Fine structure of synapses in the cerebral cortex

Summary The synaptolemma of axo-dendritic synapses in the rabbit striate and peristriate cortex has been examined in the electron microscope. Observations are reported of 1) a subsynaptic organelle, in the juxta-membranous part of the dendrite cytoplasm; 2) a typical attenuation of the synaptic cleft, located at the circumference or at the center of the synaptolemma; 3) interlemmal elements of two types, designated A and B.The possible functional implications of the findings are discussed in the light of present-day knowledge of synaptic transmission.The support by the Joseph P. Kennedy, Jr., Memorial Foundation and the U.S. Public Health Service (Grant B. 4012) is gratefully acknowledged.The author is Joseph P. Kennedy, Jr., Memorial Foundation Senior Research Scholar.     

Fine structure of the mammalian egg cortex

The cortices of a number of mammalian eggs are not structurally homogeneous but are polarized. In mouse ova the plasma membrane is a mosaic; the cytoplasm overlying the meiotic spindle is devoid of cortical granules and consists of a filamentous layer containing actin. Functionally, this cortical polarity may be related to the restriction of sperm-egg interaction and fusion to a specific region of the ovum cortex and to dynamic changes of the egg cortex during fertilization, including cortical granule exocytosis, polar body formation, and fertilization cone development. The origin of cortical polarity in mammalian oocytes and its possible relation to components of the cytoskeletal system and meiotic apparatus are discussed and compared with cortical features of eggs of other vertebrates and invertebrates.     

Pipecolic acid receptors in rat cerebral cortex

Identification of [¹⁴C]pipecolic acid (PA) receptors was attempted in the solubilized membrane fraction from rat cerebral cortex. Specific binding proteins for both PA and muscimol, a potent -aminobutyric acid (GABA) agonist, were detected in the same preparation. Separation of labeled PA and GABA binding proteins by glycerol gradient centrifugation has shown labeled protein bands of similar sedimentation rates, suggesting that PA and GABA may be binding to identical proteins. It seems likely that the PA binding receptor either may possess the same sedimentation characteristics as that of the GABA receptor, or both GABA and PA which is an endogenous and weak GABA agonist may bind to the same receptor complex, if not to the same binding site.     

The diglyceride kinase of rat cerebral cortex

1. Formation of phosphatidic acid by diglyceride kinase (EC 2.7.1.-) in the presence of ATP and Mg(2+) was shown in a homogenate and subcellular fractions of rat cerebral cortex. 2. The kinase was activated by Mg(2+). Ca(2+) activated to a smaller extent but was inhibitory in the presence of optimum concentration of Mg(2+). Activity was greatly increased in the presence of added 1,2-diglyceride. 3. Sodium deoxycholate markedly stimulated the reaction, but other detergents (Cutscum and Triton X-100) did not. 4. Diglyceride kinase was concentrated in the supernatant and microsomal fractions from rat cerebral cortex. The distribution of the kinase in the particulate fractions resembled that of acetylcholinesterase and 5'-nucleotidase. 5. The rate of phosphatidic acid synthesis by the diglyceride kinase route was much greater than reported rates for acylation of 3-glycerophosphate and was also very rapid in comparison with the rates of other steps in the synthesis of phosphoinositides. 6. Acetylcholine had no stimulatory effect on diglyceride kinase of isolated intact nerve-ending particles or of nerve-ending membranes obtained after osmotic shock.     

Fine structure of trypsin-dissociated rat adrenal cells

Summary Electron micrographs of trypsin-dissociated rat adrenal showed predominantly intact rounded cells without internal damage. The population contained cells from the glomerular, intermediary and fascicular zones with cells from the zona fasciculata predominant. The presence or absence of cells from the reticular zone could not be determined. Cells from the medullary zone were absent. The addition of adrenocorticotropic hormone (ACTH) to the cellular suspension for 2 hours produced corticosterone. However, these stimulated cells did not display any significant ultrastructural change.Supported by research grants from the U.S.P.H.S. (No. GM15872), and the Research Council of Rutgers University, and the National Science Foundation (No. GB7427 to Dr. George Sayers). We are indebted to Mrs. Jean A. Gibney, Miss Rose-Marie Ma, and Mrs. Mary Vegh for their excellent technical assistance.Postdoctoral Trainee, U.S.P.H.S. Training Grant No. 5T01-GM00899-11.     

Acetylcholinesterase in neurons of the rat cerebral cortex.

AChE-containing neurons have been demonstrated by electron-microscopical histochemistry in the neocortex of the rat. These cells are mainly located in layer VI. AChE activity is seen in the cisternae of the RER, in subsurface cisternae and in the dendritic membranes.     

Ultrastructural demonstration of dehydrogenases in rat cerebral cortex.

Techniques for the ultrastructural demonstration of dehydrogenases in cerebral cortex are described. The best fixation for good fine structural preservation and retention of LDH and NADH-diphorase was obtained by perfusion with a misture of formaldehyde and glutaraldehyde and for SDH by perfusion with formaldehyde. Comparison of incubation conditions showed that consistent results were obtained using enzyme markers NBT and DS-NBT for LDH and NADH-diaphorase: DS-NBT was more satisfactory than NBT and BSPT for SDH. Penetration of incubation media was improved by Triton X-100: DMSO and ultrasonic treatment were less effective. The techniques enabled the first electron cytochemical demonstration of dehydrogenases in different elements of prefixed cerebral cortex. Ultrastructural sites of enzyme activities were localized within cristae and inter-membrane spaces of mitochondria in nerve cell cytoplasm and its processes, oligodendrocytes and astrocytes. Authenticity of the ultrastructural sites was confirmed by four different control experiments.     

Day-night cycle of lipidic composition in rat cerebral cortex

A study of the lipidic pattern of the cerebral cortex of the normal adult rat during the daynight cycle was carried out. The changes observed were the following: phosphatidylcholine, phosphatidylethanolamine, phosphatidylinositol and phosphatidylserine plus phosphatidic acid showed a peak at 16:00 hr possibly due to a general increase in phospholipid biosynthesis. During the nocturanl period the variations of phosphatidylcholine and phosphatidylethanolamine were not clearly observe, they might be due to an increase in the interconversion or exchange reaction, since the ratio phosphatidylcholine/phosphatidylethanolamine showed a significative change at 04:00 hr. This occurred because small but opposite changes in both phospholipids were observed, suggesting an increase in the methylation reactions of phospholipids. Cardiolipin showed a significant peak at 04:00 hr. Plasmalogens exhibited significative changes, an important diminution at 16:00 hr and a prominent peak at 24:00 hr. Cholesterol levels were high during the light period and low in the dark one. Cerebrosides and gangliosides showed no day-night variations. The changes observed indicate a phenomenon of biological rhythmicity synchronized by the photoperiod, suggesting that these fluctuations could act as physiological modulators of the properties and functions of the nerve cell membrane.     

Freeze-etching study of synaptosomes from rat cerebral cortex

Synaptosomes from rat cerebral cortex were studied using the freeze-etching technique. The intra-membranous structure of the pre- and postsynaptic membranes was examined. Particles with an electron-dense spot on their apex are reported from all fracture faces. Most probably these are related to transmembrane channels whose significance in the synaptic transmission is discussed.     

Aging and benzodiazepine binding in the rat cerebral cortex

The specific binding of ³H-diazepam in the cerebral cortex was investigated in membrane preparations from 6 and 30 month old Fischer-344 rats. No age-related differences in the association, equilibrium, or dissociation binding characteristics were observed. The increased sensitivity of the elderly to the central sedative effects of the benzodiazepines does not, therefore, appear to involve changes in binding to the receptor site located in the cortex.     

Fine structural localization of potassium-stimulated rho nitrophenylphosphatase activity in denrites of the cerebral cortex

A histochemical technique for the demonstration of K+-rho-nitrophenylphosphatase (K+-rhoNPPase) activity, a component of the Na+,K+-ATPase, has been applied at the fine structural level in the somatosensory cortex of the rat. Reaction product was consistenly found in dendrites and in association with the cytoplasmic aspect of the dendritic plasmalemma. Reaction product often filled portions of the tubular smooth endoplasmic reticulum in these processes. The results of these studies are interpreted to indicate that enzymatic activity is associated with large-and small-diameter dendrites. No convincing evidence of high activity was found in glial profiles. The importance of neurons and their dendrites in active transport of sodium and potassium ions in the cerebral cortex may be more significant than indicated by studies with isolated neurons and glia.     

Diminution of preprosomatostatin-mRNA in cerebral cortex of the aged rat

The aim of the present study was to examine the effect of normal aging on somatostatin neurotransmission. Somatostatin gene expression was analysed in several regions of the cerebral cortex and hippocampus in 3, 7 and 21 month-old Sprague-Dawley rats using quantitative in situ hybridization with a 48mer oligodeoxynucleotide antisense probe. Furthermore the distribution of ¹²⁵I-Tyr¹¹ somatostatin receptor binding sites was studied using quantitative receptor autoradiography. The results demonstrated a significant reduction of preprosomatostatin-mRNA in the frontal cortex of the aged (21 month) group compared with the young (3 month) and the middle-aged (7 month) groups. The receptor binding densities of the aged (21 month) group tended to be lower, compared to the other groups although no significant region-specific changes were evident. These results indicate neurochemical changes in somatostatin-containing neurons in the frontal cortex during aging.     

An intralaminar analysis of beta-glucuronidase in cerebral cortex of rat

Abstract— Activities of β-glucuronidase were measured microchemically in the rat within cortical layers and subcortical white matter of somatosensory and visual cortex and the dorsal hippocampus. Distributions were related to histological composition, densities of myelinated fibers, and lysosome content as indicated by acid phosphatase staining. Three zones of relatively high activities were noted. The first corresponded to the pia-arachnoid and has been related to lysosmal particles within pericytes and macrophages of the meninges and in the pial cells. A second peak appeared in layer V and correlated well with the presence of neuronal lysosomes as detected by histochemical reaction. A third contribution was related to the presence of myelinated fibre bundles and white matter. Data from the literature and from unpublished results were cited to support the conclusion that nonlysosomal sources of enzyme in white matter included a major component from particles sedimenting with the microsomal fraction and a small component from myelin.