Immunocytochemical Study on the Distribution Pattern of Corticotropin Releasing Factor and Norepinephrine in the Middle Lobe of Monkey Cerebellum

Immunocytochemical methods, employing a specific antiserum against human corticotropin releasing factor (CRF) and dopamine beta hydroxylase, were applied to investigate the distribution pattern of CRF and norepinephrine fibers in the cerebellar cortex of squirrel monkey. CRF fibers were present mainly in the molecular layer throughout the major regions of cerebellar cortex. Howeeer, the most intensely labeled axons were strikingly clustered within particular regions and parasagittal domains. In the vermis and intermediate zone, intensely labeled axons were present only within parasagittal zones similar in location to those defined by climbing fiber innervation from the medial accessory olive, Intensely labeled axons were also densely but uniformly distributed within the uvula, the medial region of the dorsal paraflocculus, and the dorsal region of the pyramis, areas that recetee their climbing input primarily from the medial accessory olive. Labeled fibers were much less dense and were not clustered in the lateral hemispheres. Norepinephrine fibers were found throughout the cerebellar cortex, and the prominent population of norepinephrine fibers in cerebellar cortex was localized within the granular layer and Purkinje cell layer. In the vermis, the great density is seen in posterior lobules, especially lobules VII-X In the hemispheric region, a dense plexus of norepinephrine fibers was present throughout the granule cell layer, and the immunoreactive density in this region was greater than the density in the vermis. These results indicate that (l) CRF is the main neurotransmitter in the molecular layer and norepinephrine is the important transmitter in the granular layer (2) there were significant differences in the laminar distribution in different lobules of the cerebellum between CRF and norepinephrine

Corticotropin-releasing Factor (CRF) and Urocortin Promote the Survival of Cultured Cerebellar GABAergic Neurons Through the Type 1 CRF Receptor

Corticotropin releasing factor (CRF) is known to be involved in the stress response and in some degenerative brain disorders. In addition, CRF has a role as a neuromodulator in adult cerebellar circuits. Data from developmental studies suggest a putative role for CRF as a trophic factor during cerebellar development. In this study, we investigated the trophic role for CRF family of peptides by culturing cerebellar neurons in the presence of CRF, urocortin or urocortin II. Primary cell cultures of cerebella from embryonic day 18 mice were established, and cells were treated for either 1, 5 or 9 days with Basal Medium Eagles complete medium alone or complete medium with 1 µM CRF, urocortin, or urocortin II. The number of GABA-positive neurons in each treatment condition was counted at each culture age for monitoring the changes in neuronal survival. Treatment with 1 µM CRF or 1 µM urocortin increased the survival of GABAergic neurons at 6 days in vitro and 10 days in vitro, and this survival promoting effect was abolished by treatment with astressin in the presence of those peptides. Based on these data, we suggest that CRF or urocortin has a trophic role promoting the survival of cerebellar GABAergic neurons in cultures

원숭이 소뇌핵의 Corticotropin Releasing Factor와 Serotonin 분포에 대한 면역조직화학적 연구

Immunohistochemical methods, employing a specific antiserum against human corticotropin releasing factor (CRF) and serotonin, were applied to determine the distribution of corticotropin releasing factor-immunoreactivity (CRF-IR) and serotonin- IR in the deep cerebellar nuclei of the Squirrel monkey. CRF-IR labeled fibers were demonstrated in three all deep cerebellar nuclei, including the dentate, interposed and fastigial nucleus. All these fibers were varicose and greater density was observed in the dentate and the interposed nuclei. The serotonin-IR fibers were observed in all deep cerebellar nuclei. But the distribution of these fibers was denser than that of CRF-IR fibers. Labeled fibers were varicose, and there was no evidence of greater density within deep cerebellar nuclei

Substance P immunoreactive cell reductions in cerebral cortex of Niemann-Pick disease type C mouse

Niemann-Pick disease type C (NPC) is characterized by progressive neurodegeneration and arises from mutations in the NPC1 gene. Cholesterol has received most attention in the pathogenesis of NPC, but normalizing lipid levels in humans or mouse does not prevent neurodegeneration. In NPC mouse, neuronal degeneration in the cerebellum is the most commonly detected change, and thus previous studies have tended to focus on the cerebellum, especially Purkinje cells. Although numerous peptides have been found in the mammalian central nervous system, little data on neurotransmitters in NPC are available, and information on neurotransmitter system abnormalities could explain the complex and characteristic deficits of NPC. Thus, we performed an immunohistochemical study on NPC mouse cortices to compare cell numbers exhibiting vasoactive intestinal polypeptide (VIP), neuropeptide Y (NPY), and substance P (SP) immunoreactivity. In terms of VIP and NPY-immunoreactive (ir) cell numbers in the cerebral cortex, SP-ir cells were significantly reduced by about 90% in NPC (-/-) versus NPC (+/+) mouse, and were also mildly decreased in frontal and parietal NPC (+/-) versus NPC (+/+) mouse cortex. This study demonstrates for the first time, reduced number of SP-ir cells in the NPC mouse cortex

Reduced immunoreactivities of a vasoactive intestinal peptide and pituitary adenylate cyclase-activating polypeptide receptor (VPAC1 receptor) in the cerebral cortex, hippocampal region, and amygdala of aged rats

In this study, we examined expressional changes of VPAC1 receptor in aged rat brains using an immunohistochemical approach and found that its immunoreactivities are significantly reduced in the cerebral cortex, hippocampal region, and amygdala of aged rats. These results suggest that this reduction could underlie aging-associated memory/learning deficits and several other age-induced functional changes in these areas. However, the functional consequences of these down-regulations require further elucidation

면역조직화학적 방법을 이용한 흰쥐 갑상선에서의 C-cell의 발생 및 성장에 관한 연구

This study was performed to investigate the developmental change of the C-cells in the thyroid glands of the rats. The animal groups used in this study included 17.5-, 18.5-, 19.5-, 20.5-, 21.5-day-old rat fetuses and 3-, 7-, 15-day-old neonatal rats and adult rats weighing about 200 gm. The specimens were fixed with Souin's fluid and embedded in paraffin. Transverse serial sections of the thyroid were cut at 5 p. m from the superior to the inferior pole and eleven representative sections per thyroid lobe were selected with same intervals for immunohistochemical staining. The sections were subjected to the immunoperoxidase staining using anti-synthetic human calcitonin antibody for the detection of C-cells and counterstained with hematoxylin. Then, they were observed under the light microscope and the results were as follows: 1. Immunoreactive C-cells were first observed in the thyroid from the 20.5-day-old fetuses, but in only one and in all thyroids from 21.5-day-old fetuses. 2. The area of the thyroid gland containing C-cells, which was the posterocentral area of the lobe in 21.5-day-old fetuses, increased with aging. It occupied almost entire thyroid except for a small peripheral zone and the polar regions in the adult rats. 3. The numbers of C-cells per unit area (in this study, defined as 227 p. m X 227 fl m) of the thyroid from 21.5-day-old fetuses, 3-,7-, 15-day-old rats and adult rats were 17.8±4.8, 16.2±4.9, 20.8±4.4, 22.6±6.1, 54.7±9.3, respectively. 4. The ratios of intrafollicullar C-cells to total C-cells were 18.3%, 20.0%, 29.5%, 30.3% in the specimens from 3-, 7-, 15-day-old rats and adult rats, respectively. 5. The ratio of C-cells which appeared singly to total C-cells was decreased with aging from 88.8% in the 21.5-day-old fetus thyroids to 33.6% in the adult thyroids. 6. C-cells in the adult rat thyroids were polymorphic in shape; round, ovoid, triangular and spindle-shaped. They were relatively uniform in size with the diameter of 13.2± 2.3 p. m. C-cells in the fetal thyroid were variable in size ranging from 5.6 p.m to 11.3 p.m in diameter and had less cytoplasm than that of adult rat

Immunohistochemical study of p47Phox and gp91Phox distributions in rat brain

NADPH oxidase is multi-component enzyme, which comprises the cytosolic proteins p40Phox, p47Phox, and p67Phox and the two membrane proteins, gp91Phox and p22Phox, and which is well characterized in phagocytic cells. NADPH oxidase is a primary source of reactive oxygen species (ROS), and recent studies indicate that free radicals and ROS might be causative factors of several brain degenerative diseases and dysfunctions. However, though previous studies have shown the presence of NADPH oxidase subunits in cell culture and mouse brain, they have not provided detailed high power resolution data. Therefore, we investigated the distributions of the p47Phox and gp91Phox subunits in rat brain using immunohistochemical approach. Cortex, hippocampus, and Purkinje cells of cerebellum were prominently stained by p47Phox and gp91Phox antibodies. As compared with the distributions of p47Phox, gp91Phox in mouse, some differences in the rat brain were observed in the hippocampus, thalamus, amygdala, reticular nucleus, and basal ganglia. Additionally, at the cellular level, most p47Phox immunoreactivity was largely confined to cell bodies and proximal portions of the dendritic tree. Taken together, the widespread observed distributions of p47Phox and gp91Phox subunits indicate that they are probably needed to maintain normal brain function

Decreased expression of calretinin in the cerebral cortex and hippocampus of SOD1G93A transgenic mice

In the present study, we investigated the changes of calretinin (CR) expression in the central nervous system of SOD1G93A transgenic mice as an in vivo model of amyotrophic lateral sclerosis (ALS). In wild-type SOD1 (wtSOD1) transgenic mice, many CR-immunoreactive neurons were found in all cortical regions. In the cerebral cortex of SOD1G93A transgenic mice, the number and staining intensity of CR-positive neurons were decreased. In the hippocampal formation, layer-specific alterations in the staining intensity of CR-immunoreactive neurons were observed in the CA1-3 areas and dentate gyrus. In wtSOD1 transgenic mice, CR-immunoreactive neurons with long processes were found in the stratum oriens and stratum radiatum of CA1-3 areas, and heavily stained band-like molecular layer was prominent in the dentate gyrus. CR immunoreactivity was decreased in each layer of CA1-3 areas and dentate gyrus of SOD1G93A transgenic mice. The first demonstration of decreased immunoreactivity for CR in the cerebral cortex and hippocampus of SOD1G93A transgenic mice may provide insights into the pathogenesis of motor neuron degeneration in human ALS although further quantitative studies are needed

Region-specific changes in the immunoreactivity of vasoactive intestinal peptide and pituitary adenylate cyclase-activating polypeptide receptors (VPAC(2), and PAC(1) receptor) in the aged rat brains

Pituitary adenylate cyclase-activating peptide (PACAP) and vasoactive intestinal peptide (VIP) have been implicated in a large array of physiological and patho-physiological processes through their receptors (VPAC(1), VPAC(2), and PAC(1) receptor) in the central nervous system. Previously, we demonstrated age-related decreases in VPAC(1) receptor expression in the rat brain providing a possible basis of several age-induced functional changes in the aged brain. In the current study, we also examined age-related changes in PAC(1) and VPAC(2) receptors in aged rat brains using an immunohistochemical approach. We found that PAC(1) immunoreactivity was significantly increased in the hippocampal formation, hypothalamus, thalamus, midbrain septal nuclei, and white matter of aged rats compared with young control rats although its distribution pattern was not altered. In contrast, both distribution pattern and immunoreactivity of VPAC(2) receptor remained unchanged in aged rat brains. These results suggest that the PACAP/VIP receptors exhibit specific expressional changes in the aged brain and that these specific changes could underlie age-associated memory and cognitive functional declines as well as several other age-induced functional changes in the brain. However, the exact regulatory mechanism and its functional significance require further elucidation.Peters A, 2009, FRONT NEUROANAT, V3, DOI 10.3389/neuro.05.011.2009Hansson E, 2009, NEUROREPORT, V20, P957, DOI 10.1097/WNR.0b013e32832ca201Downs JL, 2009, MOL CELL ENDOCRINOL, V299, P32, DOI 10.1016/j.mce.2008.11.012JOLIVEL V, 2009, NEUROSCIENCE, V60, P434Dejda A, 2008, J MOL NEUROSCI, V36, P26, DOI 10.1007/s12031-008-9087-1Peters A, 2008, GLIA, V56, P1151, DOI 10.1002/glia.20686Guillot TS, 2008, NEUROPEPTIDES, V42, P423, DOI 10.1016/j.npep.2008.04.003Brenneman DE, 2007, PEPTIDES, V28, P1720, DOI 10.1016/j.peptides.2007.04.002Reglodi D, 2006, NEUROPEPTIDES, V40, P265, DOI 10.1016/j.npep.2006.06.001Aujard F, 2006, CHRONOBIOL INT, V23, P451, DOI 10.1080/07420520500482090Gomariz RP, 2006, ANN NY ACAD SCI, V1070, P51, DOI 10.1196/annals.1317.031Dejda A, 2006, ANN NY ACAD SCI, V1070, P220, DOI 10.1196/annals.1317.018VAUDRY H, 2006, ANN NY ACAD SCI, V1070, P1Joo KM, 2005, BRAIN RES, V1064, P166, DOI 10.1016/j.brainres.2005.09.006Peters A, 2004, CEREB CORTEX, V14, P995Joo KM, 2004, J COMP NEUROL, V476, P388, DOI 10.1002/cne.20231Bartzokis G, 2004, NEUROBIOL AGING, V25, P5, DOI 10.1016/j.neurobiolaging.2003.03.001Dewar D, 2003, J CEREBR BLOOD F MET, V23, P263, DOI 10.1097/01.WCB.0000053472.41007.F9Nonaka N, 2002, PEPTIDES, V23, P2197Krajnak K, 2002, BRAIN RES, V950, P297Vaudry D, 2002, EUR J NEUROSCI, V15, P1451, DOI 10.1046/j.1460-9568.2002.01981.xLaburthe M, 2002, RECEPTOR CHANNEL, V8, P137, DOI 10.1080/10606820290005191Wiemelt AP, 2001, J NEUROSCI RES, V65, P165Lee M, 2001, J NEUROSCI, V21, P3849Duncan MJ, 2001, MOL BRAIN RES, V87, P196Sherwood NM, 2000, ENDOCR REV, V21, P619Krajnak K, 1998, J NEUROSCI, V18, P4767Said SI, 1998, ANN NY ACAD SCI, V865, P226Gozes I, 1997, J NEUROBIOL, V33, P329Shioda S, 1997, BRAIN RES, V765, P81Cha CI, 1997, BRAIN RES, V753, P235ZHOU JN, 1995, NEUROBIOL AGING, V16, P571HARMAR T, 1994, TRENDS PHARMACOL SCI, V15, P97LUTZ EM, 1993, FEBS LETT, V334, P3PISEGNA JR, 1993, P NATL ACAD SCI USA, V90, P6345SREEDHARAN SP, 1993, BIOCHEM BIOPH RES CO, V193, P546ISHIHARA T, 1992, NEURON, V8, P811ZBUZEK V, 1988, NEUROENDOCRINOLOGY, V48, P619GOZES I, 1988, NEUROENDOCRINOLOGY, V47, P27