Immunohistochemical analysis of EGF in epiphyseal growth plate from normal,hypophysectomized, and growth hormone-treated hypophysectomized rats

Epiphyseal growth plate cartilages from the proximal tibia of normal, hypophysectomized, and growth hormone (GH)-treated hypophysectomized rats were subjected to immunohistochemistry for detection of epidermal growth factor (EGF). In the normal growth plate, EGF was distributed mainly in the proliferative zone. Hypophysectomy resulted in considerable atrophy of the chondrocytes and the cartilage matrix (a decreased number of mature-type chondrocytes and a decreased ratio of proliferating to hypertrophic chondrocytes) and a significant diminution of EGF immunoreactivity. Treatment with GH reversed these effects of hypophysectomy, causing an increased thickness of the growth plate and EGF-reactive sites in all chondrocyte layers. The most intense immunostaining for EGF, however, was frequently seen in the nuclei of chondrocytes with flattened appearance. It appears that EGF could be incorporated or synthesized in chondrocytes having marked mitogenic activity. The present results, taken with previous data on EGF involvement in growth of cartilaginous tissue in vivo and in vitro, strongly suggest that EGF-immunoreactive chondrocytes are involved in cartilage proliferation and growth under the specific influence of GH.     

Mammary gland development and alpha-lactalbumin production in hypophysectomized, pregnant mice

Swiss Webster mice were hypophysectomized on Day 10 of pregnancy and the effect of the ablation on mammary gland development was estimated by measuring the total weight and the DNA, RNA, and alpha-lactalbumin contents and concentrations of the mammary gland on Days 14 and 18 of gestation. Although a significant increase in mammary gland weight occurred in the hypophysectomized animals between Days 10 and 18 of pregnancy, the mammary gland weight of the hypophysectomized mice was significantly reduced when compared with intact and sham-operated mice on both Days 14 and 18 of pregnancy. The total RNA content of the mammary gland was also reduced in the hypophysectomized mice, although it increased significantly from Day 10 to Day 18. The alpha-lactalbumin content of the mammary gland increased only slightly between Days 10 and 14 of gestation in the intact and sham-operated mice, but a large increase was found on Day 18 in both groups. There was, on the other hand, no increment in the alpha-lactalbumin content of the mammary gland in the hypophysectomized mice either on Day 14 or 18 of gestation. The DNA content of the mammary gland was not affected by hypophysectomy when estimated on Days 14 and 18 of pregnancy. The effects of hypophysectomy on the concentrations of mouse placental lactogen II (mPL-II), progesterone, corticosterone, and thyroxine in the maternal serum were also assessed. The concentration of mPL-II was significantly elevated in the hypophysectomized mice, whereas the circulating concentrations of both corticosterone and thyroxine were greatly reduced. The serum progesterone concentration was not significantly altered by hypophysectomy. (ABSTRACT TRUNCATED AT 250 WORDS)     

Corticotropin-like immunoreactivity in the brain of intact,hypophysectomized, cortisol- and metopirone-treated eels

Summary An ACTH-like peptidergic system was demonstrated in the brain of three teleost species by immunocytochemistry. In order to investigate the origin of brain ACTH and factors modulating its synthesis, similar techniques were applied to the brain of eels (1) submitted to hypothysectomy in order to suppress pituitary ACTH and plasma cortisol, (2) injected with cortisol to inhibit pituitary ACTH synthesis and release, and (3) injected with metopirone to block cortisol synthesis and stimulate ACTH synthesis and release. Hypophysectomized eels showed a normal distribution of immunoreactive perikarya in the ventral hypothalamus and fibers in the brain, suggesting that brain ACTH does not arise from the pituitary. In cortisol-treated eels immunostaining was markedly reduced in brain perikarya and pituitary corticotropes, suggesting a reduced synthesis. In metopirone-injected eels, one third of the animals showed an increased immunostaining in perikarya and a dense network of immunoreactive fibers, suggesting that ACTH synthesis was increased. Brain ACTH was not affected in other animals. Pituitary corticotropes were rapidly degranulated. Responses of ACTH in the brain and pituitary occur independently when cortisol synthesis is inhibited. These responses are compared to those of the corticotropin-releasing factor system in the same eels.     

Bombesin produces hypothermia in hypophysectomized rats

Intracerebroventricular administration of bombesin, a naturally-occurring peptide, produces hypothermia in the rat. To determine whether a pituitary-dependent step is necessary for this effect, the thermoregulatory response was followed in hypophysectomized and intact rats maintained at room temperature. Significant hypothermia was produced in both experimental groups. This study supports an extra-pituitary mechanism for bombesin-induced hypothermia.     

The hypothalamo-hypophysial system of hypophysectomized rats

Summary The median eminence (ME) of hypophysectomized rats was studied by means of light and electron microscopy. Paraldehyde-fuchsin (PAF)-positive material is seen in the external zone (EZ) of the ME 2–5 days after the operation. Its amount gradually increases especially in the caudal part of the ME during the following few days. Some PAF-positive fibers make contact with the subependymally located blood capillaries. In the most caudal region of the recessus infundibuli they penetrate into the third ventricle. PAF-positive material decreases markedly from the ME of rats two months after hypophysectomy and exposure to a 1% salt load. Fibers of types A₁, A₂ and B containing granules of 120–220 nm, 100–150 nm and 80–100 nm in diameter, respectively, are seen in the EZ of the ME in hypophysectomized rats, although almost exclusively A₂- and B-type structures make contact with the primary portal capillaries in intact animals. All types of neurosecretory fibers establish contact with the subependymal nonfenestrated blood capillaries and penetrate the recessus infundibuli. Some neurosecretory terminals of different types make direct contact with the glandular cells of the pars tuberalis or are separated from them by a thin basal lamina. It is assumed that mainly neurosecretory fibers of types A₂ and B are permanently connected with the primary portal capillaries in the EZ of the ME in intact mammals, while the overwhelming majority of fibers of A₁-type shows ingrowth during the course of postoperative reparation. The possible physiological significance of the described changes is discussed.