaging, diffusion-weighted imaging, and diffusion tensor imaging (DTI). RESULTS: We identified DTI MRI by spin-echo sequence plus a special kidney attachment as the best option for evaluation of renal UUO fibrosis, compared with normal kidney on the opposite side. To confirm these results, we applied this technique to a rat UUO therapeutic model with the anti-fibrotic reagent Fasudil. Fractional anisotropy values calculated from DTI MRI showed statistically significant linear correlation with the RF area measured by use of Sirius red or Masson trichrome staining of the positive area [cortex (r = 0.6397, P = .0283) and outer stripe of the outer medulla (r = 0.7810, P = .0039)]. CONCLUSIONS: By use of the DTI MRI with spin-echo sequence, it may be possible to accurately evaluate RF in CKD.

Induction of aquaporin-4 water channel mRNA after focal cerebral ischemia in rat.

Aquaporin-4 (AQP4) is a member of a water-selective channel aquaporin-family and mainly expressed in the several structures of the brain and in the collecting duct of the kidney. Here we show its functional involvement in the water homeostasis of the ischemic brain. The expression of AQP4-mRNA is increased in the peri-infarcted cortex during the observation period ( approximately 7 days) after MCA-occlusion, maximally on day 3. The change corresponds to the generation and resolution of brain edema monitored by MRI. The signals for the mRNA are predominantly observed in glial cells in the molecular and outer granular layer of the peri-infarcted cortex. These results indicate that AQP4 plays a role in post-ischemic edema formation.

Synthesis and blood compatibility evaluation of segmented polyurethanes based on cholesterol and phosphatidylcholine analogous moieties.

New segmented polyurethanes based on cholesterol and phosphatidylcholine analogous moieties were synthesized. The soft segments used in this study were the poly(butadiene), poly(isoprene) or hydrogenated poly(isoprene) glycols; the hard segments of these segmented polyurethanes were composed of 4,4'-methylenediphenyl diisocyanate, 2-[bis(2-hydroxyethyl)methylammonio]ethyl 5-cholesten-3 beta-yl phosphate and 1,4-butanediol. The blood compatibilities of synthesized segmented polyurethanes were evaluated by platelet-rich plasma contact studies and scanning electron microscopy observation using glass as the reference. The results show that the blood compatibilities of the synthesized segmented polyurethanes have great difference between the glass contact side and air exposed side for the same cast films. Generally, the hydrogenated poly(isoprene)-based segmented polyurethane is the best surface in terms of platelet adhesion, and the morphology of adhered platelets undergoes the lowest degree of variation among the segmented polyurethanes investigated in this study.

Interaction between excitatory and inhibitory amino acids in the ventromedial nucleus of the hypothalamus in inducing hyper-running.

We have previously shown that blockade of neurotransmission mediated by gamma-aminobutyric acid (GABA) in the ventromedial nucleus of the hypothalamus (VMH) by the microinjection of a GABAA receptor antagonist, bicuculline methiodide (BM), exclusively induced running activity in the rat. The purpose of the present study was to examine the role of receptors for excitatory amino acids (EAAs) in the VMH in inducing hyper-running and to clarify the interaction between GABA and EAAs in the VMH in controlling running activity. Although the injection of neither N-methyl-D-aspartate (NMDA) nor quisqualate into the VMH induced hyper-running, kainate (KA) produced running activity in a dose-dependent manner with similar features to that induced by BM. This effect of KA was blocked by a non-NMDA receptor antagonist, 6,7-dinitroquinoxaline-2,3-dione (DNQX). GABA injected simultaneously with KA into the VMH failed to affect hyper-running induced by KA. On the other hand, DNQX significantly suppressed the BM-induced running activity. These results suggest that endogenous EAAs acting on the KA-type receptor in the VMH facilitates running activity and that the release of such EAAs from the nerve terminal is presynaptically inhibited by GABA.

Hyper-running activity originating from the hypothalamus is blocked by GABA.

Our previous study showed that injection of the hydrogel of a water-absorbent polymer (WAPGEL) into the ventromedial nucleus of the hypothalamus (VMH) induced stereotyped long-lasting running activity without association of any aggressive reactions, and suggested that swelling of the polymer at the site of injection affected the inhibitory neuron system, resulting in the expression of running activity. In the present study, we injected WAPGEL impregnated with different doses of GABA. By this treatment, the expression of running activity was inhibited in a dose-dependent manner. Contrarily, microinjection of a GABA-A receptor blocker, bicuculline methiodide (BM), released the running reaction in a dose-dependent manner with similar features to that induced by WAPGEL. It is suggested that the running-related neurons in the VMH are inhibited by GABA when the animals are at rest.

Attenuation of the expression of circadian rhythms by chronic outputs from the VMH in rats.

To alter neural networks in a restricted area of the brain, we previously developed the hydro-polymer gel (PG) implantation technique. In this study, we found that bilateral or unilateral injection of PG (1.6 microliters) into the ventromedial nucleus of the hypothalamus (VMH), contrary to injections into the preoptic area (POA) or posterior hypothalamic area (PHA), severely attenuated circadian changes of locomotor activity in blinded female rats. In addition, PG injection into the VMH also suppressed circadian changes in serum melatonin levels and induced persistent estrus. The effect of PG injection into the VMH on free-running rhythm was blocked by a complete cut around the VMH or a dorsal cut of the VMH but not by anterior or posterior cuts of the VMH. These results suggest that PG injection into the VMH induces some form of neural output via a dorsal route of the VMH that affects the generation of circadian activity rhythm.

The ventromedial nucleus of the hypothalamus outputs long-lasting running in rats.

We have developed a new technique for altering neural construction by implanting self-swelling, the hydrogel of water-absorbent polymer (WAPGEL) into restricted areas of the brain. Using this technique, we found that the VMH is responsible for expression of running activity. Implantation of 1.5 microliters of WAPGEL into the dorsomedial portion of the ventromedial nucleus of the hypothalamus (VMH) of the rat specifically induced bursts of running, whereas implantation into other sites of hypothalamus, e.g., the anterior, lateral and posterior areas, and the dorsomedial nucleus, caused no significant running activity. The WAPGEL-injected rat showed less aversive reactions than the sham-operated rat. Since deafferentation of the VMH after WAPGEL injection prevented the induction of running activity, the output from the VMH appeared to be responsible for the running activity.

Effects of continuous supply of light or carbachol to the SCN on the rat free-running rhythm.

Exposure to continuous light (LL) results in elongation of the free-running period of the rat circadian rhythm. However, the LL may not always mean constant intensity of light for the suprachiasmatic nucleus (SCN), since rats may regulate contrast of illumination by their eyelids which are closed during sleep phase. The removal of eyelids caused shortening of the free-running period or arrhythmicity under LL. In addition, implantation of carbachol--acetylcholine agonist--near the SCN also resulted in similar effects.

Effect of constant light on the circadian rhythm in locomotor activity in intact or eyelids-removed rats.

According to the Aschoff's role, exposure to continuous light (LL) results in the elongation of the free-running period of the rat circadian rhythm. However, the LL may not always mean the constant intensity of the light for the suprachiasmatic nucleus, since the rat may regulate the contrast of the illumination by their eyelids which are closed during the sleep phase. In this study, the surgical removal of the eyelids under the LL caused arrhythmicity of the locomotor activity in 7 of 10 rats. The remaining 3 rats maintained the free-running rhythm after the removal of the eyelids. These results suggest that constant light may affect the free-running rhythm of the rat with or without eyelids in the different manner.

Recognition and neural plasticity responding to deficient nutrient intake scanned by a functional MRI in the brain of rats with L-lysine deficiency.

Each L-amino acid (AA) in plasma and brain remains unchanged while normal diet is available. Once L-lysine (Lys) deficient diet was offered to rats, Lys in plasma and brain declined, and anorexia occurred. When solutions of AAs were offered, they selected the Lys solution, and their food intake and growth normalized. The single neuron activity in the lateral hypothalamic area of these rats suggested that neural plasticity occurred, specifically responding to Lys, both by iontophoretic application and during ingestion of AA. The recognition site for deficient nutrient intake in the brain of rats with Lys deficiency was identified by non-invasive magnetic resonance imaging (MRI 4.7 tesla, 40 cm bore in diameter) developed to monitor changes in cerebral blood flow and oxygenation in rat brain. Wistar strain young male rats fed with Lys deficient diet for 4 days, were adapted to be settled in the center of the bore. When they received a Lys injection intraperitoneally (0.2 M, 10 mL/kg), a signal intensity decrease in the medial and lateral hypothalamus appeared 30 minutes later in T2 weighted images, reflecting increased oxygenation which lasted for 30 minutes, and then gradually recovered. These changes never occurred in any other areas of the brain of rats with Lys deficiency, i.e., the thalamus, the cortex, the hippocampus, etc. There were no changes in the signal intensity with control injection of saline. In addition, oxygen consumption in the brain of rats without Lys deficiency was not altered by intraperitoneal Lys injection. The present results suggest that in essential AA deficiency, the medial and lateral hypothalamus may play important roles in recognition responses to particular deficient nutrients in order to maintain homeostasis.

Recognition of deficient nutrient intake in the brain of rat withL-lysine deficiency monitored by functional magnetic resonance imaging, electrophysiologically and behaviorally.

EachL-amino acid (AA) in plasma and brain remains unchanged all day long while normal diet is available. But once restriction ofL-lysine (Lys) was introduced, strong anorexia happened. When Lys deficient diet was offered to rats, their growth were decreased depending upon dietary Lys intake, and they ingested Lys solution in choice quantitatively and both appetite and growth normalized. The recognition site for the deficit in rat's brain was identified by brain oxygenation using a functional MRI that higher signals in the ventromedial hypothalamus and lateral hypothalamic area (LHA) appeared, at 30-50 minutes after Lys injection i.p. and then recovered. Degree of Lys hunger, assayed by bar-pressing (50mg pellet of normal diet/30 presses), was suppressed by Lys micro-injection into the LHA, similar to free Lys ingestion but any other AA never did, suggesting the LHA as recognition site for Lys deficit in rats with Lys deficiency due to AA homeostasis.