The effects of slight pressure oscillations in the far infrasound frequency range on the pars flaccida in gerbil and rabbit ears

This study was designed to clarify whether the pars flaccida (PF) as a flexible part of the tympanic membrane is capable of reacting to pressure oscillations (PO) with amplitudes and frequencies typical for natural atmospheric pressure fluctuations in the far infrasound frequency range (APF). If so, the PF mechanical reactions to APF might be involved in the overall physiologic regulation processes, which make organisms susceptible to APF. The displacements of the PF in response to PO were measured in vitro in ears of gerbils and rabbits by means of laser Doppler vibrometry. The index of the PF reactivity (R_a) was determined as the ratio of the amplitude of the PF oscillations (PFO) to the amplitude of the PO. All kinds of PO applied caused PFO. The amplitude of the PFO increased when the amplitude of the PO was increased. In gerbils, a decrease in R_a with the increase in amplitude of the PO was observed. In the range of PO lowest amplitudes (4–20 Pa) R_a proved to be 1.4 times higher than in the range of highest amplitudes (90–105 Pa). Considering that the natural APF are usually within the range of ±20 Pa, this fact points to an important contribution of the PF to the pressure dynamics in the middle ear (ME) of gerbils. In rabbit ears, R_a was lower and recovery from plastic deformation was slower than in gerbils. Our findings are in line with the suggestion that the PF might play an important role in respect of adaptation to natural APF.     

Peptide immunocytochemistry of neurons projecting to the retrocerebral complex in the blow fly, <Emphasis Type="Italic">Protophormia terraenovae</Emphasis>

Antisera against a variety of vertebrate and invertebrate neuropeptides were used to characterize neurons with somata in the pars intercerebralis (PI), pars lateralis (PL), and subesophageal ganglion (SEG), designated as PI neurons, PL neurons, and SEG neurons, respectively, all of which project to the retrocerebral complex in the blow fly, Protophormia terraenovae. Immunocytochemistry combined with backfills through the cardiac-recurrent nerve revealed that at least two pairs of PI and SEG neurons for each were FMRFamide-immunoreactive. Immunoreactivity against [Arg7]-corazonin, beta-pigment-dispersing hormone (beta-PDH), cholecystokinin8, or FMRFamide was observed in PL neurons. Immunoreactive colocalization of [Arg7]-corazonin with beta-PDH, [Arg7]-corazonin with cholecystokinin8, or beta-PDH with FMRFamide was found in two to three somata in the PL of a hemisphere. Based on their anatomical and immunocytochemical characteristics, PI neurons were classified into two types, PL neurons into six types, and SEG neurons into two types. Fibers in the retrocerebral complex showed [Arg7]-corazonin, beta-PDH, cholecystokinin8, and FMRFamide immunoreactivity. Cholecystokinin8 immunoreactivity was also detected in intrinsic cells of the corpus cardiacum. The corpus allatum was densely innervated by FMRFamide-immunoreactive varicose fibers. These results suggest that PI, PL, and SEG neurons release [Arg7]-corazonin, beta-PDH, cholecystokinin8, or FMRFamide-like peptides from the corpus cardiacum or corpus allatum into the hemolymph, and that some PL neurons may simultaneously release several neuropeptides.     

SIFamide and SIFamide Receptor Define a Novel Neuropeptide Signaling to Promote Sleep in Drosophila

SIFamide receptor (SIFR) is a Drosophila G protein-coupled receptor for the neuropeptide SIFamide (SIFa). Although the sequence and spatial expression of SIFa are evolutionarily conserved among insect species, the physiological function of SIFa/SIFR signaling remains elusive. Here, we provide genetic evidence that SIFa and SIFR promote sleep in Drosophila. Either genetic ablation of SIFa-expressing neurons in the pars intercerebralis (PI) or pan-neuronal depletion of SIFa expression shortened baseline sleep and reduced sleep-bout length, suggesting that it caused sleep fragmentation. Consistently, RNA interference-mediated knockdown of SIFR expression caused short sleep phenotypes as observed in SIFa-ablated or depleted flies. Using a panel of neuron-specific Gal4 drivers, we further mapped SIFR effects to subsets of PI neurons. Taken together, these results reveal a novel physiological role of the neuropeptide SIFa/SIFR pathway to regulate sleep through sleep-promoting neural circuits in the PI of adult fly brains.     

Immunocytochemical identification of growth hormote (GH) cells in the pituitary of three anuran species using an antiserum against purified bullforg GH

An antiserum was prepared against the recently purified bullfrog (bf) growth hormone (GH); it was applied to sections of brain and pituitary of three urodele (Ambystoma, Pleurodeles and Cynops) and three anuran (Xenopus, Bufo vulgaris and B. japonicus) species. No immunostaining was obtained in the urodele pituitary, being consistent with the results of immunoblot analysis of the pituitary homogenate. In the three anuran species, strong immunoreactivity was observed in GH cells that were concentrated in the posterodorsal region of the pars distalis. No GH-like immunoreactivity was detectable in the brain of any of the species. A comparison using adjacent sections stained with anti-bf prolactin (PRL) confirmed the anteroventral localization of PRL cells. Colocalization of GH and PRL was not apparent. These data suggest that the molecular structure of amphibian GHs is considerably different between anurans and urodeles. The antiserum used in the present work shows a high species specificity, recognizing only anuran GHs. In contrast anti-bfPRLlabeled PRL cells in all the amphibian species studied in the present work, suggesting that PRLs possess common amino acid sequences recognized by the anti-bfPRL.     

The hypothalamo-hypophysial system of the wild carp,Cyprinus carpio L.

Summary The posterior neurohypophysis (PNH)-pars intermedia complex of the wild and pond carp, Cyprinus carpio L., has been studied by light, fluorescence and electron microscopy. Gomori-positive neurosecretory fibres are abundant in the main trunk of the neurohypophysis as well as its roots penetrating the pars intermedia. Terminals of these fibres are in contact with capillaries of the general circulation and with glandular cells of the pars intermedia. Monoaminergic fibres with a weak green fluorescence, somewhat increasing after injection of nialamide into the pond carp, have largely the same distribution. Three types of neurosecretory fibres and their terminals have been recognized in the PNH-pars intermedia complex. Types-A₁ and -A₂ fibres, containing granules of 140–180 nm and 100–160 nm in diameter respectively, are peptidergic Gomori-positive. Type-A₂ fibres predominate in the PNH. The least frequent monoaminergic type-B fibres have granules of 60–100 nm in diameter. Numerous peptidergic and few monoaminergic neurosecretory terminals make contact with the capillaries located within the roots of the PNH as well as at the border between them and the pars intermedia. Both peptidergic and monoaminergic terminals make direct synaptoid contacts with the gland cells or end close to connective tissue septa, basal lamina or pituicytes. The PAS-positive gland cells and to a lesser degree the leadhaematoxylin-positive gland cells show these relationships with neurosecretory terminals. The question concerning the mode of interaction between peptidergic and monoaminergic structures in the dual control of the gland cells of the pars intermedia of teleosts is discussed.     

Ultrastructure of the nongranulated cells and morphology of the extracellular spaces in the pars distalis of adult and pouch-young tammar wallabies (Macropus eugenii)

Summary Differentiated nongranulated (folliculo-stellate) cells were observed in the centre and periphery of the cords of cells in the pars distalis of the adult tammar wallaby (Macropus eugenii). The nongranulated cells formed follicles containing a small lumen; the cell apices were joined by junctional complexes including zonulae adhaerentes and maculae adhaerentes (desmosomes). Follicles of granulated cells were also occasionally found. Follicles were rarely observed in the adult, but were numerous and generally larger in pouch-young wallabies. Moreover, whereas the involvement of granulated cells in the follicles of the adult was infrequent, they were common components of follicular structures in the pouch-young. The marginal cells at the periphery of the pars distalis in contact with the hypophysial cleft had many of the cytological characteristics of the nongranulated cells in the central pars distalis. In both adult and pouch-young wallabies, nongranulated cells at the periphery of the cell cords were juxtaposed such that they formed sinusoid-like spaces or lacunae, presumably as part of the extravascular channel system. The observations are discussed in light of the proposed phagocytotic, metabolite and/or hormone transport and stem cell roles attributed to these cells.This work was supported by a travel grant and grant-in-aid of research from the N.S.E.R.C. (J.F.L.) and from the A.R.G.C. and the N.I.H. (M.B.R.)     

Fine structure of the chromophobe in the pars distalis of the common snapping turtle,Chelydra serpentina

Summary Chromophobes of the pars distalis in young Chelydra serpentina have sparse cytoplasm with no specific granules; however, many cytoplasmic filaments are present. The chromophobes are connected to the other cell types by desmosomes, while different types of junctional specializations occur between adjacent chromophobes. Cytoplasmic filaments traverse the cytoplasm in a random manner and terminate on both the junctional complexes and the nuclear envelope. It is proposed that, in addition to providing a structural framework, the chromophobes may be involved in integrating cellular responses of the parenchyma to changes in the endocrine milieu.Supported in part by a General Research Support Grant RR05402, NIH to Dr. Tseng and a Human Development Grant HD-03484, NIH to Dr. Yntema.     

Morphometric classification of the neurosecretory granules in the rat pars nervosa

Summary The distribution of the diameters of the neurosecretory granules in the rat pars nervosa (measured from electron micrographs taken at 40 000 × ) was compared among axons by nonparametric statistical methods and the axons were classified into five groups with median granule diameters of 143, 155, 167, 180 and 193 nm. We suggested that these five axon types carried different secretory substances contained in the pars nervosa. This investigation is supported by a grant from the Population Council, New York and grant from the Ministry of Education. Authors are grateful to Japan Electron Optics Laboratory Company for their technical assistance with the electron microscopy and to Miss Kazue Yamamoto for her help in preparing the figures.     

A light and electron microscopic study of the pre- and postnatal development and secretory differentiation of the pars tuberalis of the rat hypophysis

Summary The development of the pars tuberalis was studied in the rat fetus from 13 days of gestation to 6 weeks after birth. After the closure of Rathke's pouch, the pars tuberalis anlage is clearly distinguishable from the anlagen of the partes intermedia and distalis. It comprises the entire basal portion of the adenohypophysial anlage; the limit between the anlagen of the pars tuberalis and the pars distalis is defined by Atwell's recess, i.e. the pathway taken by the hypophysial vessels coming from the vascular plexus of the median eminence.At 14 days the pars tuberalis cells are characterized by the presence of glycogen which persists in the adult. Their secretory differentiation (elaboration of granules with a diameter of 100–120 nm) is obvious at 15 days of gestation. It therefore, clearly precedes that of the other hypophysial cell types. Its functional differentiation takes place well before its adhesion to the primary vascular plexus of the portal system. Cystic formations appear just before birth in the pars tuberalis, much later than those of the pars distalis.These observations on the development of the pars tuberalis, together with previous observations on the adult PT in various species, showing that the specific glandular cells of the pars tuberalis are cytologically different from all known adenohypophysial cell types, seem to indicate a specific endocrine function of this lobe.     

The neurohypophysis of the crested newt

Summary In the crested newt, the ultrastructural organization of the pars nervosa is analogous to that already known in non-mammal tetrapods. An orderly array of ependymal cells makes up the inner limiting layer while less abundant pituicytes are irregularly distributed within this organ. Light and dark pituicytes can be distinguished on the basis of the relative density of the cytoplasmic matrix and the distribution of the cell organelles.Both the ependymal cells and pituicytes are rich in dense bodies and possess extensive processes which ramify among the nerve fibers, often reaching the pericapillary space which they can line for long distances.The main components of the pars nervosa are nerve fibers and nerve terminals (type A), containing electron dense granules 1200–2000 Å in diameter together with clear vesicles averaging 250–400 Å. These fibers are likely to correspond to the aldehyde fuchsin positive neurosecretory fibers revealed by light microscopy. Differences in the granule size within the fibers and terminals lead to further recognition of two subgroups (A₁ and A₂).Other fibers and terminals (type B) containing clear vesicles and granular vesicles 600 to 1000 Å in diameter, possibly of aminergic type, are also encountered. These fibers are rare and can be seen only in the portion of the pars nervosa near the pars intermedia of the adenohypophysis.Lastly, fibers and terminals containing only clear vesicles ranging from 250 to 400 Å (type C) are occasionally found.Nerve endings are often formed by type A fibers on the perivascular space and on the perivascular processes of the ependymal cells and pituicytes. In agreement with recent findings available in the literature, the occurrence of synaptoid contacts between these terminals and both pituicytes and ependymal cells may confirm the active role of these cells in transport and release of neurosecretion.Work supported by a grant from the Consiglio Nazionale delle Ricerche.We are gratefully indebted to Dr. G. Gendusa and P. Balbi for technical assistance, dr. G. E. Andreoletti for statistical analysis.