Enhancement of resting-state fcMRI networks by prior sensory stimulation.

It is important to consider the effect of a previous experimental condition when analyzing resting-state functional connectivity magnetic resonance imaging (fcMRI) data. In this work, a simple sensory stimulation functional MRI (fMRI) experiment was conducted between two resting-state fcMRI acquisitions in anesthetized rats using a high-field small-animal MR scanner. Previous human studies have reported fcMRI network alteration by prior task/stimulus utilizing similar experimental paradigms. An anesthetized rat preparation was used to test whether brain regions with higher level functions are involved in post-task/stimulus fcMRI network alteration. We demonstrate significant fcMRI enhancement poststimulation in the sensory cortical, limbic, and insular brain regions in rats. These brain regions have been previously implicated in vigilance and anesthetic arousal networks. We tested their experimental paradigm in several inbred strains of rats with known phenotypic differences in anesthetic susceptibility and cerebral vascular function. Brown Norway (BN), Dahl Salt-Sensitive (SS), and consomic SSBN13 strains were tested. We have previously shown significant differences in blood oxygen level-dependent fMRI activity and fcMRI networks across these strains. Here we report statistically significant interstrain differences in regional fcMRI poststimulation enhancement. In the SS strain, poststimulation enhancement occurred in posterior sensory and limbic cortical brain regions. In the BN strain, poststimulation enhancement appeared in anterior cingulate and subcortical limbic brain regions. These results imply that a prior condition has a significant impact on fcMRI networks that depend on intersubject difference in genetics and physiology.

Akt-dependent phosphorylation of endothelial nitric-oxide synthase mediates penile erection.

In the penis, nitric oxide (NO) can be formed by both neuronal NO synthase and endothelial NOS (eNOS). eNOS is activated by viscous drag/shear stress in blood vessels to produce NO continuously, a process mediated by the phosphatidylinositol 3-kinase (PI3kinase)/Akt pathway. Here we show that PI3-kinase/Akt physiologically mediates erection. Both electrical stimulation of the cavernous nerve and direct intracavernosal injection of the vasorelaxant drug papaverine cause rapid increases in phosphorylated (activated) Akt and eNOS. Phosphorylation is diminished by wortmannin and LY294002, inhibitors of PI3-kinase, the upstream activator of Akt. The two drugs also reduce erection. Penile erection elicited by papaverine is reduced profoundly in mice with targeted deletion of eNOS. Our findings support a model in which rapid, brief activation of neuronal NOS initiates the erectile process, whereas PI3-kinase/Akt-dependent phosphorylation and activation of eNOS leads to sustained NO production and maximal erection.

Effects of endothelial impairment by saponin on the responses to vasodilators and nitrergic nerve stimulation in isolated canine corpus cavernosum.

1. Responsiveness to EDRF-releasing substances and inhibitory nerve stimulation of canine isolated penile corpus cavernosum with and without saponin treatment were investigated. 2. Histological studies demonstrated that saponin did not detach endothelial cells from underlying tissues, but induced degenerative changes in the endothelial cells selectively. 3. In the cavernous strips contracted with phenylephrine, addition of acetylcholine, sodium nitroprusside, ATP and Ca2+ ionophore A23187 induced relaxations, but substance P and bradykinin did not change the muscle tone. 4. Acetylcholine-induced relaxation was significantly attenuated but not abolished by NG-nitro-L-arginine (L-NOARG). L-arginine restored the response inhibited by L-NOARG. The L-NOARG resistant relaxation was not influenced by 1H[1,2,4]oxadiazole[4,3-a]quinoxalin-1-one (ODQ) but was suppressed in the strips contracted with K+. Treatment with saponin abolished the relaxation elicited by acetylcholine and A23187 but did not influence the response to nitroprusside and ATP. The ATP-induced relaxation was attenuated by aminophylline. 5. Transmural electrical stimulation at 2-20 Hz produced endothelium-independent relaxations which were abolished by tetrodotoxin and L-NOARG but unaffected by treatment with saponin. In saponin-treated cavernous strips, the neurogenic relaxation was not affected by acetylcholine, physostigmine, atropine and vasoactive intestinal peptide (VIP) but was abolished by ODQ. 6. It is concluded that acetylcholine-induced relaxations are endothelium-dependent and mediated partly by NO and also by other substances from the endothelium. The endothelium-independent relaxation to ATP is likely to be mediated by P1 purinoceptors. The function of nitrergic nerve does not seem to be prejunctionally modulated by acetylcholine and VIP.

Ultrastructural evidence for innervation of the endothelium and interstitial cells in the atrioventricular valves of the Japanese monkey.

BACKGROUND: A rich supply of nerves to the atrioventricular valve has been demonstrated. The role of the valvular nerves is still controversial because the target sites of the nerves have not been confirmed. METHODS: The innervation of the atrioventricular valves of the Japanese monkey (Macaca fuscata) was examined by acetylcholinesterase staining and electron microscopy. Immunoreactivity for neuropeptide Y (NPY) was also investigated by a post-embedding immunogold method. RESULTS: The valvular nerve elements were clearly concentrated between the endothelium and interstitial cells on the atrial side of cusps. Naked axon terminals were observed to make direct contact (20-nm gaps) with interstitial cells and also to be in close proximity (approximately 200-nm cleft) to the endothelium. NPY immunoreactivity was clearly detected on the large granular vesicles in some terminals that were in close proximity to interstitial cells and/or the endothelium. CONCLUSION: The present study suggests that the extensive innervation of the atrioventricular valve, which includes NPY-containing nerves, might affect valvular function via interstitial cells and/or the endothelium.

Nerve fibres containing neuropeptide Y in the atrioventricular valves of Japanese monkey and rat; a light and electron microscopic study.

Dense distribution of varicose fibres containing neuropeptide Y-like immunoreactivity (NPY-LI) was found in the atrioventricular valves of the Japanese monkey, and moderately in the rat. The immunoelectron microscopy using immunogolds resulted in the localization of NPY-LI within the dense-cored vesicles which existed with the small clear vesicles in the unmyelinated axons near the endocardium. These NPY-LI-containing fibres may participate in regulation of vasomotor role or other functions of the atrioventricular valves.

Nerve terminals associated with the central lacteal lymphatics in the duodenal and ileal villi of the monkey.

An intimate association of nerve fibers with the central lacteal endothelium was demonstrated in the duodenum and ileum of the monkey by immunohistochemistry and transmission electron microscopy. In the basal portion of the central lacteal, nerve fibers containing large cored vesicles and small clear vesicles were located closely beneath the lacteal endothelium. Identification of nerves was performed by immunohistochemistry using antisera against substance P (SP), calcitonin gene-related peptide (CGRP), vasoactive intestinal polypeptide, gastrin-releasing peptide and neuropeptide Y. These nerves contained immunoreactivities for SP and CGRP only. Some of the nerves, either singly or in a dense bundle, indented the endothelial cells to form a conspicuous cushion protruding into the lumen. The attenuated endothelium covering the cushion occasionally was failing, and the nerves were exposed to the lumen. Tight of occasionally subendothelial nerve terminals bundles formed a synapse-like association between themselves: a swollen axonal profile was invaginated by a finger-like projection of another axon, the latter being filled with synaptic vesicles. These results suggest that the central lacteal lymphatics might be afferently monitored, presumably with regard to the luminal pressure, and, at the same time, efferently modulated by these nerves.

Immunoelectron-microscopic localization of peptidergic nerve fibers around lymphatic capillaries in the rat liver.

The localization of neuropeptide Y (NPY)-, substance P (SP)- and calcitonin gene-related peptide (CGRP)-containing nerve fibers around lymphatic capillaries (initial lymphatics) in the interlobular connective tissue of the rat liver was investigated by preembedding immunoelectron-microscopy. Nerve terminals with NPY were frequently seen in close apposition to the abluminal surface of lymphatic endothelium. A small number of NPY fibers without a glial (Schwann cell) covering at the tip ran toward lymphatic capillaries in the interlobular connective tissue. Nerve fibers immunoreactive for SP were present within unmyelinated fiber bundles that ran close to lymphatic capillaries in the interlobular connective tissue. Besides these immunoreactive nerve fibers, many of which appeared to pass through the subendothelial regions of lymphatic capillaries, scattered SP nerve endings were seen in areas contiguous to lymphatic endothelium. CGRP terminals were rarely found around lymphatic capillaries, although nerve fiber bundles containing CGRP components traversed close to some lymphatic capillaries. These findings suggest that NPY and SP, if released from nerve terminals into the subendothelial areas of adjacent lymphatic capillaries, are more likely to affect the metabolic activity of lymphatic endothelium and the flow (or formation) of lymph than CGRP. SP and CGRP, as possible mediators of sensory transmission, might be involved in the conveyance of information on the hydrostatic pressures of hepatic lymphatics and surrounding tissue fluid to the central nervous system.