Cholinesterases in the hypothalamo-hypophysial neurosecretory system of the White-crowned Sparrow,Zonotrichia leucophrys gambelii

Summary The distribution of cholinesterases in hypothalamo-hypophysial neurosecretory system of the White-crowned Sparrow has been examined histochemically. The perikarya of the neurosecretory cells of the paraventricular and supraoptic nuclei have a high acetylcholinesterase activity. Acetylcholinesterase activity also occurs in the cells of the infundibular nucleus. The proximal parts of the axons of the cells of the neurosecretory and infundibular nuclei have strong acetylcholinesterase activity and weak non-specific cholinesterase activity. In the median eminence, the activity of acetylcholinesterase is strongest in the palisade layer. In the pars nervosa, there is definite, although weak, acetylcholinesterase activity.This investigation was supported by grants from the National Institutes of Health to Professor Farner (B-1353) and to Dr. Kobayashi (A-3678).     

Ultrastructure of the peptidergic and monoaminergic neurons in the hypothalamic neurosecretory system of Anuran Batracians

Summary In the toad Bufo arenarum Hensel the following regions of the hypothalamic — neurohypophyseal system were studied under the electronmicroscope: preoptic and paraventricular nuclei, median eminence and infundibular process of the neurohypophysis.Neuronal perikarya of the preoptic nucleus are loaded with typical neurosecretory granules of peptidergic nature having a mean diameter of 1660 Å. While most neurons of the winter toad are in a storage stage a few show signs of a more active synthetic activity. A distinctive feature of preoptic neurons is the presence of large lipid droplets. The paraventricular nucleus contains small neurons containing granulated vesicles with a mean diameter of 800-1000 Å. In the region extending between these two nuclei and the median eminence axons containing either neurosecretory elementary granules or granulated vesicles are observed.The inner zone of the median eminence is occupied by axons of the preoptic neurohypophyseal tract; two types of axons, according to the size and density of the neurosecretory granules, may be recognized. The outer zone of the median eminence contains mainly axons and nerve terminals containing granulated vesicles of probable monoaminergic nature and only a few with granules of peptidergic type.The neurohypophysis contains two kinds of axons: one with more dense granules of 1800 Å and the other with granules of lesser electron density and 2100 Å. At the ending proper small clear vesicles of synaptic type are found.A progressive increase in volume of the peptidergic granules along the axon is demonstrated. This is of the order of 218% from the preoptic perikarya down to the infundibular process. The physiological significance of the two neurosecretory systems — i.e. the monoaminergic and the peptidergic — and the probable nature of the two types of peptidergic axons is discussed.Supported by grants from the Consejo Nacional de Investigaciones Científicas y Técnicas and by the Air Force Office of Scientific Research (AF-AFOSR 963-67).The authors want to express their gratitude to Mrs. Defilippi-Novoa and Mr. Alberto Sáenz for their skillful assistance.     

Histochemical demonstration of acetylcholinesterase in the hypothalamus of the female guinea-pig

Summary The distribution of acetylcholinesterase (AChE) in the hypothalamus of the female guinea pig has been examined histochemically.Activity was found in neurones of the supraoptic, paraventricular and infundibular nuclei; in the lateral hypothalamic area, in cells dorsomedial to the fornix at the level of the paraventricular nucleus and in a large group of cells surrounding the fornix at the level of the in-fundibular nucleus. A small well-stained group of cells, not identified in histological preparations, was present ventral to the paraventricular nucleus. The neuropil stained at the lateral border of the paraventricular nucleus and in the medial mammillary, suprachiasmatic and dorsomedial nuclei. The walls of some blood vessels stained; activity was particularly strong in vessels in the preoptic area and at the base of the median eminence. Nerve fibres on blood vessels ventral to the hypothalamus also stained.The degree of enzyme activity in the cell groups was compared in immature, pregnant, lactating, ovariectomized and hysterectomized animals. The greatest variation occurred in the infundibular nucleus. Hypophysectomy markedly reduced staining in the supraoptic nucleus. The possibility that AChE may be involved in the elaboration or transport of releasing factors is discussed.The authors are indebted to Dr. J. S. Perry for doing the surgery involved in this work. They are also grateful to him and to Dr. R. B. Heap for helpful discussions and to Miss M. Hamon for excellent technical assistance.     

The development of the hypophysial portal system in the White-crowned Sparrow,Zonotrichia leucophrys gambelii

Summary The development of the hypophysial portal system has been studied in 35 embryos and 45 nestlings of the White-crowned Sparrow. The primordium of the hypophysis is vascularized by the infundibular (primary) capillary plexus, supplied by the right and left infundibular arteries, which, in the embryo, are constant branches of the right and left internal carotid arteries.The cellular proliferation and differentiation of the pars distalis into rostral and caudal lobes is accompanied by a penetration of portal vessels from the infundibular (primary) capillary plexus into these lobes beginning on the fifth day of incubation. The cellular proliferation of the rostral lobe of the pars distalis and development of the rostral group of the portal vessels precedes that of the caudal lobe of the pars distalis and the development of the caudal group of the portal vessels.The periglandular vessels, which originate in younger embryos from the infundibular (primary) capillary plexus, apparently become a part of the portal vessels.The portal vessels are the sole blood supply to the developing pars distalis of the White-crowned Sparrow; there is no evidence of a direct arterial supply at anytime during embryonic development. The neural-lobe artery appears at the end of incubation as a secondary branch of the right and left infundibular arteries. The rostral and caudal groups of the portal vessels are well-developed at the end of incubation (17–29 mm CRL) when aldehyde-fuchsin positive neurosecretory material first appears in the supraoptic and paraventricular nuclei, in the median eminence and in the neural lobe.The differentiation of the median eminence into rostral and caudal divisions begins at the end of the nestling period although its adult form is not achieved until later. The formation of the portal zone begins at the end of incubation (17–29 mm CRL) and is completed by the time of fledging.Dedicated to Professor Dr. W. Bargmann in honor of his 60th birthday.The investigations reported herein were supported by a research grant (HE 07240 NEUA) from the National Institutes of Health to Professor Vitums, by funds for biological and medical research made available by State of Washington Initiative Measure No 171 to Professor Vitums, by a research grant from the Deutsche Forschungsgemeinschaft to Professor Oksche, by aresearch grant (NB 01353) from the National Institutes of Health to Professor Farner, and by a Research Career Development Award from the National Institute of Arthritis and Metabolic Diseases (5 K 3 AM-18,370) to Professor King. We are grateful to Professor Bargmann for his generosity in making available the facilities of the Anatomisches Institut Kiel for this investigation. We wish to thank Frau Karin Graap and Mrs. Dianne Reno for technical assistance and Miss Janice Austin for the preparation of the drawings.     

Identification,in the external region of the rat median eminence,of separate neurophysin-vasopressin and neurophysin-oxytocin containing nerve fibres

Summary Immuno-enzyme cytochemical investigations, using single and double staining techniques, showed that the external region of the rat median eminence contains separate neurophysin-vasopressin fibres and neurophysinoxytocin fibres. These neurophysin-hormone containing nerve fibres are influenced by bilateral adrenalectomy and by colchicine treatment. The external region of the median eminence of the homozygous Brattleboro rat contains neurophysin-oxytocin fibres. It does not contain immuno-reactive neurophysin-vasopressin fibres. Bilateral adrenalectomy also influences the neurophysin-vasopressin containing neurons of the suprachiasmatic nuclei. In the neurons of the parvicellular part of the rat hypothalamic paraventricular nuclei, staining for vasopressin and for oxytocin is completely absent.     

Cyto-immunological study of the somatostatin hypothalamic peptidergic system in the fox (Vulpes vulpes)]

Immunocytological investigations on the hypothalamus of the red fox with rabbit anti-SRIF showed large or small perikarya in the supra-optical and paraventricular nuclei. In the median eminence SRIF-positive nerve fibres and their endings have a characteristic distribution in areas different from those of LH-RH containing nerve fibres.     

The number of satellite cells in slow and fast fibres from human vastus lateralis muscle

The aim of this investigation was to study the distribution of satellite cells in slow (type I fibres) and fast (type II fibres) fibres from human vastus lateralis muscle. This muscle is characterised by a mixed fibre type composition and is considered as the site of choice for biopsies in research work and for clinical diagnosis. Biopsy samples were obtained from five healthy young volunteers and a total of 1,747 type I fibres and 1,760 type II fibres were assessed. Satellite cells and fibre type composition were studied on serial muscle cross-sections stained with specific monoclonal antibodies. From a total of 218 satellite cells, 116 satellite cells were found in contact with type I fibres (53.6±8% of the satellite cells associated to type I fibres) and 102 satellite cells in contact with type II fibres (46.4±8% of the satellite cells associated to type II fibres). There was no significant difference (P=0.4) between the percentages of satellite cells in contact with type I and with type II fibres. Additionally, there was no relationship between the mean number of satellite cells per fibre and the mean cross-sectional area of muscle fibres. In conclusion, our results show that there is no fibre type-specific distribution of satellite cells in a human skeletal muscle with mixed fibre type composition.     

Cytoarchitectonic pattern of the hypothalamus in the turtle,Lissemys punctata granosa

Summary In the hypothalamus of the turtle, Lissemys punctata granosa, two magnocellular and 23 parvocellular neuronal complexes can be distinguished. The magnocellular complexes include the nucleus supraopticus and the nucleus paraventricularis; paraventricular neurons are partly arranged in rows parallel to the third ventricle. Most infundibular parvocellular nuclei display neurons disposed in rows parallel to the ventricular surface. In the preoptic region, the prominent parvocellular neuronal complexes encompass the nucleus periventricularis anterior, lateral preoptic area, the nucleus of the anterior commissure and the nucleus suprachiasmaticus. The prominent nucleus periventricularis posterior extends caudad and shows neurons arranged in vertical rows parallel to the third ventricle. Other parvocellular nuclei of the rostral hypothalamus are composed of clustered subunits. The nucleus arcuatus is a fairly large nuclear entity extending from the level marked dorsally by the nucleus paraventricularis to the area occupied by the nucleus of the paraventricular organ. A well-developed ventromedial nucleus is located ventrolateral to the paraventricular organ. The prominent paraventricular organ consists of tightly arranged neurons, some of which possess apical projections into the third ventricle; it is surrounded by the nucleus of the paraventricular organ. Nucleus hypothalamicus medialis et lateralis, nucleus hypothalamicus posterior and the nuclei recessus infundibuli are further nuclear units of the tuberal region. The caudal end of the hypothalamus is marked by the nucleus mamillaris; its neurons are scattered among the fibers of the retroinfundibular commissure. The median eminence is well developed and shows a large medial and two lateral protrusions into the infundibular recess.     

The origin of the vasopressinergic and oxytocinergic fibres of the external region of the median eminence of the rat hypophysis

Summary The origin of the vasopressinergic and oxytocinergic nerve fibres of the external region of the rat median eminence was investigated by means of hypothalamic lesions, adrenalectomy and immunocytochemistry. The results obtained in bilaterally adrenalectomized animals with complete, or incomplete, destruction of the suprachiasmatic nuclei showed that, at least, the great majority of the vasopressinergic and oxytocinergic nerve fibres of the external region of the rat median eminence do not originate from the suprachiasmatic nuclei. From the observations obtained in bilaterally adrenalectomized animals with total or subtotal destruction of both paraventricular hypothalamic nuclei, it appears that the paraventricular nuclei must be the origin of (nearly) all the vasopressinergic and oxytocinergic nerve fibres of the external region of the rat median eminence. The results strongly suggest that both types of fibres originate from all parts of the paraventricular nuclei.This investigation was supported by a grant from the Belgian Nationaal Fonds voor Geneeskundig Wetenschappelijk OnderzoekThe authors are much indebted to Prof. Dr. E. Kühn (Leuven) in whose laboratory the stereotactic operations were done     

The neurohypophysis of the foetal monkey

Summary The neurohypophysis of foetal macaque monkeys has been studied by optical and electron microscopy. Abundant elementary neurosecretory granules are present in the infundibular process by the middle third of gestation. Most of these are of variable electron density, and surrounded by a membrane larger than the granule, so that they appear haloed. A few fibres contain membrane-bounded electron-dense granules which show no halo. Inclusions of synaptic vesicle size are rare. The infundibular stem contains a few fibres with typical inclusions smaller than 1000 Å in diameter; the latter resemble inclusions in fibres of the median eminence.Dedicated to Professor W. Bargmann on his 60th birthday.     

Adrenergic and cholinesterase-containing neurons of the heart

Summary The adrenergic and acetylcholinesterase-containing nerves of the hearts of mice, rats, guinea-pigs, rabbits, and cats were studied. The fluorescence technique of Falck and Hillarp was used for the demonstration of adrenergic nerves, whereas a modified Koelle cholinesterase technique was used for the cholinesterase-containing nerves. The inhibitors used were Mipafox, iso-OMPA and Nu 683. Microspectrofluorometry was used to identify the structures containing dopamine.Adrenergic as well as acetylcholinesterase-containing fibres were found in all parts of the heart, most abundantly in the atria. Dense nerve plexa supplied the sinoarial and atrioventricular nodes. There was a plexus of both fibre types in the endocardium and on the atrial side of the valves. In the valves, it could be shown that adrenergic and cholinesterase-containing fibres ran closely parallel to each other. Indirect evidence suggested that this applies also to the myocardium.No nerve fibres containing dopamine were revealed in the microspectrofluorometer. The dopamine previously found in the atria seems, instead, to be situated in so-called small intensely fluorescent cells.No adrenergic ganglion cells were found in the heart despite extensive search. The vagus of rabbits was found to contain only few adrenergic preterminals.     

Electrophoretic analysis of proteins from single bovine muscle fibres.

A number of single fibres were isolated by dissection of four bovine masseter (ma) muscles, three rectus abdominis (ra) muscles and eight sternomandibularis (sm) muscles. By histochemical criteria these muscles contain respectively, solely slow fibres (often called type I), predominantly fast fibres (type II), and a mixture of fast and slow. The fibres were analysed by conventional sodium dodecyl sulphate/polyacrylamide-gel electrophoresis and the gels stained with Coomassie Blue. Irrespective of the muscle, every fibre could be classed into one of two broad groups based on the mobility of proteins in the range 135000-170000 daltons. When zones containing myosin heavy chain were cut from the single-fibre gel tracks and 'mapped' [Cleveland, Fischer, Kirschner & Laemmli (1977) J. Biol. Chem. 252, 1102-1106] with Staphylococcus proteinase, it was found that one group always contained fast myosin heavy chain, whereas the second group always contained the slow form. Moreover, a relatively fast-migrating alpha-tropomyosin was associated with the fast myosin group and a slow-migrating form with the slow myosin group. All fibres also contained beta-tropomyosin; the coexistence of alpha- and beta-tropomyosin is at variance with evidence that alpha-tropomyosin is restricted to fast fibres [Dhoot & Perry (1979) Nature (London) 278, 714-718]. Fast fibres containing the expected fast light chains and troponins I and C fast were identified in the three ra muscles, but in only four sm muscles. In three other sm muscles, all the fast fibres contained two troponins I and an additional myosin light chain that was more typical of myosin light chain 1 slow. The remaining sm muscle contained a fast fibre type that was similar to the first type, except that its myosin light chain 1 was more typical of the slow polymorph. Troponin T was bimorphic in all fast fibres from a ra muscles and in at least some fast fibres from one sm muscle. Peptide 'mapping' revealed two forms of fast myosin heavy chain distributed among fast fibres. Each form was associated with certain other proteins. Slow myosin heavy chain was unvarying in three slow fibre types identified. Troponin I polymorphs were the principal indicator of slow fibre types. The myofibrillar polymorphs identified presumably contribute to contraction properties, but beyond cud chewing involving ma muscle, nothing is known of the conditions that gave rise to the variable fibre composites in sm and ra muscles.     

A note on the behaviour of spindle fibres at mitosis

Summary Measurements done on mitosis in plant endosperm with the use of Dr. Inoué's polarizing microscope indicate that the whole chromosomal spindle fibre is shifted during anaphase, i.e. the distance between the spindle pole and the polar end of the spindle fibre decreases during anaphase. Consequently in endosperm mitosis the chromosomes move faster to the poles than the chromosomal spindle fibres decrease in length. As the chromosomal spindle fibres in animal materials presumably extend to the spindle poles already from the beginning of anaphase, the chromosomes will here approach the poles with the same speed, as the spindle fibres contract.This paper is dedicated to Professor Franz Schrader on the occasion of his seventieth birthday.     

Corticotropin-releasing hormone-like immunoreactivity in the brain of the snake Bothrops jararaca

The distribution of corticotropin-releasing hormone in the brain of the snake Bothrops jararaca was studied immunohistochemically. Immunoreactive neurons were detected in telencephalic, diencephalic and mesencephalic areas such as dorsal cortex, subfornical organ, paraventricular nucleus, recessus infundibular nucleus, nucleus of the oculomotor nerve and nucleus of the trigeminal nerve. Immunoreactive fibres ran along the hypothalamo-hypophysial tract to end in the outer layer of the median eminence and the neural lobe of the hypophysis. In general, immunoreactive fibres occurred in the same places of immunoreactive neurons. In addition, immunoreactive fibres were observed in the septum, amygdala, lamina terminalis, supraoptic nucleus, nucleus of the paraventricular organ, ventromedial hypothalamic nucleus and interpeduncular nucleus. These results indicate that, as for other vertebrates, corticotropin-releasing hormone in B. jararaca brain, besides being a releasing hormone, may also act as a central neurotransmitter and/or neuromodulator.     

Immunohistochemical analysis of the human psoas major muscle with regards to the body side and aging

The aim of our study was to explore the age related changes of the fibre type composition of the human psoas major muscle. Moreover, we wanted to compare the fibre type composition of the left and right muscle. Muscle samples were collected from 15 young and 15 old males. Type I, IIA and IIX muscle fibres were typed using myosin heavy chain identification. The serial transverse sections were analysed using a light microscope. Results of our study showed that the age-related atrophy affected all three fibre types. Type IIA fibres were affected most profoundly while type I fibres were affected most weakly. The percentage of the different fibre types did not change during aging. There were no differences in the fibre type composition between the left and right muscle. Human psoas major muscle undergoes normal aging changes with the atrophy of all three fibre types, whereas atrophy most profoundly affects type IIA fibres. No differences in the fibre type composition between the left and right muscle point to the equal engagement of both legs in normal everyday activities of human.     

Relationship between myosin heavy chain IId isoform and fibre types in soleus muscle of the rat after hindlimb suspension

The relationship between the myosin heavy chain (HC) IId isoform and histochemically defined fibre types was investigated in the rat soleus muscle after hindlimb suspension. After 4 weeks of suspension, right and left muscles were removed and fibre type composition and total fibre number were examined by histochemical myosin adenosine triphosphatase staining sections. Myosin HC isoforms were analysed by sodium dodecyl sulphate polyacrylamide gel electrophoresis. After the suspension, there was a significant decrease in the percentage of type I fibres and a concomitant increase in that of type IIa fibres. However, the total number of fibres was not affected by suspension. The synthesis of HC IId isoform, which was not found in the control, and the decrease in the ratio of slow type myosin heavy chain isoform (HC I) were observed after suspension. These results would may suggest that the change of fibre type composition was caused by a shift from type I to IIa fibres after suspension. Furthermore, it could be suggested that the synthesis of HC IId isoform occurred during the stage of type shift from type I to IIa fibres.     

Immunocytochemical localization of neuropeptides in the hypothalamus of the Japanese long-fingered bat,Miniopterus schreibersii fuliginosus

Summary To elucidate the role of hypothalamic neuropeptides in regulation of reproductive phenomena of seasonally breeding feral mammals, we used Japanese long-fingered bats, Miniopterus schreibersii fuliginosus, for immunocytochemical study of distribution of the following neuropeptides in the hypothalamus: arginin vasopressin, oxytocin, luteinizing hormone-releasing hormone, somatostatin, corticotropin-releasing factor, and growth hormone-releasing factor. The size, shape and location of supraoptic, paraventricular, suprachiasmatic, and arcuate nuclei of the bat were determined. Arginin vasopressin-and oxytocin-immunoreactive magnocellular neurons were found in the supraoptic and paraventricular nuclei, where they exhibited separate distribution into two distinct groups. Parvocellular arginin vasopressin neurons occurred only in the suprachiasmatic nucleus. The hibernating bats exhibited slightly increased numbers of vasopressin and oxytocin neurons in the supraoptic and paraventricular nuclei. The pregnant bat displayed further increased numbers of vasopressin and oxytocin neurons in both nuclei. Somatostatin-immunoreactive neurons in the paraventricular nucleus were also immunopositive to anti-oxytocin serum, while those in the ventromedial and arcuate nuclei reacted solely to anti-somatostatin serum. They projected to the anterior median eminence and infundibular stalk. Luteinizing hormone-releasing hormone-immunoreactive perikarya were scattered throughout the basal hypothalamus, being particularly abundant in the arcuate nucleus. They were larger in size in hibernating bats than those in normal (non-pregnant) and pregnant females. They projected fibers mainly to the internal layer of the median eminence and infundibular stalk. A few luteinizing hormone-releasing hormone-reactive fibers were also observed in the organum vasculosum laminae terminalis, lateral habenular nuclei, pineal stalk, retroflexus fasciculus, and olfactory tubercle. Corticotropin releasing factor-immunoreactive perikarya were distributed in the paraventricular nucleus and medial preoptic area and projected into the external layer of the anterior median eminence, while growth hormone-releasing factor-immunoreactive perikarya occurred only in the arcuate nucleus and projected into the posterior part of the median eminence.     

The fine structure of the guinea-pig muscle spindle

Summary Nuclear bag and nuclear chain intrafusal fibres are present in guinea-pig muscle spindles. Unlike muscle spindles in other species two types of nuclear chain fibre seem to be present. The electron microscopical appearance of one type of nuclear chain fibre is similar to that of nuclear bag fibres.It is suggested that under tension the nuclei of small nuclear bag fibres become sufficiently displaced to form nuclear chain-like fibres. The frequent occurrence of fibres which combine some of the properties of both nuclear bag and nuclear chain fibres indicates the possible occurrence of a third type of intrafusal fibre.The sensory innervation of guinea-pig muscle spindles is similar to that of the cat and the rat. Three types of motor nerve ending which could be classified according to the complexity of their subneural apparatus were seen.     

Congenital muscle fibre type disproportion: clinical, morphological and biochemical findings in children

Muscle biopsies from quadriceps femoris muscle of normal subjects and subjects with symptoms of congenital myopathy, aged 1-3 years, were examined for morphological and biochemical differences. Four patients showed clinical signs of Congenital Fibre Type Disproportion (CFTD) as described originally by Brooke (1973), while the other cases showed only signs of hypotonia and diffuse weakness as described by Clancy et al. (1980). No morphological differences between patients with different clinical signs were found in any biopsy sample from the quadriceps femoris muscle, as regards fibre size; type I fibres were always smaller than type II fibres. The difference in fibre size was more than 15% in all cases, and the variability coefficient of the larger type II fibres was less than 250. Nevertheless, as regards fibre occurrence, two patients showed more type I fibres than type II fibres, four patients showed more type II fibres than type I fibres and one patient had a marked type II fibre predominance. Enzyme activities assayed in the crude mitochondrial fraction showed no abnormalities between normal subjects and patients. An increase in the activity of lactate dehydrogenase in the crude extract was found in all cases.     

Immunocytochemical localization of a galanin-like peptidergic system in the brain of two urodele and two anuran species (Amphibia).

Galanin-like immunoreactivity was localized in the brain of Urodela (Ambystoma, Pleurodeles) and Anura (Bufo, Xenopus) by immunocytochemistry with anti-porcine galanin antiserum. In the four species, immunoreactive perikarya were observed in the telencephalon (striatum, amygdala), diencephalon preoptic area mainly along the anterodorsal wall of the preoptic recessus, suprachiasmatic nucleus, lateral hypothalamus, ventral and dorsal infundibular nuclei, paraventricular organ, and rhombencephalon (nucleus of the solitary tract). Galaninergic fibres extended in similar regions and in the medial septum, ventral telencephalon, ventral hypothalamus, median eminence, and various mesencephalic and rhombencephalic regions. Contacts with the cerebrospinal fluid cavity occurred along the preoptic recessus (Ambystoma) and the ventral infundibular wall (all species). Fibres were scarce in the neurohypophysis. The distal and intermediate lobes of the pituitary were virtually devoid of immunoreactivity. The galaninergic system appeared more developed in adult amphibia than in young animals, suggesting the stimulating influence of sex steroids on the expression of galanin as previously described in Anguilla. The extensive distribution of the galanin-like immunoreactive neurons in amphibian brains suggests that this peptide may act as a neuromodulatur and/or neurotransmitter.