TY  - JOUR
TI  - Immunogold Localization of Molecular Constituents Associated with Basal Bodies,  Flagella, and Extracellular Matrices in Male Gametes of Land Plants
AU  - Lopez, Renee Anita
AU  - Mansouri, Katayoun 
AU  - Henry, Jason Scott
AU  - Flowers, Nicholas David
AU  - Vaughn, Kevin C.
AU  - Renzaglia, Karen Sue
VL  - 7
IS  - 21
PY  - 2017
DA  - 2017/11/05
SP  - e2599
C1  - Bio-protocol 2017;7:e2599
DO  - 10.21769/BioProtoc.2599
UR  - https://doi.org/10.21769/BioProtoc.2599
AB  - Male gametes (spermatozoids) are the only motile cells produced during the life cycle of land plants. While absent from flowering and most cone-bearing plants, motile cells are found in less derived taxa, including bryophytes (mosses, liverworts and hornworts), pteridophytes (lycophytes and ferns) and some seed plants (Ginkgo and cycads). During development, these cells undergo profound changes that involve the production of a locomotory apparatus, unique microtubule (MT) arrays, and a series of special cell walls that are produced in sequence and are synchronized with cellular differentiation. Immunogold labeling in the transmission electron microscope (TEM) provides information on the exact location and potential function of macromolecules involved with this developmental process. Specifically, it is possible to localize epitopes to proteins that are associated with the cellular inclusions involved in MT production and function. Spermatogenesis in these plants is also ideal for examining the differential expression of carbohydrates and glycoproteins that comprise the extracellular matrixes associated with the dramatic architectural changes in gamete shape and locomotory apparatus development. Here we provide methodologies using monoclonal antibodies (MAbs) and immunogold labeling in the TEM to localize macromolecules that are integral to spermatozoid development.
KW  - Arabinogalactan proteins
KW  - Carbohydrates
KW  - Centrin
KW  - Extracellular matrix
KW  - Flagella
KW  - Gamma tubulin
KW  - Immunogold labeling
KW  - Microtubule organizing centers
KW  - Transmission electron microscopy
JF  - Bio-protocol
SN  - 2331-8325
PB  - Bio-protocol LLC.
BIO101  - False