Somatostatin immunoreactive C cells in thyroid glands from various mammalian species

The relative distribution of somatostatin- and calcitonin-containing cells in thyroid glands from various mammalian species was investigated by immunoperoxidase staining, and the concentration of immunoreactive somatostatin by radioimmunoassay. In the thyroid glands of guinea pigs and rabbits, most of the calcitonin cells were also immunoreactive to the somatostatin antiserum, and high concentration of immunoreactive somatostatin was obtained. On the other hand, in the thyroids of other animal species--rats, dogs, pigs, cows, goats, cats, monkeys, mice, and hamsters--only a few C cells revealed the immunoreaction for somatostatin, and the concentration of somatostatin was low. In all animal species studied, the somatostatin was present in the same cells that contain calcitonin, though in guinea pigs and rats there were some C cells containing a large number of reaction products for somatostatin but very few for calcitonin. Thus, it was concluded that there was a considerable variation in somatostatin immunoreactivity of thyroid C cells from species to species.     

Localization of immunoreactive calcitonin gene-related peptide in thyroid C cells from various mammalian species

Localization of immunoreactive calcitonin gene-related peptide (CGRP) in thyroid C cells from various mammalian species was investigated by the immunoperoxidase method. In many animal species including dogs, cats, cattle, monkeys, rats, and rabbits, almost all C cells revealed an intense immunoreactivity for CGRP; the cytoplasm of C cells was filled with reaction products for CGRP. In these animal species, calcitonin and CGRP coexisted in the C cells. However, in some species including pigs, mice, hamsters, and guinea pigs, the CGRP immunoreactivity of C cells was weak or negative. It was concluded that there was a considerable variation in CGRP immunoreactivity of C cells from species to species. In rabbits and guinea pigs, almost all C cells were also intensely immunoreactive to antisomatostatin antiserum, whereas in other animal species including dogs, cats, cattle, monkeys, rats, pigs, mice, and hamsters only a few C cells were immunoreactive to somatostatin. Three peptides--calcitonin, somatostatin, and CGRP--are synthesized alone in rabbit C cells. Thus, there was no relation between CGRP and somatostatin concerning the existence of both peptides in thyroid C cells.     

Ontogeny of immunoreactive somatostatin in thyroid C cells from dogs and guinea pigs

The development of immunoreactive somatostatin in thyroid C cells of dogs and guinea pigs from early fetuses to adults was investigated by the use of immunoperoxidase histochemistry and radioimmunoassay. The time of appearance and developmental patterns of immunoreactive somatostatin in the C cells were completely different in both species. In guinea pig thyroids, the somatostatin immunoreactivity appeared later than the calcitonin immunoreactivity and the number of somatostatin-positive cells was very small during fetal periods. The somatostatin immunoreactivity rapidly increased during neonatal periods. A large population of the somatostatin cells and a high concentration of somatostatin immunoreactivity were observed in mature animals. On the other hand, in dog fetuses somatostatin immunoreactivity appeared very early, at the same time as the calcitonin immunoreactivity. The largest population of somatostatin cells was found at the stage when the primordial follicles were vigorously formed throughout whole thyroid parenchyma. At this stage almost all of calcitonin-positive cells were also somatostatin-positive. The somatostatin cells progressively decreased as the development proceeded, in contrast to the calcitonin cells which increased with gestational age. In postnatal dogs only a few C cells revealed the immunoreaction for somatostatin, and the concentration of somatostatin was very low. These findings suggest that the function of somatostatin in dog thyroid C cells may be different from that in guinea pig C cells.     

Ontogeny of immunoreactive somatostatin in thyroid C cells from dogs and guniea pigs

The development of immunoreactive somatostatin in thyroid C cells of dogs and guinea pigs from early fetuses to adults was investigated by the use of immunoperoxidase histochemistry and radioimmunoassay. The time of appearance and developmental patterns of immunoreactive somatostatin in the C cells were completely different in both species. In guinea pig thyroids, the somatostatin immunoreactivity appeared later than the calcitonin immunoreactivity and the number of somatostatin-positive cells was very small during fetal periods. The somatostatin immunoreactivity rapidly increased during neonatal periods. A large population of the somatostatin cells and a high concentration of somatostatin immunoreactivity were observed in mature animals. On the other hand, in dog fetuses somatostatin immunoreactivity appeared very early, at the same time as the calcitonin immunoreactivity. The largest population of somatostatin cells was found at the stage when the primordial follicles were vigorously formed throughout whole thyroid parenchyma. At this stage almost all of calcitonin-positive cells were also somatostatin-positive. The somatostatin cells progressively decreased as the development proceeded, in contrast to the calcitonin cells which increased with gestational age. In postnatal dogs only a few C cells revealed the immunoreaction for somatostatin, and the concentration of somatostatin was very low. These findings suggest that the function of somatostatin in dog thyroid C cells may be different from that in guinea pig C cells.     

Ontogeny of immunoreactive calcitonin gene-related peptide in thyroid C cells from dogs, rabbits, and guinea pigs

Ontogeny of immunoreactive calcitonin gene-related peptide (CGRP) in thyroid C cells of dogs, rabbits, and guinea pigs from early fetuses to adults was investigated by an immunoperoxidase method, in comparison with the development of immunoreactive calcitonin and somatostatin. The presence of immunoreactive CGRP in mature C cells was different from species to species. Dog and rabbit C cells revealed intense immunoreactivity for CGRP, whereas guinea pig C cells revealed very weak immunoreactivity or none. In dog fetuses, the appearance of immunoreactive CGRP was early. At around 35 days of gestation, when the follicular cells were not yet organized into follicles, immunoreactivities for three peptides--calcitonin, somatostatin, and CGRP--began to appear in C cells. While the highest population of somatostatin-positive cells was attained when the primordial follicles were vigorously formed throughout whole thyroid parenchyma and their frequency progressively declined thereafter, CGRP-positive cells as well as calcitonin-positive cells gradually increased in number and intensity with gestational age. The developmental pattern of immunoreactive CGRP coincided with that of immunoreactive calcitonin in dog C cells. In rabbit fetuses, at 25 days of gestation, when thyroid follicles stored large amounts of colloid and C cells already exhibited intense immunoreactivity for calcitonin, CGRP immunoreactivity as well as somatostatin immunoreactivity began to appear. Subsequently, immunoreactivities for the three peptides gradually increased with age, although calcitonin immunoreactivity was outstandingly intense among them. In guinea pig C cells, intense immunoreactivity for CGRP was not observed in any stages of development. These results indicate that there are developmental profiles of CGRP characteristic for each animal, and the ratio of CGRP and calcitonin produced from calcitonin genes in C cells seems to be fixed for life.     

Age associated increase of C cell follicles in guinea pig thyroid glands

The effect of aging on the formation of C cell follicles was examined in the thyroid gland of guinea pigs at various ages ranging from 1 to 29 months. The C cell follicles were demonstrated with the immunoperoxidase methods by using anticalcitonin and antisomatostatin antisera and with PAS reaction. They were already detected in 1-month-old guinea pigs but in low number. Thyroid glands from 1- to 14-month-old animals contained only a small number of C cell follicles and did not reveal the age-related increase. In aged guinea pigs (20- to 29-month-old), a dramatic increase of C cell follicles was found, about 13 times as high as the number of other age groups. The C cell follicles through all age groups were present in large clusters of C cells. In the aged guinea pigs, nodular large aggregates of C cells regarded as C cell hyperplasia occurred and numerous C cell follicles were formed in the large cell aggregates. Thus, the conspicuous increase of C cell follicles in aged animals was associated with a proliferative abnormality of C cells. The C cells forming follicles showed moderate to weak immunoreactivity for calcitonin, whereas they showed very intense immunoreactivity for somatostatin. In addition, the colloidlike and flocculent materials stored in the follicular lumina, which were consistently PAS-positive, were weakly immunoreactive to somatostatin but nonreactive to calcitonin.     

Alterations of lmmunoreactive somatostatin in thyroid C cells after induced hypercalcemia,hypocalcemia, and antithyroid drug treatment

In order to elucidate the functional significance of somatostatin in thyroid C cells, the alterations of immunoreactive somatostatin in the cells were investigated under various experimental conditions, i.e., hypercalcemia, hypocalcemia, and antithyroid drug treatment. Guinea pigs and rabbits, in which almost all C cells reveal the intense immunoreaction for somatostatin in addition to calcitonin, were used as experimental animals. After chronically induced hypercalcemia, somatostatin immunoreactivity conspicuously diminished coinciding with the decrease of calcitonin; somatostatin as well as calcitonin was responsive to induced hypercalcemia. After hypocalcemic tetany induced by injection of Escherichia coli Lasparaginase, C cells exhibited very intense immunoreactions for both calcitonin and somatostatin. After chronic treatment of ethylenethiourea, immunoreaction of somatostatin in C cells was the same as that of calcitonin. That is, when immunoreactivity for calcitonin remained unchanged, immunoreactivity for somatostatin was also intensive. However, when immunoreaction of calcitonin became very weak, the reaction of somatostatin was also weak. Thus, in all experimental conditions examined the alterations of immunoreactive somatostatin in C cells completely coincided with those of calcitonin. It seems likely that somatostatin in thyroid C cells exerts the synergistic effect on calcitonin action.     

Alterations of immunoreactive somatostatin in thyroid C cells after induced hypercalcemia, hypocalcemia, and antithyroid drug treatment

In order to elucidate the functional significance of somatostatin in thyroid C cells, the alterations of immunoreactive somatostatin in the cells were investigated under various experimental conditions, i.e., hypercalcemia, hypocalcemia, and antithyroid drug treatment. Guinea pigs and rabbits, in which almost all C cells reveal the intense immunoreaction for somatostatin in addition to calcitonin, were used as experimental animals. After chronically induced hypercalcemia, somatostatin immunoreactivity conspicuously diminished coinciding with the decrease of calcitonin; somatostatin as well as calcitonin was responsive to induced hypercalcemia. After hypocalcemic tetany induced by injection of Escherichia coli L-asparaginase, C cells exhibited very intense immunoreactions for both calcitonin and somatostatin. After chronic treatment of ethylenethiourea, immunoreaction of somatostatin in C cells was the same as that of calcitonin. That is, when immunoreactivity for calcitonin remained unchanged, immunoreactivity for somatostatin was also intensive. However, when immunoreaction of calcitonin became very weak, the reaction of somatostatin was also weak. Thus, in all experimental conditions examined the alterations of immunoreactive somatostatin in C cells completely coincided with those of calcitonin. It seems likely that somatostatin in thyroid C cells exerts the synergistic effect on calcitonin action.     

Calcitonin gene-related peptide immunoreactivity in afferent neurons supplying the urinary tract: combined retrograde tracing and immunohistochemistry

The innervation of rat and guinea pig urinary tract was examined using immunohistochemistry, radioimmunoassay and True Blue retrograde tracing techniques and was further assessed following both surgical and chemical denervation experiments. Substantial amounts of calcitonin gene-related peptide-like immunoreactivity (range 20-150 pmol/g) were detected in tissue extracts and localised to nerve fibres distributed throughout the urinary tract of both species, these being concentrated in the ureter and base of the bladder. In the guinea pig, the number and distribution pattern of calcitonin gene-related peptide-like immunoreactive nerves appeared to be identical to that of substance P-containing nerves, whereas in the rat the former predominated. Seven days after injection of the fluorescent dye True Blue into tissues of the urinary tract, retrogradely labelled cells were found in the dorsal root ganglia. These cells had a segmental distribution pattern which was specific for each of the injection sites. Thus, after injection of True Blue into the left kidney hilum a single group of labelled cells were found in the ipsilateral T10-L2 dorsal root ganglia. In contrast, injection into the left ureter produced labelled cells in two separate groups of ipsilateral ganglia (T11-L3 and L6-S1). Injection into the wall of the bladder and upper urethra resulted in bilateral labelling, with most labelled cells occurring in L6 and S1 ganglia. Approximately 90% of labelled cells in T10-L3 dorsal root ganglia displayed calcitonin gene-related peptide-like immunoreactivity, but only 60% of retrogradely labelled bladder neurons in L6-S1 ganglia were immunoreactive for this peptide. Adult guinea pigs and neonatal rats injected systemically with capsaicin subsequently exhibited a marked reduction both in the amount of calcitonin gene-related peptide immunostaining and the concentration of immunoreactive material in the urinary tract, dorsal root ganglia and spinal cord. In rats treated neonatally with capsaicin, there was a significant reduction in the number of retrogradely labelled cells and a hypertrophy of the bladder. Sectioning of the pelvic and hypogastric nerves in the rat also resulted in a depletion of calcitonin gene-related peptide-like immunoreactive nerves in the bladder, whereas chemical sympathectomy appeared to have no effect. The results indicate that calcitonin gene-related peptide immunoreactivity occurs in a major proportion of afferent neurons supplying the urinary tract of the rat and guinea pig.(ABSTRACT TRUNCATED AT 400 WORDS)     

Immunohistochemical study of C cell follicles in dog thyroid glands

In dog thyroid glands there are C cell follicles which are lined solely by C cells and which accumulate a colloidlike substance in the luminal cavities. In order to clarify the properties of the colloidlike substance secreted by C cells, the C cell follicles were stained with PAS reaction and immunoperoxidase method using anticalcitonin, anti-C-thyroglobulin, and anti-19S-thyroglobulin antisera, respectively. The colloidlike substance was PAS positive and revealed the strong immunoreaction for C-thyroglobulin but a faint reaction for calcitonin. It was nonreactive with anti-19-thyroglobulin antiserum. These results confirm that C cells synthesize the glycoprotein immunoreactive to anti-C-thyroglobulin antiserum in addition to calcitonin and can store it in the follicular lumens.     

Studies on localization of calcitonin gene-related peptide (CGRP) in the thyroid-parathyroid complex

Summary Calcitonin gene-related peptide (CGRP) was localized by an immunocytochemical technique in the thyroid-parathyroid complexes of rat, guinea pig, rabbit, and in normal human thyroids and parathyroids. Human medullary carcinomas and parathyroid adenomas were also studied. In man and all animal species examined CGRP was present in the parafollicular cell, however, in guinea pigs only in small amounts. Except in rabbits, presence of CGRP was demonstrated in nerves of the thyroid and parathyroid capsule as well as in the nerve fibers of the capsular blood vessels. In the thyroid of guinea pigs CGRP was also noted in nerve fibers and in blood vessel walls between follicles. CGRP was also present in the parathyroid glands of rat and man, in nerve fibers localized between parathyroid cells. In rabbit the parafollicular cells between parathyroid cells also expressed CGRP immunoreactivity. No CGRP was noted in the parathyroids of the guinea pig. The proximity of parathyroid cells and CGRP containing tissue structures suggests a role for CGRP in the modulation of parathyroid hormone secretion. The importance of these regulatory mechanisms appear to be different in individual species.M. Dietel is a visiting Professor from the Institute of Pathology, University of Hamburg, Federal Republic of Germany     

Ontogeny of chicken ultimobranchial glands studied by an immunoperoxidase method using calcitonin, somatostatin and 19S-thyroglobulin antisera

Summary The ontogeny of the ultimobranchial glands in chickens from 9-day-old embryos to adults was investigated by the immunoperoxidase method using anti-calcitonin, anti-somatostatin and anti-19S-thyroglobulin antisera. During embryonic development, the chick ultimobranchial glands consisted of solid cell clusters. Calcitonin immunoreactivity began to appear at 16 days of incubation and rapidly increased at late periods of incubation. At the time of hatching, almost all of the epithelial cells in the ultimobranchial glands exhibited the immunoreaction for calcitonin. Cyst structures showing various sizes, shapes and luminal contents were consistent features of the ultimobranchial glands after hatching. As age proceeded, the cysts and loose connective tissues gradually increased in the glands. In adult chickens, the calcitonin cells came to be interspersed among them and the number of the cells per unit area was very small, compared with that in young animals. No immunoreaction for somatostatin was found in the ultimobranchial glands of chickens of all ages examined. In the glands there were no cells immunoreactive to the 19S-thyroglobulin antiserum. Further, neither cyst epithelium nor luminal contents were stained with the antiserum.     

C cell hyperplasia and carcinoma developing in sheep with experimentally-induced lymphosarcoma.

Two cases of C cell hyperplasia and one case of C cell carcinoma of the thyroid glands were bilaterally recognized in 11 sheep with experimentally-induced lymphosarcoma. The serum calcium concentration in the C cell carcinoma case was slightly increased above the normal concentration of around 9 mg per dl. Bilateral C cell hyperplasia also developed in the thyroid lobes of the C cell carcinoma case. Immunohistochemically, hyperplastic C cells and tumour cells were positive for calcitonin, calcitonin gene-related peptide, chromogranin A and neurone-specific enolase. No amyloid deposition nor multiple endocrine neoplasia was demonstrated in any of the cases. Ultrastructurally, many secretory granules were observed in the cytoplasm of neoplastic cells constituting the C cell carcinoma and in the hyperplastic C cells.     

Somatostatin-immunoreactive cells in medullary carcinoma of the thyroid.

Ten medullary carcinomas of the thyroid were examined by immunohistochemistry using antisera against calcitonin, somatostatin, and ACTH. In addition to calcitonin cells, eight of the tumors contained somatostatin-immunoreactive cells in varying numbers. No ACTH-immunoreactive cells were found. The results indicate that thyroid medullary carcinomas often are multihormonal, and that somatostatin is common in such tumors. In contrast to calcitonin cells, somatostatin cells are not found in normal human thyroid tissue.     

Distinct sprouting responses of sympathetic and peptidergic sensory axons proximal to a sciatic nerve transection in guinea pigs and rats

Sprouting of sympathetic and peptidergic sensory neurones proximal to nerve lesions may reflect upregulation of growth factors around damaged dorsal root ganglion (DRG) cells. Axons containing noradrenaline or calcitonin gene-related peptide were visualized in DRGs and spinal roots of guinea pigs and rats. After sciatic transection in rats, varicose terminals of both types appeared around large DRG somata. These neurones were surrounded by proliferated satellite cells expressing glial fibrillary acidic protein (GFAP) and p75. This did not occur in guinea pigs. Instead, sympathetic axons grew through the DRG and centrally along the dorsal roots (which contained p75-positive glia), avoiding the DRG somata. Thus the glial reaction to peripheral injury differs between species such that, in guinea pigs, the environment in the spinal roots rather than in the DRGs favours sympathetic sprouting.     

Crinophagy in Thyroid Follicular and Parafollicular Cells of Male Obese Zucker Rat

Comparison between lean (Fa/?) and obese (fa/fa) young adult male Zucker rat thyroids reveals that obese rats display larger clusters of parafollicular cells than the lean ones with a lesser blood supply. Fa/? thyroid typically shows single or “twin” C cells in follicles; fa/fa parafollicular cells appear with three functional aspects. Crinophagy is found in the fa/fa C cells amassing numerous aberrant calcitonin-containing vesicles among which lysosomes build these autophagic bodies by capturing vesicle contents, other organelles and, fusing with each other, increase their size. Other C cells contain many secretory vesicles but show few or no crinophagic structures. Another parafollicular cell type is revealed with scant organelles and highly contrasted secretory vesicles, different from calcitonin. Hypercalcemia of fa/fa rats corresponds to increased C cells population with accrued calcitonin production but a low calcitonin plasma level – verified by others – is likely caused by crinophagy of the altered vesicles. In addition, the T thyrocytes of fa/fa rats exhibit crinophagy bodies; this can confirm their hypothyroidism. Possibly, the known leptin mutation along with other unknown paracrine secretions alter both T and C thyrocytes’ functions of the fa/fa rats, allowing high intracellular calcium and lower pH favoring autophagocytosis. Other longitudinal, interdisciplinary studies should further clarify the complex paracrine interactions existing between these endocrine structures because this animal model could be useful to understand human defects, such as the metabolic syndrome that involves obesity, cardiovascular, renal, hepatic, non-insulin dependent diabetes mellitus (NIDDM), hypothyroidism defects, as well as the etiology of thyroid medullary tumors.     

鹅胃肠内分泌细胞的免疫组织化学定位

应用６种胃肠激素抗血清，对鹅胃肠各段的内分泌细胞分布进行了免疫组织化学定位。腺胃显示有较多促胃泌素释放肽和生长抑素细胞。肌胃有较多促胃泌素释放肽、生长抑素和胃泌素细胞，少量胰多肽细胞。幽门部有大量的生长抑素细胞，密集的胃泌素细胞，偶见胰多肽细胞。小肠内有胃泌素、胰多肽和生长抑素细胞，细胞类型和数量由前段向后段逐渐减少。未检出胃动素和抑胃肽细胞。