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Actin levels fluctuate in the seminiferous epithelium at various stages during spermatogenesis in the rat

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BIOCELL 2017年第2期41卷 63-65页

作者： CRISTIAN ACOSTA MABEL FOSCOLO JUAN CARLOS CAVICCHIA Instituto de Histología y Embriología de Mendoza(ihem)-CONICET Universidad Nacional de Cuyo5500MendozaArgentina

The testis is a double gland comprised of the sperm-producing seminiferous tubules and a complex endocrine interstice.The former structures generate and release whole cells(exocrine aspect of the gland)while the latter synthesize and release androgens and related hormones.The testis also has poorly understood paracrine connexions.These connexions play an important role during spermatogenesis.A key molecule within these structures is β-actin.Therefore,the present study aims at examining the expression pattern of β-actin during the various stages of the spermatic cycle in the rat.To achieve this goal,we used a combination of trans-illumination assisted microdissection of seminiferous tubules,confocal immunofluorescence and Western blot analysis.Our experiments revealed that the levels of β-actin fluctuate significantly during spermatogenesis,peaking immediately after spermiation.

关键词： Beta actin Seminiferous epithelium Trans-illumination assisted microdissection

Cell bioenergetics:Simple logics,complex solut

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BIOCELL 2022年第10期46卷 2217-2220页

作者： LUIS S.MAYORGA Instituto de Histología y Embriología de Mendoza(ihem Universidad Nacional de CuyoCONICET)Facultad de Ciencias Exactas y NaturalesMendoza5500Argentina

Cells are open systems that exchange energy and molecules with their environment.As any material system,they perform all the complex activities required for homeostasis and reproduction,obeying the thermodynamic laws.This viewpoint will argue that the basic logic governing the energy flux required to preserve cell organization and function is simple:to decrease the activation energy(Ea)of specific processes.Almost none of the possible chemical reactions and energy transformations inside a cell occur at a measurable speed at room or body temperature.Enzymes or other macromolecular structures speed up particular transformations by decreasing the corresponding energetic barriers.However,to maintain the systems in a homeostatic state,capable of sophisticated functions based on this simple strategy requires an inconceivably complex solution.The conclusion will point to the challenging and intricate problems that cells have solved to carve the highly regulated channel through which the energy flows,fueling the work of these nanoscale machines.

关键词： Thermodynamics Cellular logics Self-organization Complexity

Coxiella burnetii:living inside the host cell

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BIOCELL 2019年第1期43卷 1-6页

作者： Jesus Sebastian DISTEL Walter BERON Rodolfo Matias ORTIZ FLORES Instituto de Histologia y Embriologia de Mendoza(ihem) ONICETUniversidad Nacional de CuyoMendozaArgentina Facultad de Odontologia Universidad Nacional de CuyoMendozaArgentina

Coxiella burnetii is an obligate intracellular pathogen and the causative agent of Q fever.In this brief review,we describe how recently described mechanisms help our understanding of C.burnetii invasion and its survival in the host cell by the formation of a replicative niche:the Coxiella-containing vacuole.We describe the actin-associated proteins involved in the internalization of C.burnetii,and we discuss the contribution of diverse degradation pathways of the cell during the formation and stabilization of the Coxiella-containing vacuole.

关键词： Actin Coxiellosis Phagocytic pathway Q fever

GTP-bound Rab3A exhibits consecutive positive and negative roles during human sperm dense-core granule exocytosis

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Journal of Molecular Cell Biology 2014年第4期8卷 286-298页

作者： Matias A. Bustos Carlos M. Roggero Paola X. De la Iglesia Luis S. Mayorga Claudia N. Tomes Instituto de Histologia y Embriologia IHEM-CONICET Facultad de Ciencias Medicas Universidad Nacional de Cuyo Mendoza Argentina Department of Biophysics UT Southwestern Medical Center Dallas TX USA Servicio de Patologla Hospital Italiano de Buenos Aires Buenos Aires Argentina

哺乳动物的精子的 Exocytosis 稠密核心的能分泌的小粒作为所有另外的能分泌的房间依靠一样的熔化分子；一个如此的分子是小 GTPase Rab3A。这里，我们由细看 streptolysin O-permeabilized 人的精子的 exocytotic 反应到很多包含 Rab3A... 详细信息

哺乳动物的精子的 Exocytosis 稠密核心的能分泌的小粒作为所有另外的能分泌的房间依靠一样的熔化分子；一个如此的分子是小 GTPase Rab3A。这里，我们由细看 streptolysin O-permeabilized 人的精子的 exocytotic 反应到很多包含 Rab3A 构造的尖锐 applicationof 并且相关报导 Rab3Ain 分泌物的角色的深入的生物化学的描述有那些的调查结果与内长的蛋白质聚在一起。Fulllength， geranylgeranylated，和活跃 Rab3A 得到了人的精子 exocytosis 本身。与 Rab3A/Rab22A 妄想的蛋白质，我们证明 Rab3A 分子的 carboxy 终端领域是足够的 necessaryand 支持 exocytosis，而它的氨基终点的阻止的被触发钙的分泌物。有趣地，当在停靠 acrosome 到血浆膜以后增加了时， fulllength Rab3A 停止了分泌物。这效果看情况到水解 GTP 的 Rab3A 的在无能。我们联合了在单个房间同时染色协议到 detectmembrane 熔化和内长的 Rab3 的激活地位的修改 immunofluorescence 和 acrosomal，并且发现那 GTP hydrolysison 内长的 Rab3 是强制的让熔化毛孔开。我们的调查结果贡献证实 Rab3 调制不同地在学习下面取决于核苷酸界限和 exocytosis 阶段的 regulatedexocytosis。

关键词： Rab3a G-蛋白胞吐过程分泌颗粒致密核精子负面作用人类

Microglia-precursor cell interactions in health and in pathology

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BIOCELL 2018年第2期42卷 41-45页

作者： Estela M.MUÑOZ Instituto de Histología y Embriología de Mendoza Dr.Mario H.Burgos(ihem) Universidad Nacional de Cuyo(UNCuyo)Consejo Nacional de Investigaciones Científicas y Técnicas(CONICET)MendozaArgentina

Until recently,microglia were mainly known as the resident phagocytes of the brain,i.***.the‘immunological warriors’of the brain.However,extensive knowledge is being accumulated about the functions of microglia beyond immunity.Nowadays,it is well accepted that microglial cells are highly dynamic and responsive,and that they intervene in a dual manner in many developmental processes that shape the central nervous system,including neurogenesis,gliogenesis,spatial patterning,synaptic formation and elimination,and neural circuit establishment and maturation.The differentiation and the pool of precursor cells were also shown to be under microglia regulation via bidirectional communication.In this concise review,I discuss our recent work in microglia-Pax6+cell interactions in one of the circumventricular organs,the pineal gland.An analogy with the rest of the central nervous system is also presented.In addition,I briefly examine mechanisms of interaction between microglia and non-microglial cells in both health and disease.New avenues are also introduced,which may lead us to better comprehend the impact of microglia in physiological and pathological conditions.

关键词： Phagocyte Phagocytosis Phagoptosis Apoptosis Autophagy

Tubulointerstitial injury and proximal tubule albumin transport in early diabetic nephropathy induced by type 1 diabetes mellitus

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BIOCELL 2017年第1期41卷 1-12页

作者： Maximiliano GIRAUD-BILLOUD Fernando EZQUER Javiera BAHAMONDE Marcelo EZQUER ihem Universidad Nacional de CuyoCONICETFacultad de Ciencias MédicasMendozaArgentina Centro de Medicina Regenerativa Facultad de Medicina Clínica AlemanaUniversidad del DesarrolloAvenida Las Condes 12438SantiagoChile

A decrease in the tubular expression of albumin endocytic transporters megalin and cubilin has been associated with diabetic nephropathy,but there are no comprehensive studies to date relating early tubulointerstitial injury and the effect of the disease on both transporters in type 1 diabetes mellitus(T1DM).We used eight-week-old male C57BL/6 mice divided into two groups;one of them received the vehicle(control group),while the other received the vehicle+200 mg/kg streptozotocin(T1DM).Ten weeks after the injection,we evaluated plasma insulin,enzymuria,urinary vitamin D-binding protein(VDBP),tubulointerstitial fibrosis and proximal tubule histology,markers of autophagy,and megalin and cubilin levels.We found a reduction in tubular protein reabsorption(albumin and VDBP as specific substances carried by both transporters)with increased tubulointerstitial injury,development of fibrosis,thickening of tubular basement membrane,and an increase in tubular cell metalloproteases.This was associated with a decrease in the renal expression of megalin and cubilin.We also observed an increase in the amount of cellular vesicles of the phagocytic system in the tubules,which could be linked to an alteration of normal intracellular trafficking of both receptors,thus affecting the normal function of transporters in early stages of diabetic nephropathy.In diabetic animals,the added effects of tubulointerstitial injury,the decreases in megalin and cubilin expression,and an altered intracellular trafficking of these receptors,seriously affect protein reabsorption.

关键词： Albuminuria Megalin Cubilin Autophagy

Freely Available Tool(FAT)for automated quantification of lipid droplets in stained cells

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BIOCELL 2017年第2期41卷 55-58页

作者： DIEGO MASONE ALDANA DGOJANOVICH YESICA R.FRONTINI-LOPEZ SAMANTA DEL VELIZ MARINA UHART DIEGO M.BUSTOS Laboratorio de Integración de Senales Celulares.ihem-CONICET Universidad Nacional de Cuyo MendozaArgentina Facultad de Ingeniería Universidad Nacional de CuyoMendozaArgentina Facultad de Ciencias Exactas y Naturales Universidad Nacional de CuyoMendozaArgentina

In this study,we propose an automatic procedure for digital image processing.We describe a method that can efficiently quantify and characterize lipid droplets distributions in different cell types in culture.Prospectively,the lipid droplets detection method described in this work could be applied to static or time-lapse data,collected with a simple visible light or fluorescence microscopy equipment.Fully automated algorithms were implemented in Octave,a freely available scientific package.

关键词： Adipogenesis 3T3-L1 cells human stem cells automatic image processing

The impact of paralog genes:detection of copy number variation in spinal muscle atrophy patients

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BIOCELL 2018年第3期42卷 87-91页

作者： Sergio LAURITO Juan A.CUETO Jimena PEREZ María ROQUé ihem Universidad Nacional de CuyoCONICET Departamento de Biología Universidad Nacional de CuyoFacultad de Ciencias Exactas y Naturales Instituto de Fisiología Universidad Nacional de CuyoFacultad de Ciencias Médicas

Spinal muscular atrophy(SMA)is caused by dysfunction of the alpha motor neurons of the spinal cord.It is an autosomal recessive disease associated to the SMN1 gene,located in the subtelomeric region of 5q13.A paralog SMN2 gene is located at the centromeric region of the same chromosome,which apparently originated by an ancestral inverted duplication occurring only in humans.The exon sequence differs in two nucleotides in exon 7 and exon 8,which leads to an SMN2 transcript that lacks exon 7 and results in a truncated protein.Part(10%)of the SMN2 transcripts avoids the splicing of exon 7 but most of the copies are dysfunctional.In a disease scenario,the more SMN2 copies the higher possibility to restore at least partly the effects of SMN1 deficiency.Some therapeutic approaches are being developed to increase the expression of SMN2.To determine the number of SMN1 and SMN2 copies,the methodology must distinguish accurately between both genes.In this work,we present the results obtained using multiplex ligation-dependent probe amplification(MLPA)in 60 SMA suspected patients/carriers derived from different regions of Argentina.In 32 of these DNA samples we found alterations in SMN1.Among these,16 presented a heterozygous deletion(carrier status)and 14 an homozygous deletion(patient status)in exon 7 and 8 of SMN1.In one case,exon 7 was found homozygously deleted but exon 8 presented a single copy,and in another case,exon 7 was found heterozygously deleted while exon 8 was normal.Almost half of the patients(7/15)presented a normal diploid number of SMN2 while the other half(8/15)presented an increased number.In this work we showed how a probe-based methodology such as MLPA was able to distinguish between the paralog genes and determine the amount of copies in DNA samples from suspected patients/carriers of SMA.

关键词： SMA CNV MLPA Paralog gene

Adipose-derived mesenchymal stem/stromal cells:from the lab bench to the basic concepts for clinical translation

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BIOCELL 2018年第3期42卷 67-77页

作者： Yesica Romina FRONTINI-LóPEZ Aldana Daniela GOJANOVICH Diego MASONE Diego Martín BUSTOS Marina UHART Laboratorio de Integración de Señles Celulares.ihem-CONICET Universidad Nacional de Cuyo MendozaArgentina Facultad de Ingeniería Universidad Nacional de CuyoMendozaArgentina Facultad de Ciencias Exactas y Naturales Universidad Nacional de CuyoMendozaArgentina

In the last years,much work has shown that the most effective repair system of the body is represented by stem cells,which are defined as undifferentiated precursors that own unlimited or prolonged self-renewal ability,which also have the potential to transform themselves into various cell types through differentiation.All tissues that form the body contain many different types of somatic cells,along with stem cells that are called‘mesenchymal stem(or stromal)cells’(MSC).In certain circumstances,some of these MSC migrate to injured tissues to replace dead cells or to undergo differentiation to repair it.The discovery of MSC has been an important step in regenerative medicine because of their high versatility.Moreover,the finding of a method to isolate MSC from adipose tissue,so called‘adipose-derived mesenchymal stem cells’(ASC),which share similar differentiation capabilities and isolation yield that is greater than other MSC,and less bioethical concerns compared to embryonic stem cells,have created self-praised publicity to procure almost any treatment with them.Here,we review the current techniques for isolation,culture and differentiation of human ASC(hASC),and describe them in detail.We also compile some advantages of the hASC over other stem cells,and provide some concepts that could help finding strategies to promote their therapeutic efficiency.

关键词： hASC Tissue regeneration Cell therapy