Biomedical subjects

Karoly Gulya

Publications and source records attributed to Karoly Gulya.

9 recordsLinked to original sources

PubMed2006

Dithranol abolishes UCH-L1 immunoreactivity in the nerve fibers of the rat orofacial skin.

Dithranol has been used to treat psoriasis for decades. Although its beneficial effect may involve the induction of cutaneous inflammation, and inflammation often leads to damages in nerve fibers, these alterations are not well documented. Therefore, we investigated the effects of dithranol on the immunohistochemical characteristics of the cutaneous nerve fibers in the rat skin. Epidermal nerve fiber staining was achieved with ubiquitin carboxyl-terminal hydrolase L1 (UCH-L1) immunohistochemistry in the orofacial skin of control rats, rats treated with (a) dithranol for 5 days, (b) corticosteroid for 5 days following dithranol treatment for 5 days, and (c) corticosteroid for 5 days. The results revealed a complete loss of UCH-L1 immunoreactivity in the dithranol-treated animals. Topical application of corticosteroid onto the inflamed skin for 5 days reversed this effect: the UCH-L1 immunoreactivity was almost completely restored. Steroid treatment for 5 days did not change the appearance of the UCH-L1-immunoreactive nerve fibers. These findings were supported by Western blot analyses. We conclude that dithranol, incidentally similarly to psoriasis, causes inflammation and abolishes UCH-L1 immunoreactivity in the rat orofacial skin in a corticosteroid-reversible manner. This phenomenon may be due to the ability of dithranol to cause oxidative damage to the UCH-L1 protein, and to the antioxidant activity of the corticosteroids countering this effect.

Animals↗

PubMed2006

Differential calmodulin gene expression in the nuclei of the rat midbrain-brain stem region.

We investigated the expression patterns of the three calmodulin (CaM) genes, using in situ hybridization techniques, to detect gene-specific [(35)S]- and digoxigenin-labeled cRNA probes complementary to the multiple CaM mRNAs in the nuclei of the midbrain-brain stem region of the adult rat. The distinct CaM genes were widely expressed throughout this region with moderate intensities. In spite of the similar general pattern, significant differences in the distributions of the multiple CaM mRNA species were found in certain areas. In general, the CaM III mRNAs were most abundant, followed by the CaM I and CaM II mRNA populations. Most of the transcripts were found in the neuronal somata comprising the medullar nuclei, while much less label was detected in the neuropil. The CaM III mRNAs were more than 2.5 times more abundant than the CaM II mRNAs in the nucleus of the trapezoid body, and more than two times more abundant in the motor trigeminal nucleus, the principal sensory trigeminal nucleus and the olivary nucleus. The CaM III mRNAs were less dominant in the medial lemniscus, the inferior colliculus and the pontine reticular nucleus than those of the other CaM gene-specific transcripts. The CaM mRNA levels were low to moderate, without significant differences, in the mesencephalic trigeminal nucleus. The differential control of the expression of the CaM genes may contribute to the regulation of the multiple neuronal functions linked to this complex brain region and regulated by different CaM-dependent mechanisms via its target proteins.

Animals↗

PubMed2005

Cloning and characterization of rat importin 9: implication for its neuronal function.

We describe the structure of the rat importin 9 gene, together with its transcripts and the encoded protein with its putative functional domains. The importin 9 gene contains 24 exons in a genomic region spanning >52,000 bp. It is transcribed into two mRNAs, generated by means of alternative polyadenylation site usage arranged in tandem. Both transcripts possess the same noncanonical polyadenylation signal (AGUAAA) in rat, this hexamer being conserved in all vertebrates examined. Additionally, intron 8 is bordered by AT-AC dinucleotides. Importin 9 is expressed throughout adult rat tissues, but the 114-kDa Importin 9 protein was detected only in the brain. The localization of the Importin 9 protein was examined by immunohistochemistry in both adult rat tissues and primary hippocampal cell cultures. The strongest labeling was detected in vivo in areas populated by neurons in high density and also in the dendritic processes emanating from these cells. This protein was clearly concentrated in the nuclei of these cells, although their cytoplasms too were heavily labeled. Strong cytoplasmic and very strong nuclear staining was found in a vast majority of the cells with neuronal morphology in vitro. Cultured cells with glial morphology generally exhibited a weaker cytoplasmic labeling. In these cells, the signal decorated the nuclear envelope without nuclear staining and gradually dwindled toward the cell periphery. These results hint at the cell- or tissue-type specific functions of this type of importin protein.

Amino Acid Sequence↗

PubMed2005

Multiple calmodulin mRNAs are selectively transported to functionally different neuronal and glial compartments in the rat hippocampus. An electron microscopic in situ hybridization study.

The ultrastructural distribution of the calmodulin (CaM) mRNAs transcribed from the three CaM genes was studied in the CA1 region of the adult rat hippocampus by means of electron microscopic in situ hybridization. Digoxigenin-labeled CaM gene-specific riboprobes were detected with nanogold-anti-digoxigenin antibody conjugate. The CaM mRNAs were differentially distributed in both the neuronal and glial cell compartments. The greatest difference in neuronal distribution of the CaM mRNAs was found in the dendrites, where the mRNAs transcribed from the CaM I and III genes were much more abundant than the CaM II mRNA. The neuronal perikarya were heavy labeled for all the CaM mRNAs. Interestingly, the myelinated axons and axon terminals also contained small amounts of nanogold particles for all the CaM mRNAs, which diminished with increasing distance from the soma. Most of the synaptic profiles, however, contained labeling only in the postsynaptic region. The CaM mRNAs were differentially distributed in the glial cells. While the glial cell somata were only lightly labeled, surprisingly concentrated labeling was present in the perisynaptic and perivascular astrocytic processes. In general, the CaM II mRNA was the least represented in the glial processes. Only a very low CaM gene expression was observed in the endothelial and resting microglial cells. These results provide ultrastructural evidence for differential targeting of the multiple CaM mRNA transcripts to the intracellular compartments and suggest their microdomain-specific regulation.

Animals↗

PubMed2004

Calmodulin, and various ways to regulate its activity.

Calmodulin (CaM), the ubiquitous calcium sensor protein, is involved in almost all intracellular events. In higher vertebrates, a single protein is encoded by multiple, co-expressed genes, and the number of discrete CaM transcripts produced by a single cell is further increased by intense alternative polyadenylation signal usage. It appears most likely that the individual transcripts possess unique intracellular fates, so that this apparent redundancy multiplies the number of challenges which the cell is able to respond to. The promoter regions of the different CaM genes have been analyzed. Several putative transcription factor binding sites have been identified; however, the elements responsible for their generally strong co-expression, and even those providing different spatial and temporal control, remain to be elucidated. Moreover, a powerful posttranscriptional control mechanism is responsible for the establishment of local intracellular CaM mRNA pools. This is mainly achieved by the selective targeting of mRNAs to various cellular domains, although regulation via mRNA stability cannot be ruled out. Finally, tailoring of the CaM protein itself offers the fastest way whereby the properties of this Ca2+-receptor protein can be changed. Indeed, several posttranslational modifications of CaM were described earlier, but their functions are not yet understood. Here, we briefly review the regulatory levels from the gene transcription to the covalent modifications of the synthesized protein.

Binding Sites↗

PubMed2003

Calmodulin gene expression in the neural retina of the adult rat.

Calmodulin (CaM) mRNAs are expressed with low abundancy in the adult rat neural retina. However, when digoxigenin (DIG)-labeled cRNA probes specific for each CaM mRNA population were hybridized at slightly alkaline pH (pH 8.0), the widespread distribution of CaM mRNA-expressing cells was revealed, with similar abundance for all three CaM genes. The CaM genes displayed a uniquely similar, layer-specific expression throughout the retina, and no significant differences were found in the distribution patterns of the CaM mRNA populations or the labeled cell types. The strongest signal for all CaM mRNAs was demonstrated in the ganglion cell layer and the inner nuclear layer, where the highest signal intensity was found within the inner sublamina. Similarly intermediate signal intensities for all CaM genes were detected in the inner and outer plexiform layers, within the vicinity of the outer limiting membrane and in the retinal pigment epithelium. A very low specific signal was characteristic in the outer nuclear layer and the photoreceptor inner segment layer, while no specific hybridization signal was observed in the photoreceptor outer segment layer. In summary, all CaM genes exhibited a similar and a characteristically layer-specific expression pattern in the adult rat retina.

Animals↗

PubMed2003

Intracellular targeting of calmodulin mRNAs in primary hippocampal cells.

We investigated the intracellular distribution of the mRNAs corresponding to the three non-allelic CaM genes in cultured hippocampal cells by in situ hybridization with digoxigenin-labeled gene-specific riboprobes. In neurons the perikaryon was heavily stained and strong dendritic mRNA targeting was detected for all three CaM genes. The color labeling exhibited a punctate distribution, suggesting that CaM mRNAs are transported in RNA granules. Immunocytochemistry for S100 demonstrated that glial cells express CaM mRNAs at a very low level. A minority of the cultured cells were negative for either labeling.

Animals↗

PubMed2002

Differential expression of multiple calmodulin genes in cells of the white matter of the rat spinal cord.

Calmodulin (CaM) displays complex cytoplasmic and synaptic functions in the nervous system. However, the very little information that is available on the gene expression of the multiple CaM genes in different glial cell types are from brain tissues of rodents, and no data have been published on their CaM gene expression in the spinal cord. Therefore, we have modified and tested a color in situ hybridization method sensitive enough to detect mRNA populations in cells with low CaM mRNA abundances in the white matter of the rat lumbar spinal cord. Morphologically, two distinct cell types expressing CaM mRNAs were detected. Differential CaM gene expression was demonstrated in medium-sized astrocyte-like cells that reside predominantly in the dorsal column of the spinal cord, where CaM I mRNA was most abundant, followed by the CaM III and CaM II mRNA populations. The oligodendrocytes displayed a less differential CaM gene expression in both the dorsal and the lateral columns, but the CaM I gene had a slightly higher expression level than those of the other CaM genes. The results indicate that the CaM gene expression profile of the spinal cord is richer and more complex than previously thought on the basis of conventional radioactive in situ hybridization techniques. Thus, when a method that is sufficiently sensitive was used, more cell types could be demonstrated to express CaM mRNAs; hence, in spite of their lower CaM expression, glial cells could also be visualized.

Animals↗

PubMed2002

Differential calmodulin gene expression in the rodent brain.

Apparently redundant members of the calmodulin (CaM) gene family encode for the same amino acid sequence. CaM, a ubiquitous cytoplasmic calcium ion receptor, regulates the function of a variety of target molecules even in a single cell. Maintenance of the fidelity of the active CaM-target interactions in different compartments of the cell requires a rather complex control of the total cellular CaM pool comprising multiple levels of regulatory circuits. Among these mechanisms, it has long been proposed that a multigene family maximizes the regulatory potentials at the level of the gene expression. CaM genes are expressed at a particularly profound level in the mammalian central nervous system (CNS), especially in the highly polarized neurons. Thus, in the search for clear evidence of the suggested differential expression of the CaM genes, much of the research has been focused on the elements of the CNS. This review aims to give a comprehensive survey on the current understanding of this field at the level of the regulation of CaM mRNA transcription and distribution in the rodent brain. The results indicate that the CaM genes are indeed expressed in a gene-specific manner in the developing and adult brain under physiological conditions. To establish local CaM pools in distant intracellular compartments (dendrites and glial processes), local protein synthesis from differentially targeted mRNAs is also employed. Moreover, the CaM genes are controlled in a unique, gene-specific fashion when responding to certain external stimuli. Additionally, putative regulatory elements have been identified on the CaM genes and mRNAs.

Animals↗