Platycarya strobilacea leaf extract protects mice brain with focal cerebral ischemia by antioxidative property / 대한해부학회지

Protective effect of Rhus verniciflua Stokes extract in an experimental model of post-menopausal osteoporosis / 대한해부학회지

Post-menopausal osteoporosis (PMO) is a major global human health concern. Owing to the need for therapeutic drugs without side effects, natural extracts containing various polyphenolic compounds that may exert estrogenic effects have been studied in depth. Rhus verniciflua Stokes (RVS), which has been used as a traditional herbal medicine for centuries in Korea, was recently revealed to exert estrogenic effects attributable to its bioactive ingredients sulfuretin and butein, which have strong estrogen receptor–binding affinities. In this study, the protective potential of RVS in PMO was evaluated by using an experimental animal model of PMO, which was established by ovariectomy (OVX) of female Sprague Dawley rats. The oral administration of RVS at 20 mg/kg or 100 mg/kg for 8 weeks markedly protected against OVX-induced atrophy of the uterine tube and reversed the elevation in the ratio of serum receptor activator of nuclear factor-κB ligand to osteoprotegerin, which is a marker of disease severity. In addition, RVS inhibited OVX-induced tibia bone loss, activated osteogenic activity, and suppressed osteoclastic activity in the tibial epiphyseal plate, a region of bone remodeling. Collectively, these factors indicated that the oral intake of RVS might be beneficial for the prevention of PMO.

Rhus verniciflua Stokes attenuates cholestatic liver cirrhosis-induced interstitial fibrosis via Smad3 down-regulation and Smad7 up-regulation / 대한해부학회지

Cholestatic liver cirrhosis (CLC) eventually proceeds to end-stage liver failure by mediating overwhelming deposition of collagen, which is produced by activated interstitial myofibroblasts. Although the beneficial effects of Rhus verniciflua Stokes (RVS) on various diseases are well-known, its therapeutic effect and possible underlying mechanism on interstitial fibrosis associated with CLC are not elucidated. This study was designed to assess the protective effects of RVS and its possible underlying mechanisms in rat models of CLC established by bile duct ligation (BDL). We demonstrated that BDL markedly elevated the serological parameters such as aspartate aminotransferase, alanine transaminase, total bilirubin, and direct bilirubin, all of which were significantly attenuated by the daily uptake of RVS (2 mg/kg/day) for 28 days (14 days before and after operation) via intragastric route. We observed that BDL drastically induced the deterioration of liver histoarchitecture and excessive deposition of extracellular matrix (ECM), both of which were significantly attenuated by RVS. In addition, we revealed that RVS inhibited BDL-induced proliferation and activation of interstitial myofibroblasts, a highly suggestive cell type for ECM production, as shown by immunohistochemical and semi-quantitative detection of α-smooth muscle actin and vimentin. Finally, we demonstrated that the anti-fibrotic effect of RVS was associated with the inactivation of Smad3, the key downstream target of a major fibrogenic cytokine, i.e., transforming growth factor β (TGF-β). Simultaneously, we also found that RVS reciprocally increased the expression of Smad7, a negative regulatory protein of the TGF-β/Smad3 pathway. Taken together, these results suggested that RVS has a therapeutic effect on CLC, and these effects are, at least partly, due to the inhibition of liver fibrosis by the downregulation of Smad3 and upregulation of Smad7.

Pes anserinus and anserine bursa: anatomical study / 대한해부학회지

This study investigated the boundary of anserine bursa with the recommended injection site and shape on the insertion area of pes anserinus (PA), with the aim of improving clinical practice. Eighty six legs from 45 Korean cadavers were investigated. The mixed gelatin solution was injected to identify the shape of anserine bursa, and then the insertion site of the PA tendons was exposed completely and carefully dissected to identify the shape of the PA. The sartorius was inserted into the superficial layer and gracilis, and the semitendinosus was inserted into the deep layer on the medial surface of the tibia. The number of the semitendinosus tendons at the insertion site varied: 1 in 66% of specimens, 2 in 31%, and 3 in 3%. The gracilis and semitendinosus tendons were connected to the deep fascia of leg. Overall, the shape of the anserine bursa was irregularly circular. Most of the anserine bursa specimens reached the proximal line of the tibia, and some of the specimens reached above the proximal line of the tibia. In the medial view of the tibia, the anserine bursa was located posteriorly and superiorly from the tibia's midline, and it followed the lines of the sartorius muscle. The injection site for anserine bursa should be carried out at 20degrees from the vertical line medially and inferiorly, 15 or 20 mm deeply, and at the point of about 20 mm medial and 12 mm superior from inferomedial point of tibial tuberosity.

Pes anserinus and anserine bursa: anatomical study / 대한해부학회지

This study investigated the boundary of anserine bursa with the recommended injection site and shape on the insertion area of pes anserinus (PA), with the aim of improving clinical practice. Eighty six legs from 45 Korean cadavers were investigated. The mixed gelatin solution was injected to identify the shape of anserine bursa, and then the insertion site of the PA tendons was exposed completely and carefully dissected to identify the shape of the PA. The sartorius was inserted into the superficial layer and gracilis, and the semitendinosus was inserted into the deep layer on the medial surface of the tibia. The number of the semitendinosus tendons at the insertion site varied: 1 in 66% of specimens, 2 in 31%, and 3 in 3%. The gracilis and semitendinosus tendons were connected to the deep fascia of leg. Overall, the shape of the anserine bursa was irregularly circular. Most of the anserine bursa specimens reached the proximal line of the tibia, and some of the specimens reached above the proximal line of the tibia. In the medial view of the tibia, the anserine bursa was located posteriorly and superiorly from the tibia's midline, and it followed the lines of the sartorius muscle. The injection site for anserine bursa should be carried out at 20degrees from the vertical line medially and inferiorly, 15 or 20 mm deeply, and at the point of about 20 mm medial and 12 mm superior from inferomedial point of tibial tuberosity.

Sex determination using upper limb bones in Korean populations / 대한해부학회지

The purpose of this research is to establish metric standards for the determination of sex from the upper limb bones of Korean. We took a set of eleven measurements on each of 175 right sides of adult skeletons chosen at Korean sample. Classification accuracy dropped only one or two individuals when only vertical head diameter of humerus is used. Variables in relation with maximal length were less accurate than head diameter of humerus. Two variables were selected by the stepwise procedure: maximal length of humerus, vertical head diameter of humerus. The combined accuracy was 87%. This study of modern Korean skeletons underscores the need for population-specific techniques, not only for medicolegal investigations, but also for the study of population affinities and factors affecting bone configurations.

TBR2-immunopsitive unipolar brush cells are associated with ectopic zebrin II-immunoreactive Purkinje cell clusters in the cerebellum of scrambler mice / 대한해부학회지

Unipolar brush cells (UBCs) are excitatory interneurons with their somata located in the granular layer. Recently, T-brain factor 2 (Tbr2) was shown to be expressed in a subset of UBCs in mouse cerebellum. Scrambler mice exhibit severe cerebellum abnormalities, including the failure of embryonic Purkinje cell dispersal and a complete absence of foliation due to a mutation in the disabled-1 adaptor protein. Since most UBC markers are expressed postnatally, it has proven difficult to identify the relationship between developing Purkinje cell clusters and migrating UBCs. Because scrambler mice closely mimic normal embryonic day 18 cerebellum, we examined whether Tbr2-positive UBCs are associated with Purkinje cell cluster markers such as zebrin II, which is the most studied compartmentation marker in the cerebellum. We investigated the distribution of Tbr2-positive UBCs in this mutant by using anti-Tbr2 immunocytochemistry. The data revealed that Tbr2 immunoreactivity was exclusively present in the nucleus of UBCs in scrambler cerebellum. Based on expression data, a Tbr2-positive UBC map was constructed. In addition, Tbr2-positive UBCs are found associated with ectopic zebrin II-immunoreactive Purkinje cell clusters in scrambler cerebellum. These data suggest that UBCs use Purkinje cell compartmentation to migrate into their final position through interactions with the embryonic array of specific Purkinje cell subtypes.

Early cerebellar granule cell migration in the mouse embryonic development / 대한해부학회지

Pax6, a paired homeobox DNA binding protein, has been found to be expressed in the cerebellum in both granule cells and their precursors in the external granular layer (EGL). In this study we have traced Pax6 expression through embryonic development in mice by using a polyclonal antibody against Pax6 and used it to study the cellular dispersal pattern of the EGL. During dispersal the EGL was thicker and Pax6 expression was more intense on the rostral side of the lateral corners of the cerebellum. Pax6 immunoreactive cells were found to be migrating from the EGL during the early stage of EGL dispersal, which suggested the early inward migration of granule cells. Double staining with various markers confirmed that the early-migrating cells are not Purkinje cells, interneurons or glia. Although the Pax6 immunoreactive cells within the cerebellum were not apparently proliferating, NeuN, a marker for postmitotic granule cells, was not expressed in these cells until E16. Furthermore, granule cells were observed migrating inwards from the EGL both during and after EGL dispersal. These early migrating granule cells populated the whole cerebellum. These findings offer novel views on specific stages of granule cell dispersal and migration.

Absence Expression of Heat Shock Protein 25 in the Cerebellum of Ataxic Mutant Pogo Mouse / 대한해부학회지

Heat shock proteins (Hsps) are generally known to be induced in response to a range of stressful stimuli such as hyperthermia, immobilization, UV radiation, arsenite, various chemicals, and drugs. In addition, these proteins have been suggested to have roles in protecting cells against apoptotic cell death. The ataxic mutant Pogo (pogo/pogo) mouse is a novel neurological ataxic mutant, which is derived from Korean wild type mouse (KJR/Mskist) strain. Pogo mutation is considered as an alleles of alpha subunit of P/Q-type calcium channel mutants such as rolling mouse Nagoya (RMN), tottering, and leaner. We investigated the topographical Hsp25 expression using immunohistochemistry and western blot analysis in several ataxic mutant mice: RMN, tottering, leaner, Pogo and Korean wild mouse. In the cerebellum of the RMN, tottering, leaner, and normal mouse including Balb/C, C57BL/6 and ICR mouse, Hsp25 was expressed in a subset of Purkinje cells that form parasagittal stripes. The Hsp25 expression is largely restricted to specific cerebellar lobules: VI /VII (the central zone: CZ), and IX/X (the nodular zone: NZ). Surprisingly, no Hsp25-immunoreactive Purkinje cells were seen in CZ and NZ of the cerebellum of Pogo (pogo/pogo), heterozygotes Pogo (pogo/+), and Korean wild mouse. Moreover, in western blot analysis, there was no cerebellar Hsp25 expression in ataxic Pogo mouse including Korean wild mouse. These data suggest that cerebellar Hsp25 expression was irrelevant with the development of ataxia in Pogo mouse.

Decreased Calbindin-immunoreactive Renshaw Cells (RCs) in the Lumbar Spinal Cord of the Ataxic Pogo Mice / 대한해부학회지

Calbindin D-28K (CALB) is one of the calcium-binding proteins which is assumed to be buffering, transport of Ca2+, and regulation of various enzyme systems. In the spinal cord, a subpopulation of calbindin-immunoreactive neurons located in the ventral portion of lamina VII, medial to the motoneuron column, has recently been proposed to be Renshaw cells (RCs), that mediate recurrent inhibition of spinal alpha-motoneurons, based on the anatomical location. In this study, we have performed to investigate the correlation between RCs containing high levels of CALB and motoneurons in the ventral horn of lumbar spinal cord of the ataxic pogo mice, that characterized by a failures of interlimb coordination, and prolonged excessive tone of hindlimb extensor muscles. We have shown that CALB immunoreactive RCs was significantly decreased in the ventral horn of lumbar spinal cord of the ataxic pogo mice (p.0.05), when compared with the control mice. Whereas, CALB immunoreactivity expression levels were no difference in the dorsal horn. Furthermore, CALB protein was significantly decreased in the lumbar spinal cord of the ataxic pogo mice (p.0.01). However, there were no difference in the cervical and thoracic spinal cord of the between control and pogo mice. These results suggest that motoneurons of ventral horn of the lumbar spinal cord might be more excited state, results in the decreased CALB immunoreactive RCs have not mediated a motoneuron excitability, in the atxic mice, pogo.

Voltage-dependent Calcium Channel (VDCC) alpha(1A) Subunit Expression in the Ataxic Mutant, Pogo Mice Cerebellum / 대한해부학회지

The pogo mouse is a new ataxic mutant derived from a Korean wild mouse. The pogo mutation is inherited as an autosomal recessive trait on chromosome 8. Mutations in gene coding for the alpha(1A)subunit of voltagegated P/Q-type Ca(2+) channel have been shown to cause phenotypes in humans and mice, i.e., tottering, leaner, rolling mouse mouse Nagoya. Using immunohistochemistry, the expression of the alpha(1A)subunit of voltage-gated P/Q-type Ca(2+) channel was examined in pogo mice cerebellum including deep cerebellar nuclei (DCN). We observed alpha(1A)immunoreactivity in the cerebellar cortex (Purkinje cell and granule cell) and DCN of ataxic pogo mice and heterozygote control mice. There was no difference in cerebellar cortical alpha(1A)immunoreactivity between ataxic pogo mice and heterozygous littermate controls (pogo/+). However, we observed alpha(1A)immunoreactivity in the Purkinje cells of control and ataxic pogo mice cerebellum and DCN. We found a significant difference between pogo and heterozygous controls in terms of alpha(1A)immunoreactivities in the DCN. alpha(1A)immunoreactivity in this nucleus in pogo was much higher than in heterozygous littermate controls. No significant differences were observed in the interposed nucleus between pogo and heterozygous controls, but we found that the alpha(1A)subunits were clearer and more abundant in the lateral and medial regions of pogo than in control mice in these regions, where only weak immunoreactivity was observed. This elevated expression of the alpha(1A)subunit in deep cerebellar neurons of pogo might be a compensation for the altered function of P/Q type calcium channel and be related with the induction of the ataxic phenotype in pogo mice.

Elevated Serotonin-immunoreactive Neurons in the Raphe Nucleus of the Ataxic Mutant Mouse, Pogo / 대한해부학회지

This study was carried out to investigate the distribution of serotonin-immunoreactive neruons in the raphe nucleus of the ataxic pogo (pogo/pogo) mice derived from a Korean wild mice. Using by immunohistochemistry, we undertook to elucidate any correlation between the serotonin expression and behavior ataxia including abnormal hindlimb extension in the ataxic pogo mice. The present study has two important findings. First, serotonin immunoreactivity was increased in the raphe nucleus of the ataxic pogo mice. Second, serotonin immunoreactivity was different with the region of raphe nucleus. In the dorsal part of dorsal raphe nucleus (DRD), ventrolateral part of dorsal raphe nucleus (DRVL) and median raphe nucleus (MR), serotonin immunoreactivity was increased, whereas the ventral part of dorsal raphe nucleus (DRV) and interfascicular part of dorsal raphe nucleus (DRI) was similar with the control mice. Therefore, elevated expression of the serotonin in the raphe nucleus of ataxic pogo mice might be a source of behavior ataxia and may be related with the induction of the ataxic phenotype including abnormal hindlimb movements.

Distribution of Calretinin-immunoreactive Unipolar Brush Cells in Ataxic Mutant Pogo Mice Cerebellum / 대한해부학회지

Unipolar brush cells (UBCs) are a class of putative interneurons found in the granular layer of mammalian cerebellum and dorsal cochlear nucleus. The unipolar brush cells (UBCs), as with granular cells, which receives afferent synaptic input from extrinsic mossy fiber and whose axons branch in the granular layer and establish a system of cortex-intrinsic mossy fibers, which synapse with granule cells and other UBCs. In general, UBCs have been identified most readily by their expression of the calcium-binding protein, calretinin. The purpose of this study was to provide information about UBCs distributions of the new ataxic animal model, pogo mouse cerebellum using anti-calretinin immunohistochemistry, immunofluorescence and its effect on calcium homeostasis. Through the examination of calretinin immunohistochemistry and immunofluorescence, we observed that many calretinin immunoreactive UBCs were distributed widely throughout the lobules IX and X of the granular layer of both group. But, we found the number of calretinin immunoreactive UBCs of ataxic pogo (pogo/pogo) mouse was decreased and distribution pattern was altered, compared to control mouse. This result also suggest that reduced calretinin expression may effect on cerebellar Ca2+/-homeostasis, and it may in turn, explain the impaired motor coordination found in the ataxic pogo mice.

Abnormal Expression of Neuropeptide Y in the Cerebellar Purkinje Cells of the Ataxic Mutant Mice, Pogo / 체질인류학회지

The pogo mouse is an autosomal recessive ataxic mutant that arose spontaneously in the inbred KJR/MsKist strain derived originally from Korean wild mice. The ataxic phenotype is characterized by difficulty in maintaining posture and the consequent inability to walk straight. In our previous study about pogo mice cerebellum, we reported the Purkinje cell abnormalities and ectopic expression of tyrosine hydroxylase (TH) in Purkinje cell. In this study, we have provided an abnormal expression of NPY in ataxic mutant pogo mice for the first time. There was increased immunoreactivity for NPY in Purkinje cell of ataxic pogo (pogo/pogo) mice compared to those of heterozygote non-ataxic pogo mice (pogo/+, control group). In our previous study, TH is also expressed abnormally in Purkinje cells of ataxic mutant pogo (pogo/pogo) mouse cerebellum. To compare the expression patterns of TH and NPY within some Purkinje cell using double immunofluorescence, most of NPY-immunoreactive Purkinje cells in the ataxic pogo mice are TH-immunoreactive Purkinje cells. However, all of TH-immunoreactive Purkinje cells are not express the NPY. These data reveal that abnormal NPY-immunoreactivity in the ataxic pogo (pogo/pogo) cerebellum is restricted to a subset of cells within the ectopic TH-immunoreactive Purkinje cell subset. These results further suggest that Purkinje cell abnormalities contribute to motor ataxia in the ataxic pogo mouse.

Glutamate and GABA concentrations in the cerebellum of novel ataxic mutant Pogo mice

The Pogo mouse is an autosomal recessive ataxic mutant that arose spontaneously in the inbred KJR/MsKist strain derived originally from Korean wild mice. The ataxic phenotype is characterized by difficulty in maintaining posture and side to side stability, faulty coordination between limbs and trunk, and the consequent inability to walk straight. In the present study, the cerebellar concentrations of glutamate and GABA were analyzed, since glutamate is a most prevalent excitatory neurotransmitter whereas gammar-aminobutyric acid (GABA) is one of the most abundant inhibitory neurotransmitters, which may be the main neurotransmitters related with the ataxia and epilepsy. The concentration of glutamate of cerebellum decreased significantly in ataxic mutant Pogo mouse compared to those of control mouse. However, GABA concentration was not decrease. These results suggested that the decrease in glutamate concentration may contribute to ataxia in mutant Pogo mouse.

Differences of Zebrin II Expression Pattern Between Normal Balb/C and Ataxic Pogo Mouse Cerebellum / 대한해부학회지

The purpose of this study is to identify the differences of zebrin II expression between ataxic pogo and normal Balb/C mouse cerebellum. Zebrin II is expressed by subsets of Purkinje cells that form an array of parasagittal bands that extend rostrocaudally throughout the cerebellar cortex, separated by similar bands of Purkinje cells that do not express zebrin II. Zebrin II immunoreactivity was localized in the perikarya of Purkinje cells, and the dendrites. Distribution of zebrin II-immunoreactive Purkinje cells were very similar pattern in pogo and Balb/C mouse cerebellum. But, in the lobule III, distribution of zebrin II expression was different between pogo and Balb/C mouse cerebellum. In lobule III of Balb/c mouse cerebellum, 10~15 zebrin II-immunoreactive Purkinje cells were observed and clustered to form a parasagittal bands. On the other hand, zebrin II expressions of lobule III in pogo mouse cerebellum showed a little different patterns. In lobule III of pogo mouse cerebellum, three bilateral zebrin II immunoreactive parasagittal band were observed. P1 band was almost same with lobule III of Balb/C mouse cerebellum. But, P2 bands were composed of 50~60 Purkinje cells which were immunoreactive with zebrin II. These kind of thickening in zebrin II expression of pogo mouse cerebellum may be due to the genetical difference. Furthermore, these results may provide useful information with further ataxic pogo mice cerebellum studies.

Immunohistochemistry of Voltage-Gated Calcium Channel alpha1B Subunit in Mouse Cerebellum

Secretion of neurotransmitters is initiated by voltagegated calcium influx through presynaptic, voltage- gated N-type calcium channels. However, little is known about their cellular distribution in the mouse cerebellum. In the cerebellum, alpha1B immunoreactivity is found mainly on the cell bodies of all Purkinje cells. In addition, the immunoreactivity was detected on a subset of Purkinje cell dendrites, clustered to form a parasagittal array of bands. In the anterior lobe vermis, immunoreactive Purkinje cell dendrites form narrow stripes separated by broad bands of unstained dendrites. Moving caudally through the vermis, these stripes become thicker as a larger fraction of the Purkinje cell dendrites become immunoreactive. This localization study of the alpha1B pore-forming subunits in mouse cerebellum may guide future investigations of the role of calcium channels in neurological pathways.

Immunohistochemical Localization of Nerve Growth Factor, Glial Fibrillary Acidic Protein and Ciliary Neurotrophic Factor in Mesencephalon, Rhombencephalon, and Spinal Cord of Developing Mongolian Gerbil

The distribution of the nerve growth factor (NGF), the glial fibrillary acidic protein (GFAP) and the ciliary neurotrohic factor (CNTF) was performed in coronal sections of the mesencephalon, rhombencephalon and spinal cord in the developing Mongolian gerbils. Generally, NGF specifically recognizes neurons with the NGF receptor, whereas GFAP does the glia, and CNTF does the motor neurons. The receptor expression was examined separately in gerbils between embryonic days 15 (E15) and postnatal weeks 3 (PNW 3). The NGF-IR was first observed in the spinal cord at E21, which might be related to the maturation. The GFAP reactivity was peaked at the postnatal days 2 (PND2), while the highest CNTF-reaction was expressed at PNW 2. The GFAP stains were observed in the aqueduct and the spinal cord, which appeared to project laterally at E19. The CNTF was observed only after the birth and found in both the neurons and neuroglia of the substantia nigra, mesencephalon, cerebellum and the spinal cord from PND1 to PNW3. These results suggest that NGF, GFAP and CNTF are important for the development of the neurons and the neuroglia in the central nervous system at the late prenatal and postnatal stages.

Regional Distribution of Zinc Transporter 3 (ZnT-3)-Immunoreactivities in the Mouse Cerebellum / 대한해부학회지

The presence of the zinc ion pump antibody ZnT-3 in zinc enriched (ZEN) cerebellar axonal terminals was detected in immunohistochemical sections of mouse cerebellum. All the ZnT-3-immunoreactive structures appeared punctate in the cerebellar cortex, while the white matter including intrinsic nuclei were void. The staining intensity varied in the different phylogenetic divisions of cerebellum. Archicortex was stained most intensively while paleo and in particular neocortex was stained only faintly. The ZnT-3 fraction was detected mainly in the upper half of the granule cell layer (GCL), although where they appeared ultrastructurally in small folded varicosities located corresponding to axon terminals of the Golgi cells. In the molecular layer (ML), the ZnT-3 staining was found to be distributed diffusely but in perpendicular radiating pattern through the layer but with the most intense staining in the upper half of the layer. Ultrastructurally the coarse patches of staining found in the granule cell layers were corresponded to presynaptic axon terminals contacting with granule cells. No ZnT-3 staining was observed in the Purkinje cell layer. The present results suggest that the Golgi cells in the mouse cerebellum are one of the ZEN neurons in the mammalian brain. In conclusion, we first showed putative ZEN terminals in the mouse cerebellum. These results will provide a valuable aid for investigation of further localization of the ZEN neurons in the mammalian cerebellum. More work is needed to further characterize the ZEN terminals, and so autometallographical studies are currently being investigated in this laboratory.

Changes In The Distribution of Oxytocin and Vasopressin-Immunoreactive Neurons In the Hypothalamic Area of Normal and Hypophysectomized Rats / 체질인류학회지

The localization and number of oxytocin- and vasopressin-immunoreactive neurons (OXY-IR & VP-IR) and their fibers in the hypothalamic areas (supraoptic nucleus, paraventricular nucleus, lateral hypothalamic area and median eminence) of the hypophysectomized rat were compared with normal rats at 6 months of survival after surgery at the light microscopic level. The number of VP-IR neurons was markedly decreased in the supraoptic nucleus (SON) and paraventricular nucleus (PVN) in the hypophysectomized rats as compared to normal rats. Moreover, The number of VP-IR fibers was decresed in the SON, PVN, lateral hypothalamic area (LHA) and median eminence in the hypophysectomized rats. The number of OXY-IR neurons and thier fibers were also decreased in the SON and PVN in the hypophysectomized rats. The present results demonstrate that hypophysectomy induces a significant decrease in the number of OXY- and VPIR neurons and fibers within hypothalamic areas (SON, PVN, and LHA at 6 months of post-hypophysectomy) are decreased.