Biomedical subjects

G Germanà

Publications and source records attributed to G Germanà.

At least 19 recordsLinked to original sources

PubMed2006

Abnormal development of pacinian corpuscles in double trkB;trkC knockout mice.

Pacinian corpuscles depend on either Aalpha or Abeta nerve fibers of the large- and intermediate-sized sensory neurons for the development and maintenance of the structural integrity. These neurons express TrkB and TrkC, two members of the family of signal transducing neurotrophin receptors, and mice lacking TrkB and TrkC lost specific neurons and the sensory corpuscles connected to them. The impact of single or double targeted mutations in trkB and trkC genes in the development of Pacinian corpuscles was investigated in 25-day-old mice using immunohistochemistry and ultrastructural techniques. Single mutations on trkB or trkC genes were without effect on the structure and S100 protein expression, and caused a slight reduction in the number of corpuscles. In mice carrying a double mutation on trkB;trkC genes most of the corpuscles were normal with a reduction of 17% in trkB-/-;trkC+/- mice, and 8% in trkB +/-;trkC -/- mice. Furthermore, a subset of the remaining Pacinian corpuscles (23% in trkB-/-;trkC+/- mice; 3% in trkB+/-;trkC-/- mice) were hypoplasic or atrophic. Present results strongly suggest that the development of a subset of murine Pacinian corpuscles is regulated by the Trk-neurotrophin system, especially TrkB, acting both at neuronal and/or peripheral level. The precise function of each member of this complex in the corpuscular morphogenesis remains to be elucidated, though.

Animals↗

PubMed2006

Effects of increased nerve growth factor plasma levels on the expression of TrkA and p75 in rat testicles.

In addition to their well-known roles within the nervous system, the neurotrophins and their receptors regulate some functions in the reproductive system. In this study we used combined morphological and immunohistochemical techniques to investigate the presence and cellular localization in the rat testicle of the two receptors of nerve growth factor (NGF), i.e. TrkA and p75(NTR). Furthermore, to evaluate whether increased plasma levels of NGF affect the ageing process, 4-methylcathechol (4-MC), an inductor of NGF synthesis, was administered. Both TrkA and p75(NTR) were expressed in rat testicles, but the pattern and intensity of immunoreaction were marginally different between them. In adult rats TrkA was expressed in spermatozoa and spermatids, and p75 was expressed in spermatogonia. In newborn rats TrkA immunoreactivity was found in the Leydig cells, whereas p75 was detected in a cellular layer that surrounds the seminiferous tubules. In adult treated animals the immunoreaction for TrkA and p75(NTR) was also localized in the spermatocytes, whereas in newborn treated rats no changes in the pattern of immunoreaction was observed. The present findings suggest a role of the NGF/TrkA/p75 system in the physiology of reproduction, but the practical relevance of this remains to be established.

Animals↗

PubMed2005

Bcl-2 immunoreactivity in human cutaneous Meissner and Pacinian corpuscles.

The occurrence and distribution of Bcl-2, a protein involved in the death-life cell pathways, was investigated in the peripheral sensory nervous system of healthy adult humans, including lumbar dorsal root ganglia, nerve trunks and glabrous skin (to analyze sensory corpuscles) using Western blot and immunohistochemistry. The antibody used labelled a protein of 26 kDa of estimated molecular weight corresponding with Bcl-2. Immunohistochemistry showed that only a neuronal population in dorsal root ganglia, some axons in peripheral nerves and the axon supplying Meissner and Pacinian corpuscles contained Bcl-2, whereas peripheral glial cells (i.e. satellite glial cells, Schwann cell, and lamellar cells of sensory corpuscles) did not. These results suggest that in normal conditions, Bcl-2 is only present in some neuronal, but not glial, elements of the sensory peripheral nervous system. The functional significance, if any, of these results remains to be determined.

Adult↗

PubMed2005

TrkA and p75NTR in the ovary of adult cow and pig.

Neurotrophins play a critical role in the development of the mammalian ovary, oogenesis and folliculogenesis. In this study we investigated the cell localization of the two main receptors for nerve growth factor (NGF), TrkA and p75NTR, in the cow and pig ovary, using immunohistochemistry. Specific immunoreactivity for TrkA and p75NTR was detected in the ovary of both species, but the pattern and intensity of immunostaining were marginally different between them. The follicular cells regularly expressed immunoreactivity for both receptors. Immunoreactivity was also detected in the oocytes, independently of the maturational stage of the follicles, with the exception of primordial and primary follicles of the pig which did not display p75NTR. Taken together, these results suggest a possible direct role of NGF on oocytes expressing TrkA and p75NTR, in addition to the well-known roles in other ovary functions. The practical relevance of these data remains to be clarified.

Animals↗

PubMed2005

Effects of neurotrophin-3 gene mutation in the expression of neurocalcin.

Neurocalcin (NC) is a neuron-specific "EF-hand" calcium-binding protein present in a non-fully characterized subpopulation of dorsal root ganglion (DRG) neurons, some kinds of mechanoreceptors and proprioceptors, and in motor end-plates. In the present study we have characterized NC expression in spinal sensory and motor neurons, and their endings in newborn mouse. Because the neurotrophic factor neurotrophin-3 (NT-3) appears to plays a major role in the development and maturation of sensory and motor neuronal populations, we have studied NC immunoreactivity in newborn NT-3 null mutant. In NT-3 deficient animals the overall number of NC-immunoreactive DRG neurons was reduced by as much as 70% including all large neurons, but subpopulations of NC expressing small and intermediate-sized neurons survived. As expected no muscle spindles were found in NT-3 mutant mice while they were present and normally innervated by NC-positive nerve fibers in wild-type animals. On the other hand, NC immunoreactivity was dramatically decreased in motoneurons of the spinal cord, ventral root nerves and motor end-plates in the absence of NT-3. The present results demonstrate that NC-containing DRG neurons include all proprioceptive, and a subset of mechanoreceptive and proprioceptive. Furthermore, they strongly suggest that NT-3 is involved in the maturation of motor end-plates.

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PubMed2004

S100 protein is a useful and specific marker for hair cells of the lateral line system in postembryonic zebrafish.

The neuromast of the lateral line system of zebrafish has become an ideal model for the study of both developmental genetics and the vertebrate auditory system. Interestingly, the hair cells of this system have been found to selectively display immunoreactivity for S100 protein in some teleosts. In order to provide a selective marker for the sensory cells of the lateral line system, we have analyzed immunohistochemically the expression of S100 protein in zebrafish from the larval to the adult stage. In larval and adult animals S100 protein immunoreactivity was detected restricted to the hair cells of both superficial and canal neuromasts. Apparently the expression of S100 protein by hair cells was independent of the age, but it was expressed heterogeneously in the hair cells of canal neuromasts. The results of this work provide a feasible method to easily identify sensory cells in the neuromasts, and may be of interest in studies regarding development, differentiation or turnover of hair cells.

Animals↗

PubMed2003

Structural and ultrastructural aspects of the pericardial sac in some sea water teleosts.

Generally, in fishes the parietal pericardium is considered to be a semi-rigid structure. It is widely known that the entire parietal pericardium in fish, in contrast to that of higher vertebrates, is fixed to the body wall which consists mainly of skeletal muscle. This structural arrangement could explain the low end-systolic intrapericardial pressure in fishes. The morphology of the parietal pericardium of 11 sea-water teleosts, either free or breeding, coming from various areas of the southern Italy was studied. Macroscopical evaluation, but even more, histological and ultrastructural studies, showed some differences among the hearts of the examined subjects, more pronounced in the European hake (Merluccius merluccius, L.), in which a true pericardial sac was demonstrated, well differentiated and distinguishable from the other structures, in contrast to the observations in other subjects. The pericardial connective tissue showed no continuity with that of the surrounding structures, mainly skeletal muscle. This difference could be retraced to the well-known pattern of life and, therefore, to other individual as well as evolutionary factors in comparison with other teleosteans examined.

Animals↗

PubMed2003

Diprosopiasis in a lamb. A case report.

Conjoined twinning has been reported in most of the domestic animal species. Among them, sheep have the highest incidence of craniofacial defects. A live male crossbreed dystocic two-headed lamb was delivered from a 2-year-old Pinzerita sheep after first mating. After 40 h of life, the lamb spontaneously died. The most important gross findings involved the head, whereas the examination of different organ and tissue sections did not reveal remarkable histomorphological changes. The lamb was classified as a conjoined twinning and, on the basis of the facial duplication, as a diprosopus tetraophtalmus.

Animals↗

PubMed2002

S-100 protein is a selective marker for sensory hair cells of the lateral line system in teleosts.

The distribution of S100 protein in the neuromast of the lateral line system (LLS) was investigated immunohistochemically in alevins of three species of teleosts (Salmo trutta, Salmo salar and Dicentrarchus labrax), using a polyclonal antibody. In both the neuromasts of the canals, as well as in the pit organs, the hair cells, regarded as the specific sensory cells, displayed cytoplasmic immunoreactivity for S100 protein. Conversely, the supporting cells, mantle cells and basal cells were devoid of immunoreaction. These results demonstrate for the first time the occurrence of S100 in the LLS of teleosts. Due to the cell specific localization, this protein might serve as a marker for sensory hair cells in neuromasts.

Animals↗

PubMed2001

Involvement of the NGF receptors (Trka and p75lngfr) in the development and maintenance of the thymus.

Nerve growth factor (NGF) and its main low- (p75LNGFR) and high-affinity (TrkA) receptors have been found in the vertebrate thymus, thus suggesting they are involved in the control of thymic function. However, its role in this organ is poorly known. In the present study we used combined morphological and immunohistochemical techniques to analyze the distribution of TrkA and p75LNGFR in the rat thymus, as well as the structural changes in the thymus of p75 LNGFR or TrkA deficient mice. In adult rats both TrkA and p75LNGFR were localized in a subset of thymic epithelial cells found primarily in the subcapsular and medullary thymic regions, regarded to be endodermal-derived cells. Consistently, animals with a non-functional TrkA, but not those lacking p75LNGFR, showed structural changes consisting of a decrease in the density of thymocytes, absence of cortico-medullary border, and large cysts lined of endodermal epithelium. These results strongly suggest a function of the TrkA-NGF system in thymic functions mediated by epithelial cells, as well as a role of TrkA in the development of the murine thymus. The function of p75LNGFR remains to be established.

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PubMed2000

Neurocalcin-immunoreactive neurons in the mammalian dorsal root ganglia, including humans.

Neurocalcin (NC) is a recently characterized EF-hand calcium-binding protein present in a discrete population of sensory neurons and their peripheral mechanoreceptors, but its presence in peripheral nervous system neurons other than in the rat is still unknown. The present study was designed to investigate the occurrence of NC in the dorsal root ganglia (DRG) of several mammalian species (horse, buffalo, cow, sheep, pig, dog, and rat), including humans. DRG were fixed, embedded in paraffin, and processed for immunohistochemistry using a polyclonal antibody against NC. The size of the immunoreactive neurons was measured. In all species examined, NC immunoreactivity (IR) was restricted to neurons but the percentage, as well as the size of the immunoreactive neurons, varied among different species. As a rule, small neurons (diameter <20 microm) lack NC IR. In some species (pig, dog, buffalo, cow), only the largest neurons showed IR, whereas in others (sheep, horse, rat, and humans) they covered the entire range of neuron sizes. The pattern of immunostaining was cytoplasmic, although in some species (cow and buffalo), it formed a peripheral "ring." The present results demonstrate that mammalian DRG contain a subpopulation of NC-positive neurons, which varies from one species to another. Based on the neuron size, the possible function of the NC-containing neurons is discussed.

Adult↗

PubMed2000

Immunohistochemical localization of neurocalcin in human sensory neurons and mechanoreceptors.

The localization of neurocalcin in the developing and adult human peripheral nervous system (dorsal root and sympathetic ganglia (DRG, SG), and enteric nervous system (ENS)) was investigated using immunohistochemistry. A subpopulation of large-sized neurons in DRG of 9 and 12 weeks old embryos showed immunoreactivity (IR), whereas the sympathetic ganglia or enteric neurons did not. In adults, neurocalcin IR was restricted to a subpopulation of large (13%) and intermediate (15%) sized neurons in DRG. The protein was also found in muscular (67%) and cutaneous (12%) nerve fibers, as well as in the axons supplying muscular (muscle spindles, Golgi's tendon organs, and perimysial Pacinian corpuscles) and cutaneous (Meissner's but not Pacinian corpuscles) mechanoreceptors, as well as motor end-plates. Present results demonstrate that neurocalcin in both developing and adult humans can be used as a specific marker for a subpopulation of sensory neurons coupled to proprioception and touch, and for axons of motoneurons forming motor end-plates.

Adult↗

PubMed1998

A neuronal subpopulation in the mammalian enteric nervous system expresses TrkA and TrkC neurotrophin receptor-like proteins.

Increasing evidence suggests that, in addition to peripheral sensory and sympathetic neurons, the enteric neurons are also under the control of neurotrophins. Recently, neurotrophin receptors have been detected in the developing and adult mammalian enteric nervous system (ENS). Nevertheless, it remains to be established whether neurotrophin receptors are expressed in all enteric neurons and/or in glial cells and whether expression is a common feature in the enteric nervous system of all mammals or if interspecific differences exist. Rabbit polyclonal antibodies against Trk proteins (regarded as essential constituents of the high-affinity signal-transducing neurotrophin receptors) and p75 protein (considered as a low-affinity pan-neurotrophin receptor) were used to investigate the cell localization of these proteins in the ENS of adult man, horse, cow, sheep, pig, rabbit, and rat. Moreover, the percentage of neurons displaying immunoreactivity (IR) for each neurotrophin receptor protein was determined. TrkA-like IR and TrkC-like IR were observed in a neuronal subpopulation in both the myenteric and submucous plexuses, from esophagus to rectum in humans, and in the jejunum-ileum of the other species. Many neurons, and apparently all glial cells, in the human and rat enteric nervous system also displayed p75 IR. TrkB-like IR was found restricted to the glial cells of all species studied, with the exception of humans, in whom IR was mainly in glial cells and a small percentage of enteric neurons (about 5%). These findings indicate that the ENS of adult mammals express neuronal TrkA and TrkC, glial TrkB, and neuronal-glial p75, this pattern of distribution being similar in all examined species. Thus, influence of specific neurotrophins on their cognate receptors may be considered in the physiology and/or pathology of the adult ENS.

Aged↗

PubMed1998

Morphological and immunohistochemical considerations on the basal ganglia in pigeon (Columba livia).

Morphological and immunohistochemical studies carried out particularly around the level of the basal ganglia (BG) in birds, are reported and commented on. Our results showed, on paraffin embedded avian BG, both the histological features and the immunohistochemical findings on immunofluorescence distribution of some neuropeptides (especially Metenkephalin) in the striatal complex. By comparing our results of Metenkephalin immunoreactivity (Menkir) with the referred analogous ones of Substance P (SP) quoted in literature, we confirmed the strikingly similar labelling at the levels of the Lobus paraolfactorius (LPO) and Paleostriatum augmentatum (PA), in contrast with the very low immunoreactivity at the Paleostriatum primitivum (PP) levels. We were able also to demonstrate the strong MEnk-ir of the neurons of the Nucleus accumbens, Nucleus septalis and paraventricularis. All these findings are also in accord to the interpretation by many AA (Wynne and Gunturkun, 1995; Reinez et al., 1983), about the anatomical correspondence between the LPO-PA complex of birds and the caudate-putamen in mammals. Some MEnk + 'wooly like fibres' described in our specimens in the PA (on the contrary reported in the Gpe segment of mammals) apparently escape these correspondence.

Animals↗

PubMed1998

Immunohistochemical localization of S100 proteins in dorsal root, sympathetic and enteric ganglia of several mammalian species, including man.

The occurrence of S100 proteins in neurons of the mammalian peripheral nervous system is still controversial. This study was designed to investigate this topic in dorsal root ganglia (DRG) and the enteric nervous system (ENS) of several mammalian species (horse, buffalo, cow, sheep, pig, dog, rabbit and rat), as well as in DRG, paravertebral sympathetic ganglia (SG) and ENS of the adult man. Rat embryos of E17 and E19 were also examined. The material was fixed in Bouin's fixative, paraffin-embedded and processed for immunohistochemistry, combined with image analysis, using a panel of mono and polyclonal antibodies against S100alpha, S100beta or S100alpha + beta (referred to here as S100) proteins. In all species examined, strong S100 protein immunoreactivity (IR) was found in satellite glial cells and Schwann cells, which also showed S100alpha and S100beta IR in humans. Furthermore, faint S100 protein IR was observed in a subpopulation of DRG intermediate- and large-sized sensory neurons in humans, buffalo, sheep, and pig. The rat was the only species showing clear S100 and S100beta in neurons, labelling in about 30-35% in adults (small, intermediate and large in size), and about 88% at E17 and 42% at E19, respectively. Weak S100alpha protein IR was observed in most of human SG neurons. In ENS, S100 protein IR was restricted to enteric glial and Schwann cells, with the exception of cow and goat in which a subset of neurons in both the myenteric and submucous plexuses displayed strong S100 protein IR. Neuronal S100alpha IR and glial S100beta IR was found in the human ENS. The present results demonstrate intra- and inter-specific differences in the expression of S100 proteins by neurons of the peripheral nervous system among mammalian species. Furthermore, they also suggest that neuronal S100 protein, at least in humans, consists of both S100alpha and S100beta.

Animals↗

PubMed1997

Sensory epithelium of the vomeronasal organ express TrkA-like and epidermal growth factor receptor in adulthood. An immunohistochemical study in the horse.

BACKGROUND: The medial wall of the vomeronasal organ (VNO) is lined with a sensory epithelium that is closely related to the olfactory epithelium, which is developed from the olfactory placode. It undergoes continuous replacement during its life span. In other sensory epithelia, cell proliferation is under the control of some trophic factors. Whether these proteins are involved in the continuous turnover of the VNO epithelium is unknown. This study approaches this topic by analyzing the occurrence of signal-transducing receptor proteins for neurotrophins (Trk proteins) and epidermal growth factor (EGFr). METHODS: VNO samples were obtained from adult horses (n = 9) and processed for Western blot or immunohistochemical detection of TrkA, TrkB, TrkC, and EGFr. For immunohistochemistry, both frozen and formalin-fixed, paraffin-embedded sections were used. Antibodies against Trk proteins were polyclonal antibodies that map within the intracytoplasmic domain. Antibodies against EGFr were monoclonal antibodies that map within the external (clone EGFR1) or the cytoplasmic (clone F4) domains. RESULTS: TrkA-like, but not TrkB- or TrkC-like, protein was detected in the VNO. By using immunoblotting, protein bands of TrkA-like protein with estimated molecular weights of 43-45, 55, and 60 kDa were found. In agreement with these findings, the sensory epithelium lining the VNO displayed strong TrkA-like immunoreactivity. On the other hand, regular protein bands with estimated molecular weights of 100 and 170 kDa, corresponding with immature and full-length EGFr, respectively, were found with the clone F4, whereas the clone EGFR1 was ineffective in detecting EGFr with Western blot analysis. Positive EGFr immunolabelling was observed regularly in the supranuclear pole of the sensory epithelial cells, and the pattern was identical with both antibodies used. CONCLUSIONS: The present results provide evidence for the occurrence of EGFr in the VNO of the adult horse, suggesting a role for their ligands (EGF and transforming growth factor-alpha) in this organ, probably in continuous cell replacement, during the adult life span. However, although immunoreactivity for TrkA-like protein was regularly observed, because the full-length protein was not found, whether or not its putative ligands (nerve growth factor and neurotrophin-3) act on these cells remains to be demonstrated.

Animals↗

PubMed1997

Neurotrophin receptor-like proteins in Peyer's patches.

BACKGROUND: The neurotrophins are a family of growth factors that act on responsive cells through specific high-affinity signal-transducing receptors called Trk (A, B, and C) proteins. The neurotrophin receptor proteins are widely distributed in both nervous and nonnervous tissues, including the lymphoid organs. The expression of these receptor proteins by a cell population is an indication of responsiveness to the respective binding neurotrophin. The present study investigated the presence and cellular localization of high-affinity neurotrophin receptor proteins in equine and bovine Peyer's patches. METHODS: Peyer's patches from horse and cow intestine were fixed in Bouin's fixative, embedded in paraffin cut 10 microns thick, and studied immunohistochemically using rabbit polyclonal antibodies against specific epitopes of the intracellular domain of the Trk receptor proteins. RESULTS: Immunoreactivity (IR) for Trk-like proteins was found in specific cell populations in Peyer's patches. TrkA-IR in the horse was localized in dendritic cells of the interfollicular T-cell zones and in follicular dendritic cells of the lymphoid follicles; in the cow, TrkA-ir was present in reticulum cells. TrkB-like IR was present in cells found inside lymphoid follicles of the horse, probably reticulum cells. Furthermore, in both species, TrkB-IR was found in interstitial dendritic cells and/or macrophages of the intestinal lamina propria. No specific TrkC-like immunostaining was found in immunocompetent cells of Peyer's patches. CONCLUSIONS: Present findings demonstrate that, as in other lymphoid organs, the accessory nonlymphoid cells express immunoreactivity for high-affinity neurotrophin receptor proteins. These results seem to favor the notion that neurotrophins, especially nerve growth factor, could have a physiological role in secondary lymphoid organs, possibly acting on accessory cells and not directly on lymphocytes.

Animals↗

PubMed1997

The architecture of the myosalpinx in the sow as revealed by scanning electron microscopy.

OBJECTIVE: To provide a definitive settlement of data on the architecture of myosalpinx in the sow in consideration of controversial data existing in literature. STUDY-DESIGN: To allow direct visualization of muscular architecture, segments of tube from fifteen sows were investigated by means of scanning electron microscopy after the removal of interstitial connective tissue with NAOH digestion. RESULTS: In the extramural portion of the tubo-uterine junction, in the isthmus and ampulla, the myosalpinx is mainly constituted by oblique bundles of variable length, which run around the tube and merge into the surrounding musculature, giving origin to a plexiform arrangement. In the ampulla the fibers join in short bundles variously oriented. CONCLUSION: The three-dimensional architecture of the sow myosalpinx consists of muscular bundles independent of one another which follow multiple spatial arrangements and form a complex network. Such a muscular structure is likely more suitable for stirring rather than pushing the embryo and gametes.

Animals↗