Search PubMed⌕ Search

Biomedical subjects

P P Graziadei

Publications and source records attributed to P P Graziadei.

At least 19 recordsLinked to original sources
PubMed1996

Lineage specification of olfactory neural precursor cells depends on continuous cell interactions.

We transplanted, as a single cell suspension, cells dissociated from the mature and immature olfactory epithelium of rats or TgR(ROSA26)26Sor mice expressing constitutively the LacZ gene into the developing brain (cerebellum, striatum, inferior colliculus, lateral ventricles) of E15 rat fetuses. Grafted cells or their descendants were still present in the central nervous system more than a month after transplantation. Transplanted cells either integrated as isolated cells or, during the first day after transplantation, reaggregated into clusters. Scattered cells, despite their placodal origin, differentiated into neuron or glial cells with a central phenotype. This was demonstrated by anatomical methods and selective amplification of cDNA encoding for neuronal specific transcripts (microtubule-associated protein 2 and middle-molecular-mass neurofilament protein) expressed by the engrafted cells. Cells in large clusters generated an epithelium containing mature olfactory neurons. Some of them were immunoreactive for the olfactory marker protein. Our findings show that cells dissociated from the developing and adult olfactory organs when transplanted into the rat fetal brain can either completely change their fate and differentiate according to their final position or generate an olfactory epithelium if they reaggregate into large clusters.

Animals↗
PubMed1996

An ultrastructural study of glomeruli associated with vomeronasal organs transplanted into the rat CNS.

Rat neonate vomeronasal organs were transplanted into the parietal cortex of littermates to examine their survival and the behavior of axon growth into the surrounding host brain parenchyma. After survival times of 10-100 days the brains were processed for ultrastructural examination. The transplanted vomeronasal organs (VNO) formed several vesicles lined with a sensory epithelium. From these sensory epithelia, VNO neurons leave the epithelium and enter the host brain. Transplant neurons grew axons that fasciculated into bundles surrounded by sheath cell processes and formed one or more fiber plexuses containing distinct globose or spherical-shaped glomerular-like structures. The glomeruli consisted of nerve terminals between which existed asymmetric synaptic contacts. Rarely did we observe clear reciprocal synapses. The glomeruli also contained terminals that showed signs of degeneration, such as increased density of the terminals, clumping of mitochondria and multivesicular bodies. The glomeruli were not partitioned or subdivided by glial septa; however, glial profiles were interspersed among the sensory terminals. Transplant glomeruli also lacked periglomerular cells and had no definitive glial envelope. These results suggest that glomerular formation is not dependent on dendrite contribution of second order neurons or glial support, but rather on a complementary population of receptor neurons.

Animals↗
PubMed1996

Synaptology of the olfactory bulb of an elasmobranch fish, Sphyrna tiburo.

The ultrastructure of the elasmobranch olfactory bulb was examined in order to determine the synaptology of the olfactory circuitry in the bonnethead shark, Sphyrna tiburo. The compartmentalization of the bulb, together with the lack of mitral cell basal dendrites, suggests a different way of performing lateral communication between mitral cells of the olfactory bulb. The results show that granule cells assume an important role by directly interlinking mitral cells. A corollary of this is the segregation of the input onto the mitral cell dendritic arborization: afferent fibers synapse onto the intraglomerular mitral terminals, whereas most local circuit interactions utilize extraglomerular synapses located on the shafts and the somas of the mitral dendrites. Therefore, the elasmobranch synaptic pattern is different from that of higher vertebrates; This might represent the use of a different neural route to achieve the same processing task.

Animals↗
PubMed1995

Eye primordium transplantation in Xenopus embryo.

A part of the eye primordium, the presumptive retinal anlage, was transplanted from stage-23/24 Xenopus borealis to replace the removed olfactory anlage of Xenopus laevis. Cells of the two species can be distinguished under fluorescence microscopy, and we used the resulting chimeras to determine whether the transplanted eye primordium would inhibit the regeneration of the olfactory anlage, whether it would connect with its usual target, the diencephalon, and whether migration of cells would occur from the transplant to the host CNS or from the host CNS to the transplant. In all cases, the olfactory anlage regenerated promptly, and normal olfactory bulbs developed. Omission of the eye stalk in the transplant resulted in failure of an optic nerve to develop from the developing retina. A cellular bridge containing the optic axons connected the transplanted retina to the diencephalon. Cells from the transplant migrated freely through the cellular bridge to several CNS regions. Their morphology, topographic arrangement, number, and relations with other host elements are consistent with the hypothesis that these cells belong to both glia and neuron types.

Animals↗
PubMed1995

Cell migration from the transplanted olfactory placode in Xenopus.

The eye vesicle of Xenopus borealis has been replaced with the transplanted olfactory primordium from Xenopus laevis in an attempt to determine whether cells from the transplant could migrate along the regrowing olfactory nerve and become incorporated into the CNS of the host. The use of X. laevis and X. borealis pairs allowed us to distinguish the cells of the host from those of the donor at the cellular level by means of the characteristic fluorescent nuclear spots (Q bands) of X. borealis. Transplantation was performed on pairs of animals at stages 23/24. The olfactory anlage was readily incorporated into the host, often fusing with the host homolateral organ and inhibiting the regrowth of the eye vesicle. An olfactory nerve developed from the transplanted organ. In the majority of cases, the nerve reached the diencephalon at the level of entrance of the optic nerve. Along the nerve originating from the transplanted organ we observed a stream of cells with the characteristics of the donor. These cells penetrated the host's CNS and became incorporated into it. The nature of these cells has not been ascertained by specific neuronal markers. However, on the basis of their morphology and disposition, the hypothesis suggested is that some of the migrating cells are neurons.

Animals↗
PubMed1995

Cell death in the olfactory epithelium.

In the nervous system of vertebrates the olfactory epithelium presents unique cytological characteristics. In the olfactory mucosa, olfactory neurons die and are replaced from undifferentiated neuroblasts over the entire life span of the animal. It remains unclear whether these neurons die as a result of a direct insult from the environment or in fulfillment of a physiological program of cell death. We have studied the distribution and the characteristics of cell death in the olfactory epithelium of normal, adult rats. The olfactory epithelium contains pycnotic bodies resembling those described for thymocytes undergoing terminal apoptotic changes. These appear at all levels in the epithelium, under both light and electron microscopes and can also be demonstrated after vital staining with acridine orange. Chromatin condensation into large blocks, often located at the nuclear periphery, is a morphological hallmark of the nuclei of mature olfactory neurons, which also present an increase in electron density of the cytoplasm. After non-radioactive in situ labeling of fragmented DNA, the nuclei of olfactory neurons are positive. Under the same reaction conditions (mild protease digestion), most of the nuclei of the supporting and basal cells are negative. In vivo incorporation of 5-bromouridine, a marker of RNA synthesis, is also lower in olfactory neurons than in basal and supporting cells. These findings suggest that olfactory neurons are committed very early to physiological cell death.

Acridine Orange↗
PubMed1995

Coculture of the vomeronasal organ and olfactory bulb of the fetal rat.

The vomeronasal organ and the olfactory bulb of the rat were cocultured from 15-day embryo siblings on collagen-coated membrane in Dulbecco's modified Eagle's medium containing fetal calf serum, horse serum, and antibiotics. At 4 days in vitro (DIV), vomeronasal axons forming two to three large fascicles were seen originating from the explants of the vomeronasal organ. Differential axonal growth was observed. Some fascicles made connections with the explants of the olfactory bulb. Twenty percent of the cocultures studied here showed the formation of connections. At 6-10 DIV many fascicles that did not connect with the olfactory bulb had degenerated, and large fascicles that were connected with the olfactory bulb survived for more than 10 DIV. The formation of connections between the vomeronasal organ and the olfactory bulb in coculture favors the survival of large nerve fascicles, but it could not be determined whether or not the presence of the olfactory bulb affects the initial orientation of the fibers and fascicles from the explants of the vomeronasal organ.

Animals↗
PubMed1995

Transplantation of postnatal vomeronasal organ in the CNS of newborn rats.

The present study was conducted to examine the survival and development of intracerebral transplanted neonatal rat vomeronasal organs (VNs). Complete neonatal (P5-P10) VNs were transplanted into the parietal cortex region of littermates and examined at 10-100 days by light microscopy. The VN survived and was organized into a series of vesicles lined by respiratory and/or sensory epithelia. Sensory neurons grew long axons that fasciculated and invaded the surrounding brain parenchyma. The newly developed axons did not prefer a specific brain region. The axons developed a complex fiber plexus either at the interface between transplant and host tissue or deep within the host brain parenchyma. Vomeronasal axons consistently formed glomerular-like structures within the fiber plexus. Our results suggest that glomerular formation is not dependent on specific target of length of axon development, but rather on a set of complementary axons that display mutual recognition.

Animals↗
PubMed1994

Vicia villosa agglutinin inhibits the fasciculation of vomeronasal axons in fetal rat vomeronasal organ culture.

The vomeronasal organ of rat was cultured from embryonic 15-day littermates. During 4-8 days in vitro, vomeronasal axons originating from the explants of vomeronasal organ formed 2-3 large fascicles. When Vicia villosa agglutinin (VVA) was added to the culture medium, fasciculation of the vomeronasal axons was inhibited. The timing of addition and the duration of the presence of VVA were related to the inhibition of fasciculation of vomeronasal axons. Glycocojugates that bind with the VVA may therefore play an important role in the fasciculation of developing vomeronasal axons.

Animals↗
PubMed1994

Monoclonal antibody (VOM2) specific for the luminal surface of the rat vomeronasal sensory epithelium.

Monoclonal antibodies were raised and selected for reactivity with the luminal surface of the rat vomeronasal organ. Among the monoclonal antibodies generated, the one named VOM2 showed specific immunoreactivity within the luminal surface of the rat vomeronasal sensory epithelium. The VOM2 antigen appeared weakly on the luminal surface at postnatal day 14 (P14). After P21, VOM2 immunoreactivity as strong as that in the adult vomeronasal organ was observed. Immunofluorescence staining using VOM2 antibody showed no reactivity on the luminal surface of the adult mouse or hamster vomeronasal organ. An immunoblotting analysis showed that the VOM2 antigen was a protein with a molecular weight of 24,500.

Animals↗
PubMed1994

Differential development of binding sites of two lectins in the vomeronasal axons of the rat accessory olfactory bulb.

Binding of fluorochrome-conjugated lectins, Bandeiraea simplicifolia lectin-I (BSL-I) and Vicia villosa agglutinin (VVA), to the vomeronasal axons was investigated in the accessory olfactory bulb (AOB) of developing rats at embryonic day (E) 16, 18, and 20, and postnatal day (P) 0, 3, 7, 14, and 28. Intense fluorescence for VVA was first observed at E18, and the position-specific binding pattern observed in adults was established at P0; intense fluorescence for VVA was observed in the posterior 2/3 of the vomeronasal nerve layer (VNL) and glomerular layer (GL) and weak fluorescence was present in the anterior 1/3 of these layers. Fluorescence for BSL-I was observed in the posterior half of VNL and GL at P0; the area bound with BSL-I was expanded to the anterior area and intensity of the fluorescence increased as the development proceeded. At P28, binding of BSL-I was observed in the entire VNL and GL as identical to adults. These results indicate that the binding sites of BSL-I and VVA in the vomeronasal axons at the level of rat AOB develop differentially during ontogeny, suggesting that rat VN axons consist of two subpopulations expressing different glycoconjugates.

Animals↗
PubMed1994

Mitral cell dendrites: a comparative approach.

Phylogenetically persistent structures such as the mitral cells of the vertebrate olfactory bulb undergo changes in their dendritic arbor in the course of evolution. The morphology of mitral cells and the main elements of the olfactory bulb circuit in all classes of vertebrates are reviewed in this paper. Most of the neuronal elements found in the mammalian olfactory bulb are present in anamniotes. However, in contrast to those of amniotes, the mitral cells of most anamniotes lack basal dendrites, and periglomerular cells are absent in fish. This suggests a different circuitry and therefore drastic changes in the processing of olfactory information within the olfactory bulb. Lateral inhibition, conferred by basal dendrites in anamniotes, must then utilize other mechanisms in anamniotes. Moreover, the marked segregation of olfactory inputs onto mammalian mitral cells is less obvious in mitral cells of anamniotes that lack basal dendrites. The general role of dendrites, including those of mitral cells, is discussed in the light of increasing evidence for dendritic excitability. The evolutionary significance of mitral cell basal dendrites is also discussed.

Animals↗
PubMed1994

Projections of the olfactory bulb in an elasmobranch fish, Sphyrna tiburo: segregation of inputs in the telencephalon.

We have previously shown that the morphological compartmentalization of the elasmobranch olfactory bulb is accompanied by a topographical arrangement of the primary olfactory projections onto the bulb. If this spatial arrangement is significant for the processing of the information, one would expect it to be preserved in the secondary olfactory centers of the telencephalon. In this paper, we describe the elasmobranch secondary projections from the olfactory bulb to the telencephalon, focusing on their spatial arrangements within the forebrain. Results show that the olfactory input onto the telencephalon are segregated. The medial olfactory tract projects rostrally onto the superficial layer of the dorsal pallium and onto the lateral pallium. The lateral olfactory tract projects caudally onto the lateral pallium, the striatum and the area superficialis basalis. Thus, the secondary olfactory projections are segregated within the telencephalon, with an overlapping of the secondary fibers in the main projection area, the lateral pallium.

Animals↗
PubMed1994

Microscopic structure of the olfactory organ of the clearnose skate, Raja eglanteria.

The olfactory organ of juvenile clearnose skates (Raja eglanteria) was studied with the light and electron microscopes. The organ is ovoid in shape, and its free surface is complicated by the presence of some 20 lamellae. Each lamella has a folded surface lined by a typical neurosensory olfactory epithelium. Bipolar olfactory receptor neurons, ciliated sustentacular cells, and basal cells are the pre-eminent cellular components of the epithelium. Two types of receptor neurons, both bearing microvilli but not cilia, were identified. The type 1 neuron is similar to that previously described in other fishes. The type 2 neuron has a characteristic morphology justifying a separate description. Its dendritic knob is larger than that of type 1, and its microvilli, which are shorter and thicker, are straight and regularly arranged. Tight bundles of filaments provide a skeleton to each microvillus, and these filament bundles reach more than 5 microns down into the dendrite. Type 2 receptor neurons have a well-developed Golgi complex and sparse rough endoplasmic reticulum (rER), whereas type 1 receptor neurons have a less well-developed Golgi complex and a conspicuous system of rER lamellae. The mucous layer on the epithelial surface is provided by the secretion of goblet cells that are situated mostly in the peripheral regions of each lamella. Secretory granules in the sustentacular cells and glands in the lamina propria were not observed.

Animals↗
PubMed1994

Influence of the olfactory organ on brain development.

The olfactory epithelium is a unique sensory structure that has the intrinsic ability to renew its receptor neurons naturally throughout the vertebrate lifetime. The olfactory epithelium is also a neurogenetic matrix that generates various cell populations during embryonic development and adulthood. Some of these cell types migrate to the forebrain and therefore contribute to brain formation. The molecules involved in cell migration and continuous reconnection of olfactory receptor neurons to the bulb are discussed. Experiments involving removal or transplants of the olfactory placode in amphibians suggest that the olfactory organ influences the development of the telencephalon. We believe that the olfactory organ has a morphogenetic influence and even possibly an inducing effect on the forebrain. This effect could be mediated by a number of organizing factors that we discuss. The profound influence of the olfactory organ on brain development underlines the importance of the olfactory organ for survival.

Aging↗
PubMed1993

A pilot study on morphological compartmentalization and heterogeneity in the elasmobranch olfactory bulb.

The olfactory bulb of many elasmobranch fishes is morphologically subdivided into distinct units or sub-bulbs immediately adjacent to the olfactory epithelium. We investigated this morphological feature in two species of shark and one species of ray in order to understand its impact on the arrangement of the primary olfactory projections onto the bulb. Using anterograde tracing methods in vitro (biocytin) as well as in fixed tissue (DiI), we observed a direct segregated projection of the olfactory afferents onto the bulb. Application of tracers to the lateral part of the olfactory epithelium resulted in staining restricted to this region of the bulb, whereas the same tracers applied to the medial part of the epithelium resulted in staining of the medial olfactory bulb. The sub-bulbs appear to be individual anatomical units that each receive input from the olfactory lamellae. Nissl and myelin staining as well as the Golgi method show that the cytoarchitecture of the sub-bulbs is not substantially different from that of other anamniotes. However, we did note the existence of two types of mitral cell, based on the morphology of their dendritic arborization. Type L cells exhibit a loose dendritic arborization, whereas type T cells are characterized by a dense, bush-like dendritic arborization. Both types of mitral cells lack basal dendrites.

Animals↗
PubMed1992

The differential staining patterns of two lectins in the accessory olfactory bulb of the rat.

We examined the binding sites of Bandeiraea simplicifolia lectin I (BSL-I) and Vicia villosa agglutinin (VVA) which bind to the vomeronasal nerve of the rat. BSL-I stained the whole vomeronasal nerve and glomerular layers. VVA strongly stained the posterior 2/3, but weakly stained the anterior 1/3 of the glomerular layer. These results indicate that the glomeruli of the rat AOB have two subdivisions revealed by lectin histochemistry using BSL-I and VVA.

Animals↗