Robert D. Yates

Effects of aging on the structural and permeability characteristics of cerebrovasculature in normotensive and hypertensive strains of rats

SummaryThis study demonstrates that markedly different patterns of age-related changes in blood pressure and body weight occur among normotensive Wistar-Kyoto (WKY) and Sprague-Dawley (SD) rats and spontaneously hypertensive rats (SHR). In addition, a variety of age-related structural alterations occurred in the walls of arterioles, capillaries, and venules of the frontal cortex. These changes include: (1) an increase in the thickness of the vascular wall by deposits of collagen and basal lamina which, in some cases, extended into the surrounding neuropil; (2) the presence of a flocculent material in the adventitia of intracerebral arterioles; (3) vesicular inclusions in perivascular macrophages, pericytes and smooth muscle cells which were labelled with i.v. administered horseradish peroxidase (HRP); (4) fragmentation of smooth muscle cells; and (5) accumulation of lipofuscin-like pigments in perivascular glial processes. The hypertensive rats exhibited these changes, but they were more advanced and more widely distributed throughout the cerebral cortex. The aged hypertensive rats occasionally had large bundles of 10 nm diameter, intermediate filaments in the endothelial cells. Whereas no change in blood-brain barrier permeability to HRP was observed in the aged normotensive rats, all age groups of the hypertensive rats exhibited increased permeability to HRP in the initial segment of penetrating arterioles in laminae I and II of the cerebral cortex.

Brain aging in normotensive and hypertensive strains of rats

SummaryA variety of age-related changes occur in the structure of neurons in the cerebral cortex of Wistar-Kyoto and spontaneously hypertensive rats. The most marked alteration associated with increasing age was the deposition of lipofuscin pigment, primarily at the bases of apical dendrites of pyramidal neurons. While no strain-related differences in the amount of lipofuscin pigment were observed in the youngest (3 months) and in the aged (22–27 months) groups of rats, it appeared that hypertensive rats had larger pigment deposits at 12 months of age. At the ultrastructural level, neurons of the aged brains exhibited numerous nuclear invaginations and filamentous nuclear inclusions, increased amounts of Golgi complex and two types of cytoplasmic inclusions. The number of degenerative structures in the neuropil (membranous whorls, dystrophic axons and alterations in myelin sheaths) was also apparently increased in the aged brains. Neurofibrillary tangles were observed in dendritic processes of a 27-month-old Wistar-Kyoto rat. Glial cells accumulated distinctive pigment granules by which the three types of glia could be identified.

Two types of glomus cell in the rat carotid body as revealed by alpha-bungarotoxin binding.

SummaryHorseradish peroxidase (HRP)-conjugated α-bungarotoxin (αBgt) was used to localize αBgt-acetylcholine receptor sites in the rat carotid body. Two types of glomus cell were differentiated on the basis of the staining of their plasma membranes by the conjugate: type A, devoid of staining or only partly stained; and type B, exhibiting staining over the entire cell surface. The parts of type A glomus and supporting cells stained were always in direct apposition to type B glomus cells. It is concluded that type B glomus cells are possibly the only cell types exhibiting specific binding sites of αBgt. Other morphological characteristics and quantitative studies indicated that the type A and type B glomus cells presented in this study were equivalent to those described in the rat carotid body by other investigators (McDonald & Mitchell, 1975). αBgt-HRP staining facilitated the observation of the distribution pattern of glomus cells in the parenchyma: type A glomus cells were arranged in groups and often showed polarity toward neural elements and sinusoidal capillaries; and clusters of type B glomus cells were frequently situated in a demilune-like fashion over groups of type A glomus cells. Because of differences in morphology, synaptology, αBgt-binding affinity, and polarity toward the blood vessels, we propose that type A and type B glomus cells in the rat carotid body represent functionally distinct cell types.

Synaptic connections of central carotid sinus afferents in the nucleus of the tractus solitarius of the rat. I. An electron microscopic study.

SummaryA transganglionic transport technique was used to study the synaptic connections of the central carotid sinus afferents in the nucleus of the tractus solitarius of the rat by electron microscopy. The caudal part of the nucleus was profusely innervated. Labelled fibres extended to the contralateral nucleus, and to the ipsilateral dorsal motor nucleus of the vagus nerve, nucleus ambiguus, spinal nucleus of the trigeminal nerve and the area postrema. The labelled terminals were densely packed with clear, predominantly spherical vesicles about 50 nm in diameter and a few often swollen mitochondria. The terminals synapsed on dendrites of various calibres, spindle- or pear-shaped somal profiles with short axes lesser than 8 μm, and axon terminals. In axo-axonal synapses, most labelled terminals appeared to be presynaptic. Frequently, profiles of labelled terminals were in direct apposition with one another. The latter may represent the morphological substrate of the interaction between baro- and chemoreceptor inputs in the nucleus of the tractus solitarius and warrants further study. The present results indicate that in addition to direct inputs, the carotid sinus afferents are able to influence second-order neurons in the nucleus of the tractus solitarius indirectly through presynaptic modulation.

Dopamine β-hydroxylase-like immunoreactivity in the rat and cat carotid body: a light and electron microscopic study

SummaryImmunocytochemical localization of dopamine β-hydroxylase (DBH) was used to study the synthesis and storage sites of norepinephrine (noradrenaline) in the rat and cat carotid bodies. In the rat carotid body some parenchymal cells exhibited strong DBH-like immunoreactivity (DBH-I), while others displayed only faint DBH-I. In a typical parenchymal cell cluster, most cells with strong DBH-I were irregular in shape and appeared to partially surround those with weak DBH-I which usually were rounded in contour. In the cat carotid body most parenchymal cells showed a strong to moderate DBH-I. In both the rat and cat carotid bodies varicose nerve fibres with DBH-I were associated primarily with blood vessels. All autonomic ganglion cells examined, which were associated with the rat carotid body, showed DBH-I. Electron microscopy revealed that most DBH-I in the strongly positive cells of the rat carotid body was associated with dense granules (possibly corresponding to dense-cored vesicles of various sizes), although some was found in other sites. In oval cells with less DBH-I, reactivity resided in some of the large granules. In the cat carotid body the glomus cells contained more granules of various sizes and shapes than did those of the rat carotid body. Most of the cat glomus cell granules exhibited DBH-I activity. Our results indicate that some of glomus cells in the rat and most of the glomus cells in the cat contain DBH and therefore may be sites of norepinephrine synthesis.

Autoradiographic localization of α-bungarotoxin-binding sites in the carotid body of the rat

SummaryRadioiodinated α-bungarotoxin (α-Bgt) was used to localize α-Bgt-acetylcholine receptors in the carotid body of the rat. The gamma spectrometer analyses indicated a high uptake of [125I] α-Bgt in carotid bodies incubated in vitro (1.51 fmole per organ). Incorporation of the isotope was effectively blocked by pretreatment of carotid bodies with d-tubocurarine and unlabeled α-Bgt, but not by atropine. Light microscopic autoradiography showed a heavy labeling of some parenchymal cells. Electron-microscopic autoradiography revealed that labeling was localized along the interface between parenchymal cells, especially where their cytoplasmic processes engage in complex interdigitations. The silver grain counts on electron-microscopic autoradiographs suggest that labelings are preferentially associated with the plasma membrane of certain Type I cells. It is suggested that these Type I cells in the rats carotid body probably are provided with nicotinic acetylcholine receptors on their plasma membranes.

Synaptic connections of central carotid sinus afferents in the nucleus of the tractus solitarius of the rat. II. Connections with substance P-immunoreactive neurons

SummaryA combined transganglionic transport and immunocytochemical technique was used to study the synaptic morphology of central carotid sinus afferents and substance P-immunoreactive neurons in the commissural subnucleus of the nucleus of the tractus solitarius in rats. A large population of substance P-immunoreactive neurons (88.32%) were seen in close association with central carotid sinus afferents by light microscopy. However, many labelled central carotid sinus afferents appeared not associated with substance P-immunoreactive neurons in the nucleus of the tractus solitarius. Substance P-immunoreactive neurons were spindle, pear, or oval-shaped with a short axis ranging from 5 to 11 μm. Their long axis was oriented predominantly in a lateral-medial direction along the path of the central carotid sinus afferents from the solitary tract to the midline. Synaptic contacts between central carotid sinus afferents and substance P-structures, including dendritic profiles of different calibers and somas, were readily found by electron microscopy. Many central carotid sinus afferents were also found in synaptic contact with non-immunoreactive dendrites and somas. Appositions between central carotid sinus afferents and unlabelled axon terminals were common, but only in a few cases were morphological manifestations of synapses revealed. In the latter, the substance P-immunoreactive terminals appeared mostly presynaptic but postsynaptic ones were also encountered. Our data provide the evidence that some of the substance P-immunoreactive cells in the nucleus of the tractus solitarius are 2nd order neurons of the carotid sinus afferent pathway. The possibility that some of the substance P-immunoreactive neurons in the nucleus of the tractus solitarius may be interneurons and mediate carotid sinus afferent inputs to catecholaminergic neurons in the nucleus of the tractus solitarius is considered. Our findings also provide an anatomical substrate for a possible presynaptic modulatory role of central carotid sinus afferents on the inputs from other brain centers to the substance P-neurons in the nucleus of the tractus solitarius.

An electron microscopic study of the baroreceptors in the internal carotid artery of the spontaneously hypertensive rat

SummaryThe carotid baroreceptor field of normotensive (NTR) and spontaneously hypertensive rats (SHR) examined in this study extends for about 0.5 mm along the length and about 1/3 to 1/2 of the circumference of the wall of the internal carotid artery opposite to the carotid body. The vascular wall of the baroreceptor field exhibits neither a marked dilation to form a carotid sinus nor histological differences in the intima and media compared to other parts of the carotid artery. Histologically the adventitia of the baroreceptor field is characterized by (1) an increased thickness and by less well developed elastic lamellae in comparison with other parts of the arterial wall, (2) a profuse blood and nerve supply, and (3) a richness of cellular elements. The presumptive baroreceptor terminals are localized in the inner 1/3 of the adventitia and display local enlargements that appear to show preferential association with the cell body or processes of the Schwann cell but not with other components of the adventitia. The enlargements are characterized by an accumulation of very densely packed mitochondria, and glycogen particles. No morphological alterations were noted in the baroreceptor terminals of SHR except for proliferated basal laminae that invest the terminals. Our work does not support the concept that resetting of the baroreceptors is due to degeneration of the terminals.

The fine structure and phosphatase cytochemistry of the golgi complex and associated structures in the sertoli cells of Syrian hamsters.

SummaryThe Golgi complex in the Sertoli cell of the Syrian hamster is well developed and consists of stacks of cisternae and associated vesicles. The inner-and outermost cisternae of the Golgi stacks are usually moderately dilated and exhibit numerous fenestrations. The middle portions of the intermediate cisternae are greatly flattened and not fenestrated, but toward the periphery these cisternae gradually become dilated and show a few fenestrations. On the inner aspect of the Golgi stacks the following structures are seen frequently: (1) one or two series of linearly arrayed circular profiles some of which are interconnected by tubules; (2) networks of anastomosing tubules with circular or oval meshes (800 to 1200 Å in diameter); and/or (3) irregularly disposed tubules. The circular profiles and tubules are approximately 450 Å in diameter. Acid phosphatase activity was localized in these anastomosing tubules when the tissues were incubated for more than one hour in a modified Gomoris medium (Barka and Anderson, 1963). Strong thiamine pyrophosphatase activity was demonstrated in the inner one to three cisternae of the Golgi stacks but not in the associated tubules. The system of the Golgi associated tubules is morphologically and histochemioally distinct from the Golgi stacks and is probably equivalent to the Golgi-endoplasmic reticulum-lysosome system (GERL) in other cell types. The three dimensional aspects of the GERL-equivalent system are discussed.

The anatomical distribution and morphology of extraadrenal chromaffin tissue (abdominal paraganglia) in the dog.

Abstract The anatomical distribution, in situ , and morphology of extraadrenal chromaffin tissue in the retroperitoneum of dogs of various ages was studied by utilizing glutaraldehyde perfusion followed by glutaraldehyde/potassium dichromate immersion. This method of study produced a gross chromaffin reaction that clearly demonstrated abundant catecholamine-containing paraganglionic tissue. The procedure also provided excellent tissue preservation for subsequent light and electron microscopic study of the dichromate-traced organs. The largest chromaffin organ consistently occurred ventral and lateral to the abdominal aorta in the mid-retroperitoneum. Extensions of this body often reached the adrenal glands, but a specific continuity between intra- and extra-adrenal chromaffin tissues was not observed. Smaller organs, probably corresponding to the Organs of Zuckerkandl in the human, occurred around the inferior mesenteric artery. Light microscopy identified a parenchyma rich in epithelial cells exhibiting cytoplasmic vesicles and prominent nuclei and nucleoli. Connective tissue and numerous blood vessels delineated the chromaffin cells into groups. Electron microscopy showed cellular detail characteristic of chromaffin cells and confirmed the cytoplasmic presence of typical catecholamine granules. The majority of granules exhibited homogeneously dense cores. However, many others appeared less dense and displayed granular cores. This study provides evidence that extraadrenal chromaffin organs in dogs are voluminous, rich in catecholamine granules, and persist into adulthood.