Manuel LeeUniversidad Autónoma De Chile · Facultad de Ciencias de la Salud
Manuel Lee
PhD in Applied Cellular and Molecular Biology
About
22
Publications
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Introduction
Skills and Expertise
Additional affiliations
August 2006 - February 2009
Education
March 2015 - July 2016
March 2015 - September 2019
July 2011 - August 2011
Publications
Publications (22)
Antifreeze proteins (AFPs) are natural biomolecules found in cold-adapted organisms
that lower the freezing point of water, allowing survival in icy conditions. These proteins have the potential to improve cryopreservation techniques by enhancing the quality of genetic material postthaw. Deschampsia antarctica, a freezing-tolerant plant, possesses...
Ca2+ is a key element in the sperm activation process of Salmo salar. However, the molecular mechanisms by which this ion enters the sperm cell have been poorly studied. In this study, we examined, for the first time, the role of the voltage-gated T-type Ca2+ channel in the activation of sperm motility of Salmo salar. Using an in vitro inhibition a...
Among all the Ca2+ channels, CatSper channels have been one of the most studied in sperm of different species due to their demonstrated role in the fertilization process. In fish sperm, the calcium channel plays a key role in sperm activation. However, the functionality of the CatSper channels has not been studied in any of the fish species. For th...
Ca2+ cations play a key role in the initiation of spermatozoa motility in Atlantic salmon (Salmo salar). In this study we assess the importance of the voltage-gated L-type calcium channels in the spermatozoa motility of Atlantic salmon by combined in vitro and in silico approaches. The results of this study showed that as in other fish species, vol...
Covalent attachment of therapeutic proteins to polyethylene glycol (PEG) is widely used for the improvement of its pharmacokinetic and pharmacological properties, as well as the reduction in reactogenicity and related side effects. This technique named PEGylation has been successfully employed in several approved drugs to treat various diseases, ev...
The development of cryopreservation techniques has led to important changes in animal reproductive biotechnology. However, these techniques are associated with cellular and molecular damage, affecting the mitochondrial function and quality of spermatozoa; moreover studies in fish are limited. In this work, the effects of cryopreservation on ultrast...
Sperm motility in fish with external fertilization is critical for reproductive efficiency in aquaculture, especially in salmonids. Gamete preservation techniques, such as cryopreservation, however, reduce sperm motility and fertilizing capacity. Very few studies have addressed cryodamage from energetic and cell signalling approaches. In this study...
The cold storage and cryopreservation of semen decrease sperm quality. Morphological and biochemical analyses of spermatozoa provide valuable information for the optimization of storage protocols to obtain a sufficient number of spermatozoa for in vitro fertilization. The aim of this study was to evaluate the morphology and lipid composition of Atl...
Patagonian blenny (E. maclovinus) is a marine species recently placed in captivity and which are potentially farmable. Understanding and improving its sperm capacity to withstand short-term storage conditions is a key element of initiating an artificial propagation program for this species. The aim of this study is to evaluate the ultrastructure an...
In this research we evaluated simple aspects of the sperm biology of Genypterus blacodes, in particular assessing the effects of pH (6, 7, 8 and 9), temperature (4, 8 and 16 °C) and osmolalities 100% sea water (1010 mOsm/kg, Control), 75% sea water (774 mOsm/kg, T 1), 50% sea water (488 mOsm/kg, T 2) and distilled water (0 mOsm/ kg, T 3)) on the mo...
La membrana de los espermatozoides juega un papel importante en la capacidad de fertilización. Sin embargo, en peces existen pocas investigaciones que se han centrado en la composición lipídica y
estructura de la membrana y el importante papel que ejerce en la interacción espermatozoide-ova. Por lo tanto, el objetivo de este proyecto es estudiar la...
The use of α-tocopherol and ascorbic acid in freezing medium prevents lipoperoxidation and damage by ROS (reactive oxygen species); however, the effects of these antioxidants in salmonid spermatozoa have not been studied. In this work the protective effects of antioxidants on the enzymatic activity and fertilization capacity of frozen Atlantic salm...
Patagonian blenny (Eleginops maclovinus) is the only member of Eleginopidae family. It has been an important resource for Chilean artisanal fishermen communities, however, capture volumes have progressively decreased over last decades likely because overfishing, and concerns for conservation of this species have rised. In this work, mitochondrial D...
Sperm motility is considered as a key factor allowing determination of semen quality and predicts fertilizing capacity. In many fish species, the spermatozoa are immotile in the testes and seminal plasma, and motility is induced when they are released in the aqueous environment. Initiation and activation of sperm motility are prerequisite processes...
There is an extraordinary diversity of reproductive modes in teleost and this variability is related to the phylogenetic relationships and adaption to very different biotopes. As in all vertebrates, sperm is produced as the end product of the process of spermatogenesis, and regarding teleost the spermatozoa lack an acrosome in almost all species an...
The Zebrafish (Danio Rerio) is a Teleost fish, which offers several practical advan-
tages concerning manipulation, maintenance and versatility as well as short life
cycle, low cost and physiological similarity with most farmed species. Zebrafish
have been used as model for several aquaculture-relevant infectious agents, since
the cost of infection...
Semen conservation methods have been extensively studied for fish species over the last 60 years. Cryopreservation techniques can be divided into two types: one with slow cooling rates like traditional freezing and the other with ultrarapid rates as used in vitrification. These protocols increase the time over which semen samples can be used for re...
CellProfiler pipeline for foci analysis in DAPI-FITC images
Applied in :
J.E. González, M. Lee, J. F. Barquinero, M. Valente, S. Roch-Lefèvre, O.García. Quantitative image analysis of gamma-H2AX foci induced by ionizing radiation applying open source programs. Analytical and quantitative cytology and histology 2012; 34(2):66-71.
To test a CellProfiler pipeline for automated counting and characterization of gamma-H2AX foci in color images of human cultured cells.
A431 cells were irradiated and stained for gamma-H2AX foci detection. Sets of color images were analyzed visually, and findings were compared with those using FociCounter and CellProfiler software.
The CellProfiler...
Epidermal growth factor receptors (EGFR) are overexpressed in a wide range of malignancies including head and neck, colon, and breast cancers. It has been identified that carcinomas with high expression levels of EGFR are more resistant to radiotherapy. Therefore, inhibiting nuclear translocation of EGFR to increase the radiosensitivity of malignan...
Questions
Questions (3)
Dear colleagues, have anyone used any commercially available antibody anti-AMPK to detect fish AMPKalpha subunit? I have tried two from different manufacturers with no success. NOTE: dot blots marked positive with controls, but no mark with samples of interest.
Thanks in advance.
I have been having trouble for detecting those proteins in fish protein extracts. I am using reductive loadding buffer for SDS-PAGE. Cell lysis solution contains protease inhibitors Leupeptin and PMSF, so protein degradation is unlikely. Transference seems to be efficient, and I also performed Dot Blot Assay with no success. Secondary antobody is Jackson HRP-conjugated whole IgG reacting against heavy and light chains of the target IgG (primary Ab). Primary Abs used so far are from Santa Cruz Biotech. Any help is appreciated.
Does it work fine for this purpose? All comments, tips and suggestions are very welcome. Thanks in advance.
