Krisztina Nikovics - Academia.edu (original) (raw)

Papers by Krisztina Nikovics

Research paper thumbnail of The Balance between the<i>MIR164A</i>and<i>CUC2</i>Genes Controls Leaf Margin Serration in<i>Arabidopsis</i>

The Plant Cell, Nov 1, 2006

CUP-SHAPED COTYLEDON1 (CUC1), CUC2, and CUC3 define the boundary domain around organs in the Arab... more CUP-SHAPED COTYLEDON1 (CUC1), CUC2, and CUC3 define the boundary domain around organs in the Arabidopsis thaliana meristem. CUC1 and CUC2 transcripts are targeted by a microRNA (miRNA), miR164, encoded by MIR164A, B, and C. We show that each MIR164 is transcribed to generate a large population of primary miRNAs of variable size with a locally conserved secondary structure around the pre-miRNA. We identified mutations in the MIR164A gene that deepen serration of the leaf margin. By contrast, leaves of plants overexpressing miR164 have smooth margins. Enhanced leaf serration was observed following the expression of an miR164-resistant CUC2 but not of an miR164-resistant CUC1. Furthermore, CUC2 inactivation abolished serration in mir164a mutants and the wild type, whereas CUC1 inactivation did not. Thus, CUC2 specifically controls leaf margin development. CUC2 and MIR164A are transcribed in overlapping domains at the margins of young leaf primordia, with transcription gradually restricted to the sinus, where the leaf margins become serrated. We suggest that leaf margin development is controlled by a two-step process in Arabidopsis. The pattern of serration is determined first, independently of CUC2 and miR164. The balance between coexpressed CUC2 and MIR164A then determines the extent of serration.

Research paper thumbnail of A conserved role for<i>CUP-SHAPED COTYLEDON</i>genes during ovule development

Plant Journal, Jul 23, 2015

The evolution of plant reproductive strategies has led to a remarkable diversity of structures, e... more The evolution of plant reproductive strategies has led to a remarkable diversity of structures, especially within the flower, a structure characteristic of the angiosperms. In flowering plants, sexual reproduction depends notably on the development of the gynoecium that produces and protects the ovules. In Arabidopsis thaliana, ovule initiation is promoted by the concerted action of auxin with CUC1 (CUP-SHAPED COTYLEDON1) and CUC2, two genes encoding transcription factors of the NAC family (NAM/ATAF1,2/CUC). Here we highlight an additional role for CUC2 and CUC3 in Arabidopsis thaliana ovule separation. While CUC1 and CUC2 are broadly expressed in the medial-tissue of the gynoecium, CUC2 and CUC3 are expressed in the placental tissue between developing ovules. Consistent with the partial overlap between CUC1, CUC2 and CUC3 expression patterns, we show that CUC proteins can physically interact, both in yeast cells and in planta. We found that the cuc2;cuc3 double mutant specifically harbors defects in ovule separation, producing fused seeds sharing the seed coat, suggesting that CUC2 and CUC3 promote in a partially redundant manner ovule separation. Functional analyses show that CUC transcription factors are also involved in ovule development in Cardamine hirsuta. Additionally we show a conserved expression pattern of CUC orthologs between ovule primordia in other phylogenetically distant species with different gynoecium architectures. Taken together these results suggest an ancient role for CUC transcription factors in ovule separation, and shed light on the conservation of mechanisms involved in the development of innovative structures. This article is protected by copyright. All rights reserved.

Research paper thumbnail of Influence of the Immune Microenvironment Provided by Implanted Biomaterials on the Biological Properties of Masquelet-Induced Membranes in Rats: Metakaolin as an Alternative Spacer

Biomedicines, Nov 23, 2022

This article is an open access article distributed under the terms and conditions of the Creative... more This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY

Research paper thumbnail of Immunolabeling Of Tgf-Β During Muscle Regeneration

Muscle regeneration after injury (as irradiation) is of great importance. However, the molecular ... more Muscle regeneration after injury (as irradiation) is of great importance. However, the molecular and cellular mechanisms are still unclear. Cytokines are believed to play fundamental role in the different stages of muscle regeneration. They are secreted by many cell populations, but the predominant producers are macrophages and helper T cells. On the other hand, it has been shown that adipose tissue derived stromal/stem cell (ASC) injection could improve muscle regeneration. Stem cells probably induce the coordinated modulations of gene expression in different macrophage cells. Therefore, we investigated the patterns and timing of changes in gene expression of different cytokines occurring upon stem cells loading. Muscle regeneration was studied in an irradiated muscle of minipig animal model in presence or absence of ASC treatment (irradiated and treated with ASCs, IRR+ASC; irradiated not-treated with ASCs, IRR; and non-irradiated no-IRR). We characterized macrophage populations by...

Research paper thumbnail of Molecular and Cellular Mechanisms of Inflammation and Tissue Regeneration

Biomedicines, May 10, 2023

This article is an open access article distributed under the terms and conditions of the Creative... more This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY

Research paper thumbnail of In situ identification of both IL-4 and IL-10 Cytokine-Receptor Interactions during tissue regeneration

Cytokines secreted by individual immune cells regulate tissue regeneration and allow communicatio... more Cytokines secreted by individual immune cells regulate tissue regeneration and allow communication between various cell types. Cytokines bind to cognate receptors and trigger the healing process. Determining the orchestration of cytokine interactions with their receptors on their cellular targets is essential to fully understand the process of inflammation and tissue regeneration. To this end, we have investigated the interactions of Interleukin-4 cytokine (IL-4)/Interleukin-4 cytokine receptor (IL-4R) and Interleukin-10 cytokine (IL-10)/Interleukin-10 cytokine receptor (IL-10R) using in situ Proximity Ligation Assays in a regenerative model of skin, muscle and lung tissues in the mini-pig. The pattern of protein-protein interactions was distinct for the two cytokines. IL-4 bound predominantly to receptors on macrophages and endothelial cells around the blood vessels while the target cells of IL-10 were mainly receptors on muscle cells. Our results show that in situ studies of cytok...

Research paper thumbnail of Molecular and cellular mechanisms of inflammation

Biochemistry (Moscow), 2016

Inflammation is one of the most fundamental and pronounced protective reactions of the organism. ... more Inflammation is one of the most fundamental and pronounced protective reactions of the organism. From ancient times to the present day, complex and diverse patterns of inflammation development and their role in various diseases have attracted attention of investigators. This issue of Biokhimiya/Biochemistry (Moscow) includes experimental studies and reviews dedicated to various aspects of this important and interesting problem.

Research paper thumbnail of Macrophages Characterization in an Injured Bone Tissue

Biomedicines

Biomaterial use is a promising approach to facilitate wound healing of the bone tissue. Biomateri... more Biomaterial use is a promising approach to facilitate wound healing of the bone tissue. Biomaterials induce the formation of membrane capsules and the recruitment of different types of macrophages. Macrophages are immune cells that produce diverse combinations of cytokines playing an important role in bone healing and regeneration, but the exact mechanism remains to be studied. Our work aimed to identify in vivo macrophages in the Masquelet induced membrane in a rat model. Most of the macrophages in the damaged area were M2-like, with smaller numbers of M1-like macrophages. In addition, high expression of IL-1β and IL-6 cytokines were detected in the membrane region by RT-qPCR. Using an innovative combination of two hybridization techniques (in situ hybridization and in situ hybridization chain reaction (in situ HCR)), M2b-like macrophages were identified for the first time in cryosections of non-decalcified bone. Our work has also demonstrated that microspectroscopical analysis is ...

Research paper thumbnail of A Novel Family in Medicago truncatula Consisting of More Than 300 Nodule-Specific Genes Coding for Small, Secreted Polypeptides with Conserved Cysteine Motifs

PLANT PHYSIOLOGY, 2003

Transcriptome analysis of Medicago truncatula nodules has led to the discovery of a gene family n... more Transcriptome analysis of Medicago truncatula nodules has led to the discovery of a gene family named NCR (nodule-specific cysteine rich) with more than 300 members. The encoded polypeptides were short (60–90 amino acids), carried a conserved signal peptide, and, except for a conserved cysteine motif, displayed otherwise extensive sequence divergence. Family members were found in pea (Pisum sativum), broad bean (Vicia faba), white clover (Trifolium repens), and Galega orientalis but not in other plants, including other legumes, suggesting that the family might be specific for galegoid legumes forming indeterminate nodules. Gene expression of all family members was restricted to nodules except for two, also expressed in mycorrhizal roots. NCR genes exhibited distinct temporal and spatial expression patterns in nodules and, thus, were coupled to different stages of development. The signal peptide targeted the polypeptides in the secretory pathway, as shown by green fluorescent protein...

Research paper thumbnail of In Situ Gene Expression in Native Cryofixed Bone Tissue

Biomedicines, 2022

Bone is a very complex tissue that is constantly changing throughout the lifespan. The precise me... more Bone is a very complex tissue that is constantly changing throughout the lifespan. The precise mechanism of bone regeneration remains poorly understood. Large bone defects can be caused by gunshot injury, trauma, accidents, congenital anomalies and tissue resection due to cancer. Therefore, understanding bone homeostasis and regeneration has considerable clinical and scientific importance in the development of bone therapy. Macrophages are well known innate immune cells secreting different combinations of cytokines and their role in bone regeneration during bone healing is essential. Here, we present a method to identify mRNA transcripts in cryosections of non-decalcified rat bone using in situ hybridization and hybridization chain reaction to explore gene expression in situ for better understanding the gene expression of the bone tissues.

Research paper thumbnail of Material and Methods

A. caerulea seeds were bought from a gardener shop (Jardiland, Maurepas, France). S. lycorpersicu... more A. caerulea seeds were bought from a gardener shop (Jardiland, Maurepas, France). S. lycorpersicum M82 wild type were received from S. Biemelt, the la mutant (accession LA0335) from the Tomato Genetics Ressource Center and the gob mutants from D. Zamir. P. sativum (cv Térèse and line NGB5839) were obtained from C. Rameau and the uni mutant (line JI2171) from J. Hofer. S. tuberosum cv Pompadour were a gift from B. Letarnec. RNA extraction and RT-PCR RNA was extract from partially dissected apices or young leaves of the different species using TRIsol reagent (Invitrogen) according to the manufacturer recommendations. Contaminating DNA was removed by DNaseI treatment (Invitrogen) and the RNA was reverse transcribed (SuperScript TM II reverse transcriptase, Invitrogen). Gene specific primers used for semiquantitative RT-PCR are listed in Supplementary Table 4. PCR products were observed on ethidium bromide-stained gels or following southern blotting (PsNAM1/2).Cloning of NAM/CUC genes. ...

Research paper thumbnail of Synchronization of Oryza sativa L.cv. Taipei-309 cell suspension culture

... Ágnes Lendvai*, Krisztina Nikovics, László Bakó, Dénes Dudits, János Györgyey ... Nikovics K,... more ... Ágnes Lendvai*, Krisztina Nikovics, László Bakó, Dénes Dudits, János Györgyey ... Nikovics K, Simidjieva J, Peres A, Ayaydin F, Pasternak T, Davies JW, Boulton MI, Dudits D, Horvath GV (2001) Cell-cycle, phase-specific activation of Maize streak virus promoters. Mol. ...

Research paper thumbnail of An Arabidopsis Cyclin Promoter Region is Active in Transgenic Maize Plants

We have produced transgenic maize carrying Arabidopsis cyclin promoter and beta-glucuronidase chi... more We have produced transgenic maize carrying Arabidopsis cyclin promoter and beta-glucuronidase chimeric fusion (cycB1;1At::GUS) by direct DNA uptake into embryogenic cell suspension protoplasts to study the activity of this cyclin promoter in a heterologous cell system. The transgenic maize lines exhibiting different levels of reporter gene expression show that regulatory elements from cycB1;1At gene as a dicot promoter can function in maize. Expression patterns revealed by fluorimetric, as well as, histochemical GUS reporter enzymatic assays indicated cell division-dependent expression of the GUS gene driven by this cycB1;At;1 promoter with elevated activity in proliferating cells and meristems. Treatment of tissues with inhibitors of mitosis increased the activity of cycB1;1At promoter, that suggests a link between mitotic phase of cell division and activity of cycB1;1At regulatory elements in transgenic maize.

Research paper thumbnail of Docking of acetyl-CoA carboxylase to the plastid envelope membrane attenuates fatty acid production in plants

Nature Communications

In plants, light-dependent activation of de novo fatty acid synthesis (FAS) is partially mediated... more In plants, light-dependent activation of de novo fatty acid synthesis (FAS) is partially mediated by acetyl-CoA carboxylase (ACCase), the first committed step for this pathway. However, it is not fully understood how plants control light-dependent FAS regulation to meet the cellular demand for acyl chains. We report here the identification of a gene family encoding for three small plastidial proteins of the envelope membrane that interact with the α-carboxyltransferase (α-CT) subunit of ACCase and participate in an original mechanism restraining FAS in the light. Light enhances the interaction between carboxyltransferase interactors (CTIs) and α-CT, which in turn attenuates carbon flux into FAS. Knockouts for CTI exhibit higher rates of FAS and marked increase in absolute triacylglycerol levels in leaves, more than 4-fold higher than in wild-type plants. Furthermore, WRINKLED1, a master transcriptional regulator of FAS, positively regulatesCTI1expression by direct binding to its pro...

Research paper thumbnail of Hybridization‐chain‐reaction is a relevant method for in situ detection of M2d‐like macrophages in a mini‐pig model

Research paper thumbnail of Characterization of macrophages, giant cells and granulomas during muscle regeneration after irradiation

Research paper thumbnail of Non-Specific Binding, a Limitation of the Immunofluorescence Method to Study Macrophages In Situ

Genes

Advances in understanding tissue regenerative mechanisms require the characterization of in vivo ... more Advances in understanding tissue regenerative mechanisms require the characterization of in vivo macrophages as those play a fundamental role in this process. This characterization can be approached using the immuno-fluorescence method with widely studied and used pan-markers such as CD206 protein. This work investigated CD206 expression in an irradiated-muscle pig model using three different antibodies. Surprisingly, the expression pattern during immunodetection differed depending on the antibody origin and could give some false results. False results are rarely described in the literature, but this information is essential for scientists who need to characterize macrophages. In this context, we showed that in situ hybridization coupled with hybridization-chain-reaction detection (HCR) is an excellent alternative method to detect macrophages in situ.

Research paper thumbnail of Macrophage Identification In Situ

Biomedicines

Understanding the processes of inflammation and tissue regeneration after injury is of great impo... more Understanding the processes of inflammation and tissue regeneration after injury is of great importance. For a long time, macrophages have been known to play a central role during different stages of inflammation and tissue regeneration. However, the molecular and cellular mechanisms by which they exert their effects are as yet mostly unknown. While in vitro macrophages have been characterized, recent progress in macrophage biology studies revealed that macrophages in vivo exhibited distinctive features. Actually, the precise characterization of the macrophages in vivo is essential to develop new healing treatments and can be approached via in situ analyses. Nowadays, the characterization of macrophages in situ has improved significantly using antigen surface markers and cytokine secretion identification resulting in specific patterns. This review aims for a comprehensive overview of different tools used for in situ macrophage identification, reporter genes, immunolabeling and in si...

Research paper thumbnail of First Insights Into the M2 Inflammatory Response After Adipose-Tissue-Derived Stem Cell Injections in Radiation-Injured Muscles

Research paper thumbnail of Material and Methods

A. caerulea seeds were bought from a gardener shop (Jardiland, Maurepas, France). S. lycorpersicu... more A. caerulea seeds were bought from a gardener shop (Jardiland, Maurepas, France). S. lycorpersicum M82 wild type were received from S. Biemelt, the la mutant (accession LA0335) from the Tomato Genetics Ressource Center and the gob mutants from D. Zamir. P. sativum (cv Térèse and line NGB5839) were obtained from C. Rameau and the uni mutant (line JI2171) from J. Hofer. S. tuberosum cv Pompadour were a gift from B. Letarnec. RNA extraction and RT-PCR RNA was extract from partially dissected apices or young leaves of the different species using TRIsol reagent (Invitrogen) according to the manufacturer recommendations. Contaminating DNA was removed by DNaseI treatment (Invitrogen) and the RNA was reverse transcribed (SuperScript TM II reverse transcriptase, Invitrogen). Gene specific primers used for semiquantitative RT-PCR are listed in Supplementary Table 4. PCR products were observed on ethidium bromide-stained gels or following southern blotting (PsNAM1/2).Cloning of NAM/CUC genes. ...

Research paper thumbnail of The Balance between the<i>MIR164A</i>and<i>CUC2</i>Genes Controls Leaf Margin Serration in<i>Arabidopsis</i>

The Plant Cell, Nov 1, 2006

CUP-SHAPED COTYLEDON1 (CUC1), CUC2, and CUC3 define the boundary domain around organs in the Arab... more CUP-SHAPED COTYLEDON1 (CUC1), CUC2, and CUC3 define the boundary domain around organs in the Arabidopsis thaliana meristem. CUC1 and CUC2 transcripts are targeted by a microRNA (miRNA), miR164, encoded by MIR164A, B, and C. We show that each MIR164 is transcribed to generate a large population of primary miRNAs of variable size with a locally conserved secondary structure around the pre-miRNA. We identified mutations in the MIR164A gene that deepen serration of the leaf margin. By contrast, leaves of plants overexpressing miR164 have smooth margins. Enhanced leaf serration was observed following the expression of an miR164-resistant CUC2 but not of an miR164-resistant CUC1. Furthermore, CUC2 inactivation abolished serration in mir164a mutants and the wild type, whereas CUC1 inactivation did not. Thus, CUC2 specifically controls leaf margin development. CUC2 and MIR164A are transcribed in overlapping domains at the margins of young leaf primordia, with transcription gradually restricted to the sinus, where the leaf margins become serrated. We suggest that leaf margin development is controlled by a two-step process in Arabidopsis. The pattern of serration is determined first, independently of CUC2 and miR164. The balance between coexpressed CUC2 and MIR164A then determines the extent of serration.

Research paper thumbnail of A conserved role for<i>CUP-SHAPED COTYLEDON</i>genes during ovule development

Plant Journal, Jul 23, 2015

The evolution of plant reproductive strategies has led to a remarkable diversity of structures, e... more The evolution of plant reproductive strategies has led to a remarkable diversity of structures, especially within the flower, a structure characteristic of the angiosperms. In flowering plants, sexual reproduction depends notably on the development of the gynoecium that produces and protects the ovules. In Arabidopsis thaliana, ovule initiation is promoted by the concerted action of auxin with CUC1 (CUP-SHAPED COTYLEDON1) and CUC2, two genes encoding transcription factors of the NAC family (NAM/ATAF1,2/CUC). Here we highlight an additional role for CUC2 and CUC3 in Arabidopsis thaliana ovule separation. While CUC1 and CUC2 are broadly expressed in the medial-tissue of the gynoecium, CUC2 and CUC3 are expressed in the placental tissue between developing ovules. Consistent with the partial overlap between CUC1, CUC2 and CUC3 expression patterns, we show that CUC proteins can physically interact, both in yeast cells and in planta. We found that the cuc2;cuc3 double mutant specifically harbors defects in ovule separation, producing fused seeds sharing the seed coat, suggesting that CUC2 and CUC3 promote in a partially redundant manner ovule separation. Functional analyses show that CUC transcription factors are also involved in ovule development in Cardamine hirsuta. Additionally we show a conserved expression pattern of CUC orthologs between ovule primordia in other phylogenetically distant species with different gynoecium architectures. Taken together these results suggest an ancient role for CUC transcription factors in ovule separation, and shed light on the conservation of mechanisms involved in the development of innovative structures. This article is protected by copyright. All rights reserved.

Research paper thumbnail of Influence of the Immune Microenvironment Provided by Implanted Biomaterials on the Biological Properties of Masquelet-Induced Membranes in Rats: Metakaolin as an Alternative Spacer

Biomedicines, Nov 23, 2022

This article is an open access article distributed under the terms and conditions of the Creative... more This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY

Research paper thumbnail of Immunolabeling Of Tgf-Β During Muscle Regeneration

Muscle regeneration after injury (as irradiation) is of great importance. However, the molecular ... more Muscle regeneration after injury (as irradiation) is of great importance. However, the molecular and cellular mechanisms are still unclear. Cytokines are believed to play fundamental role in the different stages of muscle regeneration. They are secreted by many cell populations, but the predominant producers are macrophages and helper T cells. On the other hand, it has been shown that adipose tissue derived stromal/stem cell (ASC) injection could improve muscle regeneration. Stem cells probably induce the coordinated modulations of gene expression in different macrophage cells. Therefore, we investigated the patterns and timing of changes in gene expression of different cytokines occurring upon stem cells loading. Muscle regeneration was studied in an irradiated muscle of minipig animal model in presence or absence of ASC treatment (irradiated and treated with ASCs, IRR+ASC; irradiated not-treated with ASCs, IRR; and non-irradiated no-IRR). We characterized macrophage populations by...

Research paper thumbnail of Molecular and Cellular Mechanisms of Inflammation and Tissue Regeneration

Biomedicines, May 10, 2023

This article is an open access article distributed under the terms and conditions of the Creative... more This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY

Research paper thumbnail of In situ identification of both IL-4 and IL-10 Cytokine-Receptor Interactions during tissue regeneration

Cytokines secreted by individual immune cells regulate tissue regeneration and allow communicatio... more Cytokines secreted by individual immune cells regulate tissue regeneration and allow communication between various cell types. Cytokines bind to cognate receptors and trigger the healing process. Determining the orchestration of cytokine interactions with their receptors on their cellular targets is essential to fully understand the process of inflammation and tissue regeneration. To this end, we have investigated the interactions of Interleukin-4 cytokine (IL-4)/Interleukin-4 cytokine receptor (IL-4R) and Interleukin-10 cytokine (IL-10)/Interleukin-10 cytokine receptor (IL-10R) using in situ Proximity Ligation Assays in a regenerative model of skin, muscle and lung tissues in the mini-pig. The pattern of protein-protein interactions was distinct for the two cytokines. IL-4 bound predominantly to receptors on macrophages and endothelial cells around the blood vessels while the target cells of IL-10 were mainly receptors on muscle cells. Our results show that in situ studies of cytok...

Research paper thumbnail of Molecular and cellular mechanisms of inflammation

Biochemistry (Moscow), 2016

Inflammation is one of the most fundamental and pronounced protective reactions of the organism. ... more Inflammation is one of the most fundamental and pronounced protective reactions of the organism. From ancient times to the present day, complex and diverse patterns of inflammation development and their role in various diseases have attracted attention of investigators. This issue of Biokhimiya/Biochemistry (Moscow) includes experimental studies and reviews dedicated to various aspects of this important and interesting problem.

Research paper thumbnail of Macrophages Characterization in an Injured Bone Tissue

Biomedicines

Biomaterial use is a promising approach to facilitate wound healing of the bone tissue. Biomateri... more Biomaterial use is a promising approach to facilitate wound healing of the bone tissue. Biomaterials induce the formation of membrane capsules and the recruitment of different types of macrophages. Macrophages are immune cells that produce diverse combinations of cytokines playing an important role in bone healing and regeneration, but the exact mechanism remains to be studied. Our work aimed to identify in vivo macrophages in the Masquelet induced membrane in a rat model. Most of the macrophages in the damaged area were M2-like, with smaller numbers of M1-like macrophages. In addition, high expression of IL-1β and IL-6 cytokines were detected in the membrane region by RT-qPCR. Using an innovative combination of two hybridization techniques (in situ hybridization and in situ hybridization chain reaction (in situ HCR)), M2b-like macrophages were identified for the first time in cryosections of non-decalcified bone. Our work has also demonstrated that microspectroscopical analysis is ...

Research paper thumbnail of A Novel Family in Medicago truncatula Consisting of More Than 300 Nodule-Specific Genes Coding for Small, Secreted Polypeptides with Conserved Cysteine Motifs

PLANT PHYSIOLOGY, 2003

Transcriptome analysis of Medicago truncatula nodules has led to the discovery of a gene family n... more Transcriptome analysis of Medicago truncatula nodules has led to the discovery of a gene family named NCR (nodule-specific cysteine rich) with more than 300 members. The encoded polypeptides were short (60–90 amino acids), carried a conserved signal peptide, and, except for a conserved cysteine motif, displayed otherwise extensive sequence divergence. Family members were found in pea (Pisum sativum), broad bean (Vicia faba), white clover (Trifolium repens), and Galega orientalis but not in other plants, including other legumes, suggesting that the family might be specific for galegoid legumes forming indeterminate nodules. Gene expression of all family members was restricted to nodules except for two, also expressed in mycorrhizal roots. NCR genes exhibited distinct temporal and spatial expression patterns in nodules and, thus, were coupled to different stages of development. The signal peptide targeted the polypeptides in the secretory pathway, as shown by green fluorescent protein...

Research paper thumbnail of In Situ Gene Expression in Native Cryofixed Bone Tissue

Biomedicines, 2022

Bone is a very complex tissue that is constantly changing throughout the lifespan. The precise me... more Bone is a very complex tissue that is constantly changing throughout the lifespan. The precise mechanism of bone regeneration remains poorly understood. Large bone defects can be caused by gunshot injury, trauma, accidents, congenital anomalies and tissue resection due to cancer. Therefore, understanding bone homeostasis and regeneration has considerable clinical and scientific importance in the development of bone therapy. Macrophages are well known innate immune cells secreting different combinations of cytokines and their role in bone regeneration during bone healing is essential. Here, we present a method to identify mRNA transcripts in cryosections of non-decalcified rat bone using in situ hybridization and hybridization chain reaction to explore gene expression in situ for better understanding the gene expression of the bone tissues.

Research paper thumbnail of Material and Methods

A. caerulea seeds were bought from a gardener shop (Jardiland, Maurepas, France). S. lycorpersicu... more A. caerulea seeds were bought from a gardener shop (Jardiland, Maurepas, France). S. lycorpersicum M82 wild type were received from S. Biemelt, the la mutant (accession LA0335) from the Tomato Genetics Ressource Center and the gob mutants from D. Zamir. P. sativum (cv Térèse and line NGB5839) were obtained from C. Rameau and the uni mutant (line JI2171) from J. Hofer. S. tuberosum cv Pompadour were a gift from B. Letarnec. RNA extraction and RT-PCR RNA was extract from partially dissected apices or young leaves of the different species using TRIsol reagent (Invitrogen) according to the manufacturer recommendations. Contaminating DNA was removed by DNaseI treatment (Invitrogen) and the RNA was reverse transcribed (SuperScript TM II reverse transcriptase, Invitrogen). Gene specific primers used for semiquantitative RT-PCR are listed in Supplementary Table 4. PCR products were observed on ethidium bromide-stained gels or following southern blotting (PsNAM1/2).Cloning of NAM/CUC genes. ...

Research paper thumbnail of Synchronization of Oryza sativa L.cv. Taipei-309 cell suspension culture

... Ágnes Lendvai*, Krisztina Nikovics, László Bakó, Dénes Dudits, János Györgyey ... Nikovics K,... more ... Ágnes Lendvai*, Krisztina Nikovics, László Bakó, Dénes Dudits, János Györgyey ... Nikovics K, Simidjieva J, Peres A, Ayaydin F, Pasternak T, Davies JW, Boulton MI, Dudits D, Horvath GV (2001) Cell-cycle, phase-specific activation of Maize streak virus promoters. Mol. ...

Research paper thumbnail of An Arabidopsis Cyclin Promoter Region is Active in Transgenic Maize Plants

We have produced transgenic maize carrying Arabidopsis cyclin promoter and beta-glucuronidase chi... more We have produced transgenic maize carrying Arabidopsis cyclin promoter and beta-glucuronidase chimeric fusion (cycB1;1At::GUS) by direct DNA uptake into embryogenic cell suspension protoplasts to study the activity of this cyclin promoter in a heterologous cell system. The transgenic maize lines exhibiting different levels of reporter gene expression show that regulatory elements from cycB1;1At gene as a dicot promoter can function in maize. Expression patterns revealed by fluorimetric, as well as, histochemical GUS reporter enzymatic assays indicated cell division-dependent expression of the GUS gene driven by this cycB1;At;1 promoter with elevated activity in proliferating cells and meristems. Treatment of tissues with inhibitors of mitosis increased the activity of cycB1;1At promoter, that suggests a link between mitotic phase of cell division and activity of cycB1;1At regulatory elements in transgenic maize.

Research paper thumbnail of Docking of acetyl-CoA carboxylase to the plastid envelope membrane attenuates fatty acid production in plants

Nature Communications

In plants, light-dependent activation of de novo fatty acid synthesis (FAS) is partially mediated... more In plants, light-dependent activation of de novo fatty acid synthesis (FAS) is partially mediated by acetyl-CoA carboxylase (ACCase), the first committed step for this pathway. However, it is not fully understood how plants control light-dependent FAS regulation to meet the cellular demand for acyl chains. We report here the identification of a gene family encoding for three small plastidial proteins of the envelope membrane that interact with the α-carboxyltransferase (α-CT) subunit of ACCase and participate in an original mechanism restraining FAS in the light. Light enhances the interaction between carboxyltransferase interactors (CTIs) and α-CT, which in turn attenuates carbon flux into FAS. Knockouts for CTI exhibit higher rates of FAS and marked increase in absolute triacylglycerol levels in leaves, more than 4-fold higher than in wild-type plants. Furthermore, WRINKLED1, a master transcriptional regulator of FAS, positively regulatesCTI1expression by direct binding to its pro...

Research paper thumbnail of Hybridization‐chain‐reaction is a relevant method for in situ detection of M2d‐like macrophages in a mini‐pig model

Research paper thumbnail of Characterization of macrophages, giant cells and granulomas during muscle regeneration after irradiation

Research paper thumbnail of Non-Specific Binding, a Limitation of the Immunofluorescence Method to Study Macrophages In Situ

Genes

Advances in understanding tissue regenerative mechanisms require the characterization of in vivo ... more Advances in understanding tissue regenerative mechanisms require the characterization of in vivo macrophages as those play a fundamental role in this process. This characterization can be approached using the immuno-fluorescence method with widely studied and used pan-markers such as CD206 protein. This work investigated CD206 expression in an irradiated-muscle pig model using three different antibodies. Surprisingly, the expression pattern during immunodetection differed depending on the antibody origin and could give some false results. False results are rarely described in the literature, but this information is essential for scientists who need to characterize macrophages. In this context, we showed that in situ hybridization coupled with hybridization-chain-reaction detection (HCR) is an excellent alternative method to detect macrophages in situ.

Research paper thumbnail of Macrophage Identification In Situ

Biomedicines

Understanding the processes of inflammation and tissue regeneration after injury is of great impo... more Understanding the processes of inflammation and tissue regeneration after injury is of great importance. For a long time, macrophages have been known to play a central role during different stages of inflammation and tissue regeneration. However, the molecular and cellular mechanisms by which they exert their effects are as yet mostly unknown. While in vitro macrophages have been characterized, recent progress in macrophage biology studies revealed that macrophages in vivo exhibited distinctive features. Actually, the precise characterization of the macrophages in vivo is essential to develop new healing treatments and can be approached via in situ analyses. Nowadays, the characterization of macrophages in situ has improved significantly using antigen surface markers and cytokine secretion identification resulting in specific patterns. This review aims for a comprehensive overview of different tools used for in situ macrophage identification, reporter genes, immunolabeling and in si...

Research paper thumbnail of First Insights Into the M2 Inflammatory Response After Adipose-Tissue-Derived Stem Cell Injections in Radiation-Injured Muscles

Research paper thumbnail of Material and Methods

A. caerulea seeds were bought from a gardener shop (Jardiland, Maurepas, France). S. lycorpersicu... more A. caerulea seeds were bought from a gardener shop (Jardiland, Maurepas, France). S. lycorpersicum M82 wild type were received from S. Biemelt, the la mutant (accession LA0335) from the Tomato Genetics Ressource Center and the gob mutants from D. Zamir. P. sativum (cv Térèse and line NGB5839) were obtained from C. Rameau and the uni mutant (line JI2171) from J. Hofer. S. tuberosum cv Pompadour were a gift from B. Letarnec. RNA extraction and RT-PCR RNA was extract from partially dissected apices or young leaves of the different species using TRIsol reagent (Invitrogen) according to the manufacturer recommendations. Contaminating DNA was removed by DNaseI treatment (Invitrogen) and the RNA was reverse transcribed (SuperScript TM II reverse transcriptase, Invitrogen). Gene specific primers used for semiquantitative RT-PCR are listed in Supplementary Table 4. PCR products were observed on ethidium bromide-stained gels or following southern blotting (PsNAM1/2).Cloning of NAM/CUC genes. ...