Ultrastructural changes of the hypothalamo-hypophysial neurosecretory system in the magnesium deficient rats.

A magnesium deficient diet caused transient but marked degenerative changes in the rat hypothalamo-hypophysial neurosecretory system which strongly resembled in many ultrastructural respects those induced by a prolonged administration of aldosterone as previously reported by us. The possible mechanism for this selective alteration in the neurosecretory neurons has been briefly discussed with regard to aldosterone secretion.     

Cholinesterases in the hypothalamo-hypophysial neurosecretory system of the White-crowned Sparrow,Zonotrichia leucophrys gambelii

Summary The distribution of cholinesterases in hypothalamo-hypophysial neurosecretory system of the White-crowned Sparrow has been examined histochemically. The perikarya of the neurosecretory cells of the paraventricular and supraoptic nuclei have a high acetylcholinesterase activity. Acetylcholinesterase activity also occurs in the cells of the infundibular nucleus. The proximal parts of the axons of the cells of the neurosecretory and infundibular nuclei have strong acetylcholinesterase activity and weak non-specific cholinesterase activity. In the median eminence, the activity of acetylcholinesterase is strongest in the palisade layer. In the pars nervosa, there is definite, although weak, acetylcholinesterase activity.This investigation was supported by grants from the National Institutes of Health to Professor Farner (B-1353) and to Dr. Kobayashi (A-3678).     

Adenylate cyclase activity in crude plasma membranes of subcutaneous adipocytes during ontogenetic development of rats

In order to explain the differences in the hormone stimulated lipolysis during ontogenic development of rats, the activity of adenylate cyclase was determined in crude plasma membranes of subcutaneous adipocytes of 5, 14, 21 and 45 to 55-day-old animals. Stimulatory effects of nonhormonal and hormonal agents were expressed as the increment in percentage of basal values which were not significantly changed in the age groups studied. The highest stimulatory effect was observed after sodium fluoride in 14 and 21-day-old rats. Guanylylimidodiphosphate and GTP revealed the lowest stimulatory effects in adult animals (greater than 45-day-old). The beta-adrenergic agent isoproterenol revealed the highest stimulatory effect in the 5 and 45-day-old group while in the preparation from 14-day-old rats the adenylate cyclase activity was significantly lower. On the other hand, tetracosactide (beta 1-24-corticotropin) revealed the smallest stimulatory effect on the preparation from 5-day-old rats; its stimulatory effect steadily increased and reached the highest value in adenylate cyclase preparations from adult animals. It can be concluded that the adenylate cyclase system in subcutaneous adipocytes is already basically mature at early ontogenic stages of development in rats. Nevertheless, the explanation for the small variations of the enzyme activity in different age groups requires further study.     

Cholinesterase activity during embryonic development in the blood-feeding bug Triatoma patagonica

Triatoma patagonica Del Ponte (Hemiptera: Reduviidae), a vector of Chagas' disease, is widely distributed in Argentina and is found in sylvatic and peridomiciliary ecotopes, as well as occasionally in human dwellings after the chemical control of Triatoma infestans. Anti-cholinesteratic products can be applied in peridomiciliary areas and thus knowledge of cholinesterase activity during embryonic development in this species might contribute further information relevant to effective chemical control. Cholinesterase activity was characterized by reactions to eserine 10(-5) m, to increasing concentrations of substrate and to varying centrifugal speeds. Acetylcholinesterase activity was detected on day 4 and was significant from day 5. A reduction in cholinesterase activity towards acetylthiocholine (ATC) was observed on days 9 and 10 of development. Cholinesterase activity towards ATC and butyrylthiocholine (BTC) in homogenates of eggs was inhibited by eserine 10(-5) m. The shape of the curve indicating levels of inhibition at different concentrations of ATC was typical of acetylcholinesterase. Activity towards BTC did not appear to be inhibited by excess substrate, which parallels the behaviour of butyrylcholinesterases. Cholinesterase activity towards ATC was reduced in supernatant centrifuged at 15 000 g compared with supernatant centrifuged at 1100 g. The cholinesterase system that hydrolyzes mainly ATC seems to belong to the nervous system, as indicated by its behaviour towards the substrates assayed, its greater insolubility and the fact that it evolves parallel to the development of the nervous system. Knowledge of biochemical changes associated with the development and maturation of the nervous system during embryonic development would contribute to the better understanding of anti-cholinesteratic compounds with ovicidal action that might be used in control campaigns against vectors of Chagas' disease.     

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.     

Involvement of accessory neurosecretory nuclei of hypothalamus in the formation of hypothalamohypophyseal system during prenatal and postnatal development in rats

We studied the development of direct axonal connections of the accessory neurosecretory hypothalamic nuclei with the posterior pituitary lobe on the fixed rat brain from day 15 of embryogenesis until day 10 of postnatal development using the retrograde diffusion method of the lipophilic fluorescent carbocyanine dye 1,1'-dioctadecyl-3,3,3',3'-tetramethylindocarbocyanine perchlorate along the neuronal membranes. The marker was applied onto the posterior pituitary lobe and, after incubation in a fixative, fluorescing bodies of nerve cells were visualized in the hypothalamus. Neuronal axons of the retrochiasmatic nucleus were the first of the accessory nuclei to ingrow in the posterior pituitary lobe (on days 16-17 of embryogenesis). Neurons of the circular and dorsolateral nuclei and the nuclei of the median bundle of the forebrain sent their axons to the posterior pituitary lobe starting from the first days of postnatal development. No direct connections of the anterior commissure and perifornical accessory nuclei with the posterior pituitary lobe were found in perinatal development. These facts are discussed in the light of concepts about the different functional role of accessory peptidergic hypothalamic nuclei in rats.     

The hypothalamo-hypophysial system in acipenseridae

Summary The neurohypophysis of sexually mature male and female Acipenser güldenstädti Brandt and Acipenser stellatus Pallas was studied light and electron microscopically. The recessus hypophysei lined with ependymal cells of two main types, narrow and wide, are in the center of the neurohypophysial roots. Processes from both cell types run radially to the basement membrane of the connective tissue layers abutting on the hypophysial intermediate lobe. Protrusions penetrating deep into the recessus hypophyseus are found in the apical parts of the wide cells. Pituicytes are rare in the neurohypophysis. The ultrastructure of both ependymal cell types and of the pituicytes is described. Nonmyelinated Gomori-positive (peptidergic) neurosecretory A₁ and A₂ type fibres and their terminals containing elementary neurosecretory granules (1400–1800 Å and 1000–1400 Å respectively) are the main structural elements of the neurohypophysis. Some dark and single myelinated neurosecretory fibres have been found. The adrenergic fibres (type B) were described earlier (Polenov et al., 1972a). The structural peculiarities of the neurohypophysis are discussed in functional and comparative-morphological terms.     

The hypothalamo-hypophysial system in acipenseridae

Summary The distribution of adrenergic and peptidergic (Gomori-positive) structures of the hypophysial neuro-intermediate complex in Acipenseridae has been studied by means of light, fluorescence, and electron microscopy. Adrenergic fibres (B-fibres) and their terminals have been detected in the neurohypophysis of these fishes. The terminal swellings of B-fibres as well as the terminals of the neurosecretory peptidergic fibres (A₁ and A₂) make contact with the basement membrane of the connective tissue layer separating the neurohypophysis from the intermediate lobe. Capillaries are situated within this layer and, therefore, the main part of the fibre terminals is in contact with the pericapillary space. The release of catecholamines from the adrenergic terminals into the capillaries connected with the general circulation is supposed. The diffusion of catecholamines through the connective tissue layer into the parenchyma of the intermediate lobe is also suggested. Hence, the glandular activity of the intermediate lobe seems to be under the dual control of adrenergic and peptidergic elements of the hypothalamus.The authors wish to express their deep appreciation to G. M. Persov, Dr. Sc. Biol., Head of the Laboratory of Experimental Ichthyology, Petershof Biological Institute of the University of Leningrad, for the material supplied for fluorescence microscopy, and to Mr. G. V. Sabinin for photographic services.     

The hypothalamo-hypophysial system in acipenseridae

Summary The percentage of peptidergic (A₁ and A₂) and adrenergic (B) neurosecretory terminals was studied in the neurohypophysis of sexually mature female sturgeons. Neurosecretory terminals of the A₂ type prevail in the neurohypophysis, whereas A₁ and B terminals are rare. The activity of these types of terminals was established (1) during upstream migration, (2) shortly after spawning, and (3) three to six weeks after spawning. Terminals of B type are the most active elements during all the periods studied. These elements become strongly activated in sturgeons during upstream migration, i.e., earlier than the peptidergic neurosecretory terminals. Peptidergic terminals, especially elements of type A₂, become synchronously and strongly activated in fish shortly after spawning. In the late postspawning period neurosecretory terminals of all three types become synchronously inactive, persisting in a quiescent state in comparison to the two previous periods. The appearance of neurosecretory material discharged into the intercellular clefts by exocytosis correlates on the whole with the activity level of the A₁ and A₂ terminals in each individual studied. A functional correlation exists between the activity of the peptidergic and adrenergic neurosecretory terminals in the neurohypophysis. The data obtained are discussed with reference to a concept regarding spawning in some fish species as a physiological stress (Polenov et al., 1976). A possible dual control (peptide and monoamine neurohormones) over the function of visceral organs and glandular cells of the intermediate lobe of the hypophysis is also suggested (Polenov, 1970, 1975, 1978; Polenov and Belenky, 1973).Dedicated to the memory of Professor Wolfgang Bargmann, a great scientist and a generous friend