The earliest histopathological response to hypobaric hypoxia in rabbits in the rifugio torino (3370 M) on Monte Bianco

Twelve Dutch rabbits were kept on Monte Bianco at an altitude of 3370 m. Half of the animals were killed after 3 months, the remainder after 6 months, and a further six animals maintained at sea-level acted as controls. The carotid bodies of all the rabbits were processed for light and electron microscopy and examined qualitatively and quantitatively. The lungs were processed for light microscopical assessment of small pulmonary arterial vessels; the thickness of the pulmonary trunks and aortas were measured; and the hearts were dissected to obtain ratios of the ventricular weight. There was a slight increase in the right ventricular weight in the hypoxic rabbits but no change in the thickness of the pulmonary trunk compared with that of the aorta. In particular, there was no hypoxic remodelling of the pulmonary vasculature such as muscularization of pulmonary arterioles or intimal longitudinal muscie in pulmonary arteries. The earliest histopathological response to hypoxia occurred in the carotid bodies in the form of an increase in the count of the dark variant of chief cell after 3 months which returned to normal after 6 months. It is concluded that the carotid body of the rabbit responds with a change in its population of dark chief cells to a level of hypoxia which is insufficient to affect the pulmonary arterioles. Changes in the cardiopulmonary system can no longer be considered to be the earliest histopathological response to hypobaric hypoxia.     

The carotid bodies in a case of ventricular septal defect

A woman of 62 years with Turner's syndrome died in congestive cardiac failure secondary to a large ventricular septal defect with biventricular hypertrophy. During her last few months the lifelong left-to-right shunt underwent reversal exposing her carotid bodies to hypoxaemia. The carotid bodies were not enlarged, thus demonstrating that hyperplasia of glomic tissue is not brought about by increased myocardial mass per se. They were, however, abnormally cellular with more dark cells ( a variant of chief cells), many of which were abnormally large and showed ultrastructural features of metabolic activity. These changes may represent the earliest histological response of the carotid body to hypoxaemia and later, the dark cells may mature into the more familiar and common light variant. It seems likely that this dark cell activity precedes, probably by a long period, the sustentacular cell hyperplasia and proliferation of nerve axons which we have reported elsewhere as the chronic reaction of the carotid bodies to hypoxaemia.     

Comparative pathology of the enlarged carotid body

A histological study was made of the carotid bodies of man and various animal species from low and high altitudes. The animals studied were the alpaca, llama, cattle, guinea-pig, rabbit, dog, rat and man. As well as a qualitative microscopic study, a differential cell count was carried out to determine the percentage of the light and dark variants of chief cells and of sustentacular cells present. The investigation showed that the carotid bodies enlarge in cattle, guinea-pigs and rabbits living in the hypobaric hypoxia of high altitude to which they have to acclimatize. The carotid bodies are not enlarged in llamas and alpacas which show Darwinian adaptation to high altitude. There is no single histopathological appearance to be found with enlargement of the carotid body; on the contrary, there appears to be a characteristic histological reaction for different species. Thus, man shows hyperplasia of sustentacular cells, cattle show focal dark cell proliferation and rabbits, guinea-pigs, and dogs show striking hyperplasia and vacuolation of chief cells. In the rat, the enlargement of the carotid body is not characterized by the differential proliferation of any specific element and, as a result, it does not appear to be a good model for the human organ. In man and rat, carotid body enlargement occurs in response to systemic hypertension as well as to chronic hypoxaemia and the histological response to the 2 stimuli is the same, depending on the species. The normal rabbit carotid body is more reminiscent of that of man but, in this species, the reaction of glomic tissue differs from that of human glomic tissue.     

Enlargement of the carotid bodies in cirrhosis of the liver

The carotid bodies were dissected out at necropsy and weighed in seven subjects with cirrhosis of the liver and in seven control subjects of comparable age free of liver disease. The mean combined carotid body weight of the control group was 17mg but in the cirrhotic patients it was 35 mg, a statistically significant difference ( P < 0.005). Differential counts of the various types of glomic cell (progenitor, dark and light variants of chief cells and sustentacular cells) were carried out. The enlargement of the carotid bodies in the subjects with cirrhosis was associated with increased numbers of the dark variant of chief cell. The mean number of dark cells per unit area in the control group was 361 cells/mm² but in the cirrhosis group it was 1024 cells/mm², a statistically significant difference ( P < 0.005). It is postulated that the prominence of dark cells may be associated with secretion of a natriuretic peptide in response to the hyperaldosteronism and sodium retention of cirrhosis of the liver. Alternatively, it may be a response to hypoxaemia resulting from porta-pulmonary shunts.     

The carotid bodies in bronchial asthma

A histological study was made of right-sided carotid bodies resected therapeutically from 50 patients with bronchial asthma. Also studied as controls were 10 right-sided carotid bodies from subjects coming to necropsy. Hypoxaemia was considered a contra-indication to glomectomy and only patients with a resting arterial oxygen tension exceeding 65 mm Hg were submitted to operation. It was found that in patients with a short history of bronchial asthma the carotid bodies were not enlarged, but there was hyperplasia of sustentacular cells. In cases with asthma for 5 years or more the sustentacular cell hyperplasia was more pronounced and was associated with many nerve fibrils and slight enlargement of the carotid bodies. The functional significance of the proliferation of sustentacular cells and the abundance of nerve axons is obscure. In the asthma cases there was prominence of the dark variant of chief cells, thought to be related to episodes of hypoxaemia.     

The distribution of enkephalins in human carotid bodies showing cellular proliferation and chronic glomitis

Human carotid bodies obtained at necropsy that showed prominence of either the sustentacular cell or the dark variant of chief cell or chronic carotid glomitis were studied by an immunogold labeling technique. The peptides methionine and leucine enkephalin had a similar distribution to that found in the normal human carotid body. They were localized prominently and predominantly in the dark and progenitor variants of chief (type I) cells. The sustentacular (type II) cells showed no immunoreactivity for the enkephalins. Cell counts on immunolabeled chief cells in cases of sustentacular cell hyperplasia and chronic carotid glomitis were found to be at the lower end of the normal range, whereas those in dark cell prominence occurred nearer the upper limit.     

The ultrastructure of the carotid body in chronically hypoxic rabbits

1. The ultrastructure of the carotid body in the rabbit has been examined by electron microscopy.2. A comparison was made between the ultrastructure of the carotid bodies in sea level rabbits, in rabbits which had been exposed to hypoxia equivalent to an altitude of 6000 m for 7 days and in rabbits which always had lived at an altitude of 4000-4300 m.3. We could not detect any difference in the ultrastructure between the two groups of hypoxic rabbits.4. When the hypoxic rabbits were compared with sea level rabbits there was a marked increase in the number of dense cored vesicles and mitochondria in the type I cells in the hypoxic rabbits. The Golgi region also appeared to be enlarged in the type I cells in the hypoxic rabbits.5. The finding suggests that in the rabbit the production of amines, probably dopamine, within the type I cells is increased during prolonged hypoxia which might explain the lowered ventilatory response to hypoxia observed in human high altitude residents.6. If the carotid bodies are organs of internal secretion the finding is compatible with an increased production of a hormone produced within the type I cells.     

Dark cell proliferation in carotid body hyperplasia

The histopathology of the enlarged carotid bodies, in both qualitative and quantitative terms, is described in two cases. One was in a woman of 80 years with systemic hypertension and the other in a man of 72 years with pan-acinar emphysema complicated by chronic hypoxaemia. In both instances there was generalized hyperplasia of sustentacular cells, perhaps with involvement of Schwann cells and fibrocytes, with superimposed focal proliferation of dark or 'pyknotic' variants of chief cells. The functional significance of both froms of tissue response in the carotid bodies is speculated upon.     

Histopathology of the carotid bodies in neonates and infants

A histological study was made of the carotid bodies obtained at necropsy in 40 fetuses, neonates, infants and young children, ranging in age from 23 weeks of gestation to 11 years of age. Prominence of the dark variant of chief cells was found in diseases such as bronchiolitis which are associated with sustained hypoxaemia. Such cells have copious cytoplasm known to be rich in peptides such as enkephalins. The same histological features were found in two cases clinically diagnosed as sudden infant death syndrome but in which there was associated bronchiolitis and tracheo-bronchitis respectively, thought to have given rise to hypoxaemia. In a case of sudden infant death syndrome without infection there was overgrowth of sustentacular cells with predominant progenitor cells in the absence of dark cells. We associate this appearance with inability of the carotid bodies to respond adequately to changes in arterial oxygen tension.     

Light,fluorescence and electron microscopic studies of rabbit subclavian glomera

The subclavian glomera (aortic bodies) of young New Zealand white rabbits were studied with the light, fluorescence, and electron microscopes. Two cell types were identified: type I, granule-containing (chief) cells, and type II, agranular (sustentacular) cells. The type I cells possessed large nuclei, the normal complement of cytoplasmic organelles and numerous electron-opaque cytoplasmic granules. The type II cells were agranular with attenuated cytoplasmic processes which partially or completely ensheathed the type I cells. The glomera were well vascularized. Capillary endothelial cells contained numerous pinocytotic vesicles, but few fenestrae. Two profiles of nerve terminals were observed. One, apposing the type I cells, contained numerous electron-lucent vesicles, several dense-cored vesicles, mitochondria and possessed membrane specializations resembling those usually observed in synaptic zones. The other profile contained abundant mitochondria and a few electron-lucent and densecored vesicles. Structural specializations were not observed on the apposed membranes of these terminals or adjacent to type II cells. Fluorescence histochemistry revealed an intense yellow-green fluorescence in the glomera, which indicated the presence of biogenic amines, possibly primary catecholamines or an indolamine. The electron-opaque granules observed in the type I cells were believed to be the storage sites for these amines. The subclavian glomera were found to be morphologically similar to the carotid body which is a known chemoreceptor.     

Histological changes associated with ageing of the human carotid body

A qualitative and quantitative histological study was made of the carotid bodies obtained at necropsy from 47 subjects ranging in age from 14 to 100 years. With increasing age there is a loss of more than a third of the amount of glomic tissue with a diminution in the size of the lobules. In young people the basic histological pattern of clusters, composed of cores of chief cells with surrounding rims of sustentacular cells, has commonly superimposed on it prominence of the dark variant of chief cells. In middle-aged subjects there is proliferation of sustentacular cells which appear to merge imperceptibly into fibrosis of the lobules and becomes associated with diffuse or focal infiltrates of lymphocytes.     

The paraganglia within the carotid bifurcation regions of young and old spontaneously hypertensive rats (SHR) after exposure to chronic hypobaric hypoxia. I. The carotid bodies

Carotid body volumes and the histological appearance of these chemoreceptors were studied using light microscopic methods in 10 groups of spontaneously hypertensive rats (SHR). The aim of this study was to clarify the influence of chronic hypobaric hypoxia on the carotid bodies of SHR depending on the age of the rats, on the duration of exposure to hypoxia, and on different salt intake, respectively different blood pressure. We found that: 1. The carotid bodies of chronically hypoxic SHR are enlarged. 2. The degree of carotid body enlargement is dependent on the duration of exposure to hypoxia. 3. In old SHR the increase of carotid body volume was smaller than in young SHR. 4. Old chronically hypoxic SHR exhibited more distinct vascular changes in the carotid bodies than age-matched normoxic controls as well as younger chronically hypoxic and normoxic SHR. 5. The influence of different levels of systemic arterial blood pressure on the carotid body volumes was rather small compared with the effects of chronic hypobaric hypoxia.     

Fine structure of the rat carotid body transplanted into the anterior chamber of the eye

Summary Rat carotid bodies transplanted into the anterior chamber of the eye were studied by electron microscopy. Chief and sustentacular cells and a few ganglion cells survived for 3 months and maintained cytological characteristics similar to those in the intact carotid body. The transplant contained many fenestrated capillaries. Chief cells at the periphery of the cell cluster had long cytoplasmic processes which projected into the stroma of the iris. The cell processes became incorporated into bundles containing nerve fibres, which were enveloped by a perineurial sheath. Three types of nerve fibres were identified in the explant. Type I and type II nerve fibres (presumptive cholinergic and adrenergic, respectively) were enclosed by sustentacular and satellite cells. Most of the nerve fibres were completely separated from chief cells and ganglion cells by sustentacular and satellite cells. A few nerve fibres made direct apposition to chief cells and ganglion cells, where some nerves were presynaptic to them. Type III nerve fibres derived from myelinated nerve fibres were also enclosed by sustentacular and satellite cells.     

ULTRASTRUCTURAL STUDIES ON A CAROTID BODY TUMOR

Light and electron microscopic studies were made on a large carotid tumor. Light microscopy revealed adenomatous formation with dark granules giving a positive Grimelius silver reaction in the lumen and cytoplasm of chief cells. Electron microscopy showed that the tumor consisted mainly of clear cells and a few dark cells. Electron dense granules were observed both in the cytoplasm of the tumor cells and in the lumen. Microfilaments were prominent in the cytoplasm of both types of tumor cells.     

An ultrastructural and cytochemical investigation of the development of inclusions in gastric chief cells and parietal cells of mice with the Chediak-Higashi syndrome

Gastric epithelium of the beige mouse, with a mutation thought to be analogous to that in Chediak-Higashi syndrome of man, has been examined by ultrastructural morphologic and cytochemical methods. The gastric chief cell in beige mice at 2 months of age or older disclosed two types of abnormal inclusion bodies each having distinctive morphologic and cytochemical features and a different distribution pattern and relationship to other organelles. On the basis of these findings, the first type of inclusion was thought to originate from zymogen granules, in a process of crinophagy, and the second type was interpreted as arising from the maturing face of the Golgi lamellae by the route for genesis of secondary lysosomes or lipofuscins. Each type of inclusion showed evidence both for participating in autophagic processes and for fusing with each other to produce giant inclusions. Additional observations in this study provided evidence for a role of Golgi endoplasmic reticulum lysosome in genesis of secretory granules and of the mature face of the Golgi complex in development of secondary lysosomes in chief cells. The findings also afforded evidence of migration of chief cells toward the bottom of the gland in the course of their maturation. The gastric parietal cell of control black mice disclosed secondary lysosomes, thought to arise from fusion between multivesicular bodies and mitochondria. These autophagic secondary lysosomes were enlarged in beige mice.     

慢性缺氧和炎性细胞因子刺激对大鼠颈动脉体中IL-1β表达的影响

采用免疫组织化学染色和图像分析方法观察慢性低压性缺氧和腹腔注射重组鼠IL-1β(rmIL-1β)对颈动脉体中IL-1β表达的影响。雄性SD大鼠20只,随机分为4组:T1组缺氧并腹腔注射rmIL-1β;T2组单纯缺氧;T3组仅腹腔注射rmIL-1β;T4组不缺氧也未腹腔注射rmIL-1β。T1组和T2组采用慢性低压性缺氧模型,大鼠在低压氧舱内连续缺氧2周,每天9h(PB=375Torr)。T3组和T4组也每天放入低压氧舱内9h,但不给予低压缺氧。T1组和T3组在第13d放入低压氧舱前,给予腹腔注射rmIL-1β1000ng/kg,T2组和T4组给予腹腔注射相同体积的生理盐水。第14d四组动物分别在低压缺氧舱中缺氧或常氧处理6h后,注射与前一天相同量的rmIL-1β或生理盐水,然后继续低氧或对照处理3h。处理完毕后麻醉处死动物。结果显示:与未给予缺氧和rmIL-1β刺激的T4组相比,T2组(单纯缺氧组)或T3组(单纯注射rmIL-1β)颈动脉体中IL-1β表达均上调(P<0.05);双因素方差分析的结果表明,缺氧和腹腔注射rmIL-1β之间无交互作用(P>0.05)。以上结果提示:慢性缺氧和炎性细胞因子刺激均可导致颈动脉体内IL-1β的上调,IL-1β可能与颈动脉体的功能调节有关。     

In vitro recording of chemoreceptor activity in catecholamine-depleted rabbit carotid bodies

Carotid bodies, together with Hering's nerves, were excised from anesthetized rabbits 24, 48 or 72 h after single reserpine injections (5 mg kg-1, i.v. or i.p.) and were superfused in vitro. Some carotid bodies were processed for formaldehyde-induced fluorescence microscopy to assess catecholamine depletion. Twenty-four hours after reserpine treatment, most of the type I cell islets had lost their fluorescence and the number of spontaneously active chemoafferent units was dramatically reduced. Forty-eight hours after reserpine injection, both the fluorescence of type I cells had partially recovered and the number of chemoreceptor units was almost normal. A significant reduction of both the normoxic and hypoxic frequencies of discharge was demonstrated in carotid bodies examined 24 or 48 h after reserpine pretreatment. Superfusions with dopamine (1, 10, 100 microM) transiently restored the response to hypoxia. It is proposed that catecholamines contained in type I cells play a prominent role in the genesis of chemoafferent activity and in the chemoreceptor response to hypoxia.     

A quantitative morphological study of the carotid bodies of rats living at a simulated altitude of 4300 metres.

A quantitative histological study was carried out on the carotid bodies of 10 normal rats and 10 rats living in a hypobaric chamber at a pressure of 460 mm Hg from 25 to 96 days. In the chronically hypoxic rats there was a four-fold increase in the mean combined volume of the carotid bodies. Morphometric analysis disclosed a three-fold increase in the mean volume of specialised glomic cells and a ten-fold increase in the mean volume of capillaries, although the proportion of glomic cells was actually significantly decreased. In all our hypoxic rats there was evidence of both right and left ventricular hypertrophy. However, there was no linear relation between total carotid body volume or volume of glomic cells on one hand and the right and left ventricular weight, on the other hand. Although there was no linear relation between combined total carotid body volume and duration of hypoxia, the linear relation between glomic cell volume and duration of hypoxia was significant at the 5 per cent. level. The increase in vascularity of the hypoxic carotid body may be a mechanism to increase blood flow and thus oxygen transport to a hypoxic organ with increased metabolic activity. Small quantities of an amorphous hyaline material of unknown nature were found in relation to capillaries and type I cells in all the hypoxic rats.     

Investigations on the ultrastructural cytology of branchiomeric paragangliomas (author's transl)

The paragangliomas of the jugular and carotid bodies are growth processes which show varying histological and cytological patterns. The real nature of these lesions is still a matter of discussion. In this study 6 jugular and 3 carotid body tumours were investigated using the electron microscope. The main components were irregular cells with short and long cytoplasmic processes closely packed and mostly arranged in complexes. The ultrastructural cytology of the jugular and carotid bodies showed no striking differences. The great majority of the cells exhibits a clear similarity to the cells of normal paraganglia, especially to chief cells of carotid bodies (type I cells). But there were some modifications in the cellular structure including the abnormal structure of mitochondria and a greater variation in the morphology of the dense cored granules. The cytoplasm was dominated by differing populations of mitochondria comprising such of the crista type and such showing tubulus-like structures with a dense matrix and very often intramitochondrial bodies. The characteristic feature was the occurrence of randomly distributed dense cored granules measuring 700-800 A in diameter. These granules display a homogeneous electron dense or moderately dense somewhat granular core which is separated from limiting membrane by a clear electron lucent halo. A further population of granules was of a greater dimension (3,000-4,000 A) revealing an oval or cigar-like shape. In contrast to the former type the homogeneous dense cores of these granules showed a smooth contour and the lucent halo was very narrow. In general, singular exocytotic phenomena could be supposed. More frequently, however, signs of granulolysis were encountered. Free ribosomes, arranged often as polysomes, sparse, disorderly distributed microfilaments of 60-80 A diameter, few 240 A-microtubules and some lysosomal structures are also observed. Usually the cell clusters abutted directly upon the vascular connective tissue. In other cases cytoplasmic processes are intercalated resembling sustentacular cells. Fenestrated capillaries known to be typical for the paraganglionic tissues were only seldom detected. In the interstitial tissue typical collagen fibres are found showing a periodicity of 600-700 A, but occasionally fibres with a periodicity of about 1,200 A could be seen resembling fibrous-long-spacing collagen. Synaptic junctions and nerves or axons were always lacking.     

Morphological findings at the carotid bodies of humans suffering from different types of systemic hypertension or severe lung diseases

Post-mortem studies of the carotid bodies of 62 humans were carried out using light microscopic and morphometric methods. According to the clinical and autopsic data the subjects were divided into 5 groups: normotensives, essential hypertensives, renal hypertensives, chronically hypoxic persons with severe lung diseases, and people who suffered from both lung diseases and essential hypertension. Carotid body volume showed age-dependence in the normotensive group; the biggest glomera carotici were found at an age of 40-60 years, whereas younger and older people exhibited smaller carotid bodies. In the group with the essential hypertensives only old patients exhibited enlarged carotid bodies. In younger essential hypertensives but also in the renal hypertensives an increase of carotid body size was not demonstrable. The people with severe lung diseases regularly had greater carotid bodies when compared with age-matched normotensive subjects. In addition, chronically hypoxic patients had a proliferation of type II cells, perhaps with involvement of Schwann cells and fibrocytes. This increases of elongated cells was only seldom observed in the other groups. The results are discussed with respect to the alterations known so far of arterial chemoreceptor function and reflex effects in systemic arterial hypertension.