Shin-ichi Mikami

Localization of avian LHRH-immunoreactive neurons in the hypothalamus of the domestic fowl, Gallus domesticus, and the Japanese quail, Coturnix coturnix

SummaryThe localization of LHRH-containing perikarya and nerve fibers in the hypothalami of the domestic fowl and Japanese quail was investigated by means of the specific immunoperoxidase ABC method, using antisera against chicken LHRH-I ([Gln8]-LHRH), chicken GnRH-II ([His5-Trp7-Tyr8]-LHRH [2–10]) and mammalian LHRH ([Arg8]-LHRH). Chicken LHRH-I-immunoreactive perikarya were sparsely scattered in the nucleus preopticus periventricularis (POP), nucleus filiformis (FIL) and nucleus septalis medialis (SM), and in bilateral bands extending from these nuclei into the septal area in both species. A few reactive perikarya were also observed in the nucleus accumbens (Ac) and lobus parolfactorius (LPO). Numerous cLHRH-I-immunoreactive fibers were widely scattered in the preoptic, septal and tuberal areas, and were densely concentrated in the external layer of the median eminence and in organum vasculosum of the lamina terminalis (OVLT) in both species. Anti-mammalian LHRH serum cross-reacted weakly with perikarya and fibers immunoreactive to anti-cLHRH-I serum in normal chicken and quail. Anti-cGnRH-II[2–10] serum immunoreacted with magnocellular neurons distributed in the rostral end of the mesencephalon along the midline close to the nervus oculomotorius (N III). These perikarya were apparently different from cLHRH-I immunoreactive neurons. No immunoreactive cells and fibers against anti-cGnRH-II[2–10] were observed in the hypothalamus and median eminence of the chicken or quail. Anti-cGnRH-II[2–10] bound specifically with cGnRH-II. The morphological evidence suggests that cGnRH-II may not be secreted into the portal circulation to act as hypothalamic hormone.

Immunocytochemistry of the Avian Hypothalamus and Adenohypophysis

Publisher Summary This chapter discusses the immunocytochemistry of the avian hypothalamus and adenohypophysis. The localization of immunoreactive neuropeptides—such as vasotocin, mesotocin, luteinizing-hormone-releasing hormone (LHRH), somatostatin, corticotropin-releasing factor (CRF), met-enkephalin, substance P, vasoactive intestinal peptide (VIP), and glucagon in the ventral hypothalamus—has been studied by means of immunocytochemistry. Each peptide-producing neuron system represents its own peculiar distribution pattern in the hypothalamus, while many nuclei of the hypothalamus—for example, the medial preoptic, paraventricular, and infundibular nuclei—contain many kinds of neuropeptide-producing neuron systems that show an overlapping or mosaic-like distribution. Also, many kinds of peptidergic fibers form dense networks surrounding other types of parvocellular neurons and are closely intermingled with each other in many nuclei of the hypothalamus. Comparative mapping studies show that gross overlaps occur in the distribution of substance P, enkephalin, somatostatin, and vasotocin neurons, while these overlaps are not due to costorage of the peptides in the same neurons: each peptide occurs in separate neurons and fibers. The structural relationships between different peptidergic systems suggest the presence of functional correlations or interactions among these systems. Therefore, the area of overlap must be considered as a nodal point of information exchange within the central nervous system.

Immunohistochemical localization of vasoactive intestinal polypeptide(VIP)-containing neurons in the hypothalamus of the Japanese quail, Coturnix coturnix

SummaryThe localization of vasoactive intestinal polypeptide (VIP) in the hypothalamus of the quail has been studied by means of light- and electron-microscopic immunohistochemistry. Numerous VIP-immunoreactive perikarya are distributed in the caudal portion of the nucleus infundibularis (n. tuberis) and nucleus mamillaris lateralis, and sparse in the preoptic area, nucleus supraopticus and nucleus paraventricularis. Dense localization of immunoreactive-VIP fibers is observed in the external layer of the median eminence, in close contact with the primary portal capillaries. The main origins of these fiber terminals are VIP-immunoreactive perikarya of the nucleus infundibularis. These neurons are spindle or bipolar and extend one process to the ventricular surface and another to the external layer of median eminence. They are CSF-contacting neurons and apparently constitute the tubero-hypophysial tract that links the third ventricle and the hypophysial portal circulation. VIP-reactive neurons in the nucleus mamillaris lateralis also project axons to the external layer of the median eminence, constituting the posterior bundle of the tuberohypophysial tract. Numerous VIP-immunoreactive perikarya occur also in the nucleus accumbens/pars posterior close to the lateral ventricle. They are also CSF-contacting neurons extending a process to the lateral ventricle. There are moderate distributions of VIP-reactive fibers in the area ventralis and in the area septalis.Ultrastructurally, the immunoreactive products against VIP are found in the elementary granules, 75–115 nm in diameter, within the nerve fibers in the median eminence.

Immunocytochemical studies on the pituitary pars distalis of the Japanese long-fingered bat, Miniopterus schreibersii fuliginosus

SummaryImmunocytochemical studies were performed to describe the characteristics of cell types and their distribution in the pars distalis of Japanese long-fingered bat, Miniopterus schreibersii fuliginosus, collected at various stages of the reproductive cycle. Six distinct cell types have been identified in the pars distalis by the unlabeled immunoperoxidase technique and by the ABC method. Growth hormone (GH) and prolactin (PRL) cells were immunostained with antisera against chicken GH and ovine PRL. The GH-immunoreactive cells were round or oval orangeophilic cells distributed throughout the pars distalis with prominent aggregation in the posterolateral region. The PRL cells were pleomorphic carminophilic cells that occurred in small groups within the central and dorsocaudal regions of the pars distalis. They were sparsely distributed in the central region of the pars distalis in the hibernating bats, but increased significantly in the pregnant and lactating bats. The adrenocorticotropic (ACTH) cells were large round or polygonal amphophilic cells in the rostroventral and ventrolateral regions of the pars distalis. The thyrotropic (TSH) cells were small rounded or polygonal and distributed mainly in the ventrolateral region of the pars distalis. Luteinizing hormone (LH) and follicle-stimulating hormone (FSH) cells were identified immunocytochemically with antisera against the specific beta subunits of ovine LH and rat FSH. There were two populations of LH and FSH cells, one aggregated in the zona tuberalis and the other scattered singly throughout the rest of the pars distalis. The aggregated cells were immunoreactive with both antisera directed to LH and FSH, while scattered cells were reactive solely with antiserum to either LHβ or FSH and exhibited seasonal variations. In females, the proportional volume of the pars distalis occupied by LH cells was significantly reduced during pregnancy and lactation. No evidence of involution was observed in pars distalis cells except for PRL cells in males or females during hibernation.

Cytodifferentiation of the adenohypophysis of the domestic fowl

SummaryIn the chick embryo the first membrane-bound secretory granules occur in the cytoplasm of occasional cells in the cephalic lobe of pars distalis at the 7th day of incubation. On the 8th day most of the cells in both the cephalic and caudal lobes contain secretory granules that are variable in size, form and density.On the 9th day at least two types of glandular cells are distinguishable in the cephalic and in the caudal lobes; however, these cells are not comparable with those of the adult gland. Differentiation of acidophils and basophils occurs, apparently simultaneously, in 11-day embryos.The cells of the cephalic and caudal lobes are morphologically distinct from their first appearance. Thus it is concluded that these two lobes develop independently and differently from an early stage of ontogenesis.The secretory granules are formed in the Golgi area of the hypophysial cells after the 8th day of incubation. However, secretory material may be synthesized also by a process not involving the Golgi apparatus.Nerve fibers containing granules first appear in the superficial layer of the median eminence on the 8th embryonic day and by the 12th day three types of granules and two types of clear vesicles are identifiable.

Fine structure of the epithelia of the vomeronasal organ of horse and cattle

SummaryThe vomeronasal organ of both horses and cattle is a tubular structure situated bilaterally at the base of the nasal septum. In frontal plane the shape of its lumen is semilunar to crescent. The sensory epithelium lining the medial wall of the lumen contains receptor, supporting and basal cells with some surface modifications in both species. In the horse, a structure similar to a microprocess was observed among the microvilli of receptor cells. In cattle, a large mass of the cytoplasm of the receptor cell occasionally protrudes to form a bleb-like structure. The supranuclear cytoplasm of the receptor cells contain mitochondria, free ribosomes, rough endoplasmic reticula, Golgi apparatus, lysosomes and multivesicular bodies. Some receptor cells were pyknotic. In both species the respiratory epithelia of the lateral wall of the lumen contain ciliated, non-ciliated and basal cells. In the horse, this epithelium differs from that of other species in evidence of prominent secretory function.

Light and electron microscopic investigations of six types of glandular cells of the bovine adenohypophysis.

SummaryTwelve bovine adenohypophyses were prepared for light and electron microscopy of the cell types of pars distalis. Correlation between the light and electron microscopy was effected by use of alternate thin and thick sections. Cytological changes in the experimental animals were used as criteria for the identification of six different types of secretory cells.Two types of acidophils, alpha and epsilon cells, are recognized in peripheral area of the pars distalis by light and electron microscopy. The alpha cells contain orangeophilic secretory granules of a maximum diameter of 400–450 mμ and correspond to ordinary acidophils (STH cells). The second type, epsilon cells, contains larger, fuchsinophilic granules of 600 to 900 mμ in diameter, increase in number and granulation after pregnancy and thyroidectomy, and are thought to be prolactin cells (LTH cells).Two types of amphophils, zeta and delta 1 cells, were found in the central area of the pars distalis. The zeta cells contain smaller numbers of amphophilic, cored granules (200 mμ maximum diameter) and based on the comparison with literature on other species of animals, are designated as ACTH cells. The delta 1 cells are round or oval and contain very dense, spherical granules (250–300 mμ) which are stained red or reddish purple with PAS, aldehyde thionin and PAS-methyl blue methods. They show extreme enlargement and bizarre cytoplasmic appearance after castration and are designated tentatively as LH gonadotrophs or LH cells.Two types of basophils, beta and delta 2 cells, were also identified by correlative light and electron microscopy. The beta cells are polygonal in outline, distributed exclusively in the zona tuberalis and contain large, less dense secretory granules (300–400 mμ) which are stained selectively with Gomoris aldehyde fuchsin. After thyroidectomy, they lose their secretory granules and are transformed into large, vacuolated thyroidectomy cells. They are therefore, identified as thyrotrophs or TSH cells. The delta 2 cells are round, oval or polygonal in shape and contain basophilic granules ranging from 220 to 300 mμ in diameter. They show extreme enlargement and vacuolization due to the dilation of endoplasmic reticulum, after castration, and are designated tentatively as FSH gonadotrophs or FSH cells.

Light- and electron-microscopic studies on the secretory cytology of the adenohypophysis of the Japanese quail, Coturnix coturnix japonica.

SummaryThe effects of thyroidectomy, adrenalectomy, and castration on the pars distalis of male Japanese quail, and of injection of LH-RH on sexually inactive females, were investigated by light and electron microscopy. Correlation between light and electron microscopy was attained by use of alternate thin and thick sections. Six types of secretory cells were identified and the ultrastructural characteristics described. Putative endocrine functions have been designated on the basis of responses to experimental interventions and on other criteria.The putative STH cells are characterized by the presence of large dense secretory granules (250–300 nm) that are stained with orange-G by the trichrome method. They occur only in the caudal lobe and appear to be unchanged by castration, thyroidectomy, adrenalectomy and LH-RH injection.The putative prolactin cells are characterized by large (400–600 nm), spherical or polymorphic, dense secretory granules stainable with acid fuchsin and aniline blue; prominent Golgi apparatus and well developed endoplasmic reticulum with densely packed, regularly parallel lamellae. They are found mainly in the cephalic lobe. The prolactin cells develop some vacuolization after adrenalectomy and undergo some degeneration after castration. The ACTH cells, which are restricted to the cephalic lobe, are identified by the dense, spherical granules (250–300 nm) that are stained with acid fuchsin. After adrenalectomy, they lose their secretory granules and are transformed into large, chromophobic adrenalectomy cells.TSH cells are so designated by their response to thyroidectomy including loss of their fine secretory granules and transformation to large, vacuolated thyroidectomy cells. We have found TSH cells and thyroidectomy cells only in the cephalic lobe.Basophilic cells, considered to be gonadotropes, occur in both the cephalic and caudal lobes. The gonadotropes of the cephalic lobe appear to have slightly larger (120–200 nm) granules than the caudal lobe (120–150 nm). However, after castration, the gonadotropes in both lobes become hypertrophied and vacuolated and are transformed into mutually indistinguishable castration cells. Twenty minutes after injection with LH-RH, the gonadotropes of both lobes increase in size and number, degranulate, develop vacuoles in the cytoplasm, and appear very similar to castration cells.

The hypothalamic neurosecretory systems of the Japanese quail as revealed by retrograde transport of horseradish peroxidase.

SummaryWithin 24–48 h after injection of horseradish peroxidase (HRP) into the neural lobe or into the median eminence of adult Japanese quail dense accumulations of its reaction product (HRP-RP) can be demonstrated in axons of the hypothalamo-hypophysial tract and in the magnocellular neurosecretory perikarya of the supraoptic and paraventricular nuclei as well as in scattered neurons of the accessory hypothalamic neurosecretory nuclei. The HRP-RP-containing nerve fibers, which are beaded in appearance, occur prominently in the internal zone of the median eminence. They turn dorsally at its anterior border to become widely distributed in the retrochiasmatic region and extend to the paraventricular, supraoptic and preoptic areas. These observations confirm more directly conclusions drawn earlier from Gomori-type preparations and from immunologic demonstration of arginine vasotocin, mesotocin and neurophysin.HRP-RP was also found in perikarya of parvocellular secretory neurons in the infundibular nucleus 48 h after injection of HRP into the median eminence but not after injection into the pars nervosa. This provides direct evidence that a conspicuous component of the tubero-infundibular tract is formed by axons of tuberal neurons that originate from the infundibular nucleus and pass directly into the median eminence.

Immunohistochemical localization of corticotropin-releasing factor (CRF)-containing neurons in the hypothalamus of the Japanese quail, Coturnix coturnix

SummaryImmunohistochemical localization of corticotropin-releasing factor (CRF)-like immunoreactivity in the brain of the Japanese quail was studied by means of the peroxidase anti-peroxidase (PAP) method. CRF-immunopositive perikarya of parvocellular neurons were observed mainly in the nucleus praeopticus medialis and nucleus paraventricularis. Additional perikarya were also detected in the nucleus hypothalamicus posterior medialis in the hypothalamus and in the non-hypothalamic nucleus accumbens, nucleus septalis lateralis and nucleus dorsomedialis and dorsolateralis thalami. No CRF immunoreaction was found to coexist with the vasotocin (Vt)-containing system in comparative examination of consecutive sections treated with anti-vasopressin (Vp) serum.The CRF-immunoreactive fibers were detected mainly in the external layer of the anterior median eminence but not in its posterior division. Unilateral adrenalectomy induced the marked reduction in number of the CRF immunopositive fibers in the anterior median eminence.