Tendon cells in vivo form a three dimensional network of cell processes linked by gap junctions.

Tendons respond to mechanical load by modifying their extracellular matrix. The cells therefore sense mechanical load and coordinate an appropriate response to it. We show that tendon cells have the potential to communicate with one another via cell processes and gap junctions and thus could use direct cell/cell communication to detect and/or coordinate their load responses. Unfixed cryosections of adult rat digital flexor tendons were stained with the fluorescent membrane dye DiI to demonstrate cell shape. Similar sections were immunolabelled with monoclonal antibodies to rat connexin 32 or connexin 43 to demonstrate gap junctions and counterstained with propidium iodide to show nuclei, or the membrane stain DiOC7 to show cell membranes. Sections were examined with a laser scanning confocal microscope and 3-dimensional reconstructions were prepared from optical section series to demonstrate cell shape and the position of connexin immunolabel. Cells had a complex interconnected morphology with gap junctions at points of contact with other cells. Cell bodies contained the nucleus and extended broad flat lateral cell processes that enclosed collagen bundles and interacted with similar processes from adjacent cells. They also had long thin longitudinal processes interacting with the cell process network further along the tendon. Connexin 43 occurred where cell processes met and between cell bodies, whereas connexin 32 was only found between cell bodies. The results indicate the presence of a 3-dimensional communicating network of cell processes within tendons. The intimate relationship between cell processes and collagen fibril bundles suggests that the cell process network could be involved in load sensing and coordination of response to load. The presence of 2 different types of connexins suggests that there could be at least 2 distinct communicating networks

Development and evaluation of a recombinant DNA vaccine candidate expressing porcine circovirus 2 structural protein

Porcine circovirus 2 (PCV2) is generally associated with the porcine circovirosis syndrome, which is considered an important disease of swine and has potentially serious economic impact on the swine industry worldwide. This article describes the construction of a recombinant plasmid expressing the PCV2 structural protein and the evaluation of cellular and humoral immune responses produced by this recombinant vaccine in BALB/c mice. The vaccine candidate was obtained and analyzed in vivo, in an effort to determine the ability to induce a specific immune response in mice. DNA was extracted from a Brazilian PCV2 isolate and the gene coding for Cap protein was amplified by PCR and inserted into an expression plasmid. Groups of BALB/c mice were inoculated intra-muscularly and intradermally in a 15-day interval, with 100 µg and 50 µg of the vaccine construct, respectively. Another group was inoculated intramuscularly with 100 µg of empty plasmid, corresponding to the control group. Seroconversion and cellular response in BALB/c mice were compared and used for vaccine evaluation. Seroconversion was analyzed by ELISA. After a series of 3 immunizations the spleen cells of the immunized animals were used to perform lymphocyte proliferation assays. Seroconversion to PCV2 was detected by ELISA in the animals inoculated with the vaccine construct when compared with control groups. Lymphocyte proliferation assays showed a stronger cell proliferation in the inoculated animals compared with the control group. Thus, the vaccine candidate construct demonstrated to be able to induce both humoral and cellular responses in inoculated mice.O circovírus suíno 2 (PCV2) é geralmente associado à síndrome da circovirose suína, que é considerada uma importante doença de suínos e possui um sério impacto econômico na suinocultura mundial. Este trabalho descreve a construção de um plasmídeo recombinante que expressa a proteína estrutural do PCV2 e a avaliação das respostas imune humoral e celular por meio de vacinação em camundongos BALB/c. O candidato vacinal foi submetido a análises in vivo, determinando a capacidade de induzir resposta imune específica em camundongos. O DNA de um isolado brasileiro de PCV2 foi extraído e o gene que codifica para a proteína do capsídeo foi amplificado por PCR e inserido num plasmídeo de expressão. Grupos de camundongos BALB/c foram inoculados por via intramuscular e intradérmica a cada 15 dias, com 100µg e 50µg da construção vacinal, respectivamente. Outro grupo foi inoculado com 100µg do plasmídeo original, correspondente ao grupo controle. A soroconversão e a resposta celular dos grupos de camundongos BALB/c vacinados foram comparados como parâmetros de avaliação vacinal. A soroconversão foi avaliada por um teste de ELISA. Após 3 imunizações, as células esplênicas dos animais imunizados foram utilizadas nos ensaios de linfoproliferação. A soroconversão para o PCV2 foi detectada por ELISA nos animais inoculados com a construção vacinal quando comparados com o grupo controle. Nos ensaios de linfoproliferação foi observada uma grande proliferação celular nos animais inoculados comparados ao grupo controle. Portanto, o candidato vacinal demonstrou ser capaz de induzir tanto uma resposta humoral e celular nos camundongos inoculados

The differential secretion of FSH and LH : regulation through genes, feedback and packaging

International audienc

BMP-4 interacts with activin and GnRH to modulate gonadotropin secretion in LbetaT2 gonadotropes

International audienc

Metformin decreases GnRH- and activin-induced gonadotropin secretion in rat pituitary cells: Potential involvement of adenosine 5' monophosphate-activated proteine kinase (PRKA)

 absen

Physiological prediction of genetic merit for female reproduction in the sheep.

National audienc

Loss of CRMP2 O-GlcNAcylation leads to reduced novel object recognition performance in mice

O-GlcNAcylation is an abundant post-translational modification in the nervous system, linked to both neurodevelopmental and neurodegenerative disease. However, the mechanistic links between these phenotypes and site-specific O-GlcNAcylation remain largely unexplored. Here, we show that Ser517 O-GlcNAcylation of the microtubule-binding protein Collapsin Response Mediator Protein-2 (CRMP2) increases with age. By generating and characterizing a Crmp2S517A knock-in mouse model, we demonstrate that loss of O-GlcNAcylation leads to a small decrease in body weight and mild memory impairment, suggesting that Ser517 O-GlcNAcylation has a small but detectable impact on mouse physiology and cognitive function. © 2019 The Authors. Published by the Royal Society under the terms of the Creative Commons Attribution License http://creativecommons.org/licenses/by/4.0/, which permits unrestricted use, provided the original author and source are credited.Peer reviewe