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1.
Nat Commun ; 15(1): 2861, 2024 Apr 03.
Artigo em Inglês | MEDLINE | ID: mdl-38570505

RESUMO

Tissue integrity is sensitive to temperature, tension, age, and is sustained throughout life by adaptive cell-autonomous or extrinsic mechanisms. Safeguarding the remarkably-complex architectures of neurons and glia ensures age-dependent integrity of functional circuits. Here, we report mechanisms sustaining the integrity of C. elegans CEPsh astrocyte-like glia. We combine large-scale genetics with manipulation of genes, cells, and their environment, quantitative imaging of cellular/ subcellular features, tissue material properties and extracellular matrix (ECM). We identify mutants with age-progressive, environment-dependent defects in glial architecture, consequent disruption of neuronal architecture, and abnormal aging. Functional loss of epithelial Hsp70/Hsc70-cochaperone BAG2 causes ECM disruption, altered tissue biomechanics, and hypersensitivity of glia to environmental temperature and mechanics. Glial-cell junctions ensure epithelia-ECM-CEPsh glia association. Modifying glial junctions or ECM mechanics safeguards glial integrity against disrupted BAG2-proteostasis. Overall, we present a finely-regulated interplay of proteostasis-ECM and cell junctions with conserved components that ensures age-progressive robustness of glial architecture.


Assuntos
Caenorhabditis elegans , Neuroglia , Animais , Caenorhabditis elegans/genética , Astrócitos , Fenômenos Biomecânicos , Proteostase , Matriz Extracelular/metabolismo , Junções Intercelulares
2.
Life Sci Alliance ; 2(1): e201800170, 2019 02.
Artigo em Inglês | MEDLINE | ID: mdl-30599047

RESUMO

Stable cell fate is an essential feature for multicellular organisms in which individual cells achieve specialized functions. Caenorhabditis elegans is a great model to analyze the determinants of cell fate stability because of its invariant lineage. We present a tractable cell fate challenge system that uses the induction of fate-specifying transcription factors. We show that wild-type differentiated animals are highly resistant to fate challenge. Removal of heterochromatin marks showed marked differences: the absence of histone 3 lysine 9 methylation (H3K9) has no effect on fate stability, whereas Polycomb homolog mes-2 mutants lacking H3K27 methylation terminally arrest larval development upon fate challenge. Unexpectedly, the arrest correlated with widespread cell proliferation rather than transdifferentiation. Using a candidate RNAi larval arrest-rescue screen, we show that the LIN-12Notch pathway is essential for hyperplasia induction. Moreover, Notch signaling appears downstream of food-sensing pathways, as dauers and first larval stage diapause animals are resistant to fate challenge. Our results demonstrate an equilibrium between proliferation and differentiation regulated by Polycomb and Notch signaling in the soma during the nematode life cycle.


Assuntos
Proteínas de Caenorhabditis elegans/metabolismo , Caenorhabditis elegans/crescimento & desenvolvimento , Proliferação de Células/fisiologia , Larva/fisiologia , Proteínas do Grupo Polycomb/metabolismo , Receptores Notch/metabolismo , Animais , Proteínas de Caenorhabditis elegans/genética , Diferenciação Celular/fisiologia , Linhagem da Célula/fisiologia , Plasticidade Celular/fisiologia , Técnicas de Silenciamento de Genes , Hiperplasia/metabolismo , Proteínas de Membrana/metabolismo , Metilação , Fenótipo , Interferência de RNA , Receptores Notch/genética , Inanição/fisiopatologia
3.
Development ; 143(6): 910-23, 2016 Mar 15.
Artigo em Inglês | MEDLINE | ID: mdl-26980791

RESUMO

The existence of different domains within the nucleus has been clear from the time, in the late 1920s, that heterochromatin and euchromatin were discovered. The observation that heterochromatin is less transcribed than euchromatin suggested that microscopically identifiable structures might correspond to functionally different domains of the nucleus. Until 15 years ago, studies linking gene expression and subnuclear localization were limited to a few genes. As we discuss in this Review, new genome-wide techniques have now radically changed the way nuclear organization is analyzed. These have provided a much more detailed view of functional nuclear architecture, leading to the emergence of a number of new paradigms of chromatin folding and how this folding evolves during development.


Assuntos
Núcleo Celular/genética , Genes , Genoma , Animais , Linhagem da Célula/genética , Núcleo Celular/ultraestrutura , Heterocromatina/metabolismo , Humanos , Microscopia
4.
J Exp Zool A Ecol Genet Physiol ; 323(10): 714-721, 2015 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-26351243

RESUMO

Vasoactive intestinal peptide (VIP) is a pleiotropic neuropeptide involved in different functions, including testosterone synthesis. Recently, we reported the presence of VIP in the testis of Podarcis sicula, throughout the reproductive cycle. Now, we investigated the effects of the VIP on steroidogenesis in significant periods of the Podarcis reproductive cycle: winter stasis, reproductive period, and summer stasis. Using VIP treatments in testis culture in absence or presence of receptors antagonists, we demonstrated for the first time that in P. sicula, VIP is involved not only in testosterone synthesis, as in mammals, but in 17ß-estradiol synthesis too. J. Exp. Zool. 323A: 714-721, 2015. © 2015 Wiley Periodicals, Inc.

5.
Gen Comp Endocrinol ; 205: 102-8, 2014 Sep 01.
Artigo em Inglês | MEDLINE | ID: mdl-24852351

RESUMO

Pituitary adenylate cyclase-activating peptide (PACAP) is a neuropeptide involved in multiple functions, including vertebrate reproduction. Recently, we reported the presence of PACAP in the testis of Italian wall lizard Podarcis sicula during reproductive period (May-June). Herein we investigated the PACAP mRNA expression and the localization of PACAP/PACAP receptor system, in the other periods of the Podarcis reproductive cycle, namely in summer stasis, early autumnal resumption, mid-autumnal resumption, winter stasis, and spring resumption. Using biomolecular and immunohistochemical investigations, we demonstrated that PACAP mRNA was widely expressed in all germ and somatic cells; in summer stasis (July-August) and early autumnal resumption (September) in particular, the mRNA was always found in Sertoli cells while was transiently expressed in germ and in Leydig cells. Differently from the mRNA, the protein was always present in germ and somatic cells independently from the reproductive cycle phase. As PACAP, the PAC1 receptor was always present in the testis, except for the summer stasis (July-August) and the early autumnal resumption (September), when PACAP was lacking in germ and somatic cells (Leydig and Sertoli cells). The present results strongly suggest that PACAP/PAC1 receptor system is widely represented during the reproductive cycle of male lizard. The possible involvement of PACAP/PACAP receptor system in the control of spermatogenesis is discussed.


Assuntos
Lagartos/fisiologia , Polipeptídeo Hipofisário Ativador de Adenilato Ciclase/metabolismo , Receptores de Polipeptídeo Hipofisário Ativador de Adenilato Ciclase/metabolismo , Animais , Humanos , Masculino , Polipeptídeo Hipofisário Ativador de Adenilato Ciclase/genética , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Reação em Cadeia da Polimerase em Tempo Real , Receptores de Polipeptídeo Hipofisário Ativador de Adenilato Ciclase/genética , Reprodução , Testículo/citologia , Testículo/metabolismo
6.
J Exp Zool A Ecol Genet Physiol ; 321(6): 334-47, 2014 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-24753326

RESUMO

Using molecular, biochemical, and cytological tools, we studied the nucleotide and the deduced amino acid sequence of PHI/VIP and the distribution of VIP/VPAC receptor system in the testis of the Italian wall lizard Podarcis sicula to evaluate the involvement of such a neuropeptide in the spermatogenesis control. We demonstrated that (1) Podarcis sicula VIP had a high identity with other vertebrate VIP sequences, (2) differently from mammals, VIP was synthesized directly in the testis, and (3) VIP and its receptor VPAC2 were widely distributed in germ and somatic cells, while the VPAC1 R had a distribution limited to Leydig cells. Our results demonstrated that in Podarcis sicula the VIP sequence is highly preserved and that this neuropeptide is involved in lizard spermatogenesis and steroidogenesis.


Assuntos
Lagartos/fisiologia , Receptores de Peptídeo Intestinal Vasoativo/fisiologia , Testículo/fisiologia , Peptídeo Intestinal Vasoativo/fisiologia , Sequência de Aminoácidos , Animais , Sequência de Bases , Clonagem Molecular , Sequência Conservada , Hibridização In Situ , Células Intersticiais do Testículo/química , Células Intersticiais do Testículo/fisiologia , Masculino , Dados de Sequência Molecular , Reação em Cadeia da Polimerase em Tempo Real , Receptores de Peptídeo Intestinal Vasoativo/análise , Receptores de Peptídeo Intestinal Vasoativo/genética , Receptores Tipo II de Peptídeo Intestinal Vasoativo/análise , Receptores Tipo II de Peptídeo Intestinal Vasoativo/genética , Receptores Tipo II de Peptídeo Intestinal Vasoativo/fisiologia , Receptores Tipo I de Polipeptídeo Intestinal Vasoativo/análise , Receptores Tipo I de Polipeptídeo Intestinal Vasoativo/genética , Receptores Tipo I de Polipeptídeo Intestinal Vasoativo/fisiologia , Alinhamento de Sequência , Testículo/química , Peptídeo Intestinal Vasoativo/análise , Peptídeo Intestinal Vasoativo/genética
7.
Gen Comp Endocrinol ; 205: 94-101, 2014 Sep 01.
Artigo em Inglês | MEDLINE | ID: mdl-24694517

RESUMO

Starting from the knowledge that in the reproductive period the Vasoactive Intestinal Peptide (VIP) is widely distributed in Podarcis sicula testis, we studied VIP expression and the localization of the neuropeptide and its receptors in the testis of the Italian wall lizard P. sicula in the other phases of its reproductive cycle (summer stasis, autumnal resumption, winter stasis, spring resumption). By Real Time-PCR, we demonstrated that testicular VIP mRNA levels change during the reproductive cycle, showing a cyclic trend with two peaks, one in the mid-autumnal resumption and the other in the reproductive period. By in situ hybridization and immunohistochemistry, we demonstrated that both VIP mRNA and protein were widely distributed in the testis in almost all the phases of the cycle, except in the early autumnal resumption. As regards the receptors, the VPAC1R was localized mainly in Leydig cells, while the VPAC2R showed the same distribution of VIP. Our results demonstrate that, differently from mammals, where VIP is present only in nerve fibres innerving the testis, an endotesticular synthesis takes place in the lizard and the VIP synthesis changes throughout the reproductive cycle. Moreover, the VIP/VPAC receptor system distribution observed in germ and somatic cells in various phases of the cycle, and particularly in the autumnal resumption and the reproductive period, strongly suggests its involvement in both spermatogenesis and steroidogenesis. Finally, the wider distribution of VIP in lizards with respect to mammals leads us to hypothesize that during the evolution the synthesis sites have been transferred from the testis to other districts, such as the brain.


Assuntos
Lagartos/fisiologia , Receptores de Peptídeo Intestinal Vasoativo/metabolismo , Peptídeo Intestinal Vasoativo/metabolismo , Animais , Humanos , Lagartos/genética , Masculino , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Reação em Cadeia da Polimerase em Tempo Real , Receptores de Peptídeo Intestinal Vasoativo/genética , Reprodução/genética , Estações do Ano , Testículo/citologia , Testículo/metabolismo , Peptídeo Intestinal Vasoativo/genética
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