Tetsuo Kadota

Differential development of TRPV1-expressing sensory nerves in peripheral organs

In mouse ontogeny, neurons immunoreactive for transient receptor potential vanilloid receptor 1 (TRPV1) were observed primarily in the dorsal root ganglia (DRG) at embryonic day 13 (E13). In the embryonic period, the number of TRPV1+ neurons decreased, but then gradually increased postnatally. Some of TRPV1+ neurons were also immunoreactive for calcitonin gene-related peptide (CGRP). At postnatal day 7 (P7), 66% of CGRP+ neurons were TRPV1+, and 55% of TRPV1+ neurons were also CGRP+ in the L4 DRG. In the peripheral organs, TRPV1-immunorective nerve fibers were transiently observed in the skin at E14. They were also observed in the urinary tract at E14, and in the rectum at E15. Many TRPV1+ nerve fibers in these organs were also CGRP+. At P1, TRPV1+ nerve fibers were observed in the respiratory organs, and to a lesser extent in the stomach, colon, skin, and skeletal muscles. The number of TRPV1+ nerve fibers on each organ gradually increased postnatally. At P7, TRPV1+ nerve fibers were also observed in the small intestine and kidneys. The percentage of total TRPV1+ nerve fibers that co-localized with CGRP was greater in most organs at P7 than at P1. The present results indicate that TRPV1 expression on peripheral processes differs among organs. The differential time course of TRPV1 expression in the cell bodies might be related to the organs to which they project. Co-localization of TRPV1 with CGRP on nerve fibers also varies among organs. This suggests that the TRPV1-mediated neuropeptide release that occurs in certain pathophysiologic conditions also varies among organs.

Nitric oxide synthase in the glossopharyngeal and vagal afferent pathway of a teleost, Takifugu niphobles

Abstract. To examine the presence of nitric oxide synthase (NOS) in the sensory system of the glossopharyngeal and vagus nerves of teleosts, nicotinamide adenine dinucleotide phosphate diaphorase (NADPHd) activity and immunoreactivity for NOS were examined in the puffer fish Takifugu niphobles. The nitrergic sensory neurons were located in the ganglia of both the glossopharyngeal and the vagal nerves. In the vagal ganglion, positive neurons were found in the subpopulations for the branchial rami and the coelomic visceral ramus, but not for the posterior ramus or the lateral line ramus. In the medulla, nitrergic afferent terminals were found in the glossopharyngeal lobe, the vagal lobe, and the commissural nucleus. In the gill structure, the nitrergic nerve fibers were seen in the nerve bundles running along the efferent branchial artery of all three gill arches. These fibers appeared to terminate in the proximal portion of the efferent filament arteries of three gill arches. On the other hand, autonomic neurons innervating the gill arches were unstained. These results suggest that nitrergic sensory neurons in the glossopharyngeal and vagal ganglia project their peripheral processes through the branchial rami to a specific portion of the branchial arteries, and they might play a role in baroreception of this fish. A possible role for nitric oxide (NO) in baroreception is also discussed.

MICROVASCULATURE OF CROTALINE SNAKE PIT ORGANS : POSSIBLE FUNCTION AS A HEAT EXCHANGE MECHANISM

The infrared sensory membranes of the pit organs of pit vipers have an extremely rich capillary vasculature, which has been noted passim in the literature, but never illustrated or studied in detail.

Gastrin/CCK-ergic innervation of cutaneous mucous gland by the supramedullary cells of the puffer fish Takifugu niphobles

The supramedullary cells (SMCs) are spinal neurons lying at the dorsal surface of teleosts. In the present study, we examined whether the SMCs of the puffer fish (Takifugu niphobles) might express gastrin/cholecystokinin-immunoreactivity, as observed in some other teleosts. All the SMCs were immunoreactive for gastrin/cholecystokinin. On the other hand, many immunoreactive varicose nerve fibers were also found terminating in the mucous glands in the skin. In addition, immunoreactive fibers were sparsely distributed in the epidermal layer. No neuronal cells other than the SMCs showed gastrin/cholecystokinin-immunoreactivity centrally or peripherally. The results suggest that gastrin/cholecystokinin-immunoreactive axons in the cutaneous mucous glands and epidermal layer are axons of the SMCs. In view of the present findings, the possible nature of SMCs was discussed.

Ultrastructure of the capillary pericytes and the expression of smooth muscle α‐actin and desmin in the snake infrared sensory organs

The infrared sensory membranes of pit organs of pit vipers have an extremely rich capillary vasculature that forms many vascular loops, each serving a small number of infrared nerve terminals. We clarified the ultrastructure of capillary pericytes in the pit membranes by scanning and transmission electron microscopy, and examined the immunoreactivity in their cytoplasm to two contractile proteins: smooth muscle α‐actin (SM α‐actin) and desmin. The capillary pericytes had two major cytoplasmic processes: thickened primary processes that radiate to embrace the endothelial tube and flattened secondary processes that are distributed widely on the endothelium. Coexpression of SM α‐actin and desmin was observed in the pericytes of entire capillary segments, and SM α‐actin was characterized by prominent filament bundles directed mainly at right angles to the capillary long axis. This expression pattern was different from that of capillary pericytes of the scales, where SM α‐actin was expressed diffusely in the cytoplasm. In a series of electron microscopic sections, we often observed the pericyte processes depressing the endothelial wall. We also observed a close relationship of the pericytes with inter‐endothelial cell junctions, and pericyte processes connected with the endothelial cells via gap junctions.

Coexistence of galanin and substance P in the mouse nasal mucosa, including the vomeronasal organ.

Immunohistochemical fluorescent double labeling revealed the coexistence of galanin and substance P in nerve fibers in the mouse nasal mucosa. At the base of and in the epithelium, all galanin fibers also contained substance P, but around the blood vessels and glands, most of them did not. Since substance P fibers in the nasal mucosa originate from the trigeminal ganglion, these results suggest that galanin fibers in the submucosal region originate from ganglia other than the trigeminal.

The preoptic nucleus: the probable location of the circadian pacemaker of the hagfish, Eptatretus burgeri

By recording the locomotor activity rhythm of hagfish, Eptatretus burgeri, in which surgical lesions of the preoptic nucleus (PON) were made, we searched for the location of the circadian pacemaker in the hypothalamus. The characteristic rhythms were lost in animals lacking the anterior part of the hypothalamus, which includes the PON. Cuts in front of the PON did not affect the rhythm, but cuts behind the PON caused the animals to loose their rhythms. Destruction of the PON with a high-frequency lesion generator also caused loss of rhythms. These findings indicate that the pacemaker may be located in the PON, which has retinofugal connections.

Differential distribution of nerve terminals immunoreactive for substance P and cholecystokinin in the sympathetic preganglionic cell column of the filefish Stephanolepis cirrhifer

Immunoreactivity for substance P and cholecystokinin‐8 was examined in the nerve fibers in the central autonomic nucleus, a cell column for sympathetic preganglionic neurons, in the filefish Stephanolepis cirrhifer. Substance P‐immunoreactive fibers were distributed throughout the entire rostrocaudal extent, but were more abundant in the caudal part of the column, where substance P‐immunoreactive varicosities sometimes made contacts with the sympathetic preganglionic neurons. Cholecystokinin‐8‐immunoreactive fibers were found almost entirely in the rostral part of the column, where a dense network of varicosities was in close apposition to a considerable number of the sympathetic preganglionic neurons. Double labeling immunohistochemistry showed that substance P fibers and cholecystokin‐8 fibers were entirely different, and distinct from serotonin‐immunoreactive fibers. By using immunoelectron microscopy, synaptic specialization was sometimes observed between the dendrites of preganglionic neurons and varicosities immunoreactive for substance P and cholecystokinin‐8. Substance P‐ and cholecystokinin‐8 fibers were seen from the descending trigeminal tract, through the dorsolateral funiculus and the ventral portion of the dorsal horn, to the central autonomic nucleus. After colchicine treatment, substance P‐immunoreactive perikarya were found in the cranial and spinal sensory ganglia. These results suggest that the sympathetic preganglionic neurons of the filefish receive innervation by substance P fibers and cholecystokinin fibers, and that the former might be of primary sensory origin. Topographical distribution of cholecystokinin‐8‐immunoreactive terminals in the central autonomic nucleus along the rostrocaudal extent might underlie the differential regulation of sympathetic activity via a distinct population of sympathetic preganglionic neurons. J. Comp. Neurol. 428:174–189, 2000.

Substance P immunoreactivity in the vagal nerve of mice

After horseradish peroxidase was applied to the main trunk of the mouse vagal nerve, anterogradely labeled cells in the vagal ganglia and fibers in the solitary complex, and retrogradely labeled cells in the dorsal motor nucleus and the ambiguous nucleus were observed. Most of the cells in the nodose ganglion were labeled, but only a few cells in the jugular ganglion were labeled. Heavily labeled nerve terminals and fibers were found in 3 areas in the solitary nucleus: i.e., the lateral half of the medial nucleus, the ventrolateral nucleus, and the commissural nucleus. There was only weak labeling in the dorsolateral nucleus, ventral nucleus, and intermediate nucleus. Substance P immunoreactive neurons in the vagal ganglia were found in the jugular ganglion and the dorsal part of the nodose ganglion, but not in the ventral part of the nodose ganglion. Substance P immunoreactivity in the solitary nucleus was moderate in the commissural nucleus and the intermediate nucleus, but was lacking or very weak in the lateral half of the medial nucleus, ventral nucleus, dorsolateral nucleus, and ventrolateral nucleus. We conclude that most substance P containing fibers in the main trunk of the vagal nerve project centrally to the commissural nucleus and peripherally to some of the thoracic viscera.

Nervous control of blood flow microkinetics in the infrared organs of pit vipers

The pit organ of pit vipers contains a membrane which serves as an infrared retina, processing infrared information by the degree to which the temperature of trigeminal nerve receptors (terminal nerve masses) is raised. The receptors are arranged in a monolayer array within the pit membrane and irrigated by a capillary network which both supplies energy to the terminal nerve masses and serves as a heat exchange mechanism. This mechanism maintains the receptors at a stable temperature level to increase or decrease their sensitivity and to reduce to a minimum the afterimage effect of a moving stimulus. We used a Doppler laser blood flow meter to measure the local changes in blood flow in response to a point heat source (a small soldering iron) and to direct stimuli (red and infrared lasers). Resection of any one of the trigeminal A-delta fiber trunks innervating the pit membrane abolished blood flow response in the area innervated, but resection of the main trunk between the primary neurons and the medulla left the response intact. In addition to the A-delta fibers the pit membrane contains autonomic and sensory C-fiber innervation, but preganglionic resection of parasympathetic neurons, and chemical blocking of postganglionic fibers with atropine and capsaicin had no influence on the blood flow changes. Therefore, on the basis of the rapid response time and the similarity of the blood flow curves to electrophysiological recordings from the receptors, we surmised that all blood flow changes were due to a vasomotor reaction, modulated by the terminal nerve masses directly, resulting in a change in local heat capacity that cools the stimulated receptors back to a basal temperature.