Conrad W. Reifel

Structure and carbohydrate histochemistry of the intestine in ten teleostean species

The morphology and carbohydrate histochemistry of ten teleostean intestines are compared. Although there is an absence of regional differentiation seen in higher vertebrates, specializations in some species occur in the form of intestinal swellings, pyloric ceca and recta, the latter separated by a valve. The intestinal lumen is lined by a simple columnar epithelium interspersed with goblet cells; multicellular intestinal glands are absent. Thick basement membranes seen in centrarchids and Perca flavescens closely resemble the stratum compactum found in the lamina propria of esocids. Granular cells, which vary in number from species to species, are often seen in the mucosa and submucosa but less frequently in the muscularis. In species with intestino‐rectal valves, a rectum is easily defined by the abrupt appearance of lower mucosal folds, more goblet cells and a thicker muscularis. In the remaining species the above features appear gradually in the distal intestine. Goblet cells show species variations in localization of epithelial mucosubstances, which in broad terms are recognized as sulfomucins, sialomucins and neutral mucosubstances. In both proximal and distal intestines the majority of goblet cells contain sialomucin although small amounts of sulfomucin are also often present. In species without intestino‐rectal valves, no changes in carbohydrates occur between proximal and distal intestines. The possible significance of the heterogeneous character of digestive tract mucosubstances is discussed.

Adenohypophysis Has an Inherent Property for Pulsatile Prolactin Secretion

Hypophysectomized male rats with adenohypophyses grafted under the kidney capsule (HG rat) for 3 weeks with or without estradiol implantation, were used to examine prolactin (PRL) secretion independent of hypothalamic influence. Changes in the circulating PRL concentration were monitored by taking blood samples every 2 min via an indwelling atrial cannula. The circulating PRL concentration did not remain at a constant level but fluctuated with time. When estradiol (100 microgram/kg) was administered to HG rats through the indwelling cannula the size of the PRL pulses was magnified. The ability of the grafted adenohypophyses to generate pulsatile changes in the plasma PRL indicates that the adenohypophysis possesses an inherent interlactotroph communication system to permit synchronized PRL secretion without the direct involvement of hypothalamic PRL-releasing factors or inhibiting factors.

A possible phagocytic role for folliculo-stellate cells of anterior pituitary following estrogen withdrawal from primed male rats

SummaryUltrastructural changes suggesting a phagocytic role for the nongranular folliculo-stellate cells of the anterior pituitary are investigated in estrogen-primed male rats after withdrawal of estrogen. Morphological changes in mammotropes following the removal of a subcutaneous estradiol-containing Silastic implant include the formation of intracellular lipid bodies. These lipid bodies appear to be associated with enhanced estrogen-dependent prolactin secretion in mammotropes. Seven and 24 h after estrogen withdrawal intracellular lipid within mammotropes seems to be released into the intercellular space. Seventy-two h after estrogen withdrawal, lipid droplets are almost entirely cleared from mammotropes while folliculo-stellate cells become packed with lipid globules. Folliculo-stellate cells also undergo dramatic hypertrophy 7 and 24 h after the removal of E2-containing implants. Extensive intercellular junctions including zonulae adhaerentes, desmosomes, and putative gap junctions are formed. Intercellular junctions delineate extravascular channels into which numerous microvilli project. Folliculo-stellate cells appear capable of accumulating many lipid droplets, presumably related to mammotrope metabolism. What appear to be large secondary lysosomes as well as the lipid droplets are observed within folliculostellate cells; lipid, therefore, may be degraded through a lysosomal pathway in folliculo-stellate cells.

Recycling processes of cellular ascorbate generate oxidative stress in pancreatic tissues in in vitro system.

Ascorbate is a reducing agent, which is also known to oxidize cellular compoentns. Our proposed mechanism of the oxidative action is as follows: Ascorbate is concentrated in the pancreas and is leaked in adverse conditions, and oxidized to dehydroascorbate. The dehydroascorbate is carried into cells by a glucose transporter (GLUT) and reduced back to ascorbate. The reduction processes take electrons from other cellular components. Ascorbate or dehydroascorbate treatment elevated thiobarbituric acid-reactive substance (TBARS) concentrations in pancreas. The elevation in TBARS concentrations were blocked by cytochalasin B, a GLUT inhibitor. To confirm further the prooxidative action, changes in glutathione content were quantified. Glutathione concentrations were lower in ascorbate- or dehydroascorbate-treated groups. The ascorbate-induced decrease in glutathione was blocked by cytochalasin B. To prevent oxidation of ascorbate to dehydroascorbate, glutathione was added to the medium. The ascorbate plus glutathione and dehydroascorbate plus glutathione groups showed lower TBARS concentrations than those of the ascorbate and dehydroascorbate groups, respectively. There were changes in the morphology of Langerhans islets following ascorbate treatment, which disappeared following treatment with ascorbate plus cyto-chalasin B. The observations indicate that ascorbate generates oxidative stress and affects the structure of islets.

Ultrastructural changes rapidly induced by somatostatin may inhibit prolactin release in estrogen-primed rat adenohypophysis

SummaryApart from the known hypothalamic controls, which have been well documented, a myriad of compounds both endogenous and exogenous have proved effective in influencing secretion of prolactin (PRL). Recent studies have shown that somatostatin (SRIF), when injected intra-atrially as a bolus, is able to inhibit PRL secretion in vivo. However, the inhibitory effect of SRIF occurs only in adenohypophyses previously primed with estradiol. This research was undertaken to examine the ultrastructural effects of that inhibition using male Sprague-Dawley rats primed for three weeks with subcutaneous implants of estradiol. Within 2 min of injection of SRIF (1 mg/kg), the adenohypophyses were removed and processed for electron microscopy. We found dramatic changes in the estradiol-primed mammotrophs, including 1) an apparent rearrangement of rough endoplasmic reticulum (RER) into concentric cisternae, and 2) the appearance of intracellular bodies closely associated with granules. These changes were not observed in non-estradiolprimed male rats injected with SRIF which is consistent with the fact that in the normal male rats, SRIF failed to inhibit PRL secretion. These findings suggest that SRIF causes reorganization of cellular organelles so that PRL granules are sequestered thereby inhibiting secretion of PRL.

Extensive ultrastructural changes in rat mammotrophs following administration of the dopamine agonist ergocristine-reflecting inhibition of prolactin release.

SummaryWe have demonstrated an extensive reorganization of organ elles in mammotrophs immediately following administration of ergocristine (a dopamine agonist) to estradiol-primed male rats. Our ultrastructural findings are consistent with our previous results that ergocristine can block prolactin release without any noticeable latent period. Following three-week priming of male rats with estradiol implants, ergocristine was administered by a bolus injection through an indwelling cannula. Within two min of its administration, ergocristine induced dramatic changes in the ultrastructure of mammotrophs, i.e., (1) increased numbers of secretory granules, (2) peripheral relocation of rough endoplasmic reticulum which tends to sequester secretory granules, (3) change in location of nucleus and (4) increased numbers of intracellular bodies associated with secretory granules. We suggest that the extensive ultrastructural changes that occurred in such a short period following ergocristine administration may be indications of specific factors associated with blockage of hormone release.

Investigation of a cyclopic, human, term fetus by use of magnetic resonance imaging (MRI)

Using magnetic resonance imaging (MRI), the internal neural and craniofacial malformations of a cyclopic fetus are described. External facial features were characterized by a tubular proboscis situated above a single eye slit. The brain was recognized as ‘pancake’ type alobar holoprosencephaly (a condition where the undifferentiated telencephalon partially surrounds a monoventricle). Displacement of some bones that normally contribute to the orbit could be clearly discerned. Absence of neural structures (e.g. falx cerebri, corpus callosum) and missing components of the ethmoid bone indicated a midline deficit. This correlates with proposed theories of cyclopic embryopathy, which suggest that the prechordal plate and the neural crest cells are affected during the third week of gestation in cyclopia.

Cysteamine acts immediately to inhibit prolactin release and induce cellular changes in estradiol-primed male rats

Cysteamine (CSH) has an inhibitory effect on the secretion of PRL, GH and TSH. The purpose of the present study was twofold: (1) to examine the short-term effects of CSH on PRL secretion, and (2) to elucidate the mechanism of action of CSH. CSH (100 mg/kg) was injected as an intra-atrial bolus to both normal and estradiol-primed male Sprague-Dawley rats. Blood samples were drawn every 2 min for 1 h and assayed for PRL. To determine if the actions of CSH involved PIF release or PIF receptors, a dopamine receptor blocking agent, pimozide (100 micrograms/kg) was given intra-atrially 1 h after the CSH injection and PRL concentration was monitored. Within 6 min after the CSH injection PRL levels in estradiol-primed rats began to fall, and by 30 min postinjection the levels were well below baseline values. Blocking the dopamine receptors with pimozide did not result in increased PRL levels, indicating that CSH was acting through some mechanism not directly involving dopaminergic PIF receptors. In a separate study, 6 min after the injection of CSH the rats were decapitated and their anterior pituitaries were removed and processed for electron microscopy. Only mammotrophs from estradiol-primed rats treated with CSH showed an extensive rearrangement of rough endoplasmic reticulum (RER) in concentric whorls around the periphery of the cells. These studies show that the inhibition of PRL secretion by CSH begins within 6 min, and is appreciable only when basal PRL levels are increased by estrogen priming.(ABSTRACT TRUNCATED AT 250 WORDS)

Desmosomes between Mammotrophs Suggest the Existence of a Functional Syncytium

Lactotrophs in the adenohypophysis have the apparent inherent capability to produce simultaneous secretion of prolactin from individual cells. There are two possibilities to account for this synchronized, discrete pulsatile secretion: (1) humoral communication, or (2) cell-to-cell syncytial communication. As a first step in examining the second possibility, we searched for ultrastructural evidence of intercellular contacts. The presence of desmosomes, the conventional junctions that serve to anchor adjacent cells, would be the first step in proving contact between homologous cells. In order to increase the observation of desmosomes, the population of lactotrophs in the adenohypophysis was increased by implantation of capsules containing estradiol in male Sprague-Dawley rats. The adenohypophyses were then examined by electron microscopy for indications of these intercellular bridges. Desmosomes were discovered between lactotrophs, indicating that cell-to-cell contact does exist, and suggesting that the desmosome may help the formation of a syncytium among lactotrophs, either directly or indirectly. Since an ectopic pituitary without direct help from the hypothalamus can secrete prolactin in a pulsatile fashion and since there are desmosomes between lactotrophs, it is possible that the synchronized prolactin release occurs through a direct cell-to-cell communication system which is linked by the desmosomes.

Localization of acid phosphatase in lysosomes of pituitary folliculo-stellate cells following estrogen withdrawal from primed male rats

SummaryAcid phosphatase was localized in lysosomes of non-granulated, folliculo-stellate pituitary cells 72 h after estrogen was withdrawn from primed male rats. The appearance of lysosomes and the accumulation of lipid droplets in folliculo-stellate cells coincide with the clearance of lipid from pituitary mammotropes. Positive identification of lysosomes supports our proposal in a previous study that folliculo-stellate cells are phagocytic.