Innervation of the cultured fetal rat pancreas

Summary The fetal rat pancreas, explanted at 18 days of gestation and cultured up to ten days, contains numerous acetylcholinesterase-positive neurons. These nerves usually appear in small ganglia although single nerve cells are encountered. The axons of these intrapancreatic nerves appear to terminate only in the islet tissue and not on any exocrine components of the expiant. It is concluded that the fetal rat pancreas contains an islet-specific group of cholinergic neurons.We gratefully acknowledge the skilled technical assistance of Dan Whitehead and the secretarial assistance of Mary Pat Brady     

The coxal glands of geophilomorpha (chilopoda): Organs of osmoregulation

Summary The organs terminating at the coxal pores of the tug-legs of Geophilomorpha are not repugnatorial glands, but possess typical transport epithelia with deep apical and basal infoldings of the cell membranes, between which numerous large mitochondria are located. Many transport vesicles are found in the basal region but fewer in the apical cytoplasm. The apex is characterized by bundles of longitudinally oriented microtubules, sparse endoplasmic reticulum and free ribosomes. Single neurosecretory axons with synaptoid areas are scattered among the cells. It is suggested that the coxal organs have a diuretic function in moist habitats and an antidiuretic effect in arid environments. The switch-over is evidently controlled by a neuroendocrine mechanism.     

The effect of age on the ultrastructure of the mid-gut cells of the hymenopteran Nasonia vitripennis (Walk.) (Hymenoptera,Pteromalidae)

Summary The ultrastructure of the mid-gut cells of aged female Nasonia vitripennis is described. The mid-gut is a shrunken and distorted organ in the aged animal. The individual cells are highly disorganised and the organelle components are altered. The small lipid droplets formed in the apical cell region do not coalesce to form the large central lipid inclusions characteristic of the young animal. The rough endoplasmic reticulum is reduced and some of the mitochondria enlarge. The mid- and apical cell regions also contain large numbers of cytolysosomes. The basal cell region is essentially unchanged, but the channels formed by the infolded basal plasma membranes are dilated. The changes observed are discussed in relation to previous observations on other insect species.We are indebted to Professor E.W. Knight-Jones in whose Department this work was carried out, and to the Science Research Council for financial support to one of us (I.D.)     

Morphological aspects of the hypothalamic-hypophyseal system

Summary The ultrastructure of the tanycyte ependyma in male 160–180 g Wistar albino rats was studied under normal conditions and in experiments involving long-term suppression of ACTH secretion and its long-term stimulation. The former was accomplished by daily (for 8 days) intraperitoneal administration of dexamethasone phosphate at low (5 g/100 g) and high (100 g/100 g) concentrations. The effectiveness of suppression of the hypothalamic-hypophyseal-adrenal system in the experimental animals was judged by their reaction to two-minute ether stress (determination of plasma corticosterone) and by the results of measurement of the adrenal weights. Stimulation of ACTH secretion was achieved by bilateral adrenalectomy; the animals were examined on days 8, 10, 14, and 22 following the operation. The results obtained were in agreement with the previously established fact that there is a negative correlation between tanycyte activity and hypophyseal adrenocorticotrophic function (Akmayev and Fidelina, 1974). They also testified to the predominant involvement of the median eminence tanycyte ependyma (beta-tanycytes according to the authors' nomenclature) in these relationships.It is supposed that these correlations are regulated by a feedback mechanism and attest to the involvement of beta-tanycytes in the inhibiting control of hypophyseal adrenocorticotrophic function. The mechanism of this control may be explained alternatively: either the tanycytes transport ACTH-suppressing substances (catecholamines, corticosteroids, ACTH) from the CSF to the hypophyseal portal system or they themselves secrete substances possessing ACTH-suppressive activity. The authors distinguish several types of vesicles in the beta-tanycytes, the number of which changed with experimentally induced shifts in hypophyseal adrenocorticotrophic function. These vesicles are discussed in connection with the transport and secretory activity of the tanycytes and are considered to be a possible substrate of the hypothalamic inhibiting effect on ACTH secretion.     

Two major acrosomal proteins act on different parts of the oocyte vitelline coat in the abalone, Haliotis discus

Two proteins of molecular weights 20,000 (20K) and 15,500 (15.5K) are the major soluble substances released from the acrosomal vesicle of the abalone, Haliotis discus, spermatozoon. A crude preparation of them has been shown to possess lytic activity on the oocyte vitelline coat (VC). To elucidate the role(s) of each acrosomal protein (AP) in VC lysis, oocytes were examined after treatment with various AP preparations. The VC, which is about 1 micron thick, is composed of thin outer and inner electron-dense layers and a thick main layer of a fine filamentous feltwork. When oocytes were treated with a crude preparation containing both APs, the outer layer disappeared and the feltwork of the main layer loosened extensively. A preparation containing predominantly the 20K AP dissolved the outer layer completely and the main layer to some extent, whereas another preparation containing predominantly the 15.5K AP caused loosening of the main layer without alteration of the outer layer, suggesting that the 20K AP acts on the outer layer, whereas the 15.5K AP acts on the main layer. However, when purified, each AP by itself failed to dissolve the VC, although lysis occurred in a 1:1 mixture of these preparations. Moreover, when the oocytes were pretreated with the 20K AP and thoroughly washed, the 15.5K AP alone could induce lysis. These results suggest that the lysis of the outer layer requires both APs but not simultaneously. The 15.5K AP, which is located posteriorly in the acrosomal vesicle, must be released to act on the VC following the action of the 20K AP.     

Preparation of spermatogenic cell populations at specific stages of differentiation in the human.

Studying biochemical events in human spermatogenesis requires separated populations of spermatogenic cells. Dissociation of these cells was performed by a Trypsin-DNAse method adapted from the technique used for rodents. Cell separation was performed by centrifugal elutriation. Seven populations were collected, one further purified by Percoll gradient centrifugation, giving nine different cell populations. The efficiency of the cell separation was evaluated by phase contrast microscopy, flow cytometric DNA analysis, and electron microscopy. Five populations were enriched in spermatids: two in round spermatids (87% and 73%), another in round (52%) and elongating (44%) spermatids, another constituted by 80% elongating spermatids, and the last by 90% elongated spermatids. Two of the four remaining populations were enriched in primary spermatocytes (74% and 54%); another population was the upper part of the Percoll gradient and constituted cytoplasmic lobes and residual bodies (89%); the last population was made up of various cells, with no specific enrichment. Electron microscopic observations revealed good preservation of the separated cells; only the flagella from elongated spermatids were lost. Furthermore, an unusual pattern of nucleoplasm distribution during stages 2-4 of spermatid differentiation was observed and its signification is discussed with regard to the shape of the human spermatozoon.     

Ultrastructure of cuticular exudations in parasitic juvenileHeterodera schachtii,as related to cuticle structure

Summary The development ofHeterodera schachtii inside roots of a cruciferous host plant grown under monoxenic conditions in an agar medium was observed with video-enhanced contrast light microscopy. One to 6 days after inoculation, roots were excised and processed for electron microscopic observations. Exudates were present on the cuticle surfaces of J 2 and early J 3 juveniles located at feeding sites. Fibrillar exudations were correlated with similar fibrillar patterns in the epicuticle, exocuticle, intermediate zone, and the striated endocuticle. Secretion vesicles assembled at many Golgi sites in the hypodermis, appeared to coalesce and form large electron translucent vesicles in the cytoplasm. We propose that secretion vesicles migrate toward the cuticle, contact the plasmalemma and transfer their contents by exocytosis or a similar mechanism to a secretion accumulation site. These contents are associated with cuticle structure and emerge as surface exudations.     

Changes in the lysosome structure during the formation of zoospores inTrebouxia potteri

Summary Changes in the lysosome structures were examined by electron microscopy during the formation of zoospores inTrebouxia potteri. Lysosomes in vegetative cells were homogeneously filled with electron-dense material. At the beginning of zoospore formation, lysosomes invaginated or evaginated to take up mitochondria, ER, or cytoplasmic ground plasma. The ingested organelles became disorganized within the lysosomes. During this disruption of these organelles, the lysosomal contents became heterogeneous, suggesting a decrease in the amount of enzymes within the lysosomes. Golgi bodies and ER seemed to be involved with the disruption of the organelles, probably supplying some substances necessary for the functioning of the lysosomes. Amount of electron-dense materials decreased and, finally, only one to three small spherical aggregates remained in the lysosomes. Then the lysosomes appeared to shrink via loss of watery substances or cutting off of electron-transparent regions. After these changes in lysosome structure, nuclei started to divide successively for formation of the zoospores. The possibility is proposed that the drastic cytoplasmic changes operated by lysosomes trigger the following morphogenetic events in the formation of zoospores.Abbreviations ER endoplasmic reticulum - TGN trans Golgi network