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M Sakanaka

Publications and source records attributed to M Sakanaka.

At least 109 records · Page 6Linked to original sources

Fine structures of nerve fibers with corticotropin-releasing factor-like immunoreactivity in the rat lateral septum.

The fine structures of nerve fibers with corticotropin-releasing factor (CRF)-like immunoreactivity in the rat lateral septum were investigated by means preembedding immunoelectron microscopy. A number of CRF axon terminals formed synapses with cell bodies of non-immunoreactive septal neurons. They occasionally had broad terminal bulges whose subregions showed little or no immunoreactivity for CRF. CRF axon terminals were also in synaptic contact with non-immunoreactive dendrites or dendritic spines. Some dendrites with CRF were postsynaptic to non-immunoreactive axon terminals.

Animals↗

Ontogeny and afferent connections of corticotropin releasing factor-like immunoreactivity in the rat neocortex.

The first set of the present experiments was designed to investigate the postnatal development of corticotropin releasing factor-like immunoreactivity (CRFI) in the rat cerebral cortex by means of cobalt-enhanced immunohistochemistry. Results showed the occurrence of CRFI fibres before cells in the developing rat cerebral cortex with and without colchicine treatment, suggesting that some CRFI cells in subcortical regions may project to the cerebral cortex. In the second set of experiments, ipsilateral double-labelled cells which contained both retrogradely transported horseradish peroxidase (HRP) and CRFI were observed in the zona incerta, subincertal nucleus, lateral hypothalamic area, perifornical hypothalamic area, and in the dorsal hypothalamic area after unilateral HRP injections into the cerebral cortex. These findings indicate the existence of corticopetal CRFI-containing projections arising from the above areas.

Animals↗

Corticotropin releasing factor-containing afferents to the lateral septum of the rat brain.

Corticotropin releasing factor (CRF)-containing afferents to the rat lateral septum (LS) have been determined by means of cobalt-enhanced immunohistochemistry, tracing of retrograde transport of horseradish peroxidase (HRP), and by lesioning experiments. When unilateral lesions included the rostral part of the hypothalamus, CRF-like immunoreactive (CRFI) ipsilateral fibers in the LS decreased in number. Lesions in other brain regions did not cause alterations in the septal CRFI fibers. These findings suggest that the septal CRFI fibers originate in the rostral part of the hypothalamus. Furthermore, combined HRP and immunohistochemical staining on the same sections demonstrated double-labeled cells in two discrete areas within the rostral hypothalamus: one was the perifornical hypothalamic area (PeF) at the level of the paraventricular hypothalamic nucleus, and the other was the most caudal part of the anterior hypothalamic nucleus (AHc). These findings show that a large proportion of the CRFI projections to the LS have their origins in the PeF and AHc.

Animals↗

A reliable method combining horseradish peroxidase histochemistry with immuno-beta-galactosidase staining.

A sensitive combination of horseradish peroxidase (HRP) tracing and immunohistochemistry was used by Rye et al. [J Histochem Cytochem (1984) 32:1145] in a search for the origins of neurotransmitter- and neuromodulator-containing nerve fibers in brain. In this combination, peroxidase as a marker in immunohistochemistry was thought to yield a homogeneous brown immunoreaction product of diaminobenzidine, different from the black granular reaction product of retrogradely transported HRP, which is visualized by the tetramethylbenzidine (TMB) reaction and subsequent stabilization. A neuron that exhibits both kinds of reaction products in its cytoplasm in sections subjected to combination staining is referred to as a double-labeled cell. With a combined HRP and corticotropin-releasing factor (CRF) immunoperoxidase-antiperoxidase (PAP) method, the first set of experiments showed "false" double-labeled cells in the pyramidal cell layer of rat cerebral cortex, but only rarely in the subcortical areas, possibly because of the use of one enzyme system in two different histochemical procedures. This limitation of the double-staining technique prompted us to demonstrate an alternate combination of HRP tracing and immunohistochemistry in the second set of experiments by employing two previously described independent enzyme systems: HRP as a retrograde tracer and beta-galactosidase as a marker for immunohistochemical demonstration of CRF. A homogeneous blue reaction product indicated immuno-beta-galactosidase staining, and a granular black or brown reaction product labeled retrogradely transported HRP in double-labeled cells in subcortical regions. Neither double labeling nor "false" double labeling was seen in pyramidal cells of cerebral cortex. These findings suggest that application of two independent enzyme systems in a combined HRP and immunohistochemical method may be useful for investigating in origins of peptidergic fibers in brain when the combination of HRP histochemistry and the PAP method appears to be inappropriate.

Animals↗

Corticotropin-releasing factor-containing afferents to the inferior colliculus of the rat brain.

Using a modified cobalt-glucose oxidase-diaminobenzidine (Co-GOD) method in a combination of horseradish peroxidase (HRP) retrograde tracing and immunohistochemistry, a widespread localization of corticotropin releasing factor-like immunoreactive (CRFI) structures in the rat inferior colliculus (IC), and a CRFI-containing pathway from the subthalamus and the hypothalamus to the IC have been observed. By means of the modified Co-GOD method, CRFI cells were detected in almost all the subdivisions of the IC, including the dorsomedial part of the central nucleus, the ventrolateral part of the central nucleus, the pericentral nucleus and the external nucleus. Neural processes with CRFI were observed in all of the above areas. Following HRP injection into the IC, double-labeled cells which contained a homogeneous brown immunoreaction product of CRF and a granular black reaction product of retrogradely transported HRP were identified in the lateral hypothalamic area (LH), zona incerta (ZI) and perifornical hypothalamic area (PeF). These double-labeled cells provide direct evidence for CRFI projections from the LH, ZI and PeF to the IC. Thus, the present study supports the view that CRF may act as a neurotransmitter or neuromodulator in the brain.

Animals↗

Corticotropin releasing factor-like immunoreactivity in the rat brain as revealed by a modified cobalt-glucose oxidase-diaminobenzidine method.

A cobalt-glucose-oxidase diaminobenzidine (Co-GOD) method, employing a specific antiserum against rat corticotropin releasing factor (CRF), was applied to determine immunohistochemically a widespread and detailed localization of corticotropin releasing factor-like immunoreactivity (CRFI) in the rat brain. Besides the CRFI cells in the paraventricular hypothalamic nucleus that project to the median eminence, CRFI cells were demonstrated in many brain regions, including the olfactory bulb, cerebral cortex, septal nuclei, hippocampus, amygdala, thalamic nuclei, medial hypothalamic nuclei, lateral hypothalamic area, perifornical area, central gray, cuneiform nucleus, inferior colliculus, raphe nuclei, mesencephalic reticular formation, laterodorsal tegmental nucleus, locus coeruleus, parabrachial nuclei, mesencephalic tract of the trigeminal nerve, pontine reticular formation, lateral superior olive, vestibular nuclei, prepositus hypoglossal nucleus, nucleus of the solitary tract, dorsal motor nucleus of the vagus, lateral reticular nucleus, nucleus of the spinal tract of the trigeminal nerve, external cuneate nucleus, inferior olive, and medullary reticular formation. CRFI-reacting neural processes were also detected in these same areas. In particular, the median eminence, lateral septum, bed nucleus of the stria terminalis, mesencephalic reticular formation, parabrachial nuclei, and nucleus of the solitary tract contained large numbers of CRFI fibres. The widespread localization of CRFI demonstrated in the present study strongly suggests that CRF, like many other neurohormones and peptides, may act as a neurotransmitter and/or neuromodulator in numerous extrahypothalamic circuits, as well as participate in neuroendocrine regulation.

3,3'-Diaminobenzidine↗

Urotensin I-like immunoreactivity in amacrine cells of the goldfish retina.

Urotensin I-like immunoreactivity (UILI), in different localization from that of corticotropin releasing factor-like immunoreactivity (CRFLI), in the goldfish retina has been demonstrated by means of radioimmunoassay, high-performance liquid chromatography (HPLC) and immunohistochemistry. Radioimmunoassay showed 350 +/- 40 pg/mg prot. of UILI in goldfish retina extracts. The immunoreactive material present in the retina was also characterized by reversed phase HPLC. Some of the UILI co-eluted with synthetic carp UI, though the HPLC experiments suggested the existence of other UILI substance(s) with less hydrophobicity than synthetic UI. By immunohistochemistry, UILI and CRFLI were seen in different amacrine cells of the goldfish retina. It is suggested that UI may be involved in the fish visual transmission system together with CRF and other neuropeptides.

Animals↗

Improved fixation and cobalt-glucose oxidase-diaminobenzidine intensification for immunohistochemical demonstration of corticotropin-releasing factor in rat brain.

An optimal fixation method and intensification procedure may be required in brain immunohistochemistry to obtain intense and widespread staining for a specific antigen, in cases where ordinary fixation and conventional immunohistochemistry result in only partial demonstration of the antigen. In the present study of localization of corticotropin-releasing factor immunoreactivity (CRFI) in rat brain, the importance of such intensification is shown. We describe a fixation procedure in which perfusion of rat brain with Bouin's solution is followed by a PBS wash and a further perfusion with either Zamboni's fluid or 4% paraformaldehyde in 0.1 M phosphate buffer (pH 7.4), for subsequent investigation of the detailed localization of CRFI in cerebral cortex and subcortical structures. The cobalt-glucose oxidase-diaminobenzidine (Co-GOD) intensification method has been modified to increase the sensitivity of immunostaining by reducing the concentration of glucose oxidase, which is added to the final incubation solution as a generator of hydrogen peroxide. The use of cobalt acetate instead of cobalt chloride appears to slightly suppress background staining in the Co-GOD method. Combination of the two modified procedures was applied to visualize intense and widespread CRFI in a variety of rat brain regions, including median eminence, cerebral cortex, and central amygdaloid nucleus.

3,3'-Diaminobenzidine↗

Distribution and efferent projections of corticotropin-releasing factor-like immunoreactivity in the rat amygdaloid complex.

Using cobalt-enhanced immunohistochemistry, the tracing of retrograde transport of horseradish peroxidase (HRP) and experimental manipulations, a widespread localization of corticotropin-releasing factor-like immunoreactive (CRFI) structures in the rat amygdaloid complex, and CRFI-containing pathways from the amygdala to the lower brainstem, bed nucleus of the stria terminalis (bst) and ventromedial nucleus of the hypothalamus (VMH) have been demonstrated. By means of cobalt-enhanced immunohistochemistry, CRFI cells were detected in almost all the regions of the amygdala, including the central amygdaloid nucleus (Ce), basolateral amygdaloid nucleus (B1), intra-amygdaloid bed nucleus of the stria terminalis (Abst), medial amygdaloid nucleus (Me), amygdalohippocampal area (Ahi), posterior cortical amygdaloid nucleus (Aco), lateral amygdaloid nucleus (La), anterior amygdaloid area (AAA) and basomedial amygdaloid nucleus (Bm). Neural processes with CRFI were found in all of the above areas. The greatest density of CRFI fibres was observed in the Ce, the Me and Ahi. Unilateral lesions located in the Ce and adjacent areas caused an ipsilateral decrease in CRFI fibre number in the lateral hypothalamic area (LH), mesencephalic reticular formation (RF), dorsal (Dpb) and ventral (Vpb) parabrachial nuclei, mesencephalic nucleus of the trigeminal nerve (MeV) and in the lateral division of the bst (bstl). In addition, ipsilateral CRFI fibres decreased in number in the core and shell of the VMH after unilateral lesions of the corticomedial amygdala (CoM) and ventral subiculum (S). These findings suggest that the CRFI cells in the Ce and adjacent areas innervate the Dpb, Vpb and MeV through the LH and RF; the CRFI fibres in the bstl are supplied by the Ce and adjacent areas; and the CoM and S give rise to the CRFI fibres to the VMH. The distribution of retrogradely transported HRP has confirmed these projections. Furthermore, combined HRP and immunohistochemical staining has demonstrated double labeled cells in the Ce following HRP injection into the Dpb, Vpb, MeV and bstl. This provides direct evidence for the amygdalofugal CRF-containing projections to the lower brainstem and bstl. Double-labeled cells were not seen in the CoM and S after HRP injection into the VMH.

Amygdala↗

The calcitonin gene-related peptide-containing fiber projection from the hypothalamus to the lateral septal area including its fine structures.

The afferent source of calcitonin gene-related peptide-like immunoreactive (CGRPI) fibers in the lateral septal area of the rat was sought by using indirect immunofluorescence technique. These fibers decreased markedly on the operated side after the destruction of the area between the anterior hypothalamic nucleus and the lateral hypothalamus where there were a number of CGRPI cells. We also examined the fine structure of CGRPI fibers in the lateral septal area. Many CGRPI fibers forming axosomatic synapses were identified. These facts strongly suggest that CGRPI cells in the area between the anterior hypothalamic nucleus and the lateral hypothalamus project ipsilaterally to the lateral septal area and directly influence the soma there.

Animals↗

Three-dimensional distribution of substance P-like immunoreactivity in the urinary bladder of rat.

The overall distribution of substance P-like immunoreactive (SPI) fibers in the rat urinary bladder was examined by means of frozen sections and whole mount preparations. Two types of SPI fibers entered the urinary bladder from the neck; one forming thick fiber bundles and the other around the blood vessels. These SPI fiber bundles branched into several thinner segments which often project many collaterals to the smooth muscle and submucosal layers. In the smooth muscle layer, single SPI fibers were seen running parallel to both longitudinal and circular muscle bundles. They appeared to be evenly distributed in the smooth muscle layer. SPI fiber bundles located in the circular muscle layer dissociated SPI fibers which entered the submucosal layer, where they directed to the epithelium and formed a meshwork just beneath it. SPI fibers often left the meshwork to enter the epithelium where abundant arborization of fine SPI fibers was seen. The density of SPI fibers in the submucosal layer and epithelium was not even. The highest density was identified in the neck and the trigonum area.

Animals↗

Growth of central substance P-containing neurons into superior cervical ganglia transplanted in the spinal cord of adult rats.

Superior cervical ganglia (SCG) contain substance P-like immunoreactive (SP-IR) fibers but not SP-IR neurons. In the present study, SCG were excised from adult rats and transplanted into the same animal's spinal thoracic cord (Th10). One or two weeks after the operation, SP-IR fibers from the host spinal cord or a higher level had grown and entered the transplanted SCG where they formed direct contacts with SCG neurons. However, these phenomena could not be observed when dorsal root ganglia (L4), which contained numerous SP-IR cells, were transplanted into their own spinal cord (Th10). This suggests that the SP-IR neuron system in the adult is able to grow "new axons' to the grafted tissue to form a "new SP-IR' neuronal circuit when the grafted tissue has lost its own SP-IR input.

Animals↗

Evidence for the existence of a substance P-containing pathway from the nucleus laterodorsalis tegmenti (Castaldi) to the medial frontal cortex of the rat.

A long ascending substance P (SP)-containing pathway from the nucleus laterodorsalis tegmenti (TLD) to the medial frontal cortex (MFC) has been demonstrated by means of experimental immunohistochemical procedures. The unilateral destruction of the TLD, which contains numerous SP-like immunoreactive (SPLI) cells, resulted in a marked ipsilateral decrease in the SPLI fibers in the MFC.

Afferent Pathways↗

Experimental immunohistochemical studies on the distribution and origins of substance P in the medial preoptic area of the rat.

Distribution of substance P (SP)-positive fibers in the medial preoptic area (POM) of the rat and their origins were examined using indirect immunofluorescence. A very high density of SP-positive fibers was seen in the POM throughout its entire rostro-caudal extent. However, the distribution of these fibers was not even; the highest density was detected in the medial part of the POM, with less dense but still numerous fibers in the lateral part. On the other hand, in this area a small number of SP-positive cells could be found; a few cells were scattered in the rostral part and, in the caudal part, several cells could be seen in the ventral part of the POM. The origins of SP-positive fibers in the POM were experimentally examined. Since the destruction of the ventro-lateral part of the anterior hypothalamus (vlAH), where numerous SP-positive cells were seen, resulted in a marked decrease of SP-positive fibers in the POM on the operated side, the majority of these fibers may originate from SP-positive cells in the vlAH. The fine structure of SP-positive terminals in the POM were investigated by electron-microscopic immunohistochemical techniques. Immunoreactive terminals contained a few large granular vesicles together with numerous small vesicles, and they made synaptic contacts mainly with dendrites which were devoid of immunoreactive materials. Two different synaptic contacts could be distinguished: one asymmetrical (Gray's type I) and the other symmetrical (Gray's type II), with the latter being predominant.

Animals↗

Experimental and immunohistochemical studies concerning the major origins of the substance P-containing fibers in the lateral lemniscus and lateral parabrachial area of the rat, including the fiber pathways.

The origins and pathways of substance P-containing fibers in the lateral lemniscus and lateral parabrachial area of the rat were investigated by means of the indirect immunofluorescence technique. Following the placement of various knife cuts or lesions, the following could be concluded: (1) The cells of origin of substance P-containing fibers in the lateral lemniscus and lateral parabrachial area are localized in the ventral reticular formation of the rhombencephalon from the level of P2.8 to P7.0 in the atlas of Palkovits and Jacobowitz; (2) The fibers from these origins pass the ventral reticular formation of the rhombencephalon to form a dense bundle; (3) the fibers turn dorsally at the mid-rhombencephalon junction to reach the lateral lemniscus and lateral parabrachial area. The present study demonstrates the linking of the functions of the parabrachial area and lateral lemniscus via substance P innervation.

Animals↗