Ps. Testillano et al., THE IMMUNOLOCALIZATION OF NUCLEAR ANTIGENS DURING THE POLLEN DEVELOPMENTAL PROGRAM AND THE INDUCTION OF POLLEN EMBRYOGENESIS, Experimental cell research, 221(1), 1995, pp. 41-54
The immunolocalization of nuclear antigens, combined with cytochemical
procedures as well as in situ hybridization and recent in situ molecu
lar methods, has been applied at different steps of pollen development
to characterize the functional organization of the nucleus during the
formation of the male gametophyte in an agronomically interesting pla
nt, Capsicum annuum L. Pollen embryogenesis has been induced in pepper
and the first stages of the process have been studied at the cellular
level. Low temperature processing methods including cryosections and
Lowicryl sections were very convenient for performing the various in s
itu techniques used in the pollen grains. Different molecular probes f
or localizing DNA, RNA, snRNPs, specific nucleolar proteins, various r
RNA species, and DNA/RNA hybrids provided positive results in the poll
en nuclei. The data obtained, and the changes observed in the organiza
tion of the nuclear compartments during pollen development, are relate
d to the variations in gene activity undergone by the male gametophyte
. The methodology used is proposed as a very convenient approach to lo
calize molecules and events involved in the nuclear function in both g
ametophytic and sporophytic pollen development. (C) 1995 Academic Pres
s, Inc.