Search PubMed⌕ Search

Biomedical subjects

D L Rosene

Publications and source records attributed to D L Rosene.

At least 37 records · Page 2Linked to original sources

Localization of alpha 2A- and alpha 2B-adrenergic receptor subtypes in brain.

Recent studies have shown that all three subtypes of alpha2-adrenergic receptor (alpha2-AR) are found in brain. The purpose of this study was to map the subtype localization of the alpha2A- and alpha2B-ARs in brain structures. RNase protection shows that both the alpha2A- and alpha2B-ARs are detectable in cortex, cerebellum, pons-medulla, and hypothalamus. We tested probes derived from the alpha2A- and alpha2B-AR cDNAs on cell lines that express each of the alpha2-AR subtypes to establish the subtype specificity of these probes for in situ hybridization. Then we used the alpha2A- and alpha2B-AR probes for in situ hybridization on sagittal and coronal sections of rat brain. Both alpha2A and alpha2B mRNA were detected throughout the brain. Overall, there appears to be a greater expression of message for alpha2A- than alpha2B-AR in most brain areas, with the exception of the thalamus. Developing these probes for in situ hybridization is an important step for further studies on the exact role of the alpha2-AR subtypes in neurons that modulate cardiovascular function.

Animals↗

CA3 neuronal degeneration follows chronic entorhinal cortex lesions.

Entorhinal cortex lesions are a common experimental paradigm to study memory function and neural plasticity after hippocampal deafferentation. The long term consequences of such lesions are of particular interest both in the context of these models and because pathological changes of Alzheimer's disease destroy entorhinal cortex projection neurons. We used stereological counting techniques to assess the structural integrity of the hippocampal formation 0.5-28 months after entorhinal lesion in the rhesus monkey. Surprisingly, 18-28 months after lesion the number of CA3 neurons was decreased by 57%, while neuron numbers in other subfields did not change. These results suggest that delayed transsynaptic neural degeneration can occur long after brain injury.

Animals↗

Mild age-related changes in the dentate gyrus of adult rhesus monkeys.

Memory and cognitive performance decline with advancing age in humans. Rhesus monkeys show a similar age-related memory deficit. Since the functional neuroanatomy of the temporal lobes in the two species is similar, and since the circuits of the temporal lobes are known to be involved in memory function, we undertook a study of the anatomical characteristics of synapses in the dentate gyrus of the rhesus monkey throughout the adult life span. Light- and electron-microscopic examinations were carried out on the dentate gyrus of 10 adult rhesus monkeys (4-35 years) to determine the effect of age on the thickness of the molecular layer and on axon terminals in the outer portion of the molecular layer. The thickness measurements were made on 100-microns-thick Vibratome sections and on 1 micron-thick Araldite-embedded sections. A total of 100 electron micrographs covering a test area of 3,600 microns2 for each monkey were taken in the outer portion of the molecular layer. Counts of axon terminals synapsing with dendritic spines or shafts, measurements of the cross-sectional area of these terminals, and the length of the postsynaptic density were taken on enlarged prints. The thickness of the molecular layer remained unchanged throughout adulthood. Statistical analysis revealed no overall age-associated loss of synapsing axon terminals or shrinkage of the cross-sectional areas of their profiles. Further, there was no loss in the total number of synapses (axospinous plus axodendritic) or any change in the lengths of their postsynaptic membrane densities. However, when axodendritic (shaft) synapses (which constitute 13% of the total) were considered separately, a statistically significant age-related loss was detected. Qualitative observations revealed that older monkeys had a moderate number of dystrophic myelinated axons and corpora amylacea located in astrocytic processes in the outer portion of the molecular layer, features not present in young monkeys. Also, glial cells and pericytes showed age-associated accumulation of lipofuscin-like inclusions. A single occurrence of a structured inclusion body in a dendrite was observed in a 10-year-old monkey. In conclusion, most synaptic measures in the dentate gyrus remain stable throughout adulthood of rhesus monkey and there are relatively few other age-related changes. The small age-associated loss of axodendritic synapses is only apparent following separate statistical treatment of these synapses. The functional significance of this loss is unclear since it would result in only 3% reduction in total synapses (shaft plus spinous) from 4 to 35 years of age, the maximal life span of the rhesus monkey.

Aging↗

Corticothalamic connections of auditory-related areas of the temporal lobe in the rhesus monkey.

Corticothalamic connections of auditory areas of the superior temporal regions (STR) were investigated in the rhesus monkey. These connections are organized according to the recently described architectonic parcellation of the STR. The core line regions of the supratemporal plane (STP) project to the medial geniculate nucleus (MGN). All regions except the primary auditory area also have projections to additional thalamic nuclei. The rostral STP has strong connections with the caudal part of the ventral subdivision (MGv) of MGN as well as with medial pulvinar (PM). In contrast the primary auditory area projects mainly to rostral MGv. The caudalmost STP projects mainly to the dorsal subdivision (MGd) and to the magnocellular subdivision (MGmc) as well as to the PM and the lateral (PL) and oral (PO) pulvinar, nucleus limitans (Li), and mediodorsal (MD) nucleus. The belt line regions of the superior temporal gyrus (STG) project mainly to the pulvinar but also have projections to MGd and MGmc. Specifically, rostral STG projects to the caudal part of PM, to MGmc, and to the suprageniculate (SG) nucleus, whereas caudal STG projects to the rostral part of PM and to PL, PO, MGd, MGmc, SG-Li and MD nuclei. The root line areas in the circular sulcus of the Sylvian fissure project mainly to MGmc but also to MGd, PM, and SG-Li nuclei. These connections originate mainly from neurons in cortical layer VI, with some from layer Vb. It is suggested that these connections may be involved in different aspects of auditory information processing.

Animals↗

Prenatal protein malnutrition effects on the serotonergic system in the hippocampal formation: an immunocytochemical, ligand binding, and neurochemical study.

Prenatally protein malnourished rats born to dams maintained on a 6% casein diet during pregnancy and then fostered at birth to females on a 25% casein diet show adult alterations in hippocampal kindling and long-term potentiation and behavioral changes that all suggest dysfunction of hippocampal formation (HF). In the present investigation, compared to well-nourished controls, 220 day malnourished rats exhibited a decrease in the 5-HT fiber density in the dentate gyrus (DG) and CA3 subfield and, a 15-25% decrease 5-HT uptake sites assayed with [3H]-citalopram in CA3 and CA1. In malnourished rats, 5-HT1A receptors assayed with [3H]8-OH-DPAT were decreased by 20% in CA3. Because most hippocampal subfields showed no 5-HT changes, hippocampal 5-HT levels determined via HPLC methods were similar in adult malnourished and control rats. These results suggest that there are localized changes in the 5-HT afferent system in the hippocampal formation of the 220 day prenatally protein malnourished rat. Considering the 5-HT afferent input to inhibitory intrahippocampal neurons, the decreased 5-HT plexus may result in increased inhibition within specific hippocampal subfields despite overall normal levels of 5-HT in the total HF.

8-Hydroxy-2-(di-n-propylamino)tetralin↗

Laminar termination patterns of thalamic, callosal, and association afferents in the primary auditory area of the rhesus monkey.

Thalamic, callosal, and association afferents to the primary auditory konicortex, area KA, were investigated in rhesus monkeys with the aid of anterograde tract tracing techniques (radioactively labeled amino acids, wheat germ agglutinin conjugated to horseradish peroxidase (WGA-HRP), and the Fink-Heimer silver impregnation method). Different patterns of anterograde label in KA were produced by radioactively labeled amino acid injections in the contralateral primary auditory area, the ipsilateral medial geniculate body (MGB), or auditory association cortex of the superior temporal gyrus (STG). Thalamic afferents from the MGB were found throughout KA in layers IV and III where the density waxed and waned, forming regular patches of higher density label separated by areas of less dense label. Callosal afferents from the contralateral auditory cortices were found in layers IV, III, and II in larger, more spatially separated, and irregular patches. In contrast, association afferents from the auditory association cortex of the STG were found in a continuous band in layer I. To determine the extent to which the patches of callosal afferents overlap with or interdigitate between the high-density thalamic clusters, two types of double anterograde labeling experiments were undertaken. In one set of experiments isotope injections of the MGB were made in conjunction with transection of the corpus callosum in the same animal. In another set of experiments isotope injections of the MGB were made in one hemisphere and an injection of WGA-HRP was made in the opposite supratemporal plane, including KA, in the same animal. Both approaches demonstrate that the patches of callosal afferents in KA mainly occur between the high-density patches of thalamic afferents, although there is a variable degree of overlap at their periphery. Thus it seems that thalamic and callosal inputs to KA are organized in a largely complementary fashion while input from the lateral association cortex is continuous and overlaps both. When considered in light of pertinent anatomical and physiological studies in a variety of species, it appears that the areas mainly occupied by callosal or thalamic inputs may be differentially involved in frequency analysis and sound localization, respectively.

Animals↗

The effects of hypertension on cerebral atherosclerosis in the cynomolgus monkey.

BACKGROUND AND PURPOSE: There is substantial clinical, pathological, and experimental evidence that hypertension aggravates atherosclerosis of the extracranial vessels. The present study assesses the effects of hypertension on the development of cerebral atherosclerosis in nonhuman primates fed an atherogenic diet. METHODS: The extent and severity of cerebral atherosclerosis were evaluated morphologically, morphometrically, and biochemically in atherosclerotic monkeys with and without hypertension. Atherosclerosis was induced by feeding a hypercholesterolemic diet for 12 months; hypertension was produced by surgical coarctation of the thoracic aorta. RESULTS: At autopsy, gross atherosclerotic lesions of the major cerebral arteries were observed in 15 of 16 atherosclerotic monkeys with hypertension compared with 5 of 16 atherosclerotic animals without hypertension. In the hypertensive-atherosclerotic group, 38.5% of the vessels examined showed gross involvement compared with only 3.4% of the vessels involved in the atherosclerotic group (P < .001). The lesions in the atherosclerotic group were generally mild, whereas those in the hypertensive-atherosclerotic group were severe and resulted in significant luminal narrowing and occlusion of vessels (P < .001). The small branches of the cerebral arteries also showed severe disease with luminal obstruction in the hypertensive-atherosclerotic group. The extent and severity of cerebral atherosclerosis were significantly related to the severity of the hypertension (P < .05). CONCLUSIONS: Hypertension is an important factor in cerebral atherosclerosis because of its accelerating effect on the disease. Nonhuman primate models may be useful in clarifying the role of hypertension and atherosclerosis in cerebral vascular disease.

Animals↗

A novel hinge system and incubation chamber for emulsion-coated coverslip autoradiography.

We describe an emulsion-coated coverslip autoradiographic technique for large (50 x 50 mm) sections of monkey or human brain. The technique uses adhesive-backed, teflon-reinforced aluminum foil as a flexible hinge that allows the coverslip to swing away from the slide so that the emulsion and tissue can be processed independently. We also describe a plexiglas incubation chamber that allows two slides with coverslips folded away to be processed back-to-back in 5 ml of reagent solution. In general use, the chamber minimizes the volume of solution needed to cover large slide-mounted sections and is ideal for radioligand or immunohistochemical reactions that use expensive reagents. For autoradiography, the chamber greatly facilitates independent processing of the emulsion and tissue section. Together, the foil-hinge method and incubation chamber virtually eliminate the major technical pitfalls of previous emulsion-coated coverslip methods. Moreover, they facilitate combination of histochemical or immunocytochemical staining with autoradiographic localization of neurotransmitter receptors in the same tissue section. This is demonstrated by thioflavin S staining of neuritic plaques and tangles and autoradiographic localization of serotonin 1A receptors in sections of Alzheimer disease brain.

Animals↗

Fields of origin and pathways of the interhemispheric commissures in the temporal lobe of macaques.

The interhemispheric connections of the cortical areas of the temporal lobe and some neighboring regions were investigated in monkeys (Macaca mulatta and Macaca fascicularis) by anterograde autoradiographic tracing, following injection of radioactively labeled amino acids. The results revealed that the interhemispheric projections of the temporal lobe course through three interhemispheric commissures on their way to the opposite hemisphere. The anterior commissure receives fibers from virtually the entire temporal lobe, including the temporal pole, superior and inferior temporal gyri, and parahippocampal gyrus. Moreover, area 13 of the orbitofrontal cortex, the frontal and temporal subdivisions of the prepiriform cortex, and the cortical and deep nuclei of the amygdala also contribute fibers to the anterior commissure. The heaviest projections arise in the rostral third of the temporal isocortex. These projections become progressively lighter from more caudal regions. By contrast, the corpus callosum receives fibers from the caudal two-thirds of the temporal lobe, including the temporal pole, superior and inferior temporal gyri, and parahippocampal gyrus. The heaviest projections arise in the caudal third of the temporal lobe and cross primarily in the caudal third of the corpus callosum, including the splenium. Progressively lighter projections arise more rostrally. Fibers from proisocortical and isocortical areas of the posterior parahippocampal gyrus cross in the ventralmost part of the splenium (inferior forceps), whereas cortical areas lateral to the occipitotemporal sulcus give rise to fibers that cross in the caudal part of the body of the corpus callosum and dorsal splenium. The dorsal hippocampal commissure receives fibers exclusively from the parahippocampal gyrus. The fibers of the corpus callosum, hippocampal commissure, and, to a lesser extent, the anterior commissure are intimately associated with the ventricular system as they course through the white matter of the temporal lobe. The fields of origin of the anterior commissure and corpus callosum overlap extensively over the caudal two-thirds of the temporal lobe. The posterior parahippocampal gyrus is unique in that it gives rise to fibers that cross in all three commissures.

Amygdala↗

Basal forebrain efferents to the medial dorsal thalamic nucleus in the rhesus monkey.

Thalamic efferent connections of the basal forebrain (BF); medial septal nucleus (MS), vertical limb of the diagonal band (VDB), horizontal limb of the diagonal band (HDB), nucleus basalis (NB), and ventral pallidum (VP) were investigated in twelve rhesus monkeys. In five animals, injections of radioactively labeled amino acids were placed in the BF. In four animals, the injections involved different divisions of the NB, HDB, and the most ventral part of the VDB. In those four cases, labeled fibers in the medial forebrain bundle were observed traveling caudally towards the hypothalamus where some turned dorsally to enter the inferior thalamic peduncle. These fibers terminated in the ventral half of the magnocellular part of the medial dorsal thalamic nucleus (MDmc). In a fifth case, the amino acid injection involved most of the MS and the VDB. Labeled fibers traveled caudally from the injection site and entered the stria medullaris. These fibers then traveled caudally before turning ventrally to terminate in the dorsal half of MDmc. To determine which of the diverse neuronal types in the BF gives rise to these thalamic projections, in two monkeys injections of horseradish peroxidase (HRP) were placed into MDmc. Labeled neurons were observed throughout the full extent of the NB, the VDB, the MS, and part of the VP. In order to determine the extent of the cholinergic input to MDmc from the BF, one of the HRP cases was processed for the simultaneous visualization of HRP, and acetylcholinesterase (AChE), the hydrolytic enzyme for acetylcholine, and a second case was processed for simultaneous visualization of HRP, and choline acetyltransferase (ChAT), the synthetic enzyme for acetylcholine. We observed that 30-50% of the HRP-labeled neurons were putatively cholinergic. In order to determine if the NB projection to MD is a collateral of the NB projection to orbital frontal cortex, one fluorescent retrograde tracer was injected into the orbital frontal cortex and one into MD. This case showed that approximately 5% of the BF neurons that project to MDmc also project to the orbital frontal cortex. These results confirm a significant subcortical projection by which the cholinergic system of the basal forebrain may influence higher cortical functions through the thalamus.

Amino Acids↗

A comparison of the efferents of the amygdala and the hippocampal formation in the rhesus monkey: I. Convergence in the entorhinal, prorhinal, and perirhinal cortices.

This is the first in a series of papers investigating the neuroanatomical basis for the interaction of the amygdala and the hippocampal formation in the rhesus monkey. The present report focuses on the complementary and convergent projections of the amygdala and hippocampal formation to the entorhinal and perirhinal cortices. These results were obtained from complementary experiments using injections of radioactively labeled amino acids to identify the anterograde projection patterns and injections of horseradish peroxidase and fluorescent retrograde tracers to confirm the cytoarchitectonic location of the neurons of origin for each projection. The results of this investigation demonstrate that both the hippocampal formation and the amygdala project to the entorhinal and perirhinal cortices where, with a few exceptions, the major projections of each structure generally are found in different layers of the same cytoarchitecture subdivisions of the entorhinal cortex but overlap in the same layers of the perirhinal cortex. Thus, the lateral and accessory basal nuclei of the amygdala project to layer 3 of areas Pr1, 28I, 28L, and 28S, and the accessory basal nucleus projects strongly to layer 1 of these same areas. In contrast, the subiculum, prosubiculum, and subfield CA1 of the of the hippocampal formation all have a projection to layer 5 of these same areas. In area 28M, the accessory basal nucleus of the amygdala projects to layer 1, while the subiculum, prosubiculum, and subfield CA1 of the hippocampal formation all project to layer 5, and the presubiculum projects to layer 3. In addition to these complementary laminar projections, there are a few areas of laminar overlap. Thus in area 28S, both the presubiculum and the CA1 subfield project to layer 3, where the lateral and accessory basal amygdaloid nuclei also project. Similarly, in 28I there is a major projection from the presubiculum and a lighter projection from the subiculum and CA1 to layer 3, where the lateral and accessory basal nuclei also project. There is also extensive laminar overlap in the perirhinal cortex. From the amygdala, the accessory basal nucleus projects to layers 1 and 3 and the lateral basal nucleus to layers 3, 5, and 6, while from the hippocampal formation, the prosubiculum projects to layers 3, 5, and 6, and the CA1 subfield projects to layer 5. This pattern of hippocampal and amygdaloid projections to the entorhinal and perirhinal cortices indicates that these cortices constitute a region of potentially extensive interaction between the amygdala and the hippocampus.

Amino Acids↗

Comparison of the efferents of the amygdala and the hippocampal formation in the rhesus monkey: II. Reciprocal and non-reciprocal connections.

The pattern of direct connections between the amygdala and the hippocampal formation in the rhesus monkey (Macaca mulatta) was delineated by using both anterograde and retrograde tract-tracing techniques. From the amygdala the accessory basal, medial basal, and the cortical nuclei and the cortical amygdaloid transition area send projections to the hippocampal formation. The efferents from the magnocellular part of the accessory basal nucleus and the cortical nuclei terminate in the molecular layer of subfields CA3, CA2, and CA1', and to a lesser extent in the molecular layer and the superficial part of the pyramidal cell layers of the prosubiculum. In contrast, the projections from the medial basal nucleus and the cortical amygdaloid transition area terminate in the molecular layer and the superficial part of the pyramidal cell layers of the prosubiculum only. From the hippocampal formation, subfield CA1' and the prosubiculum send efferents that terminate in the medial basal nucleus, the cortical transition area, and the ventral part of the cortical nuclei. In addition, the CA1' subfield projects to the ventral, parvicellular part of the accessory basal nucleus. The present data emphasize an important role for the prosubiculum and the CA1' subfield in medial temporal lobe area connections. Both regions, in addition to supporting direct connections between the amygdala and the hippocampal formation, also have extensive connections with the entorhinal cortex. As for the amygdala, the accessory basal nucleus sends efferents to both the hippocampal formation and the entorhinal cortex. The data demonstrate an anatomical means by which the amygdala, hippocampal formation, and the entorhinal cortex may interact. It is proposed that these connections may be important in the limbic memory system.

Amino Acids↗

Cingulate cortex of the rhesus monkey: I. Cytoarchitecture and thalamic afferents.

The cytoarchitecture and thalamic afferents of cingulate cortex were evaluated in the rhesus monkey (Macaca mulatta). Area 24 has three divisions of which area 24a is adjacent to the callosal sulcus and has the least laminar differentiation. Area 24b has more clearly defined layers II, III, and Va, and area 24c, which forms the lower bank of the anterior cingulate sulcus, has a particularly dense layer III. Area 23 also has three divisions, each of which has a distinct layer IV. Area 23a is adjacent to the callosal sulcus and has the thinnest layers II-IV, which have the same cell density as layers V and VI. Area 23b has the largest pyramids in layers IIIc and Va, and area 23c, in the depths of the posterior cingulate sulcus, has the broadest external and thinnest internal pyramidal layers. Finally, areas 29 and 30 are located in the posterior depths of the callosal sulcus. Two divisions of area 29 are apparent: one with a granular layer directly adjacent to layer I (area 29a-c) and another with differentiation of layers III and IV (area 29d). Area 30 has a dysgranular layer IV. Injections of the retrograde tracer horseradish peroxidase (HRP) were made into subdivisions of cingulate cortex in the monkey. Area 25 received thalamic input mainly from the midline parataenial (Pt), central densocellular (Cdc), and reuniens nuclei as well as from the dorsal parvicellular division of the mediodorsal nucleus (MDpc). A less dense projection also originated in the intralaminar parafascicular (Pf), central superior, and limitans (Li) nuclei as well as the medial division of the anterior nuclei (AM). Areas 24a and 24b received most thalamic afferents from fusiform and multipolar cells in the Cdc and Pf nuclei with fewer from the ventral anterior (VA) and MDpc and MD densocellular (MDdc) nuclei and only minor input from AM. Most input to premotor cingulate area 24c appeared to originate in VA, MDdc, and Li. Area 29 received the most dense input from nuclei traditionally associated with limbic cortex including the anteroventral (AV), anterodorsal (AD), and laterodorsal (LD) nuclei. Areas 23a and 23b, in contrast, did not receive AV, AD, or LD input, but the greatest proportion of their thalamic afferents arose in AM. Less-pronounced input also came from the lateroposterior (LP), medial pulvinar, and MDdc nuclei.(ABSTRACT TRUNCATED AT 400 WORDS)

Afferent Pathways↗

A cryoprotection method that facilitates cutting frozen sections of whole monkey brains for histological and histochemical processing without freezing artifact.

Cutting frozen sections of large (greater than 60 cc) blocks of monkey brain using the conventional procedures of infiltration with 30% sucrose as a cryoprotectant before freezing with pulverized dry ice often produces unacceptable levels of freezing artifact (FA) caused by displacement of tissue by ice crystals. Experiments investigating FA utilized perfusion-fixed brains from 46 monkeys and spanned combinations of cryoprotectants (glycerol, sucrose), freezing methods (dry ice or -75 degrees C isopentane), and fixatives (10% formalin, Karnovsky's or Timm's). The effects were evaluated by rating of FA severity in frozen sections of whole monkey brains. Minor FA appears as enlarged capillaries, more serious FA as large vacuoles, and both first appear midway between the periphery and center of the block. Stronger fixatives increased the severity of freezing artifact. The best method for eliminating FA was graded infiltration with up to 20% glycerol and 2% DMSO (in buffer or fixative), followed by rapid freezing in -75 degrees C isopentane. Although using a glycerol-DMSO infiltration before conventional freezing with pulverized dry ice or using conventional sucrose infiltration before freezing in isopentane gave better results than sucrose infiltration and dry-ice freezing, only the combination of glycerol-DMSO infiltration and freezing in isopentane produced consistently excellent results and virtually eliminated freezing artifact. To determine the effect of freezing with dry ice or isopentane on the rate of cooling in large blocks of CNS tissue, thermocouples were embedded in an 80-cc block of albumin-gelatin and frozen with the two methods. The rate of cooling (-3.5 degrees C/min) was twice as fast using isopentane.

Animals↗

Interhemispheric pathways of the hippocampal formation, presubiculum, and entorhinal and posterior parahippocampal cortices in the rhesus monkey: the structure and organization of the hippocampal commissures.

The interhemispheric pathways originating in the hippocampal formation, presubiculum, and entorhinal and posterior parahippocampal cortices and coursing through the fornix system were investigated by autoradiographic tracing in 29 rhesus monkeys (Macaca mulatta). The results revealed that crossing fibers are segregated into three contiguous systems. A ventral hippocampal commissure lies at the transition between the body and anterior columns of the fornix in the vicinity of the subfornical organ and the interventricular foramina of Monro; it is formed by axons arising in the most anterior (uncal and genual) subdivisions of the hippocampal formation. A dorsal hippocampal commissure lies inferior to the posterior end of the body of the corpus callosum; it is formed by axons arising in the presubiculum and entorhinal cortex of the anterior parahippocampal gyrus and the proisocortical and neocortical subdivisions of the posterior parahippocampal gyrus but not in the hippocampal formation. A hippocampal decussation lies between the ventral hippocampal commissure and dorsal hippocampal commissure; it is formed by axons arising in the body of the hippocampal formation. In contrast to the fibers of the ventral hippocampal commissure and dorsal hippocampal commissure, which terminate in contralateral cortical areas, these decussating fibers terminate in the contralateral septum. Thus, the ventral hippocampal commissure and dorsal hippocampal commissure of the rhesus monkey appear to be homologous to similarly designated structures in other mammals. To the extent that these observations also apply to the interhemispheric fibers of the human hippocampal formation and parahippocampal areas, their possible preservation must be considered when interpreting the effect of callosal transection on seizures and the results of "split-brain" studies, since callosal transection may fail to sever the hippocampal commissures in their entirety.

Animals↗

A perfusion-fixation procedure for the concurrent demonstration of Timm's, horseradish peroxidase (HRP), and acethycholinesterase (AChE) histochemistry.

The sulfide-silver method of Timm has been a widely used histochemical technique to demonstrate the presence of heavy metals in biological tissue, particularly in the central nervous system. However, the use of this method or its several modifications results in less than optimal morphological preservation and requires embedding the tissue in paraffin or freezing it and cutting it directly onto slides with a cryostat. These procedures can decrease the sensitivity and limit the application of other histochemical procedures, particularly when experiments necessitate processing large specimens or reaction procedures require techniques using free-floating sections. A perfusion-fixation protocol is described that yields sufficient fixation to cut whole frozen blocks of tissue with a sliding microtome, permits the use of free-floating sections, and allows the concurrent demonstration of horseradish peroxidase and acetylcholinesterase histochemistry without loss of sensitivity. The method consists of a short initial exposure to a sodium sulfide solution followed by a prolonged exposure to a combined sulfide-aldehyde fixative solution.

Acetylcholinesterase↗

A direct projection from the perirhinal cortex (area 35) to the subiculum in the rat.

An efferent projection from the perirhinal cortex (area 35) in the rat was studied using the anterograde transport of tritiated amino acids as well as horseradish peroxidase (HRP). Following injections of either tracer in either the dorsal or ventral parts of area 35, anterogradely transported label was observed in the molecular layer of the subiculum, adjacent prosubiculum and CAla. Regardless of the dorsoventral level of the injection, the label was most dense at mid-dorsoventral levels of the subiculum and decreased in density in both the septal and temporal directions. Small injections of the same tracers made into the surrounding entorhinal, ectorhinal or prepiriform cortices did not reproduce this pattern. While the entorhinal cortex is the main cortical source of afferent input to the molecular layer of the subiculum as well as the hippocampus and dentate gyrus, the perirhinal cortex appears to constitute a complementary cortical pathway for afferent input to the subiculum.

Amygdala↗