Cell cycle changes after denervation of the newt limb regenerate,studied in vitro

Summary The proliferation of the mesenchyme of medium bud stage blastemas ofPleurodeles, measured by³H-thymidine incorporation and mitotic index, decreases about 40–50% under denervated conditions when cultivated in vitro for 4 days; epidermis is not affected in this case. Autoradiography of blastemas after³H-thymidine long term labeling shows that 3/4 of the mesenchymal cells and 1/4 of the epidermal cells are cycling when the blastema is innervated; there is no significant change of these percentages when the blastema is denervated. The results show, contrary to in vivo experiments, that denervation does not provoke an exiting from the cell-cycle but only lengthening of the cycle of the mesenchymal cells (probably of the G₁ phase).     

Microdensitometric analysis of denervation effects on newt limb blastema cells

Summary This report examines the fate of cycling cells in normal and denervated blastemas of adult newts. Cells are found to accumulate in G₁ in blastemas which are nerve independent. No stage specific accumulation different from controls is found in limbs with nerve-dependent blastemas.     

Peptidergic innervation of the temporomandibular disk in the rat

Summary The peptidergic innervation of the temporomandibular disk was investigated in the postnatal young rat by using an indirect immunofluorescence method. Calcitonin gene-related peptide-containing nerve fibers were located around the blood vessels and terminated as free nerve endings in the disk. These nerve fibers may be of a sensory nature.     

Peptidergic innervation of the temporomandibular disk in the rat

The peptidergic innervation of the temporomandibular disk was investigated in the postnatal young rat by using an indirect immunofluorescence method. Calcitonin gene-related peptide-containing nerve fibers were located around the blood vessels and terminated as free nerve endings in the disk. These nerve fibers may be of a sensory nature.     

Cellular events in denervated limb stumps ofAmbystoma larvae during re-innervation and subsequent regeneration

Summary Amputated, denervatedAmbystoma forelimbs undergo cellular dedifferentiation and slight resorption through 12 days post-amputation. Subsequently, as limb stumps become re-innervated, there occur mitosis, blastema formation, and regeneration. The initial increase in the mitotic index in once-denervated limb stumps is closely correlated with the presence of ingrowing nerve fibres.This work was supported by grant PCM 76-11807 from the National Science Foundation.     

Morphological evidence for a direct neuroendocrine GABAergic control of the anterior pituitary in teleosts

Summary The anterior pituitary of teleosts is unique among vertebrates in receiving a direct innervation which represents the morphological support of the neuroendocrine control of pars distalis functions. The participation of GABAergic fibers in this innervation was studied by means of immunocytochemistry at the light and electron microscopic levels, using antibodies against GABA. Immunoreactive fibers, characterized by the presence of small clear and dense cored vesicles, were detected in all parts of the gland. Immunopositive terminals were found in close, sometimes synaptic-like, contact with most glandular cell types in the anterior lobe. The data strongly suggest that in teleosts, as in mammals, GABA is involved in the neuroendocrine control of anterior pituitary functions.     

Morphological evidence for a direct neuroendocrine GABAergic control of the anterior pituitary in teleosts

The anterior pituitary of teleosts is unique among vertebrates in receiving a direct innervation which represents the morphological support of the neuroendocrine control of pars distalis functions. The participation of GABAergic fibers in this innervation was studied by means of immunocytochemistry at the light and electron microscopic levels, using antibodies against GABA. Immunoreactive fibers, characterized by the presence of small clear and dense cored vesicles, were detected in all parts of the gland. Immunopositive terminals were found in close, sometimes synaptic-like, contact with most glandular cell types in the anterior lobe. The data strongly suggest that in teleosts, as in mammals, GABA is involved in the neuroendocrine control of anterior pituitary functions.     

Cholinergic nerves in the rat portal vein

Summary The innervation of the rat portal vein was studied using the cholinesterase histochemical method 2 plexuses of cholinergic nerve fibers were found; an external plexus localized at the level of the outer adventitial layers and an inner plexus at the level of the adventitial-medial transitional zone and in the outer layers of the media.Acknowledgments. We thank Dr C. Cavallotti for his kind help and criticism. The kind help of Dr W. Collier and of the staff of the laboratory of Neurobiology as well as the bibliographic assistance of Istituto Superiore di Sanità (Serv. Documentazione, Dr Magliola) are gratefully acknowledged.     

Innervation and control of the heart of a gastropod,Rapana

Summary The innervation and control of the heart of a prosobranch mollusc,Rapana thomasiana, were studied. Acetylcholine was found to be an inhibitory neurotransmitter. Both serotonin and FMRFamide (Phe-Met-Arg-Phe-NH₂) showed excitatory effects on the heart; FMRFamide had greater inotropic and more regulatory chronotropic effects than serotonin. The effects of serotonin were blocked by methysergide, while the effects of FMRFamide and of stimulating the excitatory cardiac nerves were not blocked. Stimulation of circumesophageal ganglia elicited a slow enhancement of heart beat together with body movement. This enhancement was blocked by methysergide. Serotonin was considered to act at the heart as a local neurohormone. Although the mechanism of action of FMRFamide is still not yet clarified, it is possible that FMRFamide plays a physiological role as a cardioregulatory substance, as indicated by the physiological and histological findings.     

Über neue elektronenmikroskopische Befunde zur peripheren vegetativen Synapse am mesenterialen Lymphgefäss vom Meerschweinchen (Cavia porcellus)

Summary Electron microscopic investigations of innervation of the mesenteric lymphatic vessels ofCavia porcellus demonstrate that, besides membrane contacts between Axon and smooth muscle cell, a local direct plasmatic connection from the nature of synapses between the nerve and the effector cell have to be considered. From this study it has been concluded that also this neuromuscular junction is a transitoric structure and not a static connection comparable to the synapses in the motor end-plate of the striated muscle.

---

---

Die Arbeit wurde in dem elektronenmikroskopischen Laboratorium des Pathologischen Institutes Mainz unter Leitung von Priv. Doz. Dr.R. Bässler durchgeführt.     

Immunohistochemical localization of pancreatic polypeptide (PP) in the brain of the larval instar of the hoverfly,Eristalis aenus (Diptera)

Summary 2 pancreatic polypeptide (PP) immunoreactive cells were observed in each half of the protocerebrum of the last (5th), larval instar of the hoverfly,Eristalis aenus. No PP immunoreactive nerve fibers could be detected in the brain.Support by a grant from the Swedish Medical Research Council (project No. 12X-102) and by a Swedish Institute scholarship.     

Ras proteins in the control of the cell cycle and cell differentiation

The Ras family of small GTPases includes three closely related proteins: H-, K-, and N-Ras. Ras proteins are involved in the transduction of signals elicited by activated surface receptors, acting as key components by relaying signals downstream through diverse pathways. Mutant, constitutively activated forms of Ras proteins are frequently found in cancer. While constitutive Ras activation induces oncogenic-like transformation in immortalized fibroblasts, it causes growth arrest in primary vertebrate cells. Induction of p53 and cyclin-dependent kinase inhibitors such as p15^INK4b, p16^INK4a, p19^ARF, and p21^WAF1 accounts for this response. Interestingly, while ras has usually been regarded as a transforming oncogene, the analysis of Ras function in most of the cellular systems studied so far indicates that the promotion of differentiation is the most prominent effect of Ras. While in some cell types, particularly muscle, Ras inhibits differentiation, in others such as neuronal, adipocytic, or myeloid cells, Ras induces differentiation, in some cases accompanied by growth arrest. Several possible mechanisms for the pleiotropic effects of Ras in animal cells are discussed. Received 8 March 2000; received after revision 24 May 2000; accepted 24 May 2000     

Aluminium in repair membranes and Al,Ca and Pi in the haemolymph of Al-injected shell-repairing snails (Helix pomatia L.)

Summary In AlCl₃-injected shell-repairing snails,Helix pomatia L., the Al-associated decrease of the weights of the shell-repair membranes was unrelated to the Al-concentration in the membranes. In the haemolymph the concentration of Al was related to the dose of injected Al, while the concentration of Ca was increased by the highest Al-dose only. No phosphate was detected in either controls or Al-injected snails. It is concluded that Al inhibits the growth of the CaCO₃-crystals by mechanisms other than incorporation in, or adsorption to, the crystals.     

Comparative cytochemical and pharmacological studies on the cholinergic innervation of the branchial heart of the cephalopodSepia officinalis (L.)

Summary The dense motor innervation of the obliquely striated muscle cells in the branchial heart of Coleoida is composed of activating and inhibiting parts. The inhibiting cholinergic system investigated in this study is characterized histo- and cytochemically by a high acetylcholinesterase activity (EC 3.1.1.7) in the glycocalices of the nerve fibers with transparent synaptic vesicles, the muscle cells and, to a lesser extent, the ovoid interstitial cells, the functions of which are endocytosis and storage of catabolic substances. The pharmacological results from the isolated organ indicate a more nicotinic type of Ach-receptor, which can be reversibly blocked by D-tubocurarine and -bungarotoxin, but not with the same intensity by tetraethylammonium or atropine.     

Interaction between bradykinin and voltage-sensitive sodium channels in myelinated nerve fibers

Summary The effects of externally and internally applied bradykinin on the excitability of single myelinated nerve fibers were studied. External bradykinin (10 M) slightly prolongs the action potential of a single myelinated nerve fiber; hence, when the fibers are stimulated by long-lasting pulses, this raises the frequency of repetitive firing in sensory fibers and evokes repetitive activity in motor fibers. Under voltage-clamp conditions, sodium channel inactivation is slowed, while sodium channel activation remains unaffected. Prolonged depolarization of th membrane leads to a maintained sodium current. The voltage dependence of the steady-state sodium current inactivation (h ) is shifted in the depolarized direction by 10 mV. Internally applied bradykinin produces a frequency-dependent block of the sodium current. The phenomena described here imply that more than one site on the sodium channel is modified by bradykinin.     

Note sur l'innervation des branchies chezAnguilla anguilla L.

Summary Maillet's technique demonstrated the innervation of pillar cells of the secondary gill lamellae inAnguilla anguilla L. Nervous control may work in conjunction with an humoral regulation which would make clearer the understanding of the in- and outflux of blood in the respiratory organs.     

A comparative study of the innervation of the choroid plexus in amphibia

The aminergic and cholinergic innervation of choroid plexuses in three species of amphibia was investigated. Plexuses of the Japanese toad and the bullfrog had poor innervation by adrenergic nerves of sympathetic origin, but in the clawed toad, these plexuses were heavily innervated by adrenergic axons from ganglion cells located in the plexus stroma. Nerve fibers positive for acetylcholinesterase were not found in the plexuses, except for a few fibers with very weak enzyme activity in the clawed toad.     

The structure of amoorastatone and the cytotoxic limonoid 12-hydroxyamoorastatin

Summary 2 new limonoid-type terpenes have been isolated from an aqueous extract of seeds produced by the Eastern Himalayan (India) plantAphanamixis grandifolia Bl. By interpreting principally mass spectral and nuclear magnetic resonance data, the structures of 12-hydroxyamoorastatin (2b) and amoorastatone (3) were elucidated. Unequivocal evidence for the 12-hydroxyamoorastatin structural assignment was obtained by chemical conversion to sendanin (4). Amoorastatin derivative2b was found to significantly inhibit growth of the murine P388 lymphocytic leukemia cell lines but amoorastatone in the same system was inactive. In a comparative biological study, sendanin (4) and anthothecol (7) were also found significantly to inhibit growth of the P388 cell line, while rohitukin (8) and limonin (9) were found to be inactive.Part 63 of the series Antineoplastic Agents. For the previous contribution refer to G.R. Pettit, T.S. Krupa and R.M. Reynolds, Int. J. Peptide Protein Res., in preparation. The present investigation was supported in part by Public Health Research Grant No. CA-16049-05 from the National Cancer Institute, the Fannie E. Rippel Foundation, Mrs Mary Dell Pritzlaff, the Spencer T. and Ann W. Olin Foundation, Mr John F. Schmidt, and the Phoenix Coca-Cola Bottling Company.     

Interaction between bradykinin and voltage-sensitive sodium channels in myelinated nerve fibers

The effects of externally and internally applied bradykinin on the excitability of single myelinated nerve fibers were studied. External bradykinin (10 microM) slightly prolongs the action potential of a single myelinated nerve fiber; hence, when the fibers are stimulated by long-lasting pulses, this raises the frequency of repetitive firing in sensory fibers and evokes repetitive activity in motor fibers. Under voltage-clamp conditions, sodium channel inactivation is slowed, while sodium channel activation remains unaffected. Prolonged depolarization of the membrane leads to a maintained sodium current. The voltage dependence of the steady-state sodium current inactivation (h infinity) is shifted in the depolarized direction by approximately 10 mV. Internally applied bradykinin produces a frequency-dependent block of the sodium current. The phenomena described here imply that more than one site on the sodium channel is modified by bradykinin.     

Über die Bedeutung der Innervation für die Adrenalinsynthese im Nebennierenmark

Summary In experiments on rabbits, the influence of the innervation on the synthesis of adrenaline in the suprarenals was investigated. After depleting the glands of adrenaline by repeated injections of reserpine, one splanchnic nerve was sectioned. It was found that in the denervated suprarenal gland, the resynthesis was slower than in the normal gland. This suggests that the innervation is of importance for the rate of synthesis of adrenaline.