Automatic identification and quantitative morphometry of unstained spinal nerve using molecular hyperspectral imaging technology

Quantitative observation of nerve fiber sections is often complemented by morphological analysis in both research and clinical condition. However, existing manual or semi-automated methods are tedious and labour intensive, fully automated morphometry methods are complicated as the information of color or gray images captured by traditional microscopy is limited. Moreover, most of the methods are time-consuming as the nerve sections need to be stained with some reagents before observation. To overcome these shortcomings, a molecular hyperspectral imaging system is developed and used to observe the spinal nerve sections. The molecular hyperspectral images contain both the structural and biochemical information of spinal nerve sections which is very useful for automatic identification and quantitative morphological analysis of nerve fibers. This characteristic makes it possible for researchers to observe the unstained spinal nerve and live cells in their native environment. To evaluate the performance of the new method, the molecular hyperspectral images were captured and the improved spectral angle mapper algorithm was proposed and used to segment the myelin contours. Then the morphological parameters such as myelin thickness and myelin area were calculated and evaluated. With these morphological parameters, the three dimension surface view images were drawn to help the investigators observe spinal nerve at different angles. The experiment results show that the hyperspectral based method has the potential to identify the spinal nerve more accurate than the traditional method as the new method contains both the spectral and spatial information of nerve sections.     

Investigation of human pancreatic cancer tissues by Fourier Transform Infrared Hyperspectral Imaging

Fourier‐transform infrared hyperspectral imaging (FTIR‐HSI) provides hyperspectral images containing both morphological and chemical information. It is widely applied in the biomedical field to detect tumor lesions, even at the early stage, by identifying specific spectral biomarkers. Pancreatic neoplasms present different prognoses and are not always easily classified by conventional analyses. In this study, tissue samples with diagnosis of pancreatic ductal adenocarcinoma and pancreatic neuroendocrine tumor were analyzed by FTIR‐HSI and the spectral data compared with those from healthy and dysplastic samples. Multivariate/univariate approaches were complemented to hyperspectral images, and definite spectral markers of the different lesions identified. The malignant lesions were recognizable both from healthy/dysplastic pancreatic tissues (high values of phospholipids and triglycerides with shorter, more branched and less unsaturated alkyl chains) and between each other (different amounts of total lipids, phosphates and carbohydrates). These findings highlight different metabolic pathways characterizing the different samples, well detectable by FTIR‐HSI.     

Enhanced spatial resolution for snapshot hyperspectral imaging of blood perfusion and melanin information within human tissue

We report a reconstruction method to achieve high spatial resolution for hyperspectral imaging of chromophore features in skin in vivo. The method utilizes an established structure‐adaptive normalized convolution algorithm to reconstruct high spatial resolution of hyperspectral images from snapshot low‐resolution hyperspectral image sequences captured by a snapshot spectral camera. The reconstructed images at chromophore‐sensitive wavebands are used to map the skin features of interest. We demonstrate the method experimentally by mapping the blood perfusion and melanin features (moles) on the facial skin. The method relaxes the constrains of the relatively low spatial resolution in the snapshot hyperspectral camera, making it more usable in imaging applications.     

Parallel morphological/neural processing of hyperspectral images using heterogeneous and homogeneous platforms

The wealth spatial and spectral information available from last-generation Earth observation instruments has introduced extremely high computational requirements in many applications. Most currently available parallel techniques treat remotely sensed data not as images, but as unordered listings of spectral measurements with no spatial arrangement. In thematic classification applications, however, the integration of spatial and spectral information can be greatly beneficial. Although such integrated approaches can be efficiently mapped in homogeneous commodity clusters, low-cost heterogeneous networks of computers (HNOCs) have soon become a standard tool of choice for dealing with the massive amount of image data produced by Earth observation missions. In this paper, we develop a new morphological/neural algorithm for parallel classification of high-dimensional (hyperspectral) remotely sensed image data sets. The algorithm’s accuracy and parallel performance is tested in a variety of homogeneous and heterogeneous computing platforms, using two networks of workstations distributed among different locations, and also a massively parallel Beowulf cluster at NASA’s Goddard Space Flight Center in Maryland.

Microscopic hyperspectral imaging studies of normal and diabetic retina of rats

A microscopic hyperspectral imager was developed based on the microscopic technology and the spectral imaging technology. Some microscopic hyperspectral images of retina sections of the normal, the diabetic, and the treated rats were collected by the new imager. Single-band images and pseudo-color images of each group were obtained and the typical transmittance spectrums were ex-tracted. The results showed that the transmittance of outer nuclear layer cells of the diabetic group was generally higher than that of the normal. A small absorption peak appeared near the 180th band in the spectrum of the diabetic group and this peak weakened or disappeared in the spectrum of the treated group. Our findings indicate that the microscopic hyperspectral images include wealthy information of retina sections which is helpful for the ophthalmologist to reveal the pathogenesis of diabetic reti-nopathy and explore the therapeutic effect of drugs.     

Multivariate analysis of Brillouin imaging data by supervised and unsupervised learning

Brillouin imaging relies on the reliable extraction of subtle spectral information from hyperspectral datasets. To date, the mainstream practice has been to use line fitting of spectral features to retrieve the average peak shift and linewidth parameters. Good results, however, depend heavily on sufficient signal-to-noise ratio and may not be applicable in complex samples that consist of spectral mixtures. In this work, we thus propose the use of various multivariate algorithms that can be used to perform supervised or unsupervised analysis of the hyperspectral data, with which we explore advanced image analysis applications, namely unmixing, classification and segmentation in a phantom and live cells. The resulting images are shown to provide more contrast and detail, and obtained on a timescale ∼10² faster than fitting. The estimated spectral parameters are consistent with those calculated from pure fitting.     

Fingerprint‐to‐CH stretch continuously tunable high spectral resolution stimulated Raman scattering microscope

Stimulated Raman scattering (SRS) microscopy is a label‐free method generating images based on chemical contrast within samples, and has already shown its great potential for high‐sensitivity and fast imaging of biological specimens. The capability of SRS to collect molecular vibrational signatures in bio‐samples, coupled with the availability of powerful statistical analysis methods, allows quantitative chemical imaging of live cells with sub‐cellular resolution. This application has substantially driven the development of new SRS microscopy platforms. Indeed, in recent years, there has been a constant effort on devising configurations able to rapidly collect Raman spectra from samples over a wide vibrational spectral range, as needed for quantitative analysis by using chemometric methods. In this paper, an SRS microscope which exploits spectral shaping by a narrowband and rapidly tunable acousto‐optical tunable filter (AOTF) is presented. This microscope enables spectral scanning from the Raman fingerprint region to the Carbon‐Hydrogen (CH)‐stretch region without any modification of the optical setup. Moreover, it features also a high enough spectral resolution to allow resolving Raman peaks in the crowded fingerprint region. Finally, application of the developed SRS microscope to broadband hyperspectral imaging of biological samples over a large spectral range from 800 to 3600 cm^?1, is demonstrated.     

Age-related changes of myelin basic protein in mouse and human auditory nerve

Age-related hearing loss (presbyacusis) is the most common type of hearing impairment. One of the most consistent pathological changes seen in presbyacusis is the loss of spiral ganglion neurons (SGNs). Defining the cellular and molecular basis of SGN degeneration in the human inner ear is critical to gaining a better understanding of the pathophysiology of presbyacusis. However, information on age-related cellular and molecular alterations in the human spiral ganglion remains scant, owing to the very limited availably of human specimens suitable for high resolution morphological and molecular analysis. This study aimed at defining age-related alterations in the auditory nerve in human temporal bones and determining if immunostaining for myelin basic protein (MBP) can be used as an alternative approach to electron microscopy for evaluating myelin degeneration. For comparative purposes, we evaluated ultrastructural alternations and changes in MBP immunostaining in aging CBA/CaJ mice. We then examined 13 temporal bones from 10 human donors, including 4 adults aged 38-46 years (middle-aged group) and 6 adults aged 63-91 years (older group). Similar to the mouse, intense immunostaining of MBP was present throughout the auditory nerve of the middle-aged human donors. Significant declines in MBP immunoreactivity and losses of MBP(+) auditory nerve fibers were observed in the spiral ganglia of both the older human and aged mouse ears. This study demonstrates that immunostaining for MBP in combination with confocal microscopy provides a sensitive, reliable, and efficient method for assessing alterations of myelin sheaths in the auditory nerve. The results also suggest that myelin degeneration may play a critical role in the SGN loss and the subsequent decline of the auditory nerve function in presbyacusis.     

Grayscale representation of infrared microscopy images by extended multiplicative signal correction for registration with histological images

Fourier‐transform infrared (FTIR) microspectroscopy is rounding the corner to become a label‐free routine method for cancer diagnosis. In order to build infrared‐spectral based classifiers, infrared images need to be registered with Hematoxylin and Eosin (H&E) stained histological images. While FTIR images have a deep spectral domain with thousands of channels carrying chemical and scatter information, the H&E images have only three color channels for each pixel and carry mainly morphological information. Therefore, image representations of infrared images are needed that match the morphological information in H&E images. In this paper, we propose a novel approach for representation of FTIR images based on extended multiplicative signal correction highlighting morphological features that showed to correlate well with morphological information in H&E images. Based on the obtained representations, we developed a strategy for global‐to‐local image registration for FTIR images and H&E stained histological images of parallel tissue sections.     

Hyperspectral scanning laser optical tomography

In order to study physical relationships within tissue volumes or even organism‐level systems, the spatial distribution of multiple fluorescent markers needs to be resolved efficiently in three dimensions. Here, rather than acquiring discrete spectral images sequentially using multiple emission filters, a hyperspectral scanning laser optical tomography system is developed to obtain hyperspectral volumetric data sets with 2‐nm spectral resolution of optically transparent mesoscopic (millimeter‐centimeter) specimens. This is achieved by acquiring a series of point‐scanning hyperspectral extended depth of field images at different angles and subsequently tomographically reconstructing the 3D intensity distribution for each wavelength. This technique is demonstrated to provide robust measurements via the comparison of spectral and intensity profiles of fluorescent bead phantoms. Due to its enhanced spectral resolving ability, this technique is also demonstrated to resolve largely overlapping fluorophores, as demonstrated by the 3D fluorescence hyperspectral reconstruction of a dual‐labeled mouse thymus gland sample and the ability to distinguish tumorous and normal tissues of an unlabeled mouse intestine sample.      

Fast Evaluation of Unhealthy and Healthy Neonates Using Hyperspectral Features on 700-850 Nm Wavelengths,ROI Extraction,and 3D-CNN

ObjectivesHyperspectral imaging (HSI) has great potential in detecting the health conditions of neonates as it provides diagnostic information about the tissue by avoiding tissue biopsy. HSI gives more features than thermal imaging, which can obtain images in a single wavelength, as it can obtain images in a large number of wavelengths. The data obtained with hyperspectral sensors are 3-dimensional data called hypercube including first two-dimensional spatial information and third-dimensional spectral information.Material and methodsIn this study, hyperspectral data were obtained from 19 different neonates in the Neonatal Intensive Care Unit (NICU) of Selcuk University, Medical Faculty. There are 16 hypercubes from 16 unhealthy neonates, 16 hypercubes from 3 healthy neonates in a period of three months, and 32 hypercubes in total are available. For the training of 3D-CNN model, data augmentation methods, such as rotation, height shifting, width shifting, and shearing were applied to hyperspectral data. A number of 32 hypercubes taken from neonates in NICU were augmented to 160 hypercubes. Spectral signatures were examined and 51 bands in the range of 700-850 nm with distinctive features were used for the classification. The spectral dimension was reduced by applying Principal Component Analysis (PCA) to all hypercubes. In addition, it is aimed to obtain both spectral and spatial features with the 3D-CNN. For increasing the classification efficiency, ROI extraction was made and four datasets were created in different spatial dimensions. These datasets contain 160, 640, 1440, and 5760 hypercubes, respectively.ResultsThe best result was achieved by using 5760 hypercubes of 25x25x51. As a result of the classification of the hypercubes, accuracy 98.00%, sensitivity 97.22%, and specificity 98.78% were obtained. It was determined how many PCs used to achieve the best result. Further, the proposed 3D-CNN model is compared to 2D-CNN model to evaluate the performance of the study.ConclusionIt was aimed to evaluate the health status of neonates fastly by using HSI and 3D-CNN for the first time. The obtained results are an indication that HSI and 3D-CNN are very effective for the evaluation of unhealthy and healthy neonates.     

无人机高光谱影像与冠层树种多样性监测

冠层树种多样性是自然森林生态系统功能和服务的重要基础。及时掌握冠层多样性的现状及变化趋势, 是探讨诸多重要生态学问题的前提, 更是制定合理生物多样性保护策略的基础。但受制于传统的多样性信息采集方法, 区域尺度的高精度冠层多样性监测发展较为缓慢; 许多在气候变化和人类干扰下的生物多样性分布信息得不到及时更新。近年来基于无人机的冠层高光谱影像收集与分析技术的发展, 使得冠层多样性监测迎来了新的发展契机。本文从森林冠层高光谱影像出发, 介绍了与多样性监测相关的无人机航拍和基于深度学习的图像处理技术, 并结合已有文献, 探讨了无人机高光谱应用于森林冠层树种多样性监测的研究现状、可行性、优势及缺陷等。我们认为冠层高光谱影像为多样性监测提供了不可或缺且丰富的原始信息; 而无人机与高光谱相机的结合, 使得区域化高频率(如每周)、高精度(如分米乃至厘米级)的冠层多样性信息自动化收集成为可能。然而高光谱影像数据量大、数据维度高与数据结构非线性的特点为影像处理带来了挑战, 而深度学习技术的飞跃, 使得从冠层高光谱影像中提取个体及物种信息达到了极高精度。恰当地使用这些技术将大大提升冠层树种多样性的自动化监测水平, 由此也将帮助我们在当前剧变环境下及时掌握森林冠层多样性的现状与变化, 为生物多样性研究与保护提供可靠的数据支撑。     

Histological Methods for Assessing Myelin Sheaths and Axons in Human Nerve Trunks

Although there are many histological techniques for assessing myelin sheaths and axons in paraffin embedded or frozen sections of the peripheral nervous system, modern approaches usually use plastic embedded material. Although plastic embedding is superior for small cutaneous branches, this method has limited value for histological assessment of nerve trunks. We report three methods which together yield a comprehensive approach for thorough and detailed investigation of human nerve trunks. The rapid osmication method permitted assessment of myelinated nerve fibers from frozen sections at operation, thus providing the surgeon with guidance on the extent of nerve resection. The modification presented here resulted in permanent slides, allowing comparison of results with those of the other two procedures. The new osmium-hematoxylin technique could be performed on paraffin embedded nerves. Paraffin, unlike plastic, permitted the study of the whole cross sectional area of the nerve in single sections. Moreover, the sharp image of the myelin permitted computerized morphometry. The significantly modified axonal silver impregnation technique was performed on frozen sections mounted on glass slides, as opposed to the time-consuming impregnation of free-floating sections. The latter technique had a high success rate and permitted semiquantitative assessment of axons in nerve trunks. These methods can be performed in any routine histology laboratory and resulted in greater accuracy compared to conventional methods.     

Experimental Study of Low Dose Ultrashortwave Promoting Nerve Regeneration after Acellular Nerve Allografts Repairing the Sciatic Nerve Gap of Rats

Objectives To observe the effect of ultrashortwave (USW) therapy on nerve regeneration after acellular nerve allografts(ANA) repairing the sciatic nerve gap of rats and discuss its acting mechanisms. Methods Sixteen Wistar rats weighing 180–220 g were randomly divided into four groups with four rats in each group: normal control group; acellular group (ANA, treated by hypotonic-chemical detergent, was applied for bridging a 10 mm-long sciatic nerve defect); USW group (After 24 h of ANA repairing the sciatic nerve gap, low dose USW was administrated for 7 min, once a day, 20 times a course of treatment, three courses of treatment in all); and autografts group. 12 weeks after operation, a series of examinations was performed, including electrophysiological methods, the restoring rate of tibialis anterior muscle wet weight, histopathological observation (myelinated nerve number, myelin sheath thickness, and axon diameter), vascular endothelial growth factor (VEGF) mRNA expression of spinal cord, and muscle at injury site, and analyzed statistically. Results Compared to acellular nerve allografts alone, USW therapy can increase nerve conductive velocity, the restoring rate of tibialis anterior muscle wet weight, myelinated nerve number, axon diameter, VEGF mRNA expression of spinal cord, and muscle at injury site, the difference is significant. There were no differences between USW group and autografts group except myelin sheath thickness. Conclusions USW therapy can promote nerve axon regeneration and Schwann cells proliferation after ANA repairing the sciatic nerve gap of rats, the upregulation of VEGF mRNA expression of spinal cord and muscle may play an important role.     

Evaluation of myelin sheath and collagen reorganization pattern in a model of peripheral nerve regeneration using an integrated histochemical approach

Peripheral nerves are complex histological structures that can be affected by a variety of conditions with different degree of axonal degeneration and demyelination. For the study of peripheral nerve regeneration in pathology and tissue engineering, it is necessary to evaluate the regeneration, remyelination and extracellular matrix reorganization of the neural tissue. Currently, different histochemical techniques must be used in parallel, and a correlation among their findings should be further performed. In this work, we describe a new histochemical method for myelin and collagen fibers based on luxol fast blue and picrosirius methods, for the evaluation of the morphology, the myelin sheath and the collagen fiber reorganization using a model of peripheral nerve regeneration. Whole brain, normal sciatic nerve and regenerating peripheral nerve samples were fixed in 10% neutral buffered formalin and paraffin-embedded, for the performance of the hematoxylin-eosin stain, the Luxol fast blue method and the new histochemical method for myelin and collagen. The results of this technique revealed that this new histochemical method allowed us to properly evaluate histological patterns, and simultaneously observe the histochemical reaction for myelin sheath and collagen fibers in normal tissue, and during the regeneration process. In conclusion, this new method combines morphological and histochemical properties that allowed us to determine with high accuracy the degree of remyelination and collagen fibers reorganization. For all these reasons, we hypothesize that this new histochemical method could be useful in pathology and tissue engineering.     

COMPARATIVE STUDIES ON THE MYELIN PROTEINS OF BOVINE PERIPHERAL NERVE AND SPINAL CORD

Abstract— (1) Two myelin fractions of bovine peripheral nerve and spinal cord have been studied comparatively. Cholesterol as well as cerebroside content per mg of protein in the peripheral nerve myelin was less than that in the spinal cord myelin, while no significant difference in the total phospholipid content was noted.
(2) The basic proteins in myelin fractions were quantitatively estimated by disc gel electrophoresis. Around one-fourth of the total myelin protein in the bovine peripheral nerve was a basic protein with a mobility of 1.07 relative to lysozyme by Reisfeld's disc gel electrophoresis.
(3) The myelin proteins in the peripheral nerve were less completely solubilized than those of the spinal cord by treatment with deoxycholate as well as by Triton-salt solution. The protein fractions obtained from the peripheral nerve myelin by techniques similar to that for obtaining the proteolipids from the spinal cord myelin, contained different types of protein.
(4) 2',3'-Cyclic nucleotide 3'-phosphohydrolase activity in the peripheral nerve myelin was only one tenth of that in the spinal cord myelin. The Triton-salt insoluble fraction showed remarkable high activity among subfractions of the spinal cord myelin.
(5) By immunological studies, it may be concluded that an antigenic substance for experimental allergic neuritis was localized in the peripheral nerve myelin, but not in its basic protein.     

叶冠尺度野鸭湖湿地植物群落含水量的高光谱估算模型

利用高光谱遥感技术定量估测野鸭湖湿地植被含水量,对于监测和诊断野鸭湖湿地植被的生理状况及生长趋势具有重要意义,也能够为高光谱遥感影像在野鸭湖湿地植被含水量诊断中的实际应用提供理论依据和技术支持.采用Field Spec 3野外高光谱辐射仪,获取了野鸭湖典型湿地植被冠层和叶片的光谱,并测定了对应的含水量.以上述实测数据为基础,首先以芦苇为例初步探明了不同含水量水平下典型湿地植被冠层和叶片光谱反射率的响应模式,然后采用相关性及单变量线性与非线性拟合分析技术,从冠层和叶片两种层次,对不同尺度下的含水量与“三边”参数及高光谱植被指数进行了分析拟合,并采用交叉检验中的3K-CV方法对估算模型进行了测试和检验,确立了不同尺度下野鸭湖湿地植被含水量的定量监测模型.结果表明:(1)随着含水量水平的增加,芦苇冠层与叶片光谱在可见光波段(350-760 nm)和红外波段(760-2500 nm)的反射率均呈逐渐降低趋势.(2)不同尺度含水量与选取的光谱特征参数整体上相关性较强,与“三边”参数基本上都呈极显著相关,相关系数最大达到0.906;与高光谱指数全部呈极显著相关,相关系数最小为0.455,最大达到0.919,并通过选取不同尺度上相关性最佳的光谱特征参数,分别基于“三边”参数和高光谱植被指数构建了不同尺度下的含水量估算模型.其中,冠层尺度下,黄边面积(SDy)与SRWI( Simple Ratio Water Index)的估算效果最好,估算模型分别为y=-9.462x2 -2.671x+0.608和y=0.219e1.010x;叶片尺度下,红边面积(SDr)与WI( Water Index)的估算效果最好,估算模型分别为y=0.562x+0.376和y=2.028x2 -0.476x-1.009.通过3K-CV的交叉验证,不同尺度下的含水量估算模型均取得了较为理想的预测精度,预测精度的最小值为94.92％,最大值为97.06％,表明估测模型具有较高的可靠性与普适性.(3)高光谱植被指数与含水量拟合方程的拟合度相对高于“三边”参数与之建立方程的拟合度,说明多波段组合的光谱特征参数更适合含水量的判别.     

Hyperspectral imaging: a novel approach for microscopic analysis

BACKGROUND: The usefulness of the light microscope has been dramatically enhanced by recent developments in hardware and software. However, current technologies lack the ability to capture and analyze a high-resolution image representing a broad diversity of spectral signatures in a single-pass view. We show that hyperspectral imaging offers such a technology. METHODS AND RESULTS We developed a prototype hyperspectral imaging microscope capable of collecting the complete emission spectrum from a microscope slide. A standard epifluorescence microscope was optically coupled to an imaging spectrograph, with output recorded by a CCD camera. Software was developed for image acquisition and computer display of resultant X--Y images with spectral information. Individual images were captured representing Y-wavelength planes, with the stage successively moved in the X direction, allowing an image cube to be constructed from the compilation of generated scan files. This prototype instrument was tested with samples relevant to cytogenetic, histologic, cell fusion, microarray scanning, and materials science applications. CONCLUSIONS: Hyperspectral imaging microscopy permits the capture and identification of different spectral signatures present in an optical field during a single-pass evaluation, including molecules with overlapping but distinct emission spectra. This instrument can reduce dependence on custom optical filters and, in future imaging applications, should facilitate the use of new fluorophores or the simultaneous use of similar fluorophores.     

Immunocytochemical localization of carbonic anhydrase in the spinal cords of normal and mutant (shiverer) adult mice with comparisons among fixation methods

The peroxidase-antiperoxidase technique was used for immunocytochemical localization of carbonic anhydrase in the mouse spinal cord to detect whether this antigen was normally present in myelinated fibers, in oligodendrocytes in both white and gray matter, and in astrocytes, and to determine where the carbonic anhydrase might be localized in the spinal cords of dysmyelinating mutant (shiverer) mice. The most favorable methods for treating tissue were: 1) immersion in formalin-ethanol-acetic acid followed by paraffin embedding, or 2) light fixation with paraformaldehyde and preparation of vibratome sections. Carnoy's solution, followed by paraffin embedding, extracted myelin from the tissue, while aqueous aldehydes, when used before paraffin embedding, reduced staining everywhere except at sites of compact myelin. The latter conclusion was based, in part, on the almost complete loss of this antigen from the shiverer cord, where compact myelin is known to be virtually absent but where membrane-bound carbonic anhydrase was demonstrated enzymatically. When the optimal methods were used with normal mouse cords, carbonic anhydrase was found throughout the white matter columns and in the oligodendrocytes in gray and white matter. The staining of the white matter was attributed to myelinated fibers because of the similarity in distribution to both a histological myelin stain and the immunocytochemical staining for myelin basic protein. In the mutant mice the oligodendrocyte cell bodies and processes, which were stained in all areas of the spinal cord, were particularly numerous at the periphery of the sections. In contrast to the oligodendrocytes, the fibrous astrocytes appeared to lack carbonic anhydrase, or to have lower than detectable levels, since the astrocyte marker, glial fibrillary acidic protein, had a very different distribution from that of carbonic anhydrase. Even finer localization was obtained in vibratome sections, where the antibody against carbonic anhydrase permitted visualization of the processes connecting oligodendrocytes to myelinated fibers in the normal adult spinal cord.     

Immunocytochemical localization of carbonic anhydrase in myelinated fibers in peripheral nerves of rat and mouse

Rat sciatic nerve, spinal root, and cranial nerve were immunostained with an antibody against rat brain carbonic anhydrase II (ca), to determine the localization of ca in the rat peripheral nervous system (PNS). Similar methods were applied to mouse nerves to see if that antigen could be detected in the PNS of this species. In rat nerves, intense immunostaining was observed in the axoplasm of many of the myelinated fibers, whereas others were stained less intensely or were negative. A heterogeneous pattern of immunostaining was also found in neuronal perikarya within the ganglia, and in some regions of the ganglia ca immunostaining was found in putative satellite cells and their processes. Ca in rat PNS therefore appears to occur at both neuronal and glial sites, whereas it is exclusively glial in the CNS. In longitudinal sections of some fibers within rat nerves, ca immunostaining could be detected at the inner boundaries of the myelin sheaths. In mouse nerves, axoplasmic staining was observed but it was fainter than in rat nerves. Interspecies differences were most obvious in the dorsal columns of the spinal cord. In rat, intensely stained axons proceeded through the roots into the gracilis or cuneate and often into the gray matter. In mouse, there was much less immunostaining of axons but more intense ca immunostaining in CNS myelin than in the CNS myelin in the rat cord. The implications concerning putative functions of ca in the rodent nervous system are discussed.