Metabolic profile of visual cortex in diabetic rats measured with in vivo proton MRS

The purpose of the present study was to characterize the metabolic profile of the visual cortex in streptozotocin‐induced Type 1 diabetic rats by means of in vivo proton MRS. Several metabolite concentration ratios in the visual cortex were calculated. In addition, postmortem histologic analyses for retinal ganglion cell (RGC) loss, optic nerve injury and visual cortex alterations were monitored. The results showed that diabetes induced several changes in visual cortex metabolites, such as reduced N‐acetylaspartate, glutamate, γ‐aminobutyric acid, taurine and choline‐containing compound levels. Nevertheless, myo‐inositol levels increased significantly as compared with controls. Remarkable RGC loss and optic nerve degeneration were observed by morphological analysis. Moreover, the results showed significant neuronal loss and glial activation in the visual cortex. These findings indicated that, besides vascular abnormalities, neuronal loss and degeneration in the visual pathway were induced due to disrupted glucose homeostasis in diabetes. Metabolic or functional abnormalities were induced in cerebral neurons of the visual cortex by diabetes.     

Postnatal development of the corticotectal projection from the visual cortex of the mouse.

The postnatal development of the corticotectal projection was investigated by injecting the axon tracer DiI into the visual cortex of mouse pups. It was found that DiI-labeled axons arrive at the ipsilateral superior colliculus and enter the optic nerve layer of this structure on postnatal days 3 and 4 (P3-P4). These corticotectal axons extend into the caudal end of the superior colliculus on P4 and give off small collateral branches that ascend vertically to the superficial gray layer. During the first two postnatal weeks, the collateral branches do not form a demarcated terminal zone, but rather diffusely spread within the superficial gray layer of the superior colliculus. These collateral branches continue to dichotomize and form a bright terminal zone within the superficial gray layer on P11. The terminal zone decreases in size during the second and third postnatal weeks, and appears to be of the same size when compared with the adult counterpart by P19. The terminal zone of the corticotectal axons from the visual cortex is established by P19. In parallel with the maturation of the terminal zone of the corticotectal projection, the distal segment of the corticotectal axons is lost during the second postnatal week. We conclude that the growing tips of the corticotectal axons do not strictly project to their future terminal zone within the superior colliculus, and 'misdirected' axons are eliminated during the early postnatal period.     

Impaired transport of iron-dextran complex in myelinated central fibers

The labeling of retinal ganglion cells by axonal transport of an iron-dextran complex (injected into the superior colliculus and the lateral geniculate nucleus) was compared to the previously described labeling of the dopaminergic neurons of the substantia nigra (after striatal injection), to test the validity of the method in myelinated and unmyelinated CNS pathways. It was shown that the retrograde labeling in the visual pathway was impaired, consecutive to the penetration of iron-dextran into the myelin sheaths and a subsequent myelin alteration along the optic nerve.     

Calpain and calpastatin activity in the optic pathway

The levels of the neutral proteolytic enzymes calpains and their endogenous inhibitor calpastatin were determined in the retina and in the retrobulbar optic pathway in the albino rabbit. The highest level of calpains was observed in the optic nerve with decreasing levels in the optic tract and superior colliculus. The level of calpastatin in the retina was very low compared to that in the optic nerve and tract and other parts of the nervous system.     

Anatomical correlations of intrinsic axon repair after partial optic nerve crush in rats.

About 15% of retinal ganglion cells survive diffuse axonal injury of the optic nerve in adult rats. Following initial blindness, discrimination of visual stimuli in behavioral tests recovers within three weeks. To investigate the mechanisms promoting this functional recovery the axonal transport and the neurofilaments were studied. Intraocularly applied MiniRuby is transported until the place of crush and accumulated in enlarged axon terminals. Three weeks after lesion the anterograde transport of MiniRuby recovers distal to the place of crush. At the same point in time the retrograde transport of surviving retinal ganglion cells is restored which was visualized by horseradish peroxidase injected into the superior colliculus. The heavy neurofilament was stained immunohistochemically and analyzed statistically up to three weeks after optic nerve crush. The stained filaments in the axon fibers of retinal ganglion cells appear wavelike and/or fragmented up to day 8, but first signs of heavy neurofilament restitution in the fibers of the optic nerve are seen at day 12 after axonal injury. Because these results cannot be explained by longlasting axon regeneration, the present results provide convincing evidence for intrinsic axon repair soon after diffuse axonal injury that correlates in time with recovery of vision.     

Reduced myelinated fiber size correlates with loss of axonal neurofilaments in peripheral nerve of chronically streptozotocin diabetic rats.

Peripheral sensory nerve abnormalities were investigated in long-term streptozotocin diabetic rats using quantitative analysis. To determine whether the characteristic structural changes occur with a proximodistal gradient, three levels of the sensory peripheral nervous system were investigated: the postganglionic segment of the dorsal root, the midportion of the sciatic nerve, and the distal sural nerve. Reduction of myelinated fiber size due to reduced axonal caliber was the most characteristic change at both proximal and distal levels of the peripheral nerve. The relationship between axonal size and myelin spiral length indicated a more severe axonal atrophy in the distal portion. The axonal atrophy was related to a proportional loss of axonal neurofilaments at proximal levels, whereas in the distal sural nerve the loss of neurofilaments exceeded that which would be expected for axonal size. The universal reduction of axonal size in diabetic nerve may be accounted for by impaired supply of neurofilaments or reduced neurofilament synthesis. Such cytoskeletal defects may, in turn, lead to distal axonal degeneration or contribute to the susceptibility of diabetic nerve to various external noxi, including ischemia and hypoglycemia.     

视神经切断诱导金黄地鼠脑内视觉通路小热休克蛋白27的表达

目的 观察成年金黄地鼠左侧视神经眶内切断术后,视交叉、视束、外侧膝状体以及上丘内小热休克蛋白27(small heat shock protein,HSP27)表达的变化。方法 免疫组织化学染色法及光密度测定。结果 在正常对照和手术对照组,双侧视交叉、视束、外侧膝状体和上丘未见明显免疫反应物,光密度值之间亦无显著差异。而在实验组,与左侧相比,右侧视交叉、视束、外侧膝状体和上丘免疫组织化学染色明显增强。术后1周,右侧脑内视觉通路可见免疫反应物明显沉着的阳性细胞,形态类似星形胶质细胞。统计学分析显示,视交叉、外侧膝状体和上丘左、右两侧的光密度差值在术后1周时最大,至术后第2周迅速降低,以后下降趋势减缓,到术后8周仍可见右侧光密度值高于左侧。结论 一侧视神经切断后,对侧视交叉、视束、外侧膝状体和上丘等区域均有HSP27表达的增强,并可持续至术后8周之久。提示上述区域HSP27表达增强与视觉通路的损伤有关,但其发生机理及生物学意义尚待进一步研究。     

早期糖尿病视网膜病变模型大鼠玻璃体腔注射安维汀后房水细胞因子变化

背景：关于抑制血管生成药安维汀治疗早期糖尿病视网膜病变大鼠的机制研究多数局限于血管内皮生长因子,而结缔组织生长因子和色素上皮衍生因子也在其中起重要的作用。 目的：探讨安维汀玻璃体腔注射在早期糖尿病视网膜病变模型大鼠应用后房水细胞因子的变化及意义。 方法：经链脲佐菌素诱导10周建立早期糖尿病视网膜病变模型大鼠,分别采用安维汀(1.25,2.5 mg)和生理盐水进行玻璃体腔注射。 结果与结论：ELISA检测显示,与生理盐水注射组比较,两个安维汀注射组房水中血管内皮生长因子质量浓度降低,色素上皮衍生因子和结缔组织生长因子质量浓度增高(P < 0.05),但两组间上述细胞因子的浓度差异无显著性意义(P > 0.05)。结果证实,安维汀玻璃体腔注射在早期糖尿病视网膜病变大鼠应用促进新生血管的细胞因子血管内皮生长因子水平降低,抑制新生血管的细胞因子色素上皮衍生因子质量浓度升高,促进结缔组织生长因子水平升高。 中国组织工程研究杂志出版内容重点：肾移植;肝移植;移植;心脏移植;组织移植;皮肤移植;皮瓣移植;血管移植;器官移植;组织工程全文链接：     

Effect of acute and chronic bilateral visual deafferentation on c-Fos immunoreactivity in the visual system of adult rats

In our study we examined acute and chronic changes in c-Fos expression patterns in the visual system of the rat after complete visual deafferentation. In 20 male Lewis rats, the retro-bulbar part of the optic nerve was sectioned bilaterally. Ten animals underwent c-Fos immunohistochemistry after 3 days and 10 animals after 3 weeks examining time-dependent changes. The control group consisted of 10 animals, which did not undergo any surgical manipulation. c-Fos expression in the rat visual system experienced significant changes after acute and chronic bilateral complete visual deafferentation. Acute decrease in c-Fos level was observed in the ventral lateral geniculate nucleus, intergeniculate leaflet, superficial gray layer of the superior colliculus and layers IV and V of the primary visual cortex. After chronic deafferentation, c-Fos expression was also found to be decreased in the optic and deep layers of the superior colliculus and layer VI of the primary visual cortex. No change in c-Fos expression was observed in the dorsal lateral geniculate nucleus and layers I, II and III of the primary visual cortex. This work shows that secondary complete blindness does not lead to uniform decrease in c-Fos levels in all subcortical and cortical brain regions related to vision. These findings provide important information concerning expression of the immediate-early gene product c-Fos in secondary blind rodent models. It may further serve as a relevant baseline finding when electrical stimulation of the visual system is performed, aiding the assessment of visual neuroprosthesis using c-Fos as a functional mapping tool when evaluating different stimulus parameters in blind rodent models.     

Beneficial effects of treadmill training in experimental diabetic nerve regeneration

OBJECTIVES:

We investigated the effects of treadmill training (10 weeks) on hindlimb motor function and nerve morphometric parameters in diabetic rats submitted to sciatic nerve crush.

MATERIALS AND METHOD:

Wistar rats (n = 64) were divided into the following groups: non-diabetic; trained non-diabetic; non-diabetic with sciatic nerve crush; trained non-diabetic with sciatic nerve crush; diabetic; trained diabetic; diabetic with sciatic nerve crush or trained diabetic with sciatic nerve crush. Diabetes was induced by streptozotocin injection (50 mg/kg, iv). Hindlimb motor function was evaluated weekly by assessing sciatic functional indices, and the proximal and distal portions of the sciatic nerve were used for morphometric analysis.

RESULTS:

At 13 weeks post-injury, the distal nerve portion of all injured groups and the proximal nerve portion of the diabetic with sciatic nerve crush group presented altered morphometric parameters such as decreased myelinated fiber diameter (∼7.4±0.3µm vs ∼4.8±0.2µm), axonal diameter (∼5±0.2µm vs ∼3.5±0.1µm) and myelin sheath thickness (∼1.2±0.07µm vs ∼0.65±0.07µm) and an increase in the percentage of area occupied by endoneurium (∼28±3% vs ∼60±3%). In addition, in the non-diabetic with sciatic nerve crush group the proximal nerve portion showed a decreased myelinated fiber diameter (7.4±0.3µm vs 5.8±0.3µm) and myelin sheath thickness (1.29±0.08µm vs 0.92±0.08µm). The non-diabetic with sciatic nerve crush, trained non-diabetic with sciatic nerve crush, diabetic with sciatic nerve crush and trained diabetic with sciatic nerve crush groups showed normal sciatic functional index from the 4^th, 4^th, 9^th and 7^th week post-injury, respectively. Morphometric alterations in the proximal nerve portion of the diabetic with sciatic nerve crush and non-diabetic with sciatic nerve crush groups were either prevented or reverted to values similar to the non-diabetic group by treadmill training.

CONCLUSION:

Diabetic condition promoted delay in sciatic nerve regeneration. Treadmill training is able to accelerate hindlimb motor function recovery in diabetic injured rats and prevent or revert morphometric alterations in proximal nerve portions in non-diabetic and diabetic injured rats.     

Distribution of metabotropic glutamate receptors in the superior colliculus of the adult rat, ferret and cat

The distribution of different metabotropic glutamate receptors (mGluRs 1a, 1b, 1c, 2/3, 4 and 5) has been compared in the superior colliculus of the rat, cat and ferret using immunohistochemical techniques and light microscopy. We found that although there are differences in labelling patterns between the species, there are also substantial similarities. In general, there was only light staining for the various mGluR1 splice variants, whereas labelling for the other Group I receptor, mGluR5, was heavier and with a pattern which suggested that at least some label arose from retinal afferents to the superficial superior colliculus. A further consistent feature in all species was labelling of astrocytes in the optic nerve/optic tract, superficial superior colliculus and brain at the collicular level with the antibody directed towards the Group II receptors, mGluR2 and mGluR3. Staining for the Group III receptor, mGluR4, was dense in the superficial superior colliculus in all species, with characteristics suggesting nerve fibre staining. mGluR4 staining was seen in the cat optic nerve/optic tract. One source of mGluR4 staining in the superior colliculus may thus be retinal axons, although other sources cannot be entirely excluded. These results demonstrate that distributions of mGluRs in these species have significant similarities but also some differences, suggesting that within the superior colliculus there may be some preservation of functional roles for some of the different receptor types. This is particularly so for the Group II and Group III receptors, which appear to have specific and distinct roles in the modulation of visual responses.     

Rearrangements in the retino-geniculate projections of rats following ablation of the superior colliculus in infancy

Summary The superior colliculus was bilaterally or unilaterally ablated at different early postnatal ages in rats. When adult, each rat received a unilateral eye injection of Horesradish peroxidase to reveal the crossed and uncrossed retinal terminal fields within the dorsal lateral geniculate nucleus. Collicular ablation in the first seven days after birth, but not thereafter, produced a small hole or vacancy within the contralateral retinal terminal field which was occupied by an aberrant ipsilateral retinal terminal field. These rearrangements in the retino-geniculate projections occurred in the caudal quarter of the nucleus dorso-laterally just beneath the optic tract, solely ipsilateral to the ablated colliculus. Possible causes of the formation of these rearrangements are discussed, and similarities with other aberrant retinal projections following early damage to the visual system are considered.Abbreviations dLGN Dorsal geniculate nucleus - DTN Dorsal terminal nucleus of the accessory optic tract - HRP Horseradish peroxidase - LP Latero-posterior nucleus - NOT Nucleus of the optic tract - OT Optic tract - PO Olivary pretectal nucleus - PP Posterior pretectal nucleus - SC Superior colliculus - TMB Tetramethyl benzidine     

金黄地鼠上丘和外侧膝状体之间的联系

本实验采用了顺行和逆行追踪技术,对金黄地鼠上丘与丘脑视核团的纤维联系进行了实验观察。一、用尼氏和Loyez染色法,对4只正常金黄地鼠上丘和外侧膝状体背核和腹核的正常结构做了观察。二、3~H-亮氨酸和3~H-脯氨酸注入动物上丘不同部位(4只,存活期一天)后,可见神经末梢标记于同侧的外侧膝状体背核和腹核的外侧部位。若注射部位在上丘外侧,其投射部位在外侧膝状体背核和腹核的尾外侧;注射部位移向内侧,投射部位移向吻外侧。三、将HRP注入外侧膝状体背核(4只,存活期一天)或腹核(2只,存活期一天)或后外侧核(2只,存活期一天)后,在同侧上丘浅层见有标记神经元。在上丘深层则未见。本实验说明了上丘浅层的神经元对同侧的外侧膝状体背核和腹核有局部定位的投射,并按视野和视网膜将该投射作定位排列。     

Afferent input for target survival in marsupial visual development

The influence of afferent input on the survival of target neurons in mammals has been examined by the removal of one eye of pouch young of the marsupial native cat (Dasyurus hallucatus). The ages at eye removal spanned the period of neurogenesis of the ascending visual pathway, and were earlier than the time of maximal axon number in the optic nerve. Autoradiography following the injection of tritiated proline into the intact eye of adult animals shows that the lateral geniculate nucleus contralateral to the injected eye of the earliest enucleates retains its laminated structure, despite the total absence of binocular competition throughout development. However, we find a dramatic, age-related reduction in the volume of those parts of the lateral geniculate nucleus and superior colliculus which would normally receive a contralateral-only projection from the enucleated eye. The effects of the enucleation are not restricted to the primary termination sites of the optic axons but ramify throughout a large part of the neo- and archicortex.     

Evidence of a collicular input to the dorsal lateral geniculate nucleus in rabbits — electrophysiology

Summary In anesthetized and paralyzed rabbits, unit responses of lateral geniculate nucleus (LGN) cells to focal electrical stimulation of the superior colliculus were studied. Geniculate responses to collicular stimulation (SCS) were compared with responses to optic nerve shock (ONS). A weak correlation coefficient suggested that collicular stimulation did not fire geniculate cells through collateral activation. Further differentiation between collicular and retinofugal inputs to LGN was made possible by repetitive stimulation. Geniculate cells which responded to collicular stimulation were relay cells as they were antidromically invaded from the visual cortex. This ruled out recordings from the ventral geniculate, since this area does not project to the visual cortex. A direct colliculo-geniculate pathway was revealed by antidromic activation of collicular cells by stimulation of the dorsal LGN. Finally, triggering flashes by collicular firing resulted in a marked modification of the geniculate test response. The results suggest that the superior colliculus sends fibers to the LGN and is capable of modulating the retino-cortical neuronal message at the level of the LGN.     

Early changes of LIFR and gp130 in sciatic nerve and muscle of diabetic mice

Peripheral neuropathy is a common complication of diabetes mediated by alterations of growth factors. Members of the neuropoietic cytokine family, which include IL-6, LIF, and CNTF among others, have been shown to be important regulators of peripheral nerves and the muscles that they innervate. To investigate their potential role in diabetic nerve and muscle, we studied the expression of the shared receptor subunits, LIFR and gp130 in a mouse model of streptozotocin (STZ)-induced diabetes. The results of Western blotting and densitometric analysis showed that both LIFR and gp130 protein expression were increased in diabetic sciatic nerve compared to control mice at early time points following STZ injection. In diabetic gastrocnemius muscle, LIFR and gp130 were increased from 3 days to 24 weeks following STZ injection. In contrast, both LIFR and gp130 protein expression were decreased in diabetic soleus muscle at 3-days post-injection. Our results suggest that hyperglycemia results in changes to nerve and muscle soon after the onset of diabetes and that cytokines may play a role in this process.     

Spread of Creutzfeldt-Jakob disease virus along visual pathways after intraocular inoculation

Summary We studied the targeting of spongiform lesions within the visual pathways after intraocular injection with the Fujisaki strain of Creutzfeldt-Jakob disease (CJD) virus. The first lesions were observed 18 weeks postinoculation in the most superficial layer of the superior colliculus and in the lateral geniculate body contralateral to the side of the inoculation. Asymmetrical lesions in the superior colliculus were found also in mice sacrificed at 19, 22, and 27 weeks postinoculation. These results demonstrate that CJD virus spreads within the CNS via central axons of the visual pathways following intraocular inoculation.     

Superior colliculus involvement in the locomotor effects of ambient noise and the stimulants

Partial destruction of the superior colliculus (45%) significantly decreased the normal facilitatory effect of ambient white noise on locomotor activity levels in young rats. As recovery from surgery occurred and as test experience increased, the loss observed immediately following surgery was reduced. Presumably because of the age of the rats examined, destruction of the superior colliculus failed to potentiate the stimulant effects of d-amphetamine or methylphenidate on locomotion. These data suggest that the superior colliculus is involved in changes in general activity that result from manipulation of auditory stimuli in the environment in addition to the documented involvement of the superior colliculus in alterations of general responsivity resulting from manipulations of visual stimuli in the environment. Moreover, the superior colliculus is implicated in maintaining both excitatory and inhibitory changes in response to the environment of the organism.     

视神经和视交叉冠状断层解剖与MRI

目的：研究视神经和视交叉在冠状断面上的解剖特征与MRI表现,为相关疾病的影像诊断和外科手术提供解剖学资料。方法：选取15例成人尸头标本,以冷冻切片法切制成连续冠状断层标本;应用3．0TMRI扫描仪,获取10例志愿者头部的sE序列T1WI和T2WI图像;在上述断层标本和MRI图像上,观察分析冠状断面上视神经的形态、位置和毗邻关系。结果：视神经眶内段居于眶中心偏内上方,断面呈圆形,其周围有视神经鞘包绕,蛛网膜下隙清晰可见,眼动脉位于其上方;视神经管内段居于眶内上方,断面呈水平卵圆形,周围可见视神经鞘,蛛网膜下隙不明显,眼动脉位于其下方;视交叉冠状断面呈“一”字型,分隔上方的视隐窝和后下方的漏斗隐窝,其上方有大脑前动脉A1段,下方为灰结节和垂体柄。结论：冠状断面町以清楚显示视神经、视交叉的形态特点与毗邻关系。     

Permeability of blood-brain and blood-nerve barriers in experimental diabetes mellitus in the anaesthetized rat.

Diabetes was induced with streptozotocin in rats weighing about 160 g. These were maintained with age-matched controls for up to 14 months, blood glucose being periodically monitored. Half the diabetic and control rats received the aldose reductase inhibitor, Ponalrestat, in their diet. At 3 weeks, 6-7 months and 13-14 months, the vascular permeability in regions of brain, and in optic and sciatic nerves, were measured by maintaining radiotracers in the bloodstream--125I-albumin (100 min), [14C]sucrose (60 min) and 131I-albumin (5 min)--followed by tissue sampling and counting at termination. 131I-albumin estimated residual intravascular plasma. Diabetes of up to 13-14 weeks caused no measurable increase in the sucrose permeability of microvessels in eight different brain regions, in optic or in sciatic nerve. At 3 weeks of diabetes, sucrose permeability in all brain regions and in optic nerve was reduced relative to that in controls. Extravascular albumin entry into different regions of brain and optic nerve was insignificant and insensitive to diabetes, except in the hypothalamus and optic nerves where it was raised with increasing duration of diabetes. In sciatic nerve, extravascular albumin distribution was markedly increased by diabetes, but sucrose permeability was not demonstrably affected. At the level used in the diet, Ponalrestat reduced the sorbitol content of diabetic sciatic nerve but did not protect again the increased permeability to albumin.