T. Watanabe

Sleep-related changes in human muscle and skin sympathetic nerve activities

To characterize the features of sympathetic nerve activity during non-REM sleep, we measured the muscle and skin sympathetic nerve activities (MSNA and SSNA, respectively) using a double recording microneurographic technique. Eight healthy volunteers were monitored by polysomnography (including EEG, EOG, EMG, and ECG) and received acoustic stimuli (880 Hz, 125 ms) during sleep stage 2. The specific discharge properties of MSNA during wakefulness included pulse-synchronicity, short burst duration and a non-responsiveness to arousal stimuli. These were considered to be generated by an inhibitory input from the baroreceptors to the cardiovascular center. In contrast, SSNA lacked pulse-synchronicity, had longer bursts, and was responsive to a variety of stimuli. Burst rates of MSNA and SSNA were reduced during sleep stages 1 and 2 (light sleep) vs. during a wakefulness. Both MSNA and SSNA appeared to be related to spontaneously occurring K-complexes. The baroreflex latency (from the ECG R-wave to the integrated MSNA burst peak) was constant at approximately 1.20 s during sleep, suggesting that pulse-synchronicity was maintained. The MSNA burst evolution time (interval between initiation of the burst and its peak) became longer with a transition to deeper non-REM sleep stages, whereas the SSNA burst evolution times remained constant. K-complexes induced by acoustic stimuli were frequently accompanied by MSNA and/or SSNA. MSNA responded to acoustic stimuli during light sleep, but some bursts lacked a close relation in time to cardiac rhythm.(ABSTRACT TRUNCATED AT 250 WORDS)

Enhanced muscle sympathetic nerve activity during sleep apnea in the elderly.

To examine how muscle sympathetic nerve activity (MSNA) becomes modified during sleep apnea in the elderly, we analyzed polysomnographic recording simultaneously with microneurographically recorded MSNA. Subjects were three healthy elderly males aged 72, 75, and 76. MSNA was suppressed with deeper non-REM sleep stages in these elderly subjects. In all three subjects, sleep apnea for 10 s or longer was observed during sleep of 00:00-06:00. During sleep apnea, MSNA was enhanced concomitantly with a blood pressure fall and a reduction in saturation rate of oxyhemoglobin. With the termination of sleep apnea, MSNA was maximally enhanced with a transient elevation of blood pressure. We conclude that sleep apnea induces an enhancement of MSNA, which may be responsible for hypertensive episodes during sleep.

Immunohistochemical localization of serotonin, galanin, cholecystokinin, and methionine-enkephalin in adrenal medullary cells of the chicken

The identification of adrenaline- (A) and noradrenaline- (NA) containing cells in the adrenal medulla of the chicken and colocalization of serotonin and neuropeptides with A or NA in medullary cells were investigated with the use of immunohistochemical methods. Antisera against tyrosine hydroxylase and phenylethanolamine-N-methyltransferase were used as markers for catecholamine- and A-synthesizing cells, respectively. About 70% of catecholamine-synthesizing cells also exhibited immunoreactivity for phenylethanolamine-N-methyltransferase antiserum. Therefore, these cells are A-containing ones and the rest of cells seem to be NA-containing cells. Immunoreactivity with serotonin antiserum was observed in almost all medullary cells. Galanin-immunoreactivity was also found throughout the adrenal medulla, but was stronger in A-containing cells than in NA-containing ones. Cholecystokinin-immunoreactivity was restricted to A-containing cells. Methionine-enkephalin-immunoreactivity was seen in both A- and NA-containing cells, but in about half of medullary cells. From these results, it is suggested that serotonin, galanin, cholecystokinin, and methionine-enkephalin may be co-released with A and/or NA from adrenal medullary cells of the chicken.

Immunocytochemical colocalizations of insulin, aromatic L-amino acid decarboxylase, dopamine beta-hydroxylase, S-100 protein and chromogranin A in B-cells of the chicken endocrine pancreas.

The colocalization of aromatic L-amino acid decarboxylase (AADC), dopamine beta-hydroxylase (DBH), S-100 protein and chromogranin A (CgA) in the insulin-containing B-cells of the chicken endocrine pancreas was investigated by using light microscopic immunohistochemistry and combined pre-embedding immunoperoxidase and post-embedding immunogold electron microscopic immunocytochemistry. Using the post-embedding method, immunoreactivity against the anti-insulin serum by protein A-gold technique was observed in the core of all types of B-cell granules. Immunoreaction with anti-S-100 protein serum was detected in the core of all types of B-cell granules. Immunoreaction with anti-S-100 protein serum was detected in the core of all types of B-cell granules from non-osmicated tissues even by post-embedding method, but immunoreactivities against the anti-AADC, DBH and CgA sera were only demonstrable in crystalloid granules of B-cells by pre-embedding method. Immunoreaction with the anti-CgA serum was also detected in the cytoplasmic matrix around crystalloid granules and also in the dense bodies showing immunonegative with anti-insulin serum. From these results, it seems likely that S-100 protein co-stored all types of B-cell granules involved in the maturation of granules, and AADC, DBH and CgA are related to the synthesis of noradrenaline in crystalloid granules of B-cells.

Somatostatin-14 and somatostatin-28 in chicken pancreatic islet D-cells

Somatostatin (SST)-14 and mammalian (m) SST-28[1-14] immunoreactivities of chicken pancreatic islets were investigated by using light microscopic immunohistochemistry. Chicken D-cells in both A- and B-islets showed immunoreactivity to SST-14, but not to mSST-28[1-14]. The acid-extract from both splenic and ventral lobes of pancreas was fractionated by reverse-phase high-performance liquid chromatography, and the SST-like immunoreactivity was measured in the radioimmunoassay using anti-SST-14 serum. In both lobes, the SST-like immunoreactivity was detected in the fraction which corresponded to that of SST-14 standard, but was not found in that of mSST-28 standard. Immunohistochemically, pancreatic endocrine D-cells of 1 amphibian, 4 reptiles and 12 birds showed the same immunostaining property as chicken D-cells. By contrast, both SST-14- and mSST-28[1-14]-immunoreactive D-cells were observed in the pancreatic islets of 16 mammals. From these results, we concluded that chicken islet D-cells contain only SST-14-like peptide, but not SST-28-like peptide, and that this phenomenon may be common to the avian species.

Location of Sympathetic Postganglionic and Sensory Neurons Innervating the Testis in the Male Chicken

Sympathetic postganglionic and sensory neurons were labelled by injections of horseradish peroxidase into the testis of the male chicken. The total number of labelled neurons in the paravertebral, prevertebral, dorsal root and nodose ganglia was 943 on average for five chickens. Sympathetic postganglionic neurons were located in the paravertebral ganglia T3‐LS3 (10u2003% of the total number of labelled neurons), especially in T6 and T7, and in the prevertebral ganglia adjacent to the adrenal glands and aorta (19u2003%). They were found almost ipsilaterally. No labelled neurons were observed in the dorsal motor nucleus of the vagus. Sensory neurons were found bilaterally in the dorsal root ganglia T2‐LS3 (71u2003%), especially in T5 to T7. Over a quarter of labelled sensory neurons were located in the contralateral dorsal root ganglia. In the nodose ganglia, only a few labelled sensory neurons were observed (much less than 1u2003%). These results indicate that, unlike the ovary, the testis of the chicken tends to be innervated by ipsilaterally located sympathetic postganglionic and sensory neurons, with the sensory neurons being more numerous than the sympathetic postganglionic neurons.

Cells of origin of spinal projections from the paraventricular nucleus (PVN) of the hypothalamus in the chicken

Hypothalamic neurons projecting to the lumbar (lumbar-neurons) and sacral (sacral-neurons) segments were retrogradely labeled by injections of fluorescent axonal tracers (FITC-WGA, TRITC-WGA) into both the lumbar and sacral segments. Labeled neurons were distributed in the principal part of the paraventricular nucleus (PVN), but not in the pars dispersa of this nucleus. Of these neurons, some small and oval-shaped neurons (about 10 microns in diameter) in a middle part of the PVN were observed to be labeled by both FITC-WGA and TRITC-WGA (lumbosacral-neurons) suggesting double projections to lumbar and sacral segments.

Muscle Representation within the Hypoglossal Nucleus of the Chicken Studied by Means of Horseradish Peroxidase

The distribution in the chicken of motoneurons innervating the hyolingual muscles, i.e. the Mm. hyoglossus rostralis (HR), hyoglossus obliquus (HO), ceratoglossus (CG), interceratobranchialis (CB). stylohyoideus (YH), serpihyoideus (PH) and cricohyoideus (CR), and the laryngotracheal muscles, comprising the Mm. tracheolateralis (TL), cleido‐hyoideus (CL) and sternotrachealis (ST), was examined by retrograde transport of horseradish peroxidase conjugated with wheat‐germ agglutinin. Labelled motoneurons are only found in the hypoglossal nucleus. The rostrocaudal distributions of motoneurons projecting to hyolingual muscles are restricted in the hypoglossal nucleus cranial to the obex, and those projecting to laryngotracheal muscles are distributed in the more caudal part of hypoglossal nucleus. Detailed analysis of the data showed that the most rostrally positioned motoneurons in the hypoglossal nucleus supplied to the PH, followed by the CG, CB. HR, YH, HO, CR, TL, CL and ST in that order, overlapping each other. In the hypoglossal nucleus motoneurons innervating the PH and YH have the smallest perikarya. Of the motoneurons in the hypoglossal nucleus, those supplying the laryngotracheal muscles (CL and TL) have the largest perikarya. Motoneurons innervating the other muscles are intermediate in size.

A juxtaposition between A-cells and D-cells in the chicken pancreatic islets following vagotomy.

Conventional electron microscopy and enzyme‐cytochemistry were applied to elucidate the juxtaposition between the pancreatic D‐cell and A‐cell, 1–2 weeks after abdominal vagotomy in chickens. A pancreatic D‐cell frequently encircled an A‐cell. Around this juxtaposition, several D‐cells, characterized by the occurrence of peculiar dense bodies, formed a cluster. Both the D‐cell and the A‐cell juxtaposed with each other and had an irregularly shaped nucleus with several indentations. Exocytosis of secretory granules from D‐cells encircling an A‐cell was often observed in the capillary side, but no release of secretory granules from A‐cells was detected, except on the capillary side. A few large dense bodies, resembling a multivesicular body, were observed in the A‐cell cytoplasm, showed positive acid‐phosphatase activity, and contained remnants of several types of cell organelles. They thus seemed to be secondary lysosomes. It is possible that the juxtaposition between the A‐cell and the D‐cell may be morphological evidence of the inhibitory action on the A‐cell by the D‐cell.

Somatotopic Representation within the Facial Motor Nucleus of the Chicken Studied by Means of Horseradish Peroxidase

The distribution in the chicken of motoneurons innervating the hyolingual muscles, i.e. the M. branchiomandibularis (BM), M. ceratoglossus (CG), M. interceratobranchialis (CB), M. serpihyoideus (PH), M. stylohyoideus (YH) and one of the mandibular muscles. M. depressor mandibulae (DM), was examined by retrograde transport of horseradish peroxidase conjugated with wheatgerm agglutinin. Labelled motoneurons are found in the three subnuclei of the facial (VII) nucleus, as well as hypoglossal (XII) nucleus. The distribution of motoneurons projecting to the DM are observed in the three subnuclei, those of the BM, PH and YH in the intermediate and ventral subnuclei and those of the CB and CG in the intermediate subnucleus. Motoneurons projecting to the PH, YH, CB and CG are also distributed in the XII nucleus. The ratio of labelled motoneurons of the VII to XII nuclei decreases the PH, YH, CB and CG in that order, and the ratio of labelled ones of the ventral to intermediate subnuclei decreases the BM, PH and YH in that order. The topographical and functional aspects of the subdivision of the motor nucleus are discussed.