Membrane Biology and Axonal Repair
Membrane Biology and Axonal Repair
Principal investigator: José Abad Rodríguez, Ph.D.
Tel.: (+34) 925 396 832
The Membrane Biology and Axonal Repair laboratory (LBM) investigates the organization of the plasma membrane and its role in neuronal differentiation, with special perspective on nervous system regeneration and neurodegenerative pathologies. In particular, we study the role of glycans (in glycolipids and glycoproteins) and carbohydrate-binding proteins (lectins) in axon growth, myelination and regeneration.
Relevant achievements of my scientific career before becoming PI at the Hospital Nacional de Parapléjicos (HNP) are the description of the influence of neuronal membrane cholesterol for beta-amyloid production in Alzheimer’s disease (12,13), and the role of membrane sialidase Neu3 in determination, growth and regeneration of axons both, in vitro and in vivo (8,11,14). Once at the HNP, we have established the importance of the galectins in some core functions of axonal physiology, such as the localized stimulation of axon branching by phosphorylated galectin-3 (10), and the sulfatides/galectin-4- dependent axon transport mechanism of glycoproteins (9). Recently, we have shown that galectin-4 is expressed in axon membrane segments, locally inhibiting myelination in vitro (5), and that its absence drive neurological deficits at synaptic level in mice that do not express the lectin (LGalS4-KO mice). The mechanisms underlying these deficiencies are currently under research at the level of nervous system. Nevertheless, due to the high expression of galectin-4 in the digestive tract, we are also evaluating the possible implication of the “gut-brain axis” in the above-mentioned neurological phenotype.
To reach our goals, we utilize in vitro models based either on primary cultures of rodent neurons, astrocytes, oligodendrocytes and microglia, or on cultures of established cell lines. The cellular and tissue analysis is usually carried out by conventional, confocal or time-lapse microscopy, and we are currently developing analysis methods based in “high-content screening” microscopy supported by a.i. protocols. We do this in collaboration with our Microscopy Service (Resp. Dr. José Ángel Rodríguez Alfaro). We combine microscopy analysis with standard biochemical (immunoblot, ELISA, etc.) and molecular biology (cloning, PCR, etc.) techniques. As a distinctive feature of our group, we are specialized in membrane and lipid analysis of both, intact cells (surface binding, co-patching…), tissue (Fast Blue and Black Gold staining for myelin) or extracts (membrane fractionation, raft and synaptosome purification, extraction and analysis of lipids/glycolipids). In this context, we have also started the setting-up of complex samples lipidomic analysis by LC-MS, in collaboration with the Proteomics and MS Service in our facilities (Resp. Dr. Gemma Barroso). In order to improve our in vivo experimental portfolio, we have developed several behavioural evaluation systems to measure the cognitive function of rodents, in the context of our research with the LGalS4-KO mice strain. These analyses are complemented with electrophysiology studies in vivo carried out in collaboration with the Experimental Neurophysiology laboratory of the HNP (PI. Dr. Juan de los Reyes Aguilar Lepe).
1- Habermann F.A., Kaltner H., Higuero A. M., García Caballero G., Ludwig A-K., Manning J.C. , Abad-Rodríguez J.* and Gabius H-J*. What Cyto- and Histochemistry Can Do to Crack the Sugar Code. Acta Histochem. Cytochem. 54 (2): 31–48, 2021
2- Kutzner TJ, Higuero AM, Süßmair M, Kopitz J, Hingar M, Díez-Revuelta N, Caballero GG, Kaltner H, Lindner I, Abad-Rodríguez J*, Reusch D*, Gabius HJ*. How presence of a signal peptide affects human galectins-1 and -4: Clues to explain common absence of a leader sequence among adhesion/growth-regulatory galectins. Biochim Biophys Acta Gen Subj 2020; 1864(1):129449.
3- Kaltner H, Abad-Rodríguez J, Corfield AP, Kopitz J, Gabius HJ. The sugar code: letters and vocabulary, writers, editors and readers and biosignificance of functional glycan-lectin pairing. Biochem J. 2019; 476(18):2623.
4- Ledeen RW., Kopitz J., Abad-Rodríguez J., Gabius HJ. Glycan Chains of Gangliosides: Functional Ligands for Tissue Lectins (Siglecs/Galectins). Prog Mol Biol Transl Sci. 2018;156:289-324.
5- Díez-Revuelta N.,Higuero A.M.,Velasco S, Peñas-de-la-Iglesia M., Gabius HJ, Abad-Rodríguez J*. Neurons define non-myelinated axon segments by the regulation of galectin-4-containing axon membrane domains. Sci Rep. 2017; 25;7(1):12246.
6- Higuero A.M., Díez-Revuelta N., Abad-Rodríguez J*. The sugar code in neuronal physiology. Histochem. Cell Biol. 2017; 147(2):257-267 Abad-Rodríguez J*., Díez-Revuelta N. Axon glycoprotein routing in nerve polarity, function, and repair. TIBS July 2015; 40(7):385-396
7- Oliviero A.; Carrasco-Lopez M.C.; Campolo M.; Perez-Borrego Y.A., Soto-León V., Javier Gonzalez-Rosa; Alonso M Higuero; Bryan A Strange; Jose Abad-Rodriguez; Foffani G. Safety study of transcranial static magnetic field stimulation (tSMS) of the human cortex. Brain Stimul. 2015 May-Jun;8(3):481-5
8- Kappagantula S., Andrews M.R., Cheah M., Abad-Rodríguez J., Dotti C.G., Fawcett J.W. Neu-3 Sialidase-mediated ganglioside conversion is necessary for axon regeneration and is blocked in CNS axons. J. Neurosci. 2014, 34(7):2477-2492.
9- Velasco S, Díez-Revuelta N, Hernández-Iglesias T, Kaltner H, André S, Gabius HJ, Abad-Rodríguez J. Neuronal Galectin-4 is required for axon growth and for the organization of axonal membrane L1 delivery and clustering. J Neurochem. 2013 Apr;125(1):49-62.
10- Díez-Revuelta N., Velasco S., André S., Kübler D., Gabius H.J. and Abad-Rodríguez J. Phosphorylation of adhesion/growth-regulatory human Galectin-3 leads to the induction of axonal branching by local membrane L1/ERM redistribution. J Cell Sci. 2010 Mar 1;123: 671-81.
11- Santos Da Silva J, Hasegawa T, Miyagi T, Dotti CG. and Abad Rodríguez J. Asymmetric membrane ganglioside sialidase activity specifies axonal fate. Nature Neurosci. 2005, May; 8(5): 606-15.
12- Abad-Rodríguez J, Ledesma MD, Craessaerts K, Perga S, Medina M, Delacourte A, Dingwall C, De Strooper B and Dotti CG. Neuronal membrane cholesterol loss enhances amyloid peptide generation. J. Cell Biol. 2004 Dec 6;167(5):953-60.
13- Ledesma MD*, Abad-Rodríguez J.* , Galvan C, Biondi E., Navarro P., Delacourte A, Dingwall C, and Dotti CG. Raft disorganization leads to reduced plasmin activity in Alzheimer’s disease brains. EMBO Rep. 2003 Dec; 4(12):1190-6.
14- Rodriguez JA, Piddini E, Hasegawa T, Miyagi T, Dotti CG. Plasma membrane ganglioside sialidase regulates axonal growth and regeneration in hippocampal neurons in culture. J. Neurosci. 2001 Nov 1;21(21):8387-95.
Main Research lines
1- The role of glycans and their interactions in neuronal function
We study the effect of glycan interactions in axon growth, guidance, regeneration and myelination, as well as their roles in cognitive functions such as learning and memory.
2- Lipid functions in neuronal membrane
Cholesterol is a fundamental component of biological membranes, being particularly relevant in neuronal membrane. We study its metabolism and dynamics in the nervous system , focusing synaptic function or Alzheimer’s disease among other aspects.
In addition, we have determined that gangliosides (a glycosphingolipid subfamily) modulate axon growth, and that their modification through the action of sialidases such as Neu3, is required for damaged axon regeneration. We investigate these modifications aiming for applications in the recovery of the injured nervous system.
- Molecular characterization, neuronal specificity and function of non-myelinated axon segments
Funded by: Proyectos RETOS. Agencia Estatal De Investigación MINECO SAF2017-83821-R
- Los segmentos axonales no mielinizados como reguladores de la plasticidad axonal asociada a la memoria y el aprendizaje. Caracterización molecular y funcional en el hipocampo
Funded by: Proyecto de investigación científica de la Consejería de Educación, Cultura y Deportes de Castilla La Mancha SBPLY/17/180501/000250
Jose Abad Rodriguez: PI, Ph.D. in Chemistry (Biochemistry and molecular biology)
Alonso Higuero Romero: Postdoctoral Staff Researcher, Ph.D. in Biochemistry, Molecular Biology and Biomedicine
Natalia Diez Revuelta: Postdoctoral Staff Researcher, Ph.D. in Biology
María Elvira Brocca: Postdoctoral Researcher, Ph.D. in Human Biochemistry. https://www.linkedin.com/in/maria-elvira-brocca-259964158/
Arancha Mora Rubio: Ph.D. student; BsC in Biochemistry; Master in Translational Biomedical Research.
David Martos Puñal: Laboratory technician. Superior Laboratory Technician Diploma.
Cristina de Paz Vela: Laboratory technician. Superior Laboratory Technician Diploma.
Maria Peñas de la Iglesia: Laboratory Staff technician. Superior Laboratory Technician Diploma (currently on leave).