Different types of visual cells in the photoreceptor layer of the retinae of the treeshrew (Tupaia belangeri chinensis) as revealed by scanning microscopy.

BACKGROUND: The retinae of treeshrew have never been evaluated by scanning electron microscopic studies. MATERIALS AND METHODS: This work described the visual cells in the photoreceptor layer of the retinae of treeshrew (Tupaia belangeri chinensis) living on the high plateau of Yunnan, China, via scanning electron microscopy. RESULTS: Results indicated five morphologically different types of cones, two of which contain oil droplets in their inner segments. To our knowledge, no prior studies have reported oil droplets in the visual cells of higher mammals, only in lower vertebrate and primitive mammals. In addition, this study revealed one type of degenerative visual cell without outer segments. CONCLUSIONS: The findings signal the needs for additional studies to understand the physiological functions and phylogenetic relationships of the diversity of visual cells in this group of mammal.

TUNEL and growth factor expression in the prefrontal cortex of Alzheimer patients over 80 years old.

To elucidate factors underlying the increased risk of developing Alzheimers disease (AD) in older individuals, the prefrontal cortices of younger (58-79 years) and of older (over 80 years) AD patients were examined by silver impregnation, TUNEL assay and immunohistochemistry for hyperphosphorylated tau, LDH and two growth factors (BDNF, NGF). Quantitative data were compared with those of age-matched controls. TUNEL-positive cells were mainly located in superficial cortical layers of younger and in deeper layers of older AD patients. Their density was more than 5 times higher in older AD than in younger AD (p < or = 0.05), but apoptotic cell morphology was rarely seen. Significantly more neuronal somas were contacted by degenerating fibers both in younger and older AD cortices. Density of tau-immunoreactive cells, which were virtually absent in controls, was twice as high in older AD patients as in younger AD individuals (p < or = 0.05). In younger AD, TUNEL positive cells generally lacked tau immunoreaction, whereas in older AD, most cells were double-labeled for hyperphosphorylated tau and TUNEL (p < or = 0.05). Numerical density of BDNF-immunoreactive cells was significantly reduced by 20 percent in older AD patients, compared to both control individuals and younger AD patients, whereas density of NGF-positive cells was the same in all patient groups examined. The distinct differences between younger and older AD patients suggest a faster progression of AD in older patients.

Silver impregnation of the prefrontal cortices in the brains with long postmortem delay.

Silver impregnation was performed histologically on the prefrontal parts of brains, which had long postmortem delay of one month while under 4 degrees C refrigeration. The brains were from individuals from 60+- to 80+-years old. It was evident from these specimens that, as a whole, the outer layers of the brains degenerated first while the inner layers remained to be silver stainable and had near normal morphology after silver impregnation, even in the oldest specimens. Comparatively, the postmortem degeneration was worse in the outer cortical layers of the older individual when compared with the younger.

Sex-related differences in the anteroposterior diameter of the foetal cisterna magna.

AIM: To measure the anteroposterior diameter of the foetal cisterna magna and observe whether there are differences according to sex. MATERIALS AND METHODS: Three hundred and thirty-seven Chinese women with low-risk pregnancies and a singleton foetus between 22 and 38 weeks' gestational age were included in this retrospective study. Informed consent of the volunteer subjects and hospital authority approval were first obtained. Double-blinded for gender, the anteroposterior diameter of the cisterna magna of the foetuses was measured by transabdominal sonography. Magnetic resonance imaging (MRI) was used for those foetuses with mega cisterna magna. All of the foetuses were healthy by prenatal and postnatal examination, including physical and imaging examination. RESULTS: The mean anteroposterior diameter of the cisterna magna of all foetuses was 8.01+/-1.79 mm. The anteroposterior diameter of the cisterna magna had no obvious correlation with the gestational age. The mean anteroposterior diameter of the cisterna magna of 179 male foetuses was 8.63+/-2.16 mm, and the mean anteroposterior diameter of the cisterna magna of 158 female foetuses was 7.87+/-1.74 mm. The size difference was statistically significant (p<0.001). In the 33 foetuses with mega cisterna magna, the number of male foetuses was greater than female foetuses, and the proportion of the foetuses with mega cisterna magna in the male group was significantly higher than the foetuses in the female group (p<0.05). CONCLUSION: Male foetuses had slightly larger anteroposterior diameters of the cisterna magna than female foetuses. The study would be useful for creating normal range values for the cisterna magna of male and female foetuses.

Revisit the Cavernous Sinus from Fetus to Adult-New and Old Data.

This article highlighted three advances in the study of the cavernous sinus: (1) the initial formation of the sinus reticulum in early development of the sphenoid bone before ossification (2) extension of reticulum of the sinus and connection with other venules, and (3) the cavernous sinus and the nerves evolved inside this sinus during gestation, for example, the trigeminal nerve already formed bundles of motor and parasympathetic components during fetal development. This ontogenetic study further confirmed the cavernous sinus is not a single or a dual set of sinuses, but a group of extensions of venous sinuses or sinusoids. These new insights were integrated with previous understandings of the cavernous sinus to form this review article. Anat Rec, 301:819-824, 2018. © 2017 Wiley Periodicals, Inc.

Functional magnetic resonance imaging and the brain: A brief review.

Functional magnetic resonance imaging (fMRI) is employed in many behavior analysis studies, with blood oxygen level dependent- (BOLD-) contrast imaging being the main method used to generate images. The use of BOLD-contrast imaging in fMRI has been refined over the years, for example, the inclusion of a spin echo pulse and increased magnetic strength were shown to produce better recorded images. Taking careful precautions to control variables during measurement, comparisons between different specimen groups can be illustrated by fMRI imaging using both quantitative and qualitative methods. Differences have been observed in comparisons of active and resting, developing and aging, and defective and damaged brains in various studies. However, cognitive studies using fMRI still face a number of challenges in interpretation that can only be overcome by imaging large numbers of samples. Furthermore, fMRI studies of brain cancer, lesions and other brain pathologies of both humans and animals are still to be explored.

Mutated tau, amyloid and neuroinflammation in Alzheimer disease-A brief review.

This review discussed the importance of mutated tau, amyloid and neuroinflammatory factors and microglia in Alzheimer disease. In particular tau, CD4 and TNF alpha were included in the review and the colocalizations of these factors were highlighted. It is important to realize the Alzheimer disease may result from the interactions of these factors. Some of these factors may coexist at the same region and at the same time e.g. mutated tau and amyloid in plaques. A summary scheme of etiology leading to the disease was included.

Ki67 and doublecortin positive cells in the human prefrontal cortices of normal aging and vascular dementia.

Immunohistochemical localizing of the proliferation of Ki67 nuclei and doublecortin positive cells were performed in the prefrontal cortex of normal aged and vascular dementia (multiple infarct dementia) patients. Positive Ki67 nuclei and doublecortin positive cells were observed in both groups, with slightly higher density in the prefrontal cortex of vascular dementia. When the Ki67 sites were superimposed with the neuronal specific enolase localizations, only about 5% of the cells was doubly labeled, indicating few proliferating cells were neurons. This percentage did not vary between specimens of normal aging and those of vascular dementia.

Expression of dopamine transporter in the different cerebral regions of methamphetamine-dependent rats.

OBJECTIVES: To observe the expression of the dopamine transporter (DAT) in six cerebral regions of a methamphetamine (MA)-dependent rat, which were frontal cortex, nucleus accumbens septi, striatum, hippocampus, substantia nigra and ventral tegmental area. METHODS: The rats were administrated intraperitoneally with 10 mg/kg/day of MA for 10 days consecutively; the behaviour changes were measured via the conditioned place preference (CPP), and the scores of stereotyped behaviour (SB) were used to confirm animal addiction. Then, the animals were further injected with MA respectively for 1, 2, 4 and 8 weeks to establish different periods of MA-dependent models. The expressions of DAT and DAT messenger RNA in six cerebral regions were detected. RESULTS: The results of CPP and SB scores were significant different when comparing all four experimental groups with the control group (p < 0.05). Comparing between different experimental groups, the expression of DAT mainly decreased and had dynamic changes in the same regions (p < 0.05). Comparing the different regions with each other in the same experimental group, the expression of DAT also had significant difference in several regions p < 0.05). CONCLUSIONS: The expression of DAT mainly decreased and had different in the six cerebral regions at the same MA-dependent time period as well as at different time periods in the same cerebral region. It was speculated that DAT might play a crucial role in the mechanism of MA dependence.

Immunohistochemical and ultrastructural studies of the various nuclei of the trigeminal complex in the human newborn.

The various nuclei of the trigeminal complex were studied by immunohistochemical (enkephalin localization) and ultrastructural means in the brainstems of eight newborn human babies that died within 24 h after birth. Positive enkephalin neurons were detected in the chief sensory and spinal trigeminal nuclei as well as in some fibers of the trigeminal nerve. Ultrastructurally, two morphologically distinct types of neuron were observed, respectively, in the motor nucleus, the spinal nucleus and the mesencephalic nucleus of the trigeminal complex, whereas three morphologically distinct types of neuron were observed in the chief sensory nucleus. "Glomerulus" formation was a frequently observed feature in the chief sensory nucleus. In the spinal nucleus, rolls of synaptic terminals stacking up one on top of another and synapsing onto the final synaptic element were very much in evidence. Axosomatic, axodendritic, dendrodendritic and dendroaxonic synapses were demonstrated in all the different nuclear areas of the trigeminal complex but axoaxonic synapses were absent in the mesencephalic nucleus. Some of the findings in the present human study were similar to those reported in the rats and cats.

Immunohistochemical localization of neuropeptide Y and somatostatin in human fetal retina.

The localization of neuropeptide Y and somatostatin in the retina of six prenatal human specimens was determined by the utilization of light microscopic immunoperoxidase and immunofluorescence methods. The age of the prenatal fetuses ranged from 15 to 40 weeks of gestation. At 15 weeks, round or pear-shaped neuropeptide Y-immunopositive cells were observed in the inner nuclear layer and somatostatin immunoreactivity was only detected in the ganglion cell layer. Positive neuropeptide Y ganglion cells were observed by 17 weeks of gestation and by 28 weeks neuropeptide Y-immunopositive cells were demonstrated in the ganglion cell layer and inner nuclear layer. At 26-28 weeks, neuropeptide Y-immunopositive cells exhibited approximately the same shape as at 15 weeks of gestation, but the appearance of one or two processes was detected extending from the somata. By 38-40 weeks of gestation, neuropeptide Y-immunopositive cells were detected in the ganglion cell layer, inner nuclear layer and an immunopositive fibrous configuration in the inner plexiform layer. During this same period (38-40 weeks), somatostatin-positive cells were located in the ganglion cell layer, inner nuclear layer and the inner segments layers. Due to the difficulty of obtaining fetal materials, the exact time of initiation in the expression of neuropeptide Y and somatostatin is presently hard to delineate; however, it is safe to state that the peptides appear early in development.

An immunohistochemical study of the c-fos protooncogene in the developing human retina.

The localization and distribution of the protooncogene c-fos were studied immunohistochemically in the retina of human fetuses ranging in age from 15 to 40 weeks of gestation. The highest levels of immunoreactivity were observed in the retinae of younger fetuses, decreasing in intensity with increasing age. At 15 weeks of gestation intense immunoreactivity was observed in the inner nuclear layer while the photoreceptor cells exhibited moderate staining. At 26 weeks of gestation, immunoreactivity in the inner nuclear layer was reduced. The ganglion cells, amacrine cells and photoreceptor cells showed moderate immunopositivity throughout the 26-40 weeks period. The role of c-fos in development is discussed in the light of its other known functions.

Immunohistochemical localization of enkephalin and substance P in the nucleus caudalis of the spinal trigeminal V in the medulla oblongata of the human fetus.

The distribution of enkephalin-positive neurons, substance P-positive and enkephalin-positive fibers was studied in the nucleus caudalis of the trigeminal spinal V in the medulla oblongata regions of developing humans (12 weeks gestation to 40 weeks gestation). Enkephalin-positive neurons were identified in all the subnuclei of the nucleus caudalis as early as 12 weeks of gestation and increased in number as the fetus aged. Substance P-positive neurons were absent in this area throughout development. On the other hand, substance P-positive and enkephalin-positive fibers were present in all the subnuclei, again commencing as early as 12 weeks of gestation. These fibers tended to be linked to each other in the different subnuclei and to the reticular formation in this area and to increase significantly in quantity by the latter quarter of pregnancy. These results show the early presence of these neurons and fibers in the first trimester of development.

Localization of substance P and enkephalin by immunohistochemistry in the spinal cord of human fetus.

The peroxidase-antiperoxidase method was used to study the distribution of substance P and enkephalin during development of the spinal cords of human fetuses. Thirty-seven cases were collected, ranging from 5- to 40-weeks-old (fetal ages). Both types of transmitters were present initially around the fifth week in the mantle layer of the base of the dorsal horn, around the tenth week at the anterior gray and the intermediate gray and around the sixth week at the marginal layer at the base of the ventrolateral funiculus. Substance P- and enkephalin-positive sites at the marginal layers at the base of the dorsolateral funiculus were evident in the same area at 5-6 weeks. The positive fibers in the dorsal horn were initially located in the superficial layers. By the eleventh week, the positive sites spread to other surface layers at the lateral sides of the dorsal horns bilaterally at all spinal levels above the sacral. In the sacral levels adjacent to the conus medullaris, the spreading to surface layers was not apparent bilaterally until the seventeenth week. By weeks 18-26 the positive sites penetrated deeper in the dorsal horn and by week 27 assumed the adult path. The enkephalin cell bodies were present in the Rexed layers I and II of the dorsal horn and the substance P-positive sites were apparent in the dorsal ganglia of the 28-40-week-old fetuses.

Axonal sprouting in the hemisected adult rat spinal cord.

The morphological and biochemical changes were studied in adult Sprague-Dawley rats after hemisection at the L3 spinal cord level. After survival periods of one, two and three months, fluorescent tracers, FluoroGold or rhodamine B, were implanted into the dorsal white columns of these rats at the positions of the corticospinal tract below the lesion. Following uptake of the tracer, the rats were killed and the motor cortices and spinal cords of both control and hemisected rats were analysed for positively labelled neurons. The highest number of labelled cells were found two months after hemisection. They were present in both sides of the cortices, particularly in the contralateral cortex, and also in the gray matter of the spinal cord above the hemisection. A few rats which were subjected to complete transection of the spinal cord also showed labelling of neurons in the motor cortex two months after lesion. The Protargol silver technique and the [3H]choline uptake study confirmed the presence of nerve fibres traversing the lesion site in the hemisected spinal cord. Furthermore, when the rats that had been hemisected two months earlier were subjected to a second cut at the same site, chromatolytic neurons were observed in the spinal cord as well as in the motor cortices of both sides. The hemisected rats demonstrated limited recovery in limb movement. The evidence of this study clearly shows that sprouting of nerve fibres has occurred in the lesioned adult rat spinal cord.

Effects of astrocyte implantation into the hemisected adult rat spinal cord.

Morphological and biochemical methods were applied to assess the effects of implanting cultured astrocytes into the hemisected adult rat spinal cord. Astrocytes were purified from neonatal rat cortex and introduced into the lesioned spinal cord either in suspension injection or cultured on gelfoam first. The control groups were rats which had hemisection with injection of culture media or with gelfoam grafted alone. At various time points after surgery (two weeks to two months), the spinal cord was removed and processed for routine light microscopy, immunofluorescence, gel electrophoresis and immunoblotting. As early as two weeks after surgery, a significantly smaller volume of scar tissue was consistently found in the experimental groups. This reduced scarring was also confirmed by immunofluorescence staining and immunoblotting for glial fibrillary acidic protein in the specimens two months after hemisection. Compared to the control groups, the experimental groups also had more intense staining for neurofilaments, which was confirmed by immunoblotting. However, labelling of the astrocytes with Phaseolus vulgaris leucoagglutinin conjugated with fluorescein showed that the astrocytes migrated at a rate of 0.6 mm/day from the original implanted site. The results therefore suggested that the cultured astrocytes probably exerted their effects over a short time period (less than two weeks) around the lesion site. They could have altered the microenvironment and as a result less scar tissue was formed. Hence, there was less barrier to the regrowth of nerve fibres.

Habenulo-interpeduncular descending pathways and their relationship to enkephalin- and somatostatin-immunoreactive neurons in the interpeduncular nucleus of human fetuses.

The interpeduncular nucleus of six human fetuses aged 15 (one specimen), 26 (one specimen), 38 (one specimen) and 40 (three specimens) gestation weeks was studied by immunohistochemistry for enkephalin and somatostatin localization and immunohistochemistry coupled with silver staining. Enkephalin-positive and somatostatin-positive cells were detected, the former initially at 15 weeks gestation and the latter at 26 weeks gestation. They appeared to receive long afferents from the habenular region and projected short efferents to adjacent cells devoid of enkephalin and somatostatin positivity. We postulate that these enkephalin- and somatostatin-positive neurons function as modulatory interneurons in the habenulo-interpeduncular and related pathways.

Acetylcholinesterase-containing neurons, substance P and enkephalin fibers in the ventral horns of developing human embryos and fetuses.

The presence of the acetylcholinesterase neurons and substance P-like and enkephalin-like fibers in the various nuclear columns of the ventral horns of the spinal cords was studied in the developing human by acetylcholinesterase histochemistry and substance P and enkephalin immunohistochemistry. Acetylcholinesterase-positive neurons initially appeared in the lateral neuronal columns and eventually were also observed in the medial columns as well as the median columns at various levels of the spinal cord by 10 weeks' gestation. Acetylcholinesterase-positive neurons in the lower sacral levels were not detected until 11-12 weeks' gestation. Diffused substance P- and enkephalin-like fibers were demonstrated as early as 10 weeks' gestation but did not align with any particular nuclear column until after 15 weeks' gestation. These fibers further increased in length and adopted reticular branching patterns and many of these tended to surround the cell bodies of the nuclear columns. Possible interaction of acetylcholinesterase neurons and substance P and enkephalin fibers would commence by 15 weeks' gestation.

Distribution of neuropeptide Y in the developing human spinal cord.

The distribution of neuropeptide Y at different levels of the spinal cord of 23 human fetuses aged from 10-41 weeks of gestation was studied using immunocytochemical staining. Neuropeptide Y-immunoreactive neurons were identified at all levels of the spinal cord examined as early as 10 weeks of gestation. These cells were localized in the superficial layers (laminae I and II of Rexed) of the dorsal gray matter. As the age of the fetuses increased, their cell number increased and the region containing positive neurons extended from the superficial to deep layers (laminae III and VI). Immunoreactive fibers started to appear in fetuses at 10 weeks of gestation. They were found not only in the gray and white matters, but also in the pia mater lining the spinal cord. As the fetuses aged, the neuropeptide Y-immunoreactive fibers became mostly concentrated in the intermediate zones of the thoracic and sacral segments corresponding to the developing autonomic centers. Our results suggest that neuropeptide Y may play a role in the early development of the autonomic system.

Nitric oxide synthase neurons in different areas of normal aged and Alzheimer's brains.

This study investigated the distribution of nitric oxide synthase-containing neurons in the cerebral cortex of individuals with Alzheimer's disease, and compared them with age-matched controls. Paraffin-embedded sections of the frontal (area 10), occipital (area 17) and entorhinal cortices (area 28), and hippocampal formation obtained from 13 autopsy cases were used in the study. Neurons expressing nitric oxide synthase messenger RNA and protein were identified, respectively, by in situ hybridization and immunohistochemistry. Optical densities of nitric oxide synthase-positive neurons were assessed in 50 randomly selected fields of each of the above regions of the cortices, in each case by microscopic photometry. In the frontal cortex of the Alzheimer group, while a decrease in the number of nitric oxide synthase-positive neurons was evident, the nitric oxide synthase neurons, on the other hand, showed an increased optical density in layers II-IV when compared with those of normal ageing. In the occipital cortices, no significant differences in optical density were recorded between the normal ageing and Alzheimer specimens. In the entorhinal cortex, the optical densities of nitric oxide synthase neurons were again similar between the Alzheimer and age-matched control groups. In the hippocampar formation itself, there was an increase of nitric oxide synthase staining in the Alzheimer patients. These results show that (i) nitric oxide synthase neurons are abundant in the human cortex, (ii) the distribution of nitric oxide synthase neurons differs between different cortical regions, and (iii) there are differences between normal ageing and Alzheimer patients in the frontal cortex and the hippocampus.