The enhanced expression of c-Jun immunoreactivity in the adrenalectomized gerbil hippocampus

Recent in vitro and in vivo studies have shown that glucocorticoids have a profound influence on the survival of hippocampal neurones, and that the depletion of glucocorticoids as a result of adrenalectomy (ADX) reduces nerve growth factor levels in the hippocampus. It is also believed that ADX is associated with the seizure susceptibility of the Mongolian gerbil. In the present study, the choronological changes of c-jun immunoreactivity were investigated after ADX in the hippocampal formations in the seizure-prone gerbil model. In the sham hippocampus, c-jun immunoreactivity was not observed in the neurones of the hippocampus proper and dentate gyrus. C-jun immunoreactive neurones appeared 3 h after ADX in the neurones of the CA1 area and dentate gyrus, and these immunoreactivities peaked 24 h after ADX and then gradually decreased. These results suggest that, in the adrenalectomized gerbil, c-jun may be expressed in the neurones of the hippocampus in compensation for glucocorticoid deficit. The result of enhanced c-jun expression of the hippocampal formation provides anatomical support for the hypothesis that c-jun may play a role in the reduction of seizure activity.     

Comparative studies on the distribution of glutamate transporters in the retinae of the Mongolian gerbil and the rat

Glutamate is the major excitatory amino acid transmitter in vertebrate retinae. Glutamate transporters therefore play an important role in the precise control of glutamate concentration in the synaptic cleft by regulating extracellular glutamate concentration. In the present study, we performed an analysis of the expressions of three glutamate transporters in gerbil retina using immunohistochemistry. In the gerbil retina, excitatory amino acid carrier 1 and glutamate transporter 1 immunoreactivity was predominant in the ganglion cells but not amacrine or bipolar cells. Glutamate/aspartate transporter (GLAST) immunoreactivity was observed in the radial gliocytes of which the dense network of fine processes was localized in the inner and outer plexiform layers. GLAST immunoreactivity was also detected in astrocytes in the nerve fibre layer. These results demonstrate that three glutamate transporters show specific distributions in the gerbil retina and suggest that the glutamate re-uptake system in the gerbil retina may be different from that of the rat.     

Distribution of nociceptin-like immunoreactivity in the central nervous system of the Mongolian gerbil: an immunohistochemical study

This study is designed to demonstrate the distribution of nociceptin, endogenous ORL1 receptor ligand, in the central nervous system of the Mongolian gerbil. To intensify the nociceptin-like immunoreactivity (NOC-LI), colchicine was administered into the lateral ventricle, at 48 h prior to the transcardiac perfusion. In the group without colchicine treatment, NOC-LI was observed in the fibres of the spinal dorsal horn, specifically in the superficial layers. However, the NOC-LI in the superficial layers disappeared after the administration of colchicine. In the brain, NOC-LI was prominent in the hypothalamus, hippocampus, cerebral peduncle, substantia nigra, dorsal raphe, periaqueductal grey, locus coeruleus and trapezoid nucleus. Colchicine treatment markedly intensified the NOC-LI in the somata of the central nervous system, whereas the untreated sections were too weak to observe and analyse. The distribution of NOC-LI provides informative data for studies of the neuronal circuit that nociceptin may be involved in.     

Differential effects of treadmill exercise on calretinin immunoreactivity in type 2 diabetic rats in early and chronic diabetic stages

In this study, we investigated the effects of treadmill exercise on calretinin (CR), a marker of early postmitotic neurons, immunoreactivity in the dentate gyrus (DG) of Zucker diabetic fatty (ZDF) rats, before or after diabetes onset, and Zucker lean control (ZLC) rats. For this study, 6-week-old ZLC and prediabetic ZDF rats, and 22-week-old ZLC and ZDF rats were exercised on the treadmill. Sedentary ZLC and ZDF rats of the same age were used as exercise experiment controls. The exercised prediabetic ZDF rats did not show diabetes onset, while the sedentary prediabetic ZDF rats showed significantly increased blood glucose levels. The exercised diabetic ZDF rats exhibited a decrease in their blood glucose levels compared to the sedentary diabetic ZDF rats, but the levels were still above 20 mmol/l. ZLC rats of both ages were in the normoglycemic range. CR immunoreactivity was detected throughout the DG, including the subgranular zone and the polymorphic layer. Diabetic rats exhibited a significant decrease in the number of CR-immunoreactive cells and fibers in the DG. Exercise in the prediabetic ZDF rats significantly increased the number of CR-immunoreactive cells and fibers in the subgranular zone of the DG. In the ZLC and ZDF rats of chronic diabetic phase, exercise increased CR-immunoreactive neurons in the hilar region. These results suggest that diabetes significantly reduces the number of postmitotic CR-immunoreactive neurons and the intensity of immunoreactivity and that exercise increases these CR-related parameters in a diabetic stage-dependent manner.     

Ribosomal protein s3 immunoreactivity in the young, adult and aged gerbil hippocampus

Ribosomal protein S3 (rpS3) is well known to participate in DNA repair mechanisms. In the present study, we compared rpS3 immunoreactivity and its protein levels in the hippocampus among young, adult and aged gerbils. In the postnatal month (PM) 3 group as the young, rpS3 immunoreaction was observed in pyramidal and non-pyramidal cells of the hippocampus proper and in granule and polymorphic cells of the dentate gyrus. In the PM 12 group as adult and 24 group as the aged, rpS immunoreactivity in the hippocampus was decreased compared to the PM 3 group. Western blot analysis showed that rpS3 levels were decreased with time; lowest at PM 24. These results indicate that rpS3 immunoreactivity and protein levels were markedly decreased in the aged gerbil hippocampus.     

Distribution of serotonin immunoreactivity in the main olfactory bulb of the Mongolian gerbil

The distribution of serotonin immunoreactivity in the main olfactory bulb (MOB) of the Mongolian gerbil (Meriones unguiculatus) was examined by immunohistochemistry. Seven distinct layers of the Mongolian gerbil MOB-stained with cresyl violet were identified. Serotonin-immunoreactive (IR) cell bodies were not found in the MOB. The serotonin-IR nerve fibres had a specific laminar distribution and morphology in the gerbil MOB. Serotonin-IR nerve fibres were observed in the glomerular, external plexiform and granule cell layers of the MOB. These serotonin-IR nerve fibres showed varicosities that were larger than the thickness of the axon. The highest density of serotonin-IR nerve fibres was in glomeruli of the glomerular layer. The average fibre density in the glomerular layer was more than three to four times the density in the infraglomerular layers. Glomerular serotonin-IR fibres were much more intensively stained than infraglomerular serotonin-IR fibres. This result suggests that serotonin-IR nerve fibres of Mongolian gerbil MOB are extrinsic and may act to modulate the olfactory transmission.     

Immunohistochemical study of caveolin-1 and -2 in the rat retina

The expression of caveolin-1 and -2 in the retina was examined; Western blot analysis showed that both were present. Immunohistochemistry indicated that caveolin-1 was expressed in the majority of retinal layers, including the ganglion cell layer, inner plexiform layer, outer plexiform layer, and in the vascular endothelial cells of the retina. Caveolin-2 was primarily immunostained in the vessels, but in a few other elements as well. This is the first demonstration of caveolin differential expression in the retina of rats, and suggests that caveolin plays an important role in signal transduction in glial cells and neuronal cells.     

Loss of glutamine synthetase immunoreactivity from the retina in canine primary glaucoma

Purpose Changes in retinal glutamate distribution occur in primary glaucoma (PG) in dogs. Although the redistribution resembles that induced by ischemia, decreases in glutamine synthetase (GS) activity may also induce a similar glutamate redistribution. We examined the distribution of GS, glutamate, and glial fibrillary acidic protein (GFAP), a marker for reactive glia, in PG retinas by immunohistochemistry to determine whether decreases in GS and formation of reactive glia are associated with glutamate redistribution and neuronal damage. Animals Sections from 14 control dog eyes and 22 eyes from dogs with PG. Methods Sections from 14 control and 22 glaucomatous globes were immunohistochemically stained for GS, glial fibrillary acidic protein or glutamate. Results In semiquantitative immunogold studies, decreases in GS staining density were strongly correlated with glutamate redistribution and neuronal damage. In less quantitative immunoperoxidase staining of acute (≤ 5 days after clinical signs) and chronic PG retinas, GS immunoreactivity was decreased in focal regions of some acute PG retinas, and there were widespread decreases in chronic PG retinas. GFAP immunoreactivity was increased in Müller cells primarily in severely damaged regions of chronic PG retinas. Conclusions Decreases in GS immunoreactivity were associated with glutamate redistribution. These decreases in GS occurred even in mildly damaged regions of retina before retinal thinning. Reactive Müller cells were seen primarily in chronic PG in severely damaged regions. Decreases in GS may potentiate ischemia‐induced early glutamate redistribution and neuronal damage in canine PG.     

Reduced immunoreactivity of tyrosine hydroxylase in the hypothalamic paraventricular nucleus of the seizure sensitive gerbil

We compared the immunoreactivity and numbers of tyrosine hydroxylase (TH) immunoreactive neurons and neuropil in the paraventricular nucleus (PVN) of the hypothalamus between the seizure sensitive (SS) and seizure resistant (SR) gerbils. The distributional pattern of TH immunoreactivity was similar in both groups: TH immunoreactivity was seen mainly in magnocellular neurons of the PVN. However, total TH immunoreactivity in the neurons and neuropil in the SS gerbils was significantly lower than that in the SR gerbils. In addition, the number of TH immunoreactive neurons in the SS gerbils was also much lower than those in the SR gerbils. These results indicate that SS gerbils have a low TH immunoreactivity in the hypothalamic PVN compared with that in SR gerbils.     

Effects of dizocilpine pretreatment on parvalbumin immunoreactivity and Fos expression after cerebral ischemia in the hippocampus of the Mongolian gerbil

The mechanisms of ischemic neuronal death have been focused on glutamate receptor activation and subsequent elevation of intracellular Ca2+ concentration. The purpose of this study was to evaluate the effects of dizocilpine, an NMDA receptor antagonist, pretreatment on Fos expression and parvalbumin (PV, calcium binding protein) immunoreactivity in the hippocampus of the mongolian gerbil after global ischemic insults. The number of PV-immunoreactive (PV-ir) neurons in CA1 were significantly decreased from 1 day after cerebral ischemia, while dizocilpine pretreatment completely suppressed the loss of PV-ir neurons in CA1. Dizocilpine pretreatment also protected the structural loss of microtubule-associated protein 2 immunoreactivity in CA1 after ischemic insults. In addition, dizocilpine pretreatment increased Fos expression in both hippocampal CA3 and CA4 after 3 hr ischemic reperfusion as compared to that of the saline pretreated group. Subsequently, the Fos-defined cellular activity of PV-ir neurons was slightly increased by dizocilpine pretreatment in the hippocampal area. This study demonstrated that NMDA receptor mediated calcium influx was associated with the loss of PV-ir neurons in CA1 hippocampal region, and that dizocilpine pretreatment increased Fos expression and the neuronal activity of PV-ir neurons in the non-vulnerable region of hippocampus after cerebral ischemia. Based on this data, we conclude that the protective effect of dizocilpine may be induced by the regulation of calcium overload, or by the upregulation of a neuroregenerative initiator such as Fos protein.     

Changes of calbindin D-28k immunoreactivity in the hippocampus after adrenalectomy in the seizure sensitive gerbil

Calbindin D-28k (CB), a calcium-binding protein, containing neurons in the hippocampus plays an important role in hippocampal excitability in epilepsy. In the present study, we investigated changes of CB immunoreactivity after adrenalectomy (ADX) in the hippocampus and dentate gyrus of the seizure sensitive gerbil, which is susceptible to seizure to identify roles of CB in epileptogenesis. The changes of the CB immunoreactivity after ADX were significant in the hippocampal CA1 region. By 24 h after ADX, CB-immunoreactive CA1 pyramidal cells and CB immunoreactivity increased. At this time, well-stained dendrites projected to the stratum radiatum. Thereafter, the CB immunoreactivity decreased time dependently by 96 h after ADX. In the dentate gyrus, the changes of CB-immunoreactive neurons were mainly observed in the granule cell layer. The number and immunoreactivity of CB-immunoreactive neurons was high at 24 h after ADX, thereafter, those decreased by 96 h after ADX. These results suggest that glucocorticoid has an important role in modulating the seizure activity and CB serves an inhibitory function, which regulates the seizure activity and output signals from the hippocampus.     

Age-related changes in ionized calcium-binding adapter molecule 1 immunoreactivity and protein level in the gerbil hippocampal CA1 region

Microglia are evenly distributed throughout the brain parenchyma. They respond rapidly to a variety of alterations in the microenvironment of the brain and act as sensors for pathological events in the brain. In the present study, we investigated the age-dependent changes in the immunoreactivity and protein level of ionized calcium-binding adapter molecule 1 (Iba-1), a microglial marker, in the CA1 region of the gerbil hippocampus. Iba-1 immunoreactive microglia were detected in the hippocampal CA1 region of the postnatal month 1 (PM 1) group. Iba-1 positive microglia were morphologically inactive between the PM 1 and PM 12 stages. Some Iba-1 immunoreactive microglia were present in the active form in the hippocampal CA1 region of the PM 18 and PM 24 groups. The Iba-1 protein levels in hippocampal CA1 homogenates were decreased in the PM 1 through PM 6 groups and increased in an age-dependent manner thereafter. These results suggest that Iba-1 immunoreactive microglia in the active form were detected in the hippocampal CA1 region in the PM 18 and PM 24 groups. This result may be associated with an age-dependent susceptibility to neurodegenerative diseases associated with the hippocampus.     

Histochemical study on neurodegeneration in the olfactory bulb after transient forebrain ischaemia in the Mongolian gerbil

In the present study, we investigated the ischaemia-related neurodegeneration in the main and accessory olfactory bulb (AOB) after 5 min transient forebrain ischaemia in the Mongolian gerbil using the acid fuchsin staining method. Between 5 and 15 days after ischaemia, acid fuchsin positive cells markedly increased in the external plexiform layer (EPL), mitral cell layer (ML) and glomerular layer (GL) of the main olfactory bulb (MOB), and in the mixed cell layer (MCL) and GL of the AOB. By 30 days after ischaemia reperfusion, acid fuchsin positive neurons were shrunken and showed low acidophilia in somata. Many necrotic vacuoles were found in the EPL and GL of the MOB 30 days after ischaemia. At this time, necrotic vacuoles were very few in the AOB. Therefore, our results suggest that the GL and EPL of the MOB are vulnerable to ischaemic damage at a later time after ischaemic insult, and that the AOB is more resistant to ischaemic damage as compared with the MOB.     

Effect of 7-nitroindazole, a selective neuronal nitric oxide synthase inhibitor, on parvalbumin immunoreactivity after cerebral ischaemia in the hippocampus of the Mongolian gerbil

Previous studies have demonstrated that a loss of parvalbumin-immunoreactive (PV-ir) neurones is observed in the hippocampus after transient cerebral ischaemia. However, whether the loss of parvalbumin (PV) immunoreactivity is related to the over-production of nitric oxide (NO) during cerebral ischaemia has not been evaluated. This study was designed to test the effect of 7-nitroindazole pre-treatment (7-NI, 50 mg/kg), a selective neuronal NO synthase inhibitor, on PV immunoreactivity and its cellular activity following forebrain ischaemia. PV-ir neurones in the hippocampus of the control group were widely distributed in the pyramidal cell layer and stratum oriens of CA1 and CA3, and the granular cell layer of dentate gyrus. 7-NI pre-treatment completely suppressed the reduction of PV immunoreactivity in CA1 that was observed in the ischaemia-induced group. Subsequently, 7-NI pre-treatment also protected against the structural loss of microtubule-associated protein 2 (MAP2) immunoreactivity in CA1 after ischaemic insult. In addition, the Fos-defined neuronal activity of PV-ir neurones was slightly increased by the 7-NI pre-treatment 3 h after ischaemia. Based on these data, we conclude that the neuronal toxicity of NO may be involved in the loss of PV-ir neurones after cerebral ischaemia.     

A comparative study on cardiac ganglia in midday gerbil,Egyptian spiny mouse,chinchilla laniger and pigeon

Using the thiocholine method and histological techniques, the topography and morphology of cardiac ganglia in midday gerbil, Egyptian spiny mouse, chinchilla laniger and pigeon were studied. The results demonstrated that cardiac ganglia in all investigated species are embedded in epicardial fat. They formed plexo-ganglionic structures. Each of them composed of many ganglia (from seven up to 36) different in size and shape, and interconnected by fascicles of nerve fibres. Comparative analysis showed that the density of neural network and cell aggregations was different in individual species. The richest plexo-ganglionic structure was in pigeon. It was organized in three plexo-ganglia with an average of 30 ganglia. The largest one was located along the anterior interventricular sulcus. The cardiac ganglia of investigated mammals were localized mainly on the epicardium of atria; in Egyptian spiny mouse and chinchilla laniger on the ventral surface of right atrium, but in midday gerbil on the dorsal surface of left atrium. Moreover, in midday gerbil and Egyptian spiny mouse the little plexo-ganglionic structure on the ventricle were noticed. Additionally, in midday gerbil the single nerve cells might be observed between cardiac muscle of atria. It can be said that, the strongly developed cardiac plexus in pigeon is probably connected with his behaviour and functional properties of the heart. The arrangement of neurones in cardiac ganglia of all examined mammals was uniform over the whole surface of the sections, while in the pigeon, neurones were located mainly in the peripheral part of the ganglion.     

Immunohistochemical expression of calretinin in canine testicular tumours and normal canine testicular tissue

Calretinin is a calcium-binding protein expressed abundantly in the central and peripheral neural tissues. It has been demonstrated to be a valuable marker in human testicular neoplasia. The immunohistochemical expression of calretinin has been studied in 102 samples of normal (n=25) and three different neoplastic canine testicular tumours (n=77). In normal canine testis, calretinin expression was restricted to Leydig and Sertoli cells of the testis. In tumour tissues, calretinin expression was detected in all tumours investigated (interstitial cell tumours, seminoma, and Sertoli cell tumours), with a cytoplasmic and nuclear pattern of cellular distribution. The present work reports, for the first time, calretinin immunohistochemical expression in normal and neoplastic canine testis.     

An immunohistochemical study of protein kinase C in the bovine retina

The expression of protein kinase C (PKC) was studied in the bovine retina by immunohistochemical analysis. Western blot analysis showed that PKC isoforms, including alpha, betaI, delta and theta, were detected in the bovine retina. By immunohistochemistry, both PKC alpha and betaI were expressed in all retinal layers, with an intense localization of both PKC alpha and betaI detected in bipolar cells in the inner nuclear cell layer and in some glial cells in ganglion cell layers. The immunoreactivity of both PKC delta and theta was quite weak in the retinal layers, compared with that of PKC alpha and betaI. These findings suggest that both conventional and novel PKCs are differentially expressed in the bovine retina.     

The expression of calretinin and cytokeratins in canine acanthomatous ameloblastoma and oral squamous cell carcinoma

Oral squamous cell carcinoma (OSCC) and canine acanthomatous ameloblastoma (CAA) represent two epithelium‐derived neoplasms that affect the oral cavity of dogs. The expression of cytokeratins (CKs) and calretinin has been previously established in the canine tooth bud and odontogenic tumours. The aim of this study was to characterize the CK and calretinin expression profile of OSCC in comparison to CAA and canine tooth bud tissues. Samples from 15 OSCC and 15 CAA cases, as well as 6 tooth buds and 2 normal gingival tissues were examined. OSCC CK expression was consistent with the CK expression profile of CAA and canine tooth bud tissue. Calretinin was positively expressed in 10 of 15 OSCC cases, with 5 cases demonstrating high staining intensity. Only 2 of 15 CAA cases demonstrated mild‐moderate staining intensity. The statistically significant difference in staining pattern and intensity of calretinin in OSCC and CAA can help distinguish between these two tumour types.     

不同品种白三叶蛋白质含量比较

通过对10个不同白三叶Trifolium repens品种在不同生育期内叶片和全株蛋白质含量的测定分析,结果表明：不同品种或同一品种不同生育期的白三叶蛋白质含量差异极显著（P<0.01）,蛋白质含量在生育期内呈单峰曲线变化,并以初花期为蛋白质含量降低的转折点;北京地区适宜种植的白三叶品种为海法、汉口和吉林临江,其最佳利用时期为初花期。