Pituitary response to environmental osmotic changes in the larval guppy,Lebistes reticulatus (Peters)

Summary The pituitary of the larval guppy, Lebistes reticulatus (Peters), was investigated cytologically in relation to the osmotic properties of the environment. In larvae of Stage I, removed from the mother about one week before birth, the sizes of the nuclei of the prolactin, STH, ACTH, and pars intermedia cells were small. The nuclei of these cell types became larger when Stage I larvae were transferred into fresh water (F. W.); however, these animals did not survice. In Stage-II larvae, removed from the mother just before birth, the nuclei of the abovementioned adenohypophysial cells were larger than those of the Stage-I larvae. Stage-II larvae were able to survive in F. W. Thus, it seem that osmoregulatory mechanisms in the larval guppy develop between Stages I and II.The nuclei of prolactin, STH, ACTH and pars intermedia cells decreased significantly in size after birth (0-hr). When larvae of the 0-hr group were immersed in 1/3 sea water (S. W.), only the nuclei of the ACTH cells increased in size. The neurohypophysis of Stage-I larvae contained a very small amount of aldehyde-fuchsin (AF)-positive material, while the neurohypophysis of Stage-II and 0-hr fish was rich in this material. When Stage-I larvae were immersed in F. W., the neurohypophysis showed as great an accumulation of AF-positive material as Stage-II and 0-hr fish.Supported by grants from Japan Society for Promotion of Science and from Ministry of Education of Japan to Professor Kobayashi, and by grants from the Deutsche Forschungsgemeinschaft to Professor Oksche. A part of this investigation was effected while Professor Kobayashi held a Visiting Professorship at the University of Giessen. We are indebted for histological assistance of Miss I. Woltmann, Mrs. H. Schneider and Mrs. T. Bremer.     

The rostral pars distalis of the pituitary gland of the freshwater and marine alewife (Alosa pseudoharengus)

Summary In the alewife the orohypophyseal duct, a remnant of Rathke's pouch, persists in adults as a tube passing from the rostral pars distalis to the pharyngeal region. Its lumen is not open to the buccal cavity. The prolactin cells are situated around the bifurcations of this duct in the rostral pars distalis. Contents from prolactin cells, such as granules, nuclei, mitochondria and Golgi structures were found in these bifurcations. These contents were indistinguishable from those of intact prolactin cells. Evidence of actual release into the duct was often noted. At the presumptive point of release, the cells lining the lumen separate and the contents, probably of an entire prolactin cell, are extruded. The cilia usually found at the point of extrusion arise from prolactin cells. The prolactin cells of freshwater fish were more heavily granulated than those from a marine environment. Prolactin cells of fish entering freshwater streams were not heavily granulated but showed evidence of increased activity. Granule size was not affected by salinity. The ACTH cells are arranged in bands along branches of the neurohypophysis in the rostral pars distalis. No differences in ACTH cells from fish of different salinities were noted.We would like to thank Mr. D. D. Zumwalt of the John G. Shedd Aquarium in Chicago, Dr. E. D. Warner, Mr. R. L. Flayter, Dr. J. G. Stanley of the University of Wisconsin, Milwaukee, Mr. L. Wells of the U. S. Fish and Wildlife Service, Great Lakes Fisheries Laboratory, Ann Arbor, and Mr. L. N. Flagg of the Department of Sea and Shore Fisheries, Augusta, Maine, for their assistance in obtaining the fish used in this study. Dr. T. N. Tahmisian and Mr. G. T. Chubb of Argonne National Laboratories, and Dr. L. M. Srivastava and Dr. V. Bourne of Simon Fraser University, Canada, kindly made electron microscope facilities available. Finally, we wish to thank Mr. W. Goossens and Mr. D. J. DeJong for valuable assistance. This project was supported by a grant from the National Marine Fisheries Service.     

Circadian changes in prolactin cell activity in the pituitary of the teleost Poecilia latipinna in freshwater

Summary Quantitative morphometric studies with the electron microscope were made on the prolactin cells of wild freshwater sailfin mollies taken in Florida in August at four different times of day. The results indicate a circadian rhythm in the prolactin cell, the period of highest synthetic activity being from midday to evening, as indicated by the condition of nucleus, Golgi apparatus and rough endoplasmic reticulum, and by the incidence of granule release profiles. No circadian changes were detected in the ACTH cells.Several distinct sites of prolactin granule release were recognised. However, there was no conclusive evidence of granule release by any mechanism other than classical exocytosis.We thank Mr. W. Thomson for technical assistance and Ann Grier for help in collecting the fish. T.F.C. Batten is in receipt of an S.R.C. Research Studentship.     

Fine structure of the adenohypophysis in immature sockeye salmon,Oncorhynchus nerka

Summary The ultrastructure of the secretory cells of the adenohypophysis of juvenile sockeye salmon was investigated. Pituitary glands were collected from immature fish transferred experimentally to sea water and subsequently returned to fresh water. The rostral pars distalis contained three cell types: ACTH cells, prolactin cells, and non-secretory cells. The prolactin and non-secretory cells were joined together in the form of follicles by desmosomes and they both had cilia and microvilli projecting into the follicle lumen. Various follicular structures such as lumen, multivesicular structures, and peripheral basement membrane are discussed as possible sites of prolactin cell granule release. The columnar ACTH cells were found at the junction of the rostral pars distalis and the neurohypophysis. The cytoplasmic granules in these cells were characteristically separated from their limiting membrane by a clear space. Multivesicular structures were also found in association with this cell type. The caudal pars distalis also contained three cell types: one acidophil (putative somatotrop) and two basophils (putative thyrotrops and gonadotrops), all of which were similar to those described in adult fish. The pars intermedia contained only one cell type. They appeared to be active cells and were characterized by containing membrane-bounded granules similar to those found in the ACTH cells. Changes in ambient salinity had no apparent effect on any cell type described.The work was supported by a grant in aid of research from the National Research Council of Canada. We wish to thank Mr. R. Lindsay, Mr. C. Cooper, and Mr. G. Longworth for their technical assistance. We would also like to thank Mr. S. Killick of the International Pacific Salmon Fisheries Commission for his assistance in the collection of fish and Dr. H. Cook for his helpful discussion of the project. This paper is No. 058 in the University of Guelph Migration Series.     

Immunocytochemical identification and ontogeny of adenohypophyseal cells in a cave fish,Phreatichthys andruzzii (Cypriniformes: Cyprinidae)

The morphogenesis of the pituitary gland and the chronological appearance of adenohypophyseal cells were investigated for the first time in the Somalian cave fish Phreatichthys andruzzii by immunocytochemistry. The adult adenohypophysis contained: a rostral pars distalis, with prolactin (PRL) cells arranged in follicles and adrenocorticotropic (ACTH) cells, a proximal pars distalis with somatotropic (GH), β‐thyrotropic (TSH), β‐gonadotropic type I (FSH) and type II (LH) cells and a pars intermedia with α‐somatolactin (SL), α‐melanotropic (MSH) and β‐endorphin (END) cells. All regions were deeply penetrated by neurohypophyseal branches. At hatching (24 h post‐fertilization) the pituitary was an oval cell mass, close to the ventral margin of diencephalon. The first immunoreactive cells appeared as follows: PRL at 0·5 days after hatching (dah), GH and SL at 1·5 dah, END at 2 dah, TSH, ACTH and MSH at 2·5 dah, FSH at 28 dah and LH at 90 dah. The neurohypophysis appeared at 5 dah and branched extensively inside the adenohypophysis at 130 dah, but there was no boundary between rostral pars distalis and proximal pars distalis at this stage. The potential indices of prolactin and growth hormone production increased until 28 and 60 dah, respectively. The potential index of growth hormone production correlated positively with total length. Activity of PRL and GH cells, measured as ratio of cell area to nucleus area, was significantly higher in juveniles than in larvae.     

Histogenesis of the pituitary in rainbow trout (Salmo gairdneri) in different ambient salinities with particular reference to the rostral pars distalis

Summary The pituitary gland is first evident in rainbow trout two weeks before hatching. Differentiation of prolactin and ACTH cells is not marked until 3–4 days after hatching when the follicular arrangement of the prolactin cells become apparent. There is no difference in the time of development of either prolactin or ACTH cells in larvae reared in different ambient salinities despite marked changes in tissue ion and water content. This suggests that prolactin and ACTH do not play the osmo- and iono-regulatory roles in larval rainbow trout that they are considered to play in adult salmonids.The work was supported by a grant-in-aid of research from the National Research Council of Canada     

Allometric relations of total volumes of prolactin cells and corticotropic cells to body length in the annual cyprinodont Cynolebias whitei: effects of environmental salinity,stress and ageing

Summary An analysis of the allometric relations of the total volumes occupied by prolactin (PRL) and corticotropic (ACTH) cells (PRL volume and ACTH volume, respectively) to body length and a study of the immunocytochemical staining intensity of PRL and ACTH cells were used to determine the differences in activity of PRL and ACTH cells in freshwater-reared and in saltwater-reared Cynolebias whitei during the entire lifespan of this annual cyprinodont fish. An inflection in the allometric relation of PRL volume to body length was observed in fish of one-week old. The relatively large PRL volume in younger fish may be related to PRL cell activity before hatching. No inflections were observed in the allometric relations of PRL volume and ACTH volume to body length at the onset of maturation and the onset of ageing, indicating that the increased pituitary growth in maturing and ageing C. whitei is not the result of changes in PRL or ACTH cells. The slope of the allometric relation of PRL volume to body length in freshwater-reared fish was significantly steeper than the slope in saltwater-reared fish. The PRL volume in adult freshwater-reared fish was eight times larger than that in saltwater-reared fish of the same length. The intensity of immunocytochemical staining of saltwater PRL cells was significantly reduced. These volumetric and staining differences correspond to the low functional demand put upon PRL cells in saltwater-adapted fish. In contrast, the slope of the allometric relation of ACTH volume to body length and the intensity of immunocytochemical staining of ACTH cells were similar in freshwater-reared and in saltwater-reared fish. It is concluded that the functional demand put upon ACTH cells is similar in freshwater-reared and saltwater-reared C. whitei; the involvement of ACTH cells in the osmoregulation of the fish in both environments is similar.     

Quantitative ultrastructural evidence of alterations in prolactin secretion related to external salinity in a teleost fish (Poecilia latipinna)

Summary Quantitative ultrastructural morphometric studies were made on the prolactin cells of Poecilia latipinna adapted to freshwater (FW), one-third seawater (1/3 SW) and full-strength seawater (SW), and at various times after transfers between 1/3 SW and FW.In fully-adapted fish the rates of prolactin (PRL) synthesis and PRL release are inversely related to environmental salinity. During adaptation to a new salinity the two rates are temporarily uncoordinated, with release increasing or decreasing more readily than synthesis. Synthesis appears to take 30 h or longer to come into balance with the increased release rate following transfer from 1/3 SW to FW, and 72 h or longer to adjust to the reduction in release rate that follows the reverse transfer. The excess PRL granules that accumulate in the latter situation appear to be removed by lysosomal digestion. As in other teleosts, in fish adapted to the external medium the size of the stored PRL granules is inversely related to external salinity, but this relationship breaks down during adaptation to a new salinity.The stellate cells which penetrate between the PRL cells are more prominent, more extensively ramified, and appear more metabolically active in FW-adapted fish than in the other groups. These cells seem to be closely related in function to the secretory activity of the PRL cells.We thank Mr. W.A. Thomson and Mr. D.I. Hollingworth for technical assistance and Dr. D.I.C. Pearson (Department of Physics, University of Nancy, Nancy, France) for advice on mathematical analysis and computer programs. The work was carried out during tenure of an S.R.C. Studentship by T.F.C. Batten     

Ultrastructure of the neurohypophysis of the teleost Poecilia latipinna in relation to neural control of the adenohypophysial cells

Summary The structure of the neurohypophysis of Poecilia latipinna (green molly, sailfin molly) was studied with the electron microscope. Profile diameters of neurosecretory granules in the non-myelinated neurohypophysial nerve fibres were measured and mathematically corrected for error due to section thickness. Six different types of nerve fibres could be distinguished by statistical classification of their granules and by other ultrastructural features. One fibre-type (type B) contained granules with a mean diameter of 85 nm, and the other five types (types Ala, Alb, A2, A3 and A4) all contained granules with mean diameters greater than 100 nm. Synaptic contacts were observed between type B fibres and all the adenohypophysial cell-types, although in the case of the ACTH cells the synapses were separated from the cell membrane by a continuous double basement membrane. Type A fibres were observed to contact the cells of the proximal pars distalis and pars intermedia, but did not form synapses. However, synapses occurred between type A fibres and pituicytes, and between type A fibres and the pericapillary basement membrane in the interior of the neurohypophysis. The possible roles of the different types of nerve fibres in controlling the adenohypophysial cells are discussed in the context of evidence from other teleosts.We thank Mr. W.A. Thomson and Mr. D.I. Hollingworth for technical assistance, and Dr. D.I.C. Pearson (Department of Physics, University of Nancy, Nancy, France) for advice on mathematical analysis and computer programs. The work was carried out during the tenure of an S.R.C. Research Studentship by T.F.C.B.     

The effects of osmotic experiments and prolactin on the mucous cells in the skin and the ionocytes in the gills of the teleost Cichlasoma biocellatum

Summary Adult Cichlasoma biocellatum were kept in deionized water, 25% sea-water and tap-water. The mucous cells in the epithelium of the gills and the buccal floor were not affected by the osmotic experiments. In animals kept in deionized water the mucous cells in the skin remained unaffected, but the ionocytes (chloride cells) in the gills were strongly stimulated. Keeping the animals in salt water led to a strong regression of the epidermis including the mucous cells, and of the ionocytes. The regression could be counteracted by prolactin injections. The changes in the skin and the ionocytes could be correlated with the activity of the prolactin producing cells in the adenohypophysis. It is suggested that the epidermis and its mucous cells as well as the ionocytes are under prolactin control.The authors gratefully acknowledge the interest and helpful suggestions of Prof. Dr. P. G. W. J. van Oordt. Thanks are also due to Mr. H. van Kooten for making the photographs, to Dr. L. Boomgaart for correcting the english, and to the N. I. H., Bethesda, Maryland (U.S.A.) for presenting the ovine prolactin.     

Extravascular circulation in the pituitary of Mugil cephalus (Teleostei)

Summary Extravascular circulation in the pituitary of Mugil cephalus was investigated by injecting live fish with horseradish peroxidase and studying the distribution of the enzyme in the gland. The principal components of the extravascular circulatory system are the pericapillary spaces, and, arising from them, the interlobular and circumhypophyseal spaces. Extensions of these spaces penetrate the glandular parenchyma of the pars distalis, where they merge with pericellular spaces. In the neurohypophysis, pericapillary spaces are connected to the periaxonal spaces.Capillaries penetrating from the proximal neurohypophysis into the pars distalis are accompanied by neurosecretory axons. These axons form a mass of tissue which is limited near the capillaries by the pericapillary spaces and near the adenohypophysis by the interlobular spaces. Toward the interior of the adenohypophysis the amount of nervous tissue accompanying the capillaries progressively diminishes, thus reducing the distance between pericapillary and interlobular spaces. Within the pars distalis, the neurosecretory axons accompanying the capillaries are sparse, and the secretory and stellate cells are mostly located directly adjacent to the pericapillary spaces. In the neuro-intermediate lobe, interlobular spaces outline the neuro-adenohypophyseal boundary.The relationship between extravascular spaces and hormone-secreting cells varies in the different regions of the adenohypophysis depending upon the type of neurosecretory innervation in the respective region. In the regions of prolactin and gonadotropin cells, where neurosecretory axons are in direct contact with the secretory cells, the hormone-secreting and stellate cells are adjacent to the pericapillary spaces. In the regions of ACTH and STH cells, secretory and stellate cells are found adjacent to the interlobular spaces, which are interposed between the cells and the neurosecretory axons.Abbreviations AH adenohypophysis - CH circumhypophyseal - DNH distal neurohypophysis - HRP horseradish peroxidase - NH neurohypophysis - NS neurosecretory - PD pars distalis - PI pars intermedia - PPD proximal pars distalis - RNH rostral neurohypophysis - RPD rostral pars distalis This research was supported by a grant from the National Council for Research and Development, Israel, and the GKSS Geesthacht-Tesperhude, Federal Republic of Germany     

Long-term salt loading impairs pituitary responsiveness to ACTH secretagogues and stress in rats

Male Wistar rats were allowed to drink tap water ad lib (W), 2% saline (S) or 2% saline containing dexamethasone (S + D, 1 mg/l) for 7 days. On the 8th day rats were subjected to a 3-min ether stress. Plasma ACTH, corticosterone and prolactin concentrations were determined before and after ether exposure. Prestress concentrations of plasma ACTH were low and did not vary among the three groups. In response to ether stress W rats exhibited twice as high plasma ACTH concentrations as did S rats. Rats of the S + D group exhibited a small but statistically significant ACTH response. Plasma corticosterone concentration in S rats was increased while in S + D rats was significantly decreased under resting conditions compared to that in W rats. Ether stress caused large increases in plasma corticosterone concentrations in W and S rats while a small but statistically significant increase was observed in S + D rats. Prolactin responses to ether were smaller in groups S and S + D than in group W. To test whether the decreased ACTH response to ether exposure was a result of a decreased sensitivity of corticotrope cells to corticotropin releasing factor (CRF)-41 or arginine vasopressin (AVP), adenohypophysial fragments from W, S and S + D rats were incubated in the presence of different doses of CRF-41 or AVP. Pituitary fragments obtained from W rats secreted larger amounts of ACTH than did pituitaries from S rats in response to either CRF-41 or AVP.(ABSTRACT TRUNCATED AT 250 WORDS)     

Evidence for corticotropin releasing factor (CRF) synthesis in the preoptic nucleus of Xenopus laevis tadpoles

Summary For the study of the hypothalamo-hypophysial system of Xenopus laevis tadpoles, hypothalamic lesions were made by means of the electrocoagulation technique. Lesioning of the ventral region of the preoptic nucleus resulted in a decrease of the number of ACTH cells in the pars distalis of the pituitary gland and in a diminution of the PAS-positive reaction of these cells. In addition, regeneration of the neurosecretory cells of the ventral region of the preoptic nucleus observed 6 weeks after lesioning was accompanied by the reappearance of normal PAS-positive ACTH cells in the pars distalis. It is suggested that the neurosecretory cells of the ventral region of the preoptic nucleus of Xenopus laevis tadpoles are related to the ACTH synthesizing cells, probably by producing CRF.Dedicated to Prof. Dr. med. W. Bargmann on the occasion of his 70th birthdayThe authors thank Prof. Dr. J.C. van de Kamer for his interest, Prof. Dr. P.G.W.J. van Oordt for his many helpful comments, and Messrs. H. van Kooten, E. van der Vlist, J.J. van der Vlis and M.C.A. van Pinxteren for preparing the illustrations     

Prolactin evokes lactational transmission of larvae in mice infected with Toxocara canis

We investigated the trans-lactational maternal–neonatal transmission of Toxocara canis larvae in mice, with particular interest in the role of prolactin in their migration to the mammary gland. Two female mice were infected with 300 T. canis eggs soon after delivery of 27 offspring. After 1 week of breast-feeding, seven larvae were recovered from 4 of 13 offspring. After 2 weeks of lactation, 101 larvae were recovered from all the remaining offspring. Daily prolactin administration (5 μg) was performed 2 weeks before T. canis infection and continued until 2 weeks after infection in six non-pregnant female mice, which resulted in larval accumulation in the mammary gland. Furthermore, prolactin administration in female mice that had been infected with T. canis 4 weeks prior to prolactin treatment induced migration of larvae into the mammary gland. These findings suggest that prolactin is a promoting factor contributing to lactational transmission of T. canis larvae in mice.     

Histology of the pituitary gland of the grey mullet, Mugil cephalus L.

The present communication deals with a histological study of the pituitary gland of the teleost fish Mugil cephalus , found in the estuarine waters of Cochin area. Six different cell types were identified in the pituitary gland on the basis of their grouping, distribution and staining properties. The prolactin and the TSH cells (thyroid stimulating hormone producing cells or thyrotrops) were identified in the rostral pars distalis and the ACTH cells (adrenocorticotropic hormone producing cells or corticotrops) in the interphase between the neurohypophysis and the rostral pars distalis. The STH cells (somatotropic hormone producing cells or somatotrops) and the gonadotropic cells were distinguished in the proximal pars distalis and the MSH cells (melanin stimulating hormone producing cells or melanotrops) in the pars intermedia.     

Cytological and ultrastructural changes in the pituitary pars distalis of the goldfish kept in calcium-free environments

The cytology and ultrastructure of the pars distalis, mainly that of prolactin (PRL) cells, were investigated in goldfish adapted to fresh water (FW) or deionized water (DW) for 20 and 40 days, or gradually adapted to 1/3 artificial sea water (ASW) or 1/3 Ca-free sea water. When compared to PRL cells of goldfish kept in FW, those of goldfish adapted to DW did not show signs of increased activity. The lack of exocytotic activity and the low development of various organelles suggested that cell activity was slightly reduced. In 1/3 ASW, PRL cells were smaller and less active. In 1/3 Ca-free ASW, PRL cells appeared slightly stimulated compared with those of fish in 1/3 ASW. The Golgi area was more developed and a few lamellae of endoplasmic reticulum were observed in some cell islets. However, there was no significant difference between PRL cells of goldfish kept in 1/3 Ca-free ASW and in FW. In 1/3 ASW, which is isosmotic to the blood, thyrotrophs (TSH cells) corticotrophs (ACTH cells) and somatotrophs (STH cells) were not clearly affected. In DW, these cells and their nuclei were significantly enlarged. Their stimulation was also evident in 1/3 Ca-free ASW; values for cellular and nuclear areas were maximal in this environment and significantly higher than those of fish in FW and 1/3 ASW. These data suggest that in addition to the PAS-positive cells of the pars intermedia, highly stimulated in Ca-free environments, other cell types of the pars distalis may be involved in osmoregulation, and that the role of PRL cells is not primordial in the goldfish.     

Prolactin and growth hormone cells in the human hypophysis: A study with immunoenzyme histochemistry and differential staining

Growth hormone and prolactin cells were immunostained in human hypophyses with antibody against rat growth hormone or prolactin and the peroxidase-antiperoxidase complex. Growth hormone cells were round and, in normal pituitaries, arranged in sizable groups. Prolactin cells occurred singly and were less numerous; they were often extensively branched. Only a few prolactin cells stained with carmoisine. Incubation of the antibody with an excess of the appropriate antigen greatly diminished or abolished immunostaining; absorption of anti-prolactin with growth hormone often enhanced it. Prolactin cells were somewhat hypertrophied and hyperplastic in a neonate. Many of them stained with carmoisine. An even greater hypertrophy and hyperplasia of these cells (which pushed apart the growth hormone cells) was found in a lactating woman. Immunostained giant prolactin cells were also observed. Staining of the prolactin cells with carmoisine was extensive. Upon prolonged exposure to anti-growth hormone antibody, ACTH/MSH cells also showed immunostaining which was abolished by absorption of the antiserum with growth hormone but not with synthetic 1-24ACTH. Growth hormone cells evidently correspond to the alpha acidophils of Romeis, prolactin cells in lactation to his eta cells; the relation of his epsilon cells to the pleomorphic "resting" prolactin cells is not clear.     

Immunocytochemical identification of prolactin cells in the pituitary gland of tilapia larvae (Oreochromis mossambicus: Teleostei)

Summary Using an antiserum to highly purified chum salmon prolactin, prolactin cells were identified in the putative rostral pars distalis of newly hatched tilapia larvae (Oreochromis mossambicus) by the immunogold method for the electron microscope. In the putative rostral pars distalis, some cells had another kind of secretory granule which was much less numerous, much smaller in size, and without immunoreactivity to salmon prolactin antiserum. Controls incubated with salmon prolactin-preabsorbed antiserum or normal serum showed no immunoreactive cells, confirming the specificity of the antiserum. The possible role of prolactin in the osmoregulation of tilapia larvae is discussed.     

Mate recognition inCryptomyzus aphids: copulation and insemination

The acceptability of three widely distributed Australian Menispermaceae,Tinospora smilacina Benth.,Sarcopetalum harveyanum F. Muell. andStephania japonica (Thunb.) Miers, as food for larvae of the fruitpiercing moth,Othreis fullonia (Clerck), was examined in three laboratory experiments. When larvae were presented with plant species individually total development times were shortest onT. smilacina and longest onS. japonica, despite relatively similar consumption rates within most instars.T. smilacina elicited greater (P<0.05) relative growth rates thanS. japonica in all instars except the 6th. In the second experiment, when larvae were allowed to select from each of the 3 plants, noS. japonica was chosen by 1 st instars and it represented only 3.7% of food consumed by 2nd instars. Significantly moreT. smilacina was eaten in each instar thanS. japonica, and more thanS. harveyanum except in the 2nd and 4th instars. The final experiment examined the abilities of larvae to switch hosts when forced after the 1st and 3rd instars. After the first or second food change largest average headcapsule widths were associated with feeding onT. smilacina as the most recent food. Feeding by final instars onT. smilacina also resulted in the shortest development time and highest puparial weights. While some larvae survived irrespective of plant sequence 83.3% of the recorded mortality occurred while larvae were exposed toS. japonica, principally during the 1st instar. These experiments lend support to field observations which suggest thatT. smilacina is a major host ofO. fullonia whileS. japonica is notS. harveyanum is probably an important alternate host whenT. smilacina is scarce.     

Immunohistochemical detection of ACTH and MSH cells in the hypophysis of the hermaphroditic teleost, Diplodus sargus

Hypophyseal ACTH and MSH cells were immunohistochemically characterised in the teleost fish, Diplodus sargus, using anti-ACTH (1-24) and anti alpha-MSH polyclonal antisera. ACTH cells were found both in the pars distalis and in the pars intermedia. In the former region, they appeared small, round-shaped and clustered; in the latter, they were either small or large and elongated. Moreover, a few ACTH-immunoreactive cells resembling microglia were present in the neurohypophysis. Conversely, MSH cells were found only in the pars intermedia, and were similar to the larger ACTH cells of the same region. In the pars intermedia, co-localisation of ACTH and MSH immunoreactivity in the same cell was revealed by double immunostaining, though the two hormones were also observed in distinct cell types. The distribution of ACTH cells appeared quite uniform, without any marked difference between the specimens tested. Conversely, MSH cell amount varied according to the stage of the sexual cycle of this teleost fish, which is characterised by protandrous hermaphroditism. In fact, a lower amount of MSH cells were observed in females, whereas no significant difference was found between immature and male specimens.